Author: "Niko Hildebrandt" / Topic: 01 natural sciences - Searchworks@Jio Institute Digital Library Search Results

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1. A sensitive homogeneous enzyme assay for euchromatic histone-lysine-N-methyltransferase 2 (G9a) based on terbium-to-quantum dot time-resolved FRET

Author: Niko Hildebrandt, Tooba Hallaj, and Mohammad Amjadi
Subjects: Medicine (General), Epigenetic enzymes, QH301-705.5, Inhibitor screening, Pharmaceutical Science, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, R5-920, Biology (General), Original Research, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, biology, Substrate (chemistry), General Medicine, Enzyme assay, Histone-modifying enzymes, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, Förster resonance energy transfer, Enzyme, Histone, chemistry, Biochemistry, Time-resolved FRET, Enzyme inhibitor, Acetylation, Biotinylation, biology.protein
Abstract: Introduction: Histone modifying enzymes include several classes of enzymes that are responsible for various post-translational modifications of histones such as methylation and acetylation. They are important epigenetic factors, which may involve several diseases and so their assay, as well as screening of their inhibitors, are of great importance. Herein, a bioassay based on terbium-to-quantum dot (Tb-to-QD) time-resolved Förster resonance energy transfer (TR-FRET) was developed for monitoring the activity of G9a, the euchromatic histone-lysine N-methyltransferase 2. Overexpression of G9a has been reported in some cancers such as ovarian carcinoma, lung cancer, multiple myeloma and brain cancer. Thus, inhibition of this enzyme is important for therapeutic purposes. Methods: In this assay, a biotinylated peptide was used as a G9a substrate in conjugation with streptavidin-coated ZnS/CdSe QD as FRET acceptor, and an anti-mark antibody labeled with Tb as a donor. Time-resolved fluorescence was used for measuring FRET ratios. Results: We examined three QDs, with emission wavelengths of 605, 655 and 705 nm, as FRET acceptors and investigated FRET efficiency between the Tb complex and each of them. Since the maximum FRET efficiency was obtained for Tb to QD705 (more than 50%), this pair was exploited for designing the enzyme assay. We showed that the method has excellent sensitivity and selectivity for the determination of G9a at concentrations as low as 20 pM. Furthermore, the designed assay was applied for screening of an enzyme inhibitor, S-(5’-Adenosyl)-L-homocysteine (SAH). Conclusion: It was shown that Tb-to-QD FRET is an outstanding platform for developing a homogenous assay for the G9a enzyme and its inhibitors. The obtained results confirmed that this assay was quite sensitive and could be used in the field of inhibitor screening.
Published: 2020

2. pH dependence of water anomaly temperature investigated by Eu(III) cryptate luminescence

Author: Niko Hildebrandt, Jurriaan M. Zwier, Patricia Haro-González, Daniel Jaque, Carlos Mingoes, Lucía Labrador-Páez, Herve Bazin, F. Jaque, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)
Subjects: Luminescence, Materials science, [SDV]Life Sciences [q-bio], Analytical chemistry, chemistry.chemical_element, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Biochemistry, Hydrogen bonds, Analytical Chemistry, Ph dependence, [CHIM]Chemical Sciences, Molecule, [PHYS]Physics [physics], Hydrogen bond, Water, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Highly sensitive, chemistry, 13. Climate action, Lanthanide ions, Ph range, Anomaly (physics), 0210 nano-technology, Europium
Abstract: International audience; Although water has been extensively studied, not all of its unique properties have been fully understood. There is still controversy about the temperature at which hydrogen bonds are broken or weakened, producing the anomalous temperature dependence of many water properties. Different temperatures between 23 and 48 °C have been reported, but no study has scrutinized the reasons for this discrepancy. We suggest the determining role of pH in the alteration of the water anomaly temperature. We employed a luminescent europium trisbipyridine cryptate, which is highly sensitive to changes in the arrangement of water molecules and whose luminescence intensity and lifetime are not significantly influenced by variations over a broad pH range. Our results revealed an increase of the crossover temperature from circa 35 °C at pH 3.5 to circa 45 °C at pH 7 to 9, which explains the discrepancies of previous studies. The pH dependence of water anomaly temperature is an important property for a better understanding of water and water-based systems and applications.
Published: 2019

3. Cooperative Luminescence and Cooperative Sensitisation Upconversion of Lanthanide Complexes in Solution

Author: Richard C. Knighton, Niko Hildebrandt, Guillaume Pilet, Carlos Platas-Iglesias, Lohona K. Soro, Loïc J. Charbonnière, Aurora Rodríguez-Rodríguez, Marc Lenertz, and Laura Francés-Soriano
Subjects: Lanthanide, Quenching (fluorescence), Materials science, Quantum yield, General Chemistry, 02 engineering and technology, General Medicine, Photochemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 7. Clean energy, Catalysis, Photon upconversion, 0104 chemical sciences, Upconversion nanoparticles, Excited state, Solar energy conversion, Luminescence, 0210 nano-technology
Abstract: Upconversion nanoparticles have led to various breakthrough applications in solar energy conversion, imaging, and biomedicine. One key impediment is the facilitation of such processes at the molecular scale in solution where quenching effects are much more pronounced. In this work, molecular solution-state cooperative luminescence (CL) upconversion arising from a Yb excited state is explored and the mechanistic origin behind cooperative sensitisation (CS) upconversion in Yb/ Tb systems is investigated. Counterintuitively, the best UC performances were obtained for Yb/Tb ratios close to parity, resulting in the brightest molecular upconversion complexes with a quantum yield of 2.8 × 10-6 at a low laser power density of 2.86 W/cm2.
Published: 2021

4. Discrimination of the V600E Mutation in BRAF by Rolling Circle Amplification and Förster Resonance Energy Transfer

Author: Niko Hildebrandt, Mariia O. Dekaliuk, Frédéric Troalen, Xue Qiu, Philippe Busson, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), and Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], Bioengineering, 02 engineering and technology, medicine.disease_cause, 01 natural sciences, chemistry.chemical_compound, medicine, [CHIM]Chemical Sciences, Denaturation (biochemistry), Instrumentation, Gene, [PHYS]Physics [physics], Fluid Flow and Transfer Processes, Mutation, Process Chemistry and Technology, 010401 analytical chemistry, 021001 nanoscience & nanotechnology, Molecular biology, 3. Good health, 0104 chemical sciences, Förster resonance energy transfer, chemistry, Cell-free fetal DNA, Rolling circle replication, 0210 nano-technology, DNA, V600E
Abstract: International audience; The quantification of very low concentrations of circulating tumor DNA (ctDNA) biomarkers from liquid biopsies has become an important requirement for clinical diagnostics and personalized medicine. In particular, the simultaneous detection of wild-type (WT) dsDNA and their cancer-related counterparts presenting single-point mutations with simple, sensitive, specific, and reproducible technologies is paramount for ctDNA assays in clinical practice. Here, we present the development and evaluation of an amplified dsDNA assay based on a combination of isothermal rolling circle amplification (RCA) and time-gated Förster resonance energy transfer (TG-FRET) between a Tb donor and two dye (Cy3.5 and Cy5.5) acceptors. The RCA–FRET assay is free of washing and separation steps and can quantify both WT and mutated (MT) (V600E) dsDNA in the BRAF gene from a single sample in the 75 fM to 4.5 pM (4.5 × 105 to 2.7 × 107 copies) concentration range. This assay includes all steps from denaturation of the dsDNA targets to the final duplexed quantification of WT and MT targets. High assay performance at different dsDNA sequence lengths and high target specificity even in the presence of a large excess of nonspecific cell-free DNA from human plasma samples demonstrated the applicability to clinical samples. The RCA–FRET single-point mutation sensor has the potential to become an important complementary technique for analyzing liquid biopsies in advanced cancer diagnostics.
Published: 2019

5. FRET-Modulated Multihybrid Nanoparticles for Brightness-Equalized Single-Wavelength Barcoding

Author: Liang Huang, Niko Hildebrandt, Ben Corry, Chi Chen, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)
Subjects: Brightness, Photoluminescence, [SDV]Life Sciences [q-bio], chemistry.chemical_element, Nanoparticle, Terbium, 010402 general chemistry, 01 natural sciences, Biochemistry, Catalysis, Colloid and Surface Chemistry, Quantum Dots, Fluorescence Resonance Energy Transfer, Fluorescence microscope, Spectroscopy, Fluorescent Dyes, business.industry, General Chemistry, Carbocyanines, 0104 chemical sciences, Förster resonance energy transfer, chemistry, Nanoparticles, Optoelectronics, business, Excitation
Abstract: International audience; Semiconductor quantum dots (QDs) are the most versatile fluorophores for Förster resonance energy transfer (FRET) because they can function as both donors and acceptors for a multitude of fluorophores. However, a complete understanding of multidonor-multiacceptor FRET networks on QDs and their full employment into advanced fluorescence sensing and imaging have not been accomplished. Here, we provide a holistic photophysical analysis of such multidonor-QD-multiacceptor FRET systems using time-resolved and steady-state photoluminescence (PL) spectroscopy and Monte Carlo simulations. Multiple terbium complex (Tb) donors (1-191 units) and Cy5.5 dye acceptors (1-60 units) were attached to a central QD, and the entire range of combinations of FRET pathways was investigated by Tb, QD, and Cy5.5 PL. Experimental and simulation results were in excellent agreement and could disentangle the distinct contributions of hetero-FRET, homo-FRET, and dye dimerization. The FRET efficiency was independent of the number of Tb donors and dependent on the number of Cy5.5 acceptors, which could be used to independently adapt the PL intensity by the number of Tb donors and the PL lifetime by the number of Cy5.5 acceptors. We used this unique tuning capability to prepare Tb-QD-Cy5.5 conjugates with distinct QD PL lifetimes but similar QD PL intensities. These brightness-equalized multihybrid FRET nanoparticles were applied to optical barcoding via three time-gated PL intensity detection windows, which resulted in simple RGB ratios. Direct applicability was demonstrated by an efficient RGB distinction of different nanoparticle-encoded microbeads within the same field of view with both single-wavelength excitation and detection on a standard fluorescence microscope.
Published: 2019

6. Simple, Amplified, and Multiplexed Detection of MicroRNAs Using Time-Gated FRET and Hybridization Chain Reaction

Author: Carlos Mingoes, Xue Qiu, Niko Hildebrandt, Jiajia Guo, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), and ANR-16-CE12-0030,PhenX,Base moléculaire de l'activation de la recombinaison Xer par les phages(2016)
Subjects: construction, isothermal amplification, Time Factors, [SDV]Life Sciences [q-bio], Loop-mediated isothermal amplification, 02 engineering and technology, dna, 010402 general chemistry, 01 natural sciences, Analytical Chemistry, Nucleic acid thermodynamics, chemistry.chemical_compound, diagnostic assay, Fluorescence Resonance Energy Transfer, Detection limit, Chemistry, Multiple displacement amplification, Nucleic Acid Hybridization, 021001 nanoscience & nanotechnology, 0104 chemical sciences, signal amplification, MicroRNAs, Förster resonance energy transfer, in-situ, strand displacement amplification, Biophysics, Nucleic acid, sensitive detection, 0210 nano-technology, human activities, Biosensor, DNA
Abstract: International audience; The hybridization chain reaction (HCR) is a simple and sensitive method for quantifying nucleic acids. Current approaches cannot combine a washing-free sensing format with multiplexed target quantification at low concentrations, which would be highly desirable for detection both in solution and in situ. Here, we demonstrate the implementation of time-gated Förster resonance energy transfer (TG-FRET) between terbium donors and dye acceptors into HCR for multiplexed quantification of microRNAs (miR-20a and miR-21) and their DNA analogues. HCR-TG-FRET provided washing-free nucleic acid quantification with very low limits of detection down to 240 amol (1.7 pM) of microRNA and 123 amol (0.88 pM) of DNA. Efficient distinction from very homologous microRNAs demonstrated high target specificity. Multiplexing with a single measurement, a single excitation wavelength, and a single FRET pair allowed for a simultaneous quantification of miR-20a and miR-21 at concentrations between 30 and 300 pM from the same sample. HCR-TG-FRET showed similar performance for serum-free and serum-containing samples without the use of RNase inhibitors. Our results present a significant improvement in current HCR approaches regarding simplicity, sensitivity, and multiplexing. The versatile diagnostic performance of HCR-TG-FRET even in challenging biological environments presents an important advantage for advanced nucleic acid biosensing.
Published: 2019

7. Upconversion in molecular hetero-nonanuclear lanthanide complexes in solution

Author: Niko Hildebrandt, Richard C. Knighton, Lohona K. Soro, Alexandre Lecointre, Guillaume Pilet, Alexandra Fateeva, Loïc J. Charbonnière, Laura Francés-Soriano, Laurie Pontille, Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Multimatériaux et Interfaces (LMI), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), and Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Lanthanide, Materials science, Solution state, Metals and Alloys, 02 engineering and technology, General Chemistry, Molecular systems, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, Catalysis, Photon upconversion, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Materials Chemistry, Ceramics and Composites, Physical chemistry, Molecule, [CHIM.COOR]Chemical Sciences/Coordination chemistry, 0210 nano-technology, Luminescence, Excitation
Abstract: International audience; Here we show that nonanuclear lanthanide complexes respresent a new class of solution state upconversion (UC) molecules. For a composition of one Tb per eight Yb the nonanuclear complexes display a very efficient UC phenomenon with Tb luminescence in the visible region upon 980 nm NIR excitation of Yb. An unprecedented value of 1.0 × 10−7 was obtained for the UC efficiency at only 2.86 W cm−2, demonstrating these new molecular complexes to be up to 26 times more efficient than the best current molecular systems, the UC being observed down to a concentration of 10 nM.
Published: 2021

8. Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET

Author: Anne Incamps, Shashi Bhuckory, Xue Qiu, Niko Hildebrandt, K. David Wegner, Yu-Tang Wu, Travis L. Jennings, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), BAM Bundesanstalt für Materialforschung u. -prüfung, Ocean University of China (OUC), ThermoFisher, Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB), Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), ThermoFisher Scientific, Thermofisher Scientific, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Hildebrandt, Niko, Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)
Subjects: [SDV]Life Sciences [q-bio], Pharmaceutical Science, 02 engineering and technology, 01 natural sciences, 7. Clean energy, [PHYS] Physics [physics], Analytical Chemistry, PSA, Carcinoembryonic antigen, CEA, NSE, [CHIM] Chemical Sciences, Drug Discovery, Fluorescence Resonance Energy Transfer, [PHYS]Physics [physics], Immunoassay, multiplexing, biology, medicine.diagnostic_test, Chemistry, 021001 nanoscience & nanotechnology, Fluorescence, 3. Good health, [SDV] Life Sciences [q-bio], Chemistry (miscellaneous), Molecular Medicine, Kallikreins, fluorescence, 0210 nano-technology, Plate reader, 010402 general chemistry, GPI-Linked Proteins, Article, lcsh:QD241-441, lcsh:Organic chemistry, Quantum Dots, medicine, [CHIM]Chemical Sciences, Humans, lanthanides, Physical and Theoretical Chemistry, Terbium, Detection limit, Chromatography, Organic Chemistry, Prostate-Specific Antigen, 0104 chemical sciences, Carcinoembryonic Antigen, Förster resonance energy transfer, Immunoglobulin G, biology.protein, nanoparticles, biosensing, Luminescence, Biosensor
Abstract: Time-gated F&ouml, rster resonance energy transfer (TG-FRET) between Tb complexes and luminescent semiconductor quantum dots (QDs) provides highly advantageous photophysical properties for multiplexed biosensing. Multiplexed Tb-to-QD FRET immunoassays possess a large potential for in vitro diagnostics, but their performance is often insufficient for their application under clinical conditions. Here, we developed a homogeneous TG-FRET immunoassay for the quantification of carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and prostate-specific antigen (PSA) from a single serum sample by multiplexed Tb-to-QD FRET. Tb&ndash, IgG antibody donor conjugates were combined with compact QD-F(ab&rsquo, )2 antibody acceptor conjugates with three different QDs emitting at 605, 650, and 705 nm. Upon antibody&ndash, antigen&ndash, antibody sandwich complex formation, the QD acceptors were sensitized via FRET from Tb, and the FRET ratios of QD and Tb TG luminescence intensities increased specifically with increasing antigen concentrations. Although limits of detection (LoDs: 3.6 ng/mL CEA, 3.5 ng/mL NSE, and 0.3 ng/mL PSA) for the triplexed assay were slightly higher compared to the single-antigen assays, they were still in a clinically relevant concentration range and could be quantified in 50 &micro, L serum samples on a B&middot, R&middot, A&middot, H&middot, M&middot, S KRYPTOR Compact PLUS clinical immunoassay plate reader. The simultaneous quantification of CEA, NSE, and PSA at different concentrations from the same serum sample demonstrated actual multiplexing Tb-to-QD FRET immunoassays and the potential of this technology for translation into clinical diagnostics.
Published: 2020

9. Terbium–To–Quantum Dot Förster Resonance Energy Transfer for Homogeneous and Sensitive Detection of Histone Methyltransferase Activity

Author: Kimihiro Susumu, Xue Qiu, Tooba Hallaj, Igor L. Medintz, Niko Hildebrandt, Mohammad Amjadi, Urmia University, University of Tabriz [Tabriz], Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)
Subjects: Streptavidin, Histone methyltransferase activity, [SDV]Life Sciences [q-bio], Kinetics, 010402 general chemistry, 01 natural sciences, 03 medical and health sciences, chemistry.chemical_compound, Histone H3, Quantum Dots, Fluorescence Resonance Energy Transfer, [CHIM]Chemical Sciences, General Materials Science, Enzyme kinetics, Terbium, 030304 developmental biology, [PHYS]Physics [physics], 0303 health sciences, Combinatorial chemistry, 0104 chemical sciences, 3. Good health, Förster resonance energy transfer, chemistry, Histone methyltransferase, Histone Methyltransferases, Biosensor
Abstract: International audience; The development of rapid, simple, and versatile biosensors for monitoring the activity of histone modifying enzymes (HMEs) is needed for the improvement of diagnostic assays, screening of HME inhibitors, and a better understanding of HME kinetics in different environments. Nanoparticles can play an important role in this regard by improving or complementing currently available enzyme detection technologies. Here, we present the development and application of a homogeneous methyltransferase (SET7/9) assay based on time-gated Förster resonance energy transfer (TG-FRET) between terbium complexes (Tb) and luminescent semiconductor quantum dots (QDs). Specific binding of a Tb-antibody conjugate to a SET7/9-methylated Lys4 on a histone H3(1–21) peptide substrate attached to the QD surface resulted in efficient FRET and provided the mechanism for monitoring the SET7/9 activity. Two common peptide-QD attachment strategies (biotin–streptavidin and polyhistidine-mediated self-assembly), two different QD colors (625 and 705 nm), and enzyme sensing with post- or pre-assembled QD–peptide conjugates demonstrated the broad applicability of this assay design. Limits of detection in the low picomolar concentration range, high selectivity tested against non-specific antibodies, enzymes, and co-factors, determination of the inhibition constants of the SET7/9 inhibitors SAH and (R)-PFI-2, and analysis of the co-factor (SAM) concentration-dependent enzyme kinetics of SET7/9 which followed the Michaelis–Menten model highlighted the excellent performance of this TG-FRET HME activity assay.
Published: 2020

10. Single-Measurement Multiplexed Quantification of MicroRNAs from Human Tissue Using Catalytic Hairpin Assembly and Förster Resonance Energy Transfer

Author: Nicole Golob-Schwarzl, Jingyue Xu, Jiajia Guo, Xue Qiu, Niko Hildebrandt, Johannes Haybaeck, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Medical University Graz, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE12-0030,PhenX,Base moléculaire de l'activation de la recombinaison Xer par les phages(2016)
Subjects: Bioanalysis, [SDV]Life Sciences [q-bio], Bioengineering, 02 engineering and technology, 01 natural sciences, microRNA, Fluorescence Resonance Energy Transfer, Humans, cancer, [CHIM]Chemical Sciences, Terbium, Instrumentation, clinical diagnostics, Fluid Flow and Transfer Processes, Detection limit, [PHYS]Physics [physics], catalytic hairpin assembly, Chemistry, Process Chemistry and Technology, 010401 analytical chemistry, Translation (biology), tissue, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Autofluorescence, MicroRNAs, Förster resonance energy transfer, Biochemistry, Nucleic acid, FRET, miR-21, 0210 nano-technology, Biosensor, miR-20a
Abstract: International audience; Absolute quantification of microRNAs (miRNAs) or other nucleic acid biomarkers is an important requirement for molecular and clinical biosensing. Emerging technologies with beneficial features concerning simplicity and multiplexing present an attractive route for advancing diagnostic tools toward rapid and low-cost bioanalysis. However, the actual translation into the clinic by miRNA quantification in human samples is often missing. Here, we show that implementing time-gated Förster resonance energy transfer (TG-FRET) into a catalytic hairpin assembly (CHA) can be used for the simultaneous quantification of two miRNAs with a single measurement from total RNA extracts of human tissues. A single terbium–dye FRET pair was conjugated at two specific distances within target-specific CHA hairpin probes, such that each miRNA resulted in distinct amplified photoluminescence (PL) decays that could be distinguished and quantified by TG PL intensity detection. Enzyme-free amplification in a separation-free assay format and the absence of autofluorescence background allowed for simple, specific, and sensitive detection of miR-21 and miR-20a with limits of detection down to 1.8 pM (250 amol). Reliable duplexed quantification of both miRNAs at low picomolar concentrations was confirmed by analyzing total RNA extracts from different colon and rectum tissues with single- and dual-target CHA-TG-FRET and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for comparison. These simple and multiplexed nucleic acid biomarker assays present a capable method for clinical diagnostics and biomolecular research.
Published: 2020

11. Energy transfer with nanoparticles for in vitro diagnostics

Author: Niko Hildebrandt, Jiajia Guo, Jingyue Xu, Tooba Hallaj, Laura Francés-Soriano, Xue Qiu, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Urmia University, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), and Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [PHYS]Physics [physics], Materials science, [SDV]Life Sciences [q-bio], Nanoparticle, Nanotechnology, 02 engineering and technology, Chromophore, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, Acceptor, Small molecule, Fluorescence, 0104 chemical sciences, Förster resonance energy transfer, Colloidal gold, [CHIM]Chemical Sciences, 0210 nano-technology, Luminescence, ComputingMilieux_MISCELLANEOUS
Abstract: In vitro diagnostics is an important application for nanoparticles in the life sciences and has attracted continuous research interest. While nanoparticles can be used as single luminescent probes, their unique photophysical properties provide many more advantages when they are combined with other nanoparticles or molecules to form energy transfer pairs. Energy transfer-based assays are homogeneous in nature and are often as simple as “mix and measure,” which has catalyzed their frequent use in clinical diagnostics. Most bioanalytical applications involve Forster resonance energy transfer (FRET), a distance-dependent nonradiative energy transfer from a donor to acceptor chromophore through dipole–dipole interaction. Nanosurface energy transfer (NSET) describes energy transfer to metal nanoparticles. Semiconductor quantum dots, upconversion nanoparticles, gold nanoparticles, and carbon dots are among the most applied nanoparticles in in vitro diagnostics and they have been largely explored in the analysis of disease-related nucleic acids, proteins, and small molecules. The utilizations of various nanoparticles with their intrinsic optical properties have changed the diagnostic horizon substantially by improving the assay simplicity, sensitivity, selectivity, robustness, and multiplexing capacity. In this chapter, we discuss the use of nanoparticles in energy transfer assays for in vitro diagnostics and show their strong potential for advancing the field beyond molecular fluorescent probes.
Published: 2020

12. Quantum dots for Förster Resonance Energy Transfer (FRET)

Author: Igor L. Medintz, Niko Hildebrandt, Marcelina Cardoso Dos Santos, W. Russ Algar, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), University of British Columbia (UBC), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), and Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: chemistry.chemical_classification, [PHYS]Physics [physics], Fluorescence-lifetime imaging microscopy, Materials science, Biomolecule, [SDV]Life Sciences [q-bio], 010401 analytical chemistry, technology, industry, and agriculture, Nanotechnology, [CHIM.MATE]Chemical Sciences/Material chemistry, 01 natural sciences, 0104 chemical sciences, Analytical Chemistry, Förster resonance energy transfer, chemistry, Quantum dot, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Surface modification, [CHIM]Chemical Sciences, Absorption (electromagnetic radiation), Spectroscopy
Abstract: International audience; The analysis of biomolecular interactions using quantum dots (QDs) as both FRET donors and acceptors has become an established technique in the life sciences. This development has been driven by the unique properties of QDs, which include large surfaces for the attachment of biomolecules, high brightness and photostability, strong and spectrally broad absorption, and color-tunability via QD size, shape, and material. Applications include molecular rulers for structural analysis, small-molecule sensors, immunoassays, enzyme assays, nucleic acid assays, fluorescence imaging in-vitro and in-vivo, and molecular logic gates. Here, we will explain the theory of QD-based FRET, review some aspects of QD surface functionalization that are important for FRET, and highlight and discuss the advantages and disadvantages of QDs in FRET-biosensing using both spectroscopy and imaging techniques.
Published: 2020

13. Picomolar Biosensing and Conformational Analysis Using Artificial Bidomain Proteins and Terbium-to-Quantum Dot Förster Resonance Energy Transfer

Author: Agathe Urvoas, Philippe Minard, Niko Hildebrandt, Igor L. Medintz, Corentin Leger, Kimihiro Susumu, Marie Valerio-Lepiniec, Akram Yahia-Ammar, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Modélisation et Ingénierie des Protéines (MIP), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Optical Sciences Division, Departments of Cell Biology and Chemistry, The Scripps Research Institute, Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), and Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Scaffold protein, Conformational change, [SDV]Life Sciences [q-bio], General Physics and Astronomy, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, molecular diagnostics, Protein structure, Quantum Dots, Fluorescence Resonance Energy Transfer, [CHIM]Chemical Sciences, lanthanides, General Materials Science, protein structure, Terbium, binding assay, [PHYS]Physics [physics], Chemistry, Ligand binding assay, General Engineering, molecular ruler, 021001 nanoscience & nanotechnology, Acceptor, 0104 chemical sciences, Förster resonance energy transfer, Semiconductors, Quantum dot, FRET, Biophysics, 0210 nano-technology, Biosensor
Abstract: Publisher: American Chemical Society; International audience; Although antibodies remain a primary recognition element in all forms of biosensing, functional limitations arising from their size, stability, and structure have motivated the development and production of many different artificial scaffold proteins for biological recognition. However, implementing such artificial binders into functional high-performance biosensors remains a challenging task. Here, we present the design and application of Förster resonance energy transfer (FRET) nanoprobes comprised of small artificial proteins (αRep bidomains) labeled with a Tb complex (Tb) donor on the C-terminus and a semiconductor quantum dot (QD) acceptor on the N-terminus. Specific binding of one or two protein targets to the αReps induced a conformational change that could be detected by time-resolved Tb-to-QD FRET. These single-probe FRET-switches were used in a separation-free solution-phase assay to quantify different protein targets at subnanomolar concentrations and measure the conformational changes with subnanometer resolution. Probing ligand-receptor binding under physiological conditions at very low concentrations in solution is a special feature of FRET that can be efficiently combined with other structural characterization methods to develop, understand, and optimize artificial biosensors. Our results suggest that the αRep FRET nanoprobes have a strong potential for their application in advanced diagnostics and intracellular live cell imaging of ligand-receptor interactions.
Published: 2020

14. Ultrabright Terbium Nanoparticles for FRET Biosensing and in Situ Imaging of Epidermal Growth Factor Receptors

Author: Laura Francés-Soriano, Vjona Cifliku, Cyrille Charpentier, Clémence Cheignon, Joan Goetz, Aline Nonat, Loïc J. Charbonnière, Niko Hildebrandt, Ka-Leung Wong, Marcelina Cardoso Dos Santos, Centre National de la Recherche Scientifique (CNRS), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), POLY-DTECH, Reconnaissance Ionique et Chimie de Coordination (RICC), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS), Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Hong Kong Baptist University (HKBU), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR‐11‐IDEX‐0003‐02/ Saclay Plant Sciences, ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016), ANR-11-EQPX-0029,MORPHOSCOPE 2,Imagerie et reconstruction multiéchelles de la morphogenèse. (Plateforme d'innovation technologique et méthodologique pour l'imagerie in vivo et la reconstruction des dynamiques multiéchelles de la morphogenèse)(2011), and ANR-10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010)
Subjects: EGF Family of Proteins, [SDV]Life Sciences [q-bio], Biosensing Techniques, 010402 general chemistry, 7. Clean energy, 01 natural sciences, Catalysis, Mice, Epidermal growth factor, Fluorescence Resonance Energy Transfer, Animals, [CHIM]Chemical Sciences, Terbium, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, [PHYS]Physics [physics], Bioconjugation, 010405 organic chemistry, Chemistry, Biomolecule, Organic Chemistry, General Chemistry, 0104 chemical sciences, ErbB Receptors, Förster resonance energy transfer, Biotinylation, Biophysics, NIH 3T3 Cells, Nanoparticles, Biosensor, Linker, A431 cells
Abstract: International audience; Lanthanide‐doped nanoparticles (LnNPs) have become an important class of fluorophores for advanced biosensing and bioimaging. LnNPs that are photosensitized by surface‐attached antenna ligands can possess exceptional brightness. However, their functional bioconjugation remains an important challenge for their translation into bioanalytical applications. To solve this problem, we designed a ligand that can be simultaneously applied as efficient light harvesting antenna for Tb surface ions and strong linker of biomolecules to the LnNPs surfaces. To demonstrate generic applicability of the photosensitized TbNP‐bioconjugates, we applied them in two prototypical applications for biosensing and bioimaging. First, in‐solution biorecognition was shown by time‐resolved Förster resonance energy transfer (FRET) between streptavidin‐functionalized TbNPs to biotinylated dyes (ATTO 610). Second, in‐situ detection of ligand‐receptor binding on cells was accomplished with TbNP‐antibody (Matuzumab) conjugates that could specifically bind to transmembrane epidermal growth factor receptors (EGFR). High specificity and sensitivity were demonstrated by time‐gated imaging of EGFR on both strongly (A431) and weakly (HeLa and Cos7) EGFR‐expressing cell lines, whereas non‐expressing cell lines (NIH3T3) and EGFR‐passivated A431 cells did not show any signals. Despite the relatively large size of TbNP‐antibody conjugates, they could be internalized by A431 cells upon binding to extracellular EGFR, which showed their potential as bright and stable luminescence markers for intracellular signalling.
Published: 2020

15. Er3+-to-dye energy transfer in DNA-coated core and core/shell/shell upconverting nanoparticles with 980 nm and 808 nm excitation of Yb3+ and Nd3

Author: Marta Maria Natile, Niko Hildebrandt, Nicola Peruffo, Laura Francés-Soriano, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Infrared devices, Materials science, Infrared Rays, [SDV]Life Sciences [q-bio], Kinetics, Metal Nanoparticles, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, 7. Clean energy, Biochemistry, Analytical Chemistry, Europium, Single-Stranded, Electrochemistry, Fluorescence Resonance Energy Transfer, Environmental Chemistry, Yttrium, Spectroscopy, Fluorescent Dyes, Ions, Neodymium, business.industry, Near-infrared spectroscopy, DNA, Orders of magnitude (numbers), Carbocyanines, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Förster resonance energy transfer, Energy transfer, DNA, Single-Stranded, FRET, Optoelectronics, Nanoparticles, 0210 nano-technology, Luminescence, business, Biosensor, Excitation, Upconversion, Visible spectrum
Abstract: The capability of upconverting nanoparticles (UCNPs) to convert near infrared (NIR) into visible light has become an important feature for biosensing, imaging, therapy, and their combination. While significant achievements have been accomplished during the last decade developing nanohybrids based on UCNPs as energy donors in Förster resonance energy transfer (FRET) systems, it is still challenging to understand and control FRET from UCNPs to dyes and to adapt the NIR excitation wavelength. Here, we describe the synthesis, characterization, and steady-state and time-resolved FRET analysis of UCNP–DNA nanohybrids, in which dye labelled single stranded (ss)DNA was attached to Yb–Er-co-doped core UCNPs (c-UCNPs) and c-UCNPs with a thin Nd-doped shell and a second thin undoped shell (css-UCNPs). Despite differences in sizes, compositions, donor–acceptor distances, brightness, and excitation wavelength (980 nm for Yb3+and 808 nm for Nd3+), all UCNP–DNA nanohybrids showed very similar concentration dependent FRET-quenching of UCNP luminescence with efficiencies between 0 and ∼20%. We analyzed luminescence intensities, decay times, and rise times and could show the entanglement of excitation and emission kinetics by simply changing the excitation wavelength from 980 nm to 808 nm for the same css-UCNPs. Time-gated FRET-sensitized dye luminescence showed dye-ssDNA concentration dependence over four orders of magnitude (1 nM to 10 μM), which suggested a possible application to nucleic acid biosensing for both 808 and 980 nm excitation.
Published: 2020

16. Resonance energy transfer to gold nanoparticles: NSET defeats FRET

Author: Chi Chen, Niko Hildebrandt, National University of Defense Technology [China], Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: [PHYS]Physics [physics], Materials science, Energy transfer, [SDV]Life Sciences [q-bio], 010401 analytical chemistry, Nanotechnology, 01 natural sciences, 0104 chemical sciences, Analytical Chemistry, Formalism (philosophy of mathematics), Förster resonance energy transfer, Colloidal gold, [CHIM]Chemical Sciences, Luminescence, Biosensor, Spectroscopy, ComputingMilieux_MISCELLANEOUS
Abstract: Gold nanoparticles (AuNPs) have been widely applied as highly efficient luminescence quenchers in combination with a large variety of donor fluorophores. While the application of such systems in optical biosensing has resulted in significant advances of biological, biochemical, and biophysical research, the underlying energy transfer mechanism has been much less investigated. In this review, we discuss the differences between Forster resonance energy transfer (FRET) and nanosurface energy transfer (NSET), give an overview of the literature concerning the assignment of FRET or NSET to AuNP-based luminescence quenching, show why the development of improved NSET models and their application to experimental data provided strong arguments in favor of the NSET formalism, and discuss applications of NSET in biosensing, molecular rulers, and molecular tension probes. Taking into account the existing literature, the NSET formalism, including size and wavelength-dependent issues, is superior to FRET theory for a general description of energy transfer to AuNPs.
Published: 2019

17. Single‐Nanoparticle Cell Barcoding by Tunable FRET from Lanthanides to Quantum Dots

Author: David Parker, Emmanuel Bourrier, Chi Chen, Jurriaan M. Zwier, Yu-Tang Wu, Vjona Cifliku, Martina Delbianco, Liang Huang, Niko Hildebrandt, Lijiao Ao, Marcelina Cardoso Dos Santos, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Cisbio Bioassays, DEPARTMENT OF CHEMISTRY, Durham University, Institute of Molecular Chemistry, and Universiteit van Amsterdam (UvA)
Subjects: Optical Barcoding, Materials science, Photoluminescence, resonance energy-transfer, [SDV]Life Sciences [q-bio], Nanoparticle, 02 engineering and technology, bright, 010402 general chemistry, 01 natural sciences, biomolecules, Catalysis, nanocrystals, Europium, Live cell imaging, Microscopy, Quantum Dots, Fluorescence Resonance Energy Transfer, lanthanides, Terbium, lifetime, business.industry, Communication, imaging, General Chemistry, 021001 nanoscience & nanotechnology, biosensors, europium complexes, Fluorescence, photoluminescence lifetimes, Communications, 0104 chemical sciences, microspheres, Förster resonance energy transfer, Nanocrystal, Quantum dot, microscopy, FRET, Optoelectronics, Nanoparticles, 0210 nano-technology, business
Abstract: WOS:000446306600058; International audience; Fluorescence barcoding based on nanoparticles provides many advantages for multiparameter imaging. However, creating different concentration-independent codes without mixing various nanoparticles and by using single-wavelength excitation and emission for multiplexed cellular imaging is extremely challenging. Herein, we report the development of quantum dots (QDs) with two different SiO2 shell thicknesses (6 and 12nm) that are coated with two different lanthanide complexes (Tb and Eu). FRET from the Tb or Eu donors to the QD acceptors resulted in four distinct photoluminescence (PL) decays, which were encoded by simple time-gated (TG) PL intensity detection in three individual temporal detection windows. The well-defined single-nanoparticle codes were used for live cell imaging and a one-measurement distinction of four different cells in a single field of view. This single-color barcoding strategy opens new opportunities for multiplexed labeling and tracking of cells.
Published: 2018

18. Aptamer and nanomaterial based FRET biosensors: a review on recent advances (2014–2019)

Author: Milad Torabfam, Zeki Semih Pehlivan, Niko Hildebrandt, Hasan Kurt, Cleva W. Ow-Yang, Meral Yüce, Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Materials science, [SDV]Life Sciences [q-bio], Aptamer, Nanochemistry, Nanoprobe, Carbon Dots, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, 7. Clean energy, Metal-Organic Framework, Analytical Chemistry, law.invention, Nanomaterials, law, ssDNA, Quantum Dots, Gold Nanoparticles, [CHIM]Chemical Sciences, Reduced Graphene Oxide, ComputingMilieux_MISCELLANEOUS, Graphene Quantum Dots, [PHYS]Physics [physics], Up-Converting Nanoparticles, Graphene, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Biosensors, Förster resonance energy transfer, Quantum dot, FRET, 0210 nano-technology, Biosensor
Abstract: Fluorescence resonance energy transfer, one of the most powerful phenomena for elucidating molecular interactions, has been extensively utilized as a biosensing tool to provide accurate information at the nanoscale. Numerous aptamer- and nanomaterial-based FRET bioassays has been developed for detection of a large variety of molecules. Affinity probes are widely used in biosensors, in which aptamers have emerged as advantageous biorecognition elements, due to their chemical and structural stability. Similarly, optically active nanomaterials offer significant advantages over conventional organic dyes, such as superior photophysical properties, large surface-to-volume ratios, photostability, and longer shelf life. In this report (with 175 references), the use of aptamer-modified nanomaterials as FRET couples is reviewed: quantum dots, upconverting nanoparticles, graphene, reduced graphene oxide, gold nanoparticles, molybdenum disulfide, graphene quantum dots, carbon dots, and metal-organic frameworks. Tabulated summaries provide the reader with useful information on the current state of research in the field. Graphical abstract Schematic representation of a fluorescence resonance energy transfer-based aptamer nanoprobe in the absence and presence of a given target molecule (analyte). Structures are not drawn to their original scales.
Published: 2019

19. Lanthanide-Complex-Loaded Polymer Nanoparticles for Background-Free Single-Particle and Live-Cell Imaging

Author: Andrey S. Klymchenko, Loïc J. Charbonnière, Anne Runser, Hortense Bartenlian, Niko Hildebrandt, Marcelina Cardoso Dos Santos, Andreas Reisch, Aline Nonat, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Bioimagerie et Pathologies (LBP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), ANR-16-CE09-0007,supertrack,Nanoparticules polymériques fluorescentes ultrabrillantes et biocompatibles pour l'imagerie rapide intracellulaire à haute-résolution(2016), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), Laboratoire de Biophotonique et Pharmacologie - UMR 7213 (LBP), Réseau nanophotonique et optique, and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)
Subjects: Lanthanide, General Chemical Engineering, [SDV]Life Sciences [q-bio], Nanoparticle, Nanotechnology, quantum dots, 02 engineering and technology, bright, in-vitro, 010402 general chemistry, 01 natural sciences, dyes, Live cell imaging, emission, Materials Chemistry, luminescence, Molecule, [CHIM.COOR]Chemical Sciences/Coordination chemistry, ComputingMilieux_MISCELLANEOUS, time-resolved fluorescence, chemistry.chemical_classification, General Chemistry, Polymer, tracking, 021001 nanoscience & nanotechnology, 0104 chemical sciences, agents, Autofluorescence, chemistry, Particle, photoluminescence, Time-resolved spectroscopy, 0210 nano-technology
Abstract: WOS:000471728200021; International audience; Imaging single molecules and nanoparticles in complex biological media is highly challenging notably due to autofluorescence of cells and tissues. Lanthanides and lanthanide complexes, with their particularly long luminescence lifetimes, offer the possibility to perform time-gated imaging and thus to strongly reduce the autofluorescence background. However, their very low brightness and photon flux have limited their use in single-molecule imaging. Here, we encapsulate high amounts of Europium complexes into poly(methyl methacrylate)-based particles of 10, 20, and 30 nm size. The resulting particles contain up to 5000 copies of the complex with a quantum yield of \textgreater= 0.2, resulting in a per particle brightness of up to 4 x 10(7) M-1 cm(-1). They can be imaged at the single-particle level using low illumination intensities (0.24 W cm(-2)) and low acquisition times (300 ms) and internalize well into living cells, where they can be monitored through time-gated imaging at illumination conditions compatible with living specimen. These Eu-complex-loaded nanoparticles can thus be applied for highly sensitive and autofluorescence-free imaging and have the potential to become very performant probes for fast intracellular tracking of single biomolecules.
Published: 2019
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20. Influence of the Spatial Conformation of Charged Ligands on the Optical Properties of Gold Nanoclusters

Author: Claude Zoukimian, Jean-Luc Coll, Xavier Le Guével, Franck Bertorelle, Estelle Porret, Niko Hildebrandt, Muriel Jourdan, Clothilde Comby-Zerbino, Rodolphe Antoine, Xue Qiu, Vanessa Trouillet, Didier Boturyn, Béatrice Gennaro, Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Département de Chimie Moléculaire - Ingéniérie et Intéractions BioMoléculaires (DCM - I2BM), Département de Chimie Moléculaire (DCM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Spectrométrie des biomolécules et agrégats (SPECTROBIO), Institut Lumière Matière [Villeurbanne] (ILM), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Karlsruher Institut für Technologie (KIT), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR-17-EURE-0003,CBH-EUR-GS,CBH-EUR-GS(2017), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and Boturyn, Didier
Subjects: Materials science, Quantum yield, 02 engineering and technology, Peptides and proteins, 010402 general chemistry, Ligands, 01 natural sciences, Antioxidants, Nanoclusters, Colloid, mental disorders, [CHIM]Chemical Sciences, [CHIM.COOR]Chemical Sciences/Coordination chemistry, Surface charge, Physical and Theoretical Chemistry, Spectroscopy, ComputingMilieux_MISCELLANEOUS, Optical properties, Ligand, Nuclear magnetic resonance spectroscopy, [CHIM.COOR] Chemical Sciences/Coordination chemistry, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, [CHIM.THEO] Chemical Sciences/Theoretical and/or physical chemistry, Crystallography, General Energy, Transmission electron microscopy, Gold, 0210 nano-technology
Abstract: International audience; Photoluminescent gold nanoclusters (Au NCs) were synthesized in one step in aqueous conditions using a mixture of glutathione and (mono or multivalent) glutathione-modified arginine peptides. By controlling the ratio of coligands, we investigated how the multivalency and the amount of arginines influenced the growth of Au NCs, their surface chemistry, their colloidal stability, and their optical properties. We demonstrated using two-dimensional nuclear magnetic resonance spectroscopy that the organization of the ligand on the Au surface was composed by an inner rigid layer and an outer flexible part via inter- and/or intraligand spatial proximity. This directly impacted the structure of the Au NCs, as confirmed by gel electrophoresis, high-resolution transmission electron microscopy, diffusion-ordered spectroscopy, and mass spectrometry. Increasing arginine content also induced an increase of positive surface charge and an enhancement of the near infrared emission intensity at ∼670 nm with quantum yield up to 10% validating the significant influence of the ligands to the optical properties of Au NCs.
Published: 2019

21. Quantification of Cellular Deoxyribonucleoside Triphosphates by Rolling Circle Amplification and Förster Resonance Energy Transfer

Author: Niko Hildebrandt, Olivier Guittet, Michel Lepoivre, Xue Qiu, Meng-Er Huang, Nadine El Banna, Carlos Mingoes, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Intégrité du génome, ARN et cancer, Institut Curie [Paris]-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire Bioénergétique Membranaire et Stress (LBMS), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), and Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Thioredoxin-Disulfide Reductase, Deoxyribonucleoside triphosphate, Genome integrity, DNA repair, Deoxyribonucleotides, 010402 general chemistry, Proof of Concept Study, Sensitivity and Specificity, 01 natural sciences, Analytical Chemistry, chemistry.chemical_compound, Limit of Detection, Auranofin, Cell Line, Tumor, Ribonucleotide Reductases, Fluorescence Resonance Energy Transfer, Humans, Hydroxyurea, heterocyclic compounds, Enzyme Inhibitors, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Base Sequence, 010401 analytical chemistry, 3. Good health, 0104 chemical sciences, Deoxyribonucleoside, enzymes and coenzymes (carbohydrates), Förster resonance energy transfer, chemistry, Biochemistry, Rolling circle replication, Nucleic acid, Nucleic Acid Amplification Techniques, Plate reader
Abstract: International audience; The quantification of cellular deoxyribonucleoside triphosphate (dNTP) levels is important for studying pathologies, genome integrity, DNA repair, and the efficacy of pharmacological drug treatments. Current standard methods, such as enzymatic assays or high-performance liquid chromatography, are complicated, costly, and labor-intensive, and alternative techniques that simplify dNTP quantification would present very useful complementary approaches. Here, we present a dNTP assay based on isothermal rolling circle amplification (RCA) and rapid time-gated Förster resonance energy transfer (TG-FRET), which used a commercial clinical plate reader system. Despite the relatively simple assay format, limits of detection down to a few picomoles of and excellent specificity for each dNTP against the other dNTPs, rNTPs, and dUTP evidenced the strong performance of the assay. Direct applicability of RCA-FRET to applied nucleic acid research was demonstrated by quantifying all dNTPs in CEM-SS leukemia cells with and without hydroxyurea or auranofin treatment. Both pharmacological agents could reduce the dNTP production in a time- and dose-dependent manner. RCA-FRET provides simple, rapid, sensitive, and specific quantification of intracellular dNTPs and has the potential to become an advanced tool for both fundamental and applied dNTP research.
Published: 2019

22. A new forum for upconversion research: The UPCON conference

Author: Niko Hildebrandt, Artur Bednarkiewicz, Hans H. Gorris, Julia Pérez-Prieto, Tero Soukka, Instituto de Ciencia Molecular (ICMol), Universitat de València (UV), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: upconversion, Engineering, business.industry, [SDV]Life Sciences [q-bio], 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, background-free, 01 natural sciences, Engineering physics, Atomic and Molecular Physics, and Optics, Photon upconversion, 0104 chemical sciences, luminescence, General Materials Science, 0210 nano-technology, business, Instrumentation, Spectroscopy, nanomaterials
Abstract: International audience; The Conference and Spring School on Properties, Design and Applications of Upconversion Nanomaterials (UPCON) provides a new forum for all experts and newcomers in the field of upconversion research. On the occasion of the second UPCON 2018 in Valencia (Spain), we are pleased to present a collection of 12 reviews and research articles that reflect recent advances in upconversion materials, their unique luminescent properties and many applications spanning from nanoscale thermometry to biomedicine.
Published: 2019

23. Energy Transfer with Semiconductor Quantum Dot Bioconjugates: A Versatile Platform for Biosensing, Energy Harvesting, and Other Developing Applications

Author: Niko Hildebrandt, Sebastián A. Díaz, Christopher M. Spillmann, W. Russ Algar, Igor L. Medintz, Thomas Pons, Eunkeu Oh, James B. Delehanty, Michael H. Stewart, Kimihiro Susumu, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Department of Chemistry, University of British Columbia (UBC), Laboratoire de Physique et d'Etude des Matériaux (UMR 8213) (LPEM), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Optical Sciences Division, Naval Research Laboratory (NRL), Departments of Cell Biology and Chemistry, The Scripps Research Institute, and The Scripps Research Institute [La Jolla, San Diego]
Subjects: Surface Properties, [SDV]Life Sciences [q-bio], Energy transfer, Context (language use), Nanotechnology, Biosensing Techniques, 02 engineering and technology, Ligands, 010402 general chemistry, 01 natural sciences, Nanomaterials, Quantum Dots, Bioconjugation, Chemistry, business.industry, General Chemistry, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Semiconductor, Energy Transfer, Semiconductors, Quantum dot, 0210 nano-technology, business, Energy harvesting, Biosensor
Abstract: Luminescent semiconductor quantum dots (QDs) are one of the more popular nanomaterials currently utilized within biological applications. However, what is not widely appreciated is their growing role as versatile energy transfer (ET) donors and acceptors within a similar biological context. The progress made on integrating QDs and ET in biological configurations and applications is reviewed in detail here. The goal is to provide the reader with (1) an appreciation for what QDs are capable of in this context, (2) how this field has grown over a relatively short time span, and, in particular, (3) how QDs are steadily revolutionizing the development of new biosensors along with a myriad of other photonically active nanomaterial-based bioconjugates. An initial discussion of QD materials along with key concepts surrounding their preparation and bioconjugation is provided given the defining role these aspects play in the QDs ability to succeed in subsequent ET applications. The discussion is then divided around the specific roles that QDs provide as either Förster resonance energy transfer (FRET) or charge/electron transfer donor and/or acceptor. For each QD-ET mechanism, a working explanation of the appropriate background theory and formalism is articulated before examining their biosensing and related ET utility. Other configurations such as incorporation of QDs into multistep ET processes or use of initial chemical and bioluminescent excitation are treated similarly. ET processes that are still not fully understood such as QD interactions with gold and other metal nanoparticles along with carbon allotropes are also covered. Given their maturity, some specific applications ranging from in vitro sensing assays to cellular imaging are separated and discussed in more detail. Finally a perspective on how this field will continue to evolve is provided.
Published: 2016

24. Self-Assembled Gold Nanoclusters for Bright Fluorescence Imaging and Enhanced Drug Delivery

Author: Xavier Le Guével, Akram Yahia-Ammar, Niko Hildebrandt, Daniel Sierra, Fabienne Merola, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), instituto Geofísico, Escuela Politécnica Nacional (EPN), Laboratoire de Chimie Physique D'Orsay (LCPO), Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Cancer Targets & Experimental Therapeutics, Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), and Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Subjects: Fluorescence-lifetime imaging microscopy, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Materials science, [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging, Ultraviolet Rays, Dispersity, Metal Nanoparticles, General Physics and Astronomy, Nanoparticle, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Antibodies, Cell Line, Nanoclusters, Polyamines, [CHIM]Chemical Sciences, Humans, General Materials Science, ComputingMilieux_MISCELLANEOUS, Drug Carriers, General Engineering, 021001 nanoscience & nanotechnology, Polyelectrolytes, Fluorescence, 0104 chemical sciences, Microscopy, Fluorescence, Drug delivery, [SPI.OPTI]Engineering Sciences [physics]/Optics / Photonic, Gold, Self-assembly, 0210 nano-technology, Oligopeptides, Nanogel
Abstract: Nanoparticles combining enhanced cellular drug delivery with efficient fluorescence detection are important tools for the development of theranostic agents. Here, we demonstrate this concept by a simple, fast, and robust protocol of cationic polymer-mediated gold nanocluster (Au NCs) self-assembly into nanoparticles (NPs) of ca. 120 nm diameter. An extensive characterization of the monodisperse and positively charged NPs revealed pH-dependent swelling properties, strong fluorescence enhancement, and excellent colloidal and photostability in water, buffer, and culture medium. The versatility of the preparation is demonstrated by using different Au NC surface ligands and cationic polymers. Steady-state and time-resolved fluorescence measurements give insight into the aggregation-induced emission phenomenon (AIE) by tuning the Au NC interactions in the self-assembled nanoparticles using the pH-dependent swelling. In vitro studies in human monocytic cells indicate strongly enhanced uptake of the NPs compared to free Au NCs in endocytic compartments. The NPs keep their assembly structure with quite low cytotoxicity up to 500 μg Au/mL. Enhanced drug delivery is demonstrated by loading peptides or antibodies in the NPs using a one-pot synthesis. Fluorescence microscopy and flow cytometry confirmed intracellular colocalization of the biomolecules and the NP carriers with a respective 1.7-fold and 6.5-fold enhanced cellular uptake of peptides and antibodies compared to the free biomolecules.
Published: 2016

25. Time‐Gated FRET Nanoprobes for Autofluorescence‐Free Long‐Term In Vivo Imaging of Developing Zebrafish

Author: Marcelina Cardoso Dos Santos, Kimihiro Susumu, Ingrid Colin, Michaël Demarque, Niko Hildebrandt, Gabriel Ribeiro Dos Santos, Igor L. Medintz, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), and Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: animal structures, Materials science, [SDV]Life Sciences [q-bio], Nanoprobe, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, In vivo, Microscopy, Fluorescence Resonance Energy Transfer, Animals, Nanotechnology, [CHIM]Chemical Sciences, General Materials Science, Zebrafish, [PHYS]Physics [physics], Mechanical Engineering, 021001 nanoscience & nanotechnology, Photobleaching, Molecular Imaging, 0104 chemical sciences, Autofluorescence, Förster resonance energy transfer, Mechanics of Materials, Biophysics, 0210 nano-technology, mCherry, Preclinical imaging
Abstract: International audience; The zebrafish is an important vertebrate model for disease, drug discovery, toxicity, embryogenesis, and neuroscience. In vivo fluorescence microscopy can reveal cellular and subcellular details down to the molecular level with fluorescent proteins (FPs) currently the main tool for zebrafish imaging. However, long maturation times, low brightness, photobleaching, broad emission spectra, and sample autofluorescence are disadvantages that cannot be easily overcome by FPs. Here, a bright and photostable terbium‐to‐quantum dot (QD) Förster resonance energy transfer (FRET) nanoprobe with narrow and tunable emission bands for intracellular in vivo imaging is presented. The long photoluminescence (PL) lifetime enables time‐gated (TG) detection without autofluorescence background. Intracellular four‐color multiplexing with a single excitation wavelength and in situ assembly and FRET to mCherry demonstrate the versatility of the TG‐FRET nanoprobes and the possibility of in vivo bioconjugation to FPs and combined nanoprobe‐FP FRET sensing. Upon injection at the one‐cell stage, FRET nanoprobes can be imaged in developing zebrafish embryos over seven days with toxicity similar to injected RNA and strongly improved signal‐to‐background ratios compared to non‐TG imaging. This work provides a strategy for advancing in vivo fluorescence imaging applications beyond the capabilities of FPs.
Published: 2020

26. Einzelnanopartikel-Strichkodierung von Zellen mittels durchstimmbarem FRET von Lanthanoiden auf Quantenpunkte

Author: Niko Hildebrandt, Lijiao Ao, Vjona Cifliku, Liang Huang, Chi Chen, Yu-Tang Wu, Emmanuel Bourrier, David Parker, Martina Delbianco, Jurriaan M. Zwier, Marcelina Cardoso Dos Santos, National University of Defense Technology [China], Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Cisbio Bioassays, DEPARTMENT OF CHEMISTRY, Durham University, Computing Laboratory (OUCL), University of Oxford, Institute of Molecular Chemistry, Universiteit van Amsterdam (UvA), Sichuan Provincial Center for Diseases Control and Prevention, Sichuan Government, and University of Oxford [Oxford]
Subjects: [PHYS]Physics [physics], Materials science, [SDV]Life Sciences [q-bio], 02 engineering and technology, General Medicine, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 0104 chemical sciences, Förster resonance energy transfer, Quantum dot, Biophysics, [CHIM]Chemical Sciences, 0210 nano-technology, ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2018

27. Autofluorescence-Free Live-Cell Imaging Using Terbium Nanoparticles

Author: M. Cardoso Dos Santos, Ka-Leung Wong, Loïc J. Charbonnière, Joan Goetz, Niko Hildebrandt, H. Bartenlian, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)
Subjects: Cell Survival, [SDV]Life Sciences [q-bio], Biomedical Engineering, Pharmaceutical Science, chemistry.chemical_element, Nanoparticle, Bioengineering, Terbium, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Fluorescence, Live cell imaging, Microscopy, Humans, ComputingMilieux_MISCELLANEOUS, Fluorescent Dyes, Pharmacology, Chemistry, Vesicle, Organic Chemistry, Optical Imaging, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Autofluorescence, Microscopy, Fluorescence, Biophysics, Nanoparticles, 0210 nano-technology, Biotechnology, Visible spectrum, HeLa Cells
Abstract: Fluorescent nanoparticles (NPs) have become irreplaceable tools for advanced cellular and subcellular imaging. While very bright NPs require excitation with UV or visible light, which can create strong autofluorescence of biological components, NIR-excitable NPs without autofluorescence issues exhibit much lower brightness. Here, we show the application of a new type of surface-photosensitized terbium NPs (Tb-NPs) for autofluorescence-free intracellular imaging in live HeLa cells. The combination of exceptionally high brightness, high photostability, and long photoluminecence (PL) lifetimes for highly efficient suppression of the short-lived autofluorescence allowed for time-gated PL imaging of intracellular vesicles over 72 h without toxicity and at extremely low Tb-NP concentrations down to 12 pM. Detection of highly resolved long-lifetime (ms) PL decay curves from small (∼10 μm2) areas within single cells within a few seconds emphasized the unprecedented photophysical properties of Tb-NPs for live-cell...
Published: 2018

28. A Route to Triggered Delivery via Photocontrol of Lipid Bilayer Properties Using Lanthanide Upconversion Nanoparticles

Author: Shashi Bhuckory, Carlos Mingoes, Niko Hildebrandt, Paola A. Rojas-Gutierrez, John A. Capobianco, Christine E. DeWolf, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Materials science, Biocompatibility, [SDV]Life Sciences [q-bio], photoswitching, Nanoparticle, Nanotechnology, photosensitive liposomes, 02 engineering and technology, cell-adhesion, engineering.material, azobenzene photoswitches, 010402 general chemistry, 01 natural sciences, release, Coating, General Materials Science, Soft matter, Lipid bilayer, vesicles, energy transfer, lanthanide upconversion nanoparticle, Bilayer, Vesicle, cholesterol, Nile red, 021001 nanoscience & nanotechnology, supported lipid bilayer, 0104 chemical sciences, drug-delivery, laser, upconverting nanoparticles, azobenzene, Drug delivery, engineering, decay times, 0210 nano-technology, light
Abstract: WOS:000461401000101; International audience; Soft matter shells for nanoparticles provide an alternative approach to the more traditional inorganic core-shell nanoconstructs. In the latter, functionality and biocompatibility often require the addition of a hydrophilic organic coating. Herein, a supported lipid bilayer serves as a soft matter shell that has the physicochemical properties that enable a dynamic system capable of providing biocompatibility, encapsulation, and release of small molecule active agents. Direct coupling of the conformal, supported lipid bilayer containing an azobenzene-derivatized lipid with an upconversion nanoparticle provides the means to photocontrol the bilayer properties, generating a multifunctional coating. The close contact of the bilayer with the nanoparticle surface eliminates problems associated with Brownian motion for the energy transfer; the mechanism of energy transfer was determined to be dominated by radiative reabsorption using time-resolved photoluminescence. Release of an encapsulated dye, as a hydrophobic drug model, was achieved with significantly lower proportions of azobenzene-derivatized lipid than has been reported for UV-triggered release from liposomes. This work highlights the potential of the described nanoconstruct for light controlled delivery applications, especially in a biological context and in vivo applications, for which NIR excitation is of paramount importance for low toxicity and deeper tissue penetration.
Published: 2018

29. Nanosurface Energy Transfer from Long-Lifetime Terbium Donors to Gold Nanoparticles

Author: Chi Chen, Clyde Midelet, Martinus H. V. Werts, Niko Hildebrandt, Shashi Bhuckory, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Rennes (ENS Rennes), Systèmes et Applications des Technologies de l'Information et de l'Energie (SATIE), École normale supérieure - Cachan (ENS Cachan)-Université Paris-Sud - Paris 11 (UP11)-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR)-École normale supérieure - Rennes (ENS Rennes)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM), and HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Streptavidin, Materials science, Photoluminescence, chemistry.chemical_element, Terbium, 02 engineering and technology, 010402 general chemistry, Photochemistry, 01 natural sciences, 7. Clean energy, chemistry.chemical_compound, Physical and Theoretical Chemistry, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, Spectroscopy, Quenching (fluorescence), Resonance, [CHIM.MATE]Chemical Sciences/Material chemistry, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, General Energy, Förster resonance energy transfer, chemistry, Colloidal gold, 0210 nano-technology
Abstract: International audience; Photoluminescence (PL) quenching by gold nanoparticles (AuNPs) is a frequently applied principle in nanobiosensing. The quenching is most often explained in terms of the Fö rster resonance energy-transfer (FRET) mechanism, and more rarely in terms of the nanosurface energy-transfer (NSET) mechanism. Although both consider nonradiative resonance energy transfer, there are significant differences in predictions of the strength and the distance-dependence of the quenching. Here, we investigate the energy transfer to AuNPs from a terbium(III)-complex (Tb) with a long (millisecond) PL decay time with the aim to provide a better understanding of the underlying energy-transfer process. The binding of Tb-labeled streptavidin (Tb-sAv) to biotinylated AuNPs (biot-AuNPs) was studied using light-scattering spectroscopy. Quenching of the PL of Tb-sAv upon binding to biot-AuNPs of different diameters (5, 30, 50, 80 nm) was studied by time-resolved PL spectroscopy. Energy-transfer efficiencies were found to be practically independent of the AuNP size. Analysis according to FRET theory yielded donor−acceptor distances that were inconsistent and far beyond the expected Tb−AuNP distance. In contrast, the NSET model yielded a good agreement between the Tb-to-AuNP surface distance estimated from the geometry of the Tb-sAv/biotin-AuNP assembly (4.5 nm) and those calculated from PL lifetime analysis, which range from 4.0 to 6.3 nm. Our findings strongly suggest that NSET (and not FRET) is the operational mechanism in PL quenching by AuNPs, which is important information for the development, characterization, and application of nanobiosensors based on PL quenching by AuNPs.
Published: 2018

30. Colloidal Nanoparticles for Signal Enhancement in Optical Diagnostic Assays

Author: Niko Hildebrandt, Oya Tagit, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Materials science, Cancer development and immune defence Radboud Institute for Molecular Life Sciences [Radboudumc 2], [SDV]Life Sciences [q-bio], Biomedical Engineering, Nanoparticle, Bioengineering, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Quantum Dots, Animals, Humans, General Materials Science, Colloids, Surface plasmon resonance, Metal nanoparticles, General Chemistry, Surface Plasmon Resonance, 021001 nanoscience & nanotechnology, Condensed Matter Physics, 0104 chemical sciences, Highly sensitive, Signal enhancement, Colloidal nanoparticles, Nanomedicine, Spectrometry, Fluorescence, Quantum dot, Nanoparticles, 0210 nano-technology, Biomarkers
Abstract: Item does not contain fulltext The use of nanotechnologies for the development of highly sensitive and affordable diagnostic assays has significantly improved the ability to detect and characterize multiple types of biomarkers. Semiconductor and metal nanoparticles with unique optical properties have been successfully integrated within biomarker detection schemes for the generation and enhancement of optical signals in label-based and label-free assays. Highly sensitive label-based diagnostics has been realized particularly via using quantum dots (QDs) as labeling probes. Similarly, many label-free techniques that are emerging as potential complements to label-based approaches benefit from signal enhancement strategies using e.g., metal nanoparticles. This review presents a concise overview of recent advances in diagnostic assays that utilize nanoparticles for the generation and enhancement of optical signals in fluorescence- and surface plasmon resonance-based techniques. Advanced diagnostic assays that utilize nanoparticles provide major improvements in detection sensitivity, which can potentially meet the challenging requirements of clinical diagnostics.
Published: 2018

31. Sensitivity Enhancement of Förster Resonance Energy Transfer Immunoassays by Multiple Antibody Conjugation on Quantum Dots

Author: Oya Tagit, Giacomo Annio, Travis L. Jennings, Niko Hildebrandt, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)
Subjects: Immunoconjugates, Cancer development and immune defence Radboud Institute for Molecular Life Sciences [Radboudumc 2], [SDV]Life Sciences [q-bio], Biomedical Engineering, Pharmaceutical Science, Nanoparticle, Bioengineering, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, All institutes and research themes of the Radboud University Medical Center, Quantum Dots, medicine, Fluorescence Resonance Energy Transfer, Humans, Terbium, Pharmacology, chemistry.chemical_classification, Immunoassay, Luminescent Agents, medicine.diagnostic_test, biology, Chemistry, Biomolecule, Organic Chemistry, Prostate-Specific Antigen, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Förster resonance energy transfer, Quantum dot, Homogeneous, Immunoglobulin G, biology.protein, Biophysics, Antibody, 0210 nano-technology, Biotechnology, Conjugate
Abstract: International audience; Quantum dots (QDs) are not only advantageous for color-tuning, improved brightness, and high stability, but their nanoparticle surfaces also allow for the attachment of many biomolecules. Because IgG antibodies (AB) are in the same size range of biocompatible QDs and the AB orientation after conjugation to the QD is often random, it is difficult to predict if few or many AB per QD will lead to an efficient AB-QD conjugate. This is particularly true for homogeneous Förster resonance energy transfer (FRET) sandwich immunoassays, for which the AB on the QD must bind a biomarker that needs to bind a second AB-FRET-conjugate. Here, we investigate the performance of Tb-to-QD FRET immunoassays against total prostate specific antigen (TPSA) by changing the number of AB per QD while leaving all the other assay components unchanged. We first characterize the AB-QD conjugation by various spectroscopic, microscopic, and chromatographic techniques and then quantify the TPSA immunoassay performance regarding sensitivity, limit of detection, and dynamic range. Our results show that an increasing conjugation ratio leads to significantly enhanced FRET immunoassays. These findings will be highly important for developing QD-based immunoassays in which the concentrations of both AB and QDs can significantly influence the assay performance.
Published: 2018

32. Direct conjugation of antibodies to the ZnS shell of quantum dots for FRET immunoassays with low picomolar detection limits

Author: Shashi Bhuckory, Loïc J. Charbonnière, Xue Qiu, Niko Hildebrandt, K. D. Wegner, Yu-Tang Wu, Lucia Mattera, Peter Reiss, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Electronique Moléculaire Organique et Hybride (LEMOH), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Institut de Chimie du CNRS (INC)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut Nanosciences et Cryogénie (INAC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV]Life Sciences [q-bio], chemistry.chemical_element, Nanotechnology, Terbium, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Catalysis, Materials Chemistry, Detection limit, chemistry.chemical_classification, Metals and Alloys, General Chemistry, Polymer, 021001 nanoscience & nanotechnology, Combinatorial chemistry, 0104 chemical sciences, 3. Good health, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Förster resonance energy transfer, chemistry, Nanocrystal, Quantum dot, Ceramics and Composites, 0210 nano-technology, Biosensor, Conjugate
Abstract: International audience; Compact and functional nanoparticle-antibody conjugates are of paramount importance for the development of quantum dot (QD)-based immunoassays. Here, we present a simple strategy to directly conjugate IgG, F(ab')2, and Fab antibodies via their endogenous disulfide groups directly to the inorganic ZnS shell of compact penicillamine-coated QDs. The functionality of the conjugates was demonstrated by terbium (Tb)-to-QD FRET immunoassays against prostate specific antigen in serum samples. Detection limits of 2.5 pM (0.080 ng mL(-1)) were 10 and 25 times lower compared to conjugation via maleimide-terminated ligands and polymer chains, respectively. These more compact, simple, and sensitive QD-antibody conjugates will be highly advantageous for nanocrystal-based biosensing applications.
Published: 2016

33. Hydrophobicity of Gold Nanoclusters Influences Their Interactions with Biological Barriers

Author: Jean-Luc Coll, Jean-Baptiste Fleury, Hanako Okuno, Francisca Gómez, Adriana Ariza, Estelle Porret, Xavier Le Guével, Niko Hildebrandt, Maria I. Montañez, Akram Yahia-Ammar, Lucie Sancey, Angela Martin-Serrano, Ralf Seeman, Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Biomedical Research Institute of Málaga (IBIMA), Laboratoire d'Etude des Matériaux par Microscopie Avancée (LEMMA ), Modélisation et Exploration des Matériaux (MEM), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire d'océanographie et de biogéochimie (LOB), Université de la Méditerranée - Aix-Marseille 2-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut d'électronique fondamentale (IEF), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Experimental Physics, Saarland University [Saarbrücken], Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Cancer Targets & Experimental Therapeutics, Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Subjects: Absorption (pharmacology), Materials science, Membrane insertion, General Chemical Engineering, Microfluidics, Nanotechnology, 02 engineering and technology, General Chemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 0104 chemical sciences, Nanoclusters, Metal, Colloid, Colloidal gold, visual_art, Materials Chemistry, visual_art.visual_art_medium, [PHYS.COND.CM-MS]Physics [physics]/Condensed Matter [cond-mat]/Materials Science [cond-mat.mtrl-sci], [PHYS.COND]Physics [physics]/Condensed Matter [cond-mat], 0210 nano-technology, Lipid bilayer, ComputingMilieux_MISCELLANEOUS
Abstract: Understanding how ultrasmall gold nanoparticles (metal core ∼1–1.5 nm), so-called gold nanoclusters (Au NCs), interact with biological barriers has become highly important for their future bioapplications. The properties of Au NCs with tunable hydrophobicity were extensively characterized in three different biological situations: (i) interaction with serum in solution, (ii) interaction with synthetic free-standing lipid bilayers integrated in a microfluidic device, and (iii) cell studies with two different cell types (U87MG human primary glioblastoma and A375 melanoma cell lines). Our results indicate a significant impact of the precise tailoring of the hydrophilicity/hydrophobicity balance on the Au NC surfaces, which could prevent the formation of biomolecular absorption while maintaining excellent colloidal stability in solutions with high serum contents. Increasing the surface hydrophobicity of the Au NCs enabled more efficient lipid bilayer membrane insertion and induced faster cellular uptake. We sh...
Published: 2017

34. Real-Space Investigation of Energy Transfer through Electron Tomography

Author: Niko Hildebrandt, Yohei Ishida, Thomas Pons, Ikumi Akita, Tetsu Yonezawa, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Département Biochimie, Biophysique et Biologie Structurale (B3S), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Fluorophore, Materials science, Nanostructure, [SDV]Life Sciences [q-bio], Supramolecular chemistry, 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 7. Clean energy, Fluorescence spectroscopy, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Supramolecular assembly, chemistry.chemical_compound, General Energy, Förster resonance energy transfer, Electron tomography, chemistry, Chemical physics, Quantum dot, Physical and Theoretical Chemistry, 0210 nano-technology
Abstract: We herein report the first real-space investigation of Forster resonance energy transfer (FRET) in two different types of quantum dot (QD) supramolecular assemblies by observing their three-dimensional (3D) configurations through high-resolution electron tomography. Owing to its critical role in photosynthesis, artificial light-harvesting antennas, and investigation of protein–protein interactions, the mechanism of FRET has been intensively studied by monitoring its excited-state dynamics via various spectroscopic techniques. The utilized electron tomography technique allowed the direct localization of 3D coordinates of individual QDs in self-assembled nanostructures and theoretical estimation of the FRET efficiency of a single fluorophore, domain, or supramolecular assembly. Moreover, the experimental value of the FRET efficiency determined by fluorescence spectroscopy was in good agreement with the magnitude obtained via electron tomography. We believe that the described strategy can be used in single-m...
Published: 2017

35. Fluorescence Sensing of Circulating Diagnostic Biomarkers Using Molecular Probes and Nanoparticles

Author: Oya Tagit, Niko Hildebrandt, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Materials science, Cancer development and immune defence Radboud Institute for Molecular Life Sciences [Radboudumc 2], [SDV]Life Sciences [q-bio], Nanoparticle, Bioengineering, Fluorescence sensing, Nanotechnology, quantum dots, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Fluorescence, Diagnostic biomarker, Disease biomarker, Instrumentation, clinical diagnostics, Fluid Flow and Transfer Processes, business.industry, Process Chemistry and Technology, 021001 nanoscience & nanotechnology, 3. Good health, 0104 chemical sciences, immunoassays, Förster resonance energy transfer, FRET, Nanoparticles, biosensing, Photonics, 0210 nano-technology, Molecular probe, business, Biosensor
Abstract: The interplay of photonics, nanotechnology, and biochemistry has significantly improved the identification and characterization of multiple types of biomarkers by optical biosensors. Great achievements in fluorescence-based technologies have been realized, for example, by the advancement of multiplexing techniques or the introduction of nanoparticles to biochemical and clinical research. This review presents a concise overview of recent advances in fluorescence sensing techniques for the detection of circulating disease biomarkers. Detection principles of representative approaches, including fluorescence detection using molecular fluorophores, quantum dots, and metallic and silica nanoparticles, are explained and illustrated by pertinent examples from the recent literature. Advanced detection technologies and material development play a major role in modern biosensing and consistently provide significant improvements toward robust, sensitive, and versatile platforms for early detection of circulating diagnostic biomarkers.
Published: 2017

36. Time-Gated FRET Detection for Multiplexed Biosensing

Author: Niko Hildebrandt, Jurriaan M. Zwier, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and Geddes, Chris D.
Subjects: chemistry.chemical_classification, Materials science, Photoluminescence, Biomolecule, [SDV]Life Sciences [q-bio], chemistry.chemical_element, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, 7. Clean energy, 01 natural sciences, Fluorescence, 0104 chemical sciences, Förster resonance energy transfer, chemistry, Microscopy, 0210 nano-technology, Europium, Spectroscopy, Biosensor, ComputingMilieux_MISCELLANEOUS
Abstract: Forster resonance energy transfer (FRET) has become an indispensable technique for biosensing in both microscopy and spectroscopy. FRET based on photoluminescence intensity and lifetime is both well known. However, their combination into time-gated (TG) intensity detection has been much less exploited, although many commercial diagnostic/screening assays and academic studies concerning TG FRET have demonstrated the successful application since more than two decades. In this chapter we explain the theoretical and practical background of TG FRET and discuss various applications. In particular, multiplexed detection of biomolecules and/or biomolecular interactions using photoluminescence spectroscopy and microscopy and the advantages and limitations of TG FRET compared to other technologies will be discussed.
Published: 2017

37. Facile synthesis of carbon quantum dot/silver nanocomposite and its application for colorimetric detection of methimazole

Author: Hamideh Asadollahi, Marta de Frutos, Zhenlun Song, Mohammad Amjadi, Niko Hildebrandt, Tooba Hallaj, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Materials science, Calibration curve, [SDV]Life Sciences [q-bio], chemistry.chemical_element, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Silver nanoparticle, Nano, Materials Chemistry, Electrical and Electronic Engineering, Surface plasmon resonance, Instrumentation, ComputingMilieux_MISCELLANEOUS, Detection limit, Nanocomposite, Metals and Alloys, 021001 nanoscience & nanotechnology, Condensed Matter Physics, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, chemistry, Quantum dot, 0210 nano-technology, Carbon, Nuclear chemistry
Abstract: Here we present a very simple and fast method for preparation of carbon quantum dot/silver (CQD/Ag) nanocomposites at room temperature. Glucose-derived CQDs were prepared by a facile microwave-assisted method and used as both reducing and stabilizing agents for synthesis of CQD/Ag nanocomposites. We found that a unique interaction between as-prepared nanocomposites and anti-thyroid drug methimazole occurs in the solution, which results in a sharp color change from yellow to red. At the same time, the intensity of surface plasmon resonance peak of CQD/Ag nanocomposites at 400 nm decreases and a new peak appears at higher wavelengths. This finding formed a basis for developing a new colorimetric detection method for methimazole. The calibration curve for this drug was linear in the concentration range of 2.0–40 μg L−1 with a detection limit of 1.0 μg L−1. The method was applied to the determination of methimazole in urine samples with satisfactory results.
Published: 2017

38. Multiplexed Nucleic Acid Hybridization Assays Using Single-FRET-Pair Distance-Tuning

Author: Xue Qiu, Jiajia Guo, Niko Hildebrandt, Alexandra Petreto, Igor L. Medintz, Zongwen Jin, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Materials science, Photoluminescence, business.industry, [SDV]Life Sciences [q-bio], Nanoparticle, 02 engineering and technology, General Chemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, Multiplexing, 01 natural sciences, 0104 chemical sciences, Background noise, Biomaterials, Förster resonance energy transfer, Quantum dot, Optoelectronics, General Materials Science, business, 0210 nano-technology, Biosensor, Image resolution, Biotechnology
Abstract: Multiplexed photoluminescence (PL) detection plays an important role in chemical and biological sensing. Here, it is shown that time-gated (TG) detection of a single terbium-donor-based Forster resonance energy transfer (FRET) pair can be used to selectively quantify low nanomolar concentrations of multiple DNAs or microRNAs in a single sample. This study demonstrates the applicability of single-TG-FRET-pair multiplexing for molecular (Tb-to-dye) and nanoparticle (Tb-to-quantum-dot) biosensing. Both systems use acceptor-sensitization and donor-quenching for quantifying biomolecular recognition and modification of the donor–acceptor distance for tuning the PL decays. TG intensity detection provides extremely low background noise and a quick and simple one-step assay format. Single-TG-FRET-pair multiplexing can be combined with spectral and spatial resolution, paving the way for biosensing with unprecedented high-order multiplexing capabilities.
Published: 2017

39. Core or Shell? Er3+ FRET Donors in Upconversion Nanoparticles

Author: Fiorenzo Vetrone, Shashi Bhuckory, Akram Yahia-Ammar, Eva Hemmer, Yu-Tang Wu, Niko Hildebrandt, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Énergie Matériaux Télécommunications - INRS (EMT-INRS), and Institut National de la Recherche Scientifique [Québec] (INRS)-Université du Québec à Montréal = University of Québec in Montréal (UQAM)
Subjects: Lanthanide, Photoluminescence, Chemistry, [SDV]Life Sciences [q-bio], Doping, Nanoparticle, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, 7. Clean energy, 01 natural sciences, Fluorescence, Photon upconversion, 0104 chemical sciences, Inorganic Chemistry, Förster resonance energy transfer, 0210 nano-technology, Biosensor, ComputingMilieux_MISCELLANEOUS
Abstract: Upconversion nanoparticles (UCNPs) are of high interest for biosensing because of their unique near-infrared-excitation and visible-emission features. An emerging field within UCNP biosensing is the detection of biological interactions through Forster resonance energy transfer (FRET). However, the relatively large size, the distribution of emitting lanthanide ions within the nanoparticle, the unknown photoluminescence (PL) quantum yields (QY) of these emitting ions, and the many available core–shell architectures make the interpretation of UCNP-based FRET data extremely difficult. Here, we present a detailed spectroscopic study of three types of NaGdF4:Er3+,Yb3+ UCNPs with and without shells and lanthanide-ion doping in the cores or the shells. The different architectures strongly influence the brightness and PL lifetimes of the UCNPs, which are important properties for FRET to Cy3.5 dyes attached to the UCNP surfaces through DNA. Analysis of the FRET-sensitized dye PL decays allows the determination of the FRET efficiencies, which, in turn, can be used to estimate donor–acceptor distances, Forster distances, and Er3+ donor QYs, all of which are difficult to assess by other methods.
Published: 2017

40. Three-Dimensional Solution-Phase Förster Resonance Energy Transfer Analysis of Nanomolar Quantum Dot Bioconjugates with Subnanometer Resolution

Author: Michael H. Stewart, K. David Wegner, Ramasis Goswami, Niko Hildebrandt, Frank Morgner, Kimihiro Susumu, Igor L. Medintz, Eunkeu Oh, and Publica
Subjects: Bioconjugation, Chemistry, General Chemical Engineering, Resolution (electron density), Nanotechnology, 02 engineering and technology, General Chemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, 7. Clean energy, 01 natural sciences, Solution phase, 0104 chemical sciences, Characterization (materials science), Förster resonance energy transfer, Quantum dot, Materials Chemistry, 0210 nano-technology, Luminescence, Biosensor
Abstract: Luminescent semiconductor quantum dots (QDs) play an important role in optical biosensing and, in particular, in FRET (Forster resonance energy transfer)-based luminescent probes. The QD materials that form the basis for these probes are in actuality quite heterogeneous and consist of different types of QDs with variations in material compositions, surface coatings, and available biofunctionalization strategies. To optimize their role in active sensors that rely on FRET, extensive physicochemical characterization is required. A technique that can provide precise information about size, shape, and bioconjugation properties of different QD-biomolecule conjugates from a single sample and measurement under actual experimental biosensing conditions would therefore be highly important for advancing QDs to a next generation nanobiosensing tool. Here, we present a detailed FRET study on a large set of QD-biomolecule conjugates, which allows for a homogeneous solution-phase size, shape, and bioconjugation analysis of peptide and protein self-assembled QDs at subnanomolar concentrations and with subnanometer resolution. Direct incorporation of luminescent Tb-complexes (Tb) in the peptides or proteins leads to Tb-to-QD FRET upon assembly to the different QD surfaces. Luminescence decay times and time-gated intensities, which precisely decode the FRET interactions, provide a wealth of useful information on the underlying composite structure and even biochemical functionality. In contrast to other high-resolution techniques, which require rather sophisticated instrumentation, well-defined experimental conditions, and low sample throughput, our technique uses a commercial time-resolved fluorescence plate reader for very fast and simple data acquisition of many aqueous samples in a standard microtiter plate.
Published: 2014

41. Recent developments in lanthanide-to-quantum dot FRET using time-gated fluorescence detection and photon upconversion

Author: Marcelina Cardoso Dos Santos, Niko Hildebrandt, Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: Lanthanide, [PHYS]Physics [physics], [SDV]Life Sciences [q-bio], Supramolecular chemistry, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, Fluorescence, Photon upconversion, 0104 chemical sciences, Analytical Chemistry, Förster resonance energy transfer, Quantum dot, [CHIM]Chemical Sciences, 0210 nano-technology, Luminescence, Biosensor, Spectroscopy, ComputingMilieux_MISCELLANEOUS
Abstract: Lanthanide (Ln) ions and quantum dots (QD) provide us with exceptional photophysical properties that cannot be found in any other luminescent material. Long luminescence lifetimes of supramolecular Ln complexes, combination of near infrared excitation and visible luminescence of Ln-doped upconversion nanoparticles, and color-tunability and high brightness of QDs have therefore been widely exploited for bioanalytical applications. One of the most frequently used techniques for analyzing biomolecular interactions is FRET (Forster resonance energy transfer), and the Ln-QD donor-acceptor FRET pair is one of the most versatile tools for FRET biosensing. Progress of technology development in biology, chemistry, and physics has significantly advanced Ln-to-QD FRET over the last five years, and current biosensing approaches include multiplexed detection of microRNAs, homogeneous clinical immunoassays, analysis of QD-bioconjugate morphology, and intra- and extracellular biosensing. Here, we will highlight the concepts and advantages of Ln-to-QD FRET and discuss recent progress and emerging applications.
Published: 2016

42. Six-Color Time-Resolved Förster Resonance Energy Transfer for Ultrasensitive Multiplexed Biosensing

Author: Stefan Stufler, Hans-Gerd Löhmannsröben, Niko Hildebrandt, Daniel Geißler, and Publica
Subjects: Lung Neoplasms, Time Factors, Color, Nanotechnology, Biosensing Techniques, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Biochemistry, Multiplexing, Catalysis, Colloid and Surface Chemistry, Carcinoma, Non-Small-Cell Lung, Biomarkers, Tumor, Fluorescence Resonance Energy Transfer, medicine, Humans, Terbium, Volume concentration, Fluorescent Dyes, Detection limit, medicine.diagnostic_test, business.industry, Chemistry, General Chemistry, 021001 nanoscience & nanotechnology, Small Cell Lung Carcinoma, 0104 chemical sciences, Förster resonance energy transfer, Immunoassay, Background suppression, Institut für Chemie, Optoelectronics, 0210 nano-technology, business, Fluorescent glucose biosensor, Biosensor
Abstract: Simultaneous monitoring of multiple molecular interactions and multiplexed detection of several diagnostic biomarkers at very low concentrations have become important issues in advanced biological and chemical sensing. Here we present an optically multiplexed six-color Forster resonance energy transfer (FRET) biosensor for simultaneous monitoring of five different individual binding events. We combined simultaneous FRET from one Tb complex to five different organic dyes measured in a filter-based time-resolved detection format with a sophisticated spectral crosstalk correction, which results in very efficient background suppression. The advantages and robustness of the multiplexed FRET sensor were exemplified by analyzing a 15-component lung cancer immunoassay involving 10 different antibodies and five different tumor markers in a single 50 mu L human serum sample. The multiplexed biosensor offers clinically relevant detection limits in the low picomolar (ng/mL) concentration range for all five markers, thus providing an effective early screening tool for lung cancer with the possibility of distinguishing small-cell from non-small-cell lung carcinoma. This novel technology will open new doors for multiple biomarker diagnostics as well as multiplexed real-time imaging and spectroscopy.
Published: 2013
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43. Influence of luminescence quantum yield, surface coating, and functionalization of quantum dots on the sensitivity of time-resolved FRET bioassays

Author: Nicolas Lequeux, Emerson Giovanelli, Thomas Pons, Simon M. Fairclough, Travis L. Jennings, Vaibhav Jain, Phung Thi Lanh, Niko Hildebrandt, K. David Wegner, Jason M. Smith, and Eunkeu Oh
Subjects: Materials science, Luminescence, chemistry.chemical_element, Terbium, Nanotechnology, 02 engineering and technology, Luminescence quantum yield, engineering.material, 010402 general chemistry, 01 natural sciences, Sensitivity and Specificity, Coating, Quantum Dots, Fluorescence Resonance Energy Transfer, General Materials Science, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Surface coating, Förster resonance energy transfer, chemistry, Quantum dot, engineering, Surface modification, Nanoparticles, Biological Assay, 0210 nano-technology, Europium
Abstract: In clinical diagnostics, homogeneous time-resolved (TR) FRET immunoassays are used for fast and highly sensitive detection of biomarkers in serum samples. The most common immunoassay format is based on europium chelate or cryptate donors and allophycocyanin acceptors. Replacing europium donors with terbium complexes and the acceptors with QDs offers large photophysical advantages for multiplexed diagnostics, because the Tb-complex can be used as FRET donor for QD acceptors of different colors. Water-soluble and biocompatible QDs are commercially available or can be synthesized in the laboratory using many available recipes from the literature. Apart from the semiconductor material composition, an important aspect of choosing the right QD for TR-FRET assays is the thickness of the QD coating, which will influence the photophysical properties and long-term stability as well as the donor-acceptor distance and FRET efficiency. Here we present a detailed time-resolved spectroscopic study of three different QDs with an emission maximum around 605 nm for their application as FRET acceptors (using a common Tb donor) in TR-bioassays: (i) Invitrogen/Life Technologies Qdot605, (ii) eBioscience eFluorNC605 and iii) ter-polymer stabilized CdSe/CdS/ZnS QDs synthesized in our laboratories. All FRET systems are very stable and possess large Förster distances (7.4-9.1 nm), high FRET efficiencies (0.63-0.80) and low detection limits (0.06-2.0 pM) within the FRET-bioassays. Shapes, sizes and the biotin/QD ratio of the biocompatible QDs could be determined directly in the solution phase bioassays at subnanomolar concentrations. Both commercial amphiphilic polymer/lipid encapsulated QDs and self-made ligand-exchanged QDs provide extremely low detection limits for highly sensitive TR-FRET bioassays.
Published: 2016

44. Recent developments in Förster resonance energy transfer (FRET) diagnostics using quantum dots

Author: Daniel Geißler, Niko Hildebrandt, Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Subjects: [PHYS]Physics [physics], Clinical Laboratory Techniques, Chemistry, [SDV]Life Sciences [q-bio], Nanoparticle, Nanotechnology, Biosensing Techniques, 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, Biochemistry, Fluorescence, 0104 chemical sciences, Analytical Chemistry, Förster resonance energy transfer, Semiconductor quantum dots, Quantum dot, Quantum Dots, Fluorescence Resonance Energy Transfer, [CHIM]Chemical Sciences, Light excitation, 0210 nano-technology, Biosensor, ComputingMilieux_MISCELLANEOUS
Abstract: The exceptional photophysical properties and the nanometric dimensions of colloidal semiconductor quantum dots (QD) have strongly attracted the bioanalytical community over the last approximately 20 y. In particular, the integration of QDs in the analysis of biological components and interactions, and the related diagnostics using Förster resonance energy transfer (FRET), have allowed researchers to significantly improve and diversify fluorescence-based biosensing. In this TRENDS article, we review some recent developments in QD-FRET biosensing that have implemented this technology in electronic consumer products, multiplexed analysis, and detection without light excitation for diagnostic applications. In selected examples of smartphone-based imaging, single- and multistep FRET, steady-state and time-resolved spectroscopy, and bio/chemiluminescence detection of QDs used as both FRET donors and acceptors, we highlight the advantages of QD-based FRET biosensing for multiplexed and sensitive diagnostics. Graphical Abstract Quantum dots (QDs) can be applied as donors and/or acceptors for Förster resonance energy transfer- (FRET-) based biosensing for multiplexed and sensitive diagnostics in various assay formats.
Published: 2016

45. Terbium complex to quantum dot Förster resonance energy transfer for homogeneous and multiplexed microRNA assay (Conference Presentation)

Author: Xue Qiu and Niko Hildebrandt
Subjects: Detection limit, Materials science, 010401 analytical chemistry, Stacking, Nanotechnology, 02 engineering and technology, 021001 nanoscience & nanotechnology, 01 natural sciences, Multiplexing, 0104 chemical sciences, Förster resonance energy transfer, Quantum dot, 0210 nano-technology, Luminescence, Biosensor, Plate reader
Abstract: The importance of microRNA (miRNA) dysregulation in the development and progression of diseases has made these short-length nucleic acids to next generation biomarkers. Tb-to-QD Forster resonance energy transfer (FRET) has several unique advantages over organic dye-based FRET systems for biomolecular sensing. Large Forster distances (6-11 nm) offer much high FRET efficiencies, exceptionally long Tb excited-state lifetimes (ms) enable time-gated detection void of autofluorecence background, and the narrow, symmetric, and tunable emission bands of QDs provide unrivaled potential for multiplexing. Here we report a rapid and homogeneous method to sensitively detect three different miRNAs (hsa-miR-20a-5p, hsa-miR-20b-5p, and hsa-miR-21-5p) from a single 150 µL sample based on multiplexed FRET between a luminescent Lumi4-Tb complex and three different QDs. The biosensing approach exploits both base pairing and stacking. Careful design and optimization of sequence lengths and orientations of the QD and Tb-DNA conjugates was performed to provide maximum selectivity and sensitivity for all three miRNA biomarkers. The assays work at room temperature and were designed for their application on a KRYPTOR diagnostic plate reader system.Only 30 min of sample incubation and 7.5 s of measurement are required to obtain ca. 1 nM (subpicomol) detection limits. We also demonstrate precise multiplexed measurements of these miRNAs at different and varying concentrations and the feasibility of adapting the technology to point-of-care testing (POCT) in buffer containing 10% serum. Our assay does not only demonstrate an important milestone for the integration of quantum dots to multiplexed clinical diagnostics but also a unique rapid miRNA detection technology that is complimentary to the rather complicated high-throughput and high-sensitivity approaches that are established today.
Published: 2016

46. Compact quantum dot-antibody conjugates for FRET immunoassays with subnanomolar detection limits

Author: David Djurado, Peter Reiss, K. David Wegner, Niko Hildebrandt, Loïc J. Charbonnière, Christophe Lincheneau, Shashi Bhuckory, Fabio Agnese, Xue Qiu, Lucia Mattera, Tim Senden, Laboratoire d'Electronique Moléculaire Organique et Hybride (LEMOH), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Institut de Chimie du CNRS (INC)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Institut d'électronique fondamentale (IEF), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Etude des Matériaux par Microscopie Avancée (LEMMA ), Modélisation et Exploration des Matériaux (MEM), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), ANR-12-NANO-0007,NanoFRET,Analyses Fluoro-Immunologiques Multiples par Transfert d'Energie vers des Nanocristaux Semiconducteurs(2012), AII - Amsterdam institute for Infection and Immunity, APH - Amsterdam Public Health, Global Health, Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, Serum, Photoluminescence, Immunoconjugates, InP/ZnS Nanocrystals, Analytical chemistry, 02 engineering and technology, Semiconductor Nanocrystals, 010402 general chemistry, 01 natural sciences, Antibodies, Resonance Energy-Transfer, chemistry.chemical_compound, Materials Science(all), Limit of Detection, Quantum Dots, Lanthanides, Fluorescence Resonance Energy Transfer, Humans, General Materials Science, Maleimide, chemistry.chemical_classification, Immunoassay, [PHYS]Physics [physics], Bioconjugation, Biomolecule, Prostate-Specific Antigen, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Förster resonance energy transfer, chemistry, Quantum dot, Surface modification, 0210 nano-technology, Luminescence, Time-Resolved Fluoroimmunoassay, Nuclear chemistry
Abstract: International audience; A novel two-step approach for quantum dot (QD) functionalization and bioconjugation is presented, which yields ultra-compact, stable, and highly luminescent antibody-QD conjugates suitable for use in FRET immunoassays. Hydrophobic InPZnS/ZnSe/ZnS (emission wavelength: 530 nm), CdSe/ZnS (605 nm), and CdSeTe/ZnS (705 nm) QDs were surface functionalized with zwitterionic penicillamine, enabling aqueous phase transfer under conservation of the photoluminescence properties. Post-functionalization with a heterobifunctional crosslinker, containing a lipoic acid group and a maleimide function, enabled the subsequent coupling to sulfhydryl groups of proteins. This was demonstrated by QD conjugation with fragmented antibodies (F(ab)). The obtained F(ab)-QD conjugates range among the smallest antibody-functionalized nanoprobes ever reported, with a hydrodynamic diameter 26 nm without biomolecules). The LODs of 0.8 and 3.7 ng mL(-1) obtained in 50 mu L serum samples are below the clinical cutoff level of PSA (4 ng mL(-1)) and demonstrate their direct applicability in clinical diagnostics.
Published: 2016

47. Semiconductor quantum dots for in vitro diagnostics and cellular imaging

Author: Niko Hildebrandt and Zongwen Jin
Subjects: Computer science, Point-of-Care Systems, Cytological Techniques, Bioengineering, Nanotechnology, Single sample, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, In vitro diagnostic, Semiconductor quantum dots, Quantum Dots, Animals, Humans, Volume concentration, business.industry, Cellular imaging, 021001 nanoscience & nanotechnology, 0104 chemical sciences, 3. Good health, Highly sensitive, Molecular Imaging, Molecular Diagnostic Techniques, Semiconductors, Personalized medicine, 0210 nano-technology, business, Biosensor, Biotechnology
Abstract: The need for companion diagnostics, point-of-care testing (POCT) and high-throughput screening in clinical diagnostics and personalized medicine has pushed the need for more biological information from a single sample at extremely low concentrations and volumes. Optical biosensors based on semiconductor quantum dots (QDs) can answer these requirements because their unique photophysical properties are ideally suited for highly sensitive multiplexed detection. Many different biological systems have been successfully scrutinized with a large variety of QDs over the past decade but their future as widely applied commercial biosensors is still open. In this review, we highlight recent in vitro diagnostic and cellular imaging applications of QDs and discuss milestones and obstacles on their way toward integration into real-life diagnostic and medical applications.
Published: 2012
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48. Quantum Dots as Simultaneous Acceptors and Donors in Time-Gated Förster Resonance Energy Transfer Relays: Characterization and Biosensing

Author: David K. Wegner, Anika Armstrong, Alan L. Huston, W. Russ Algar, Philip E. Dawson, Niko Hildebrandt, Juan B. Blanco-Canosa, Michael H. Stewart, and Igor L. Medintz
Subjects: Nanotechnology, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, Biochemistry, Catalysis, Colloid and Surface Chemistry, Quantum Dots, Fluorescence Resonance Energy Transfer, Terbium, Alexa Fluor, Luminescent Agents, business.industry, Chemistry, Nucleic Acid Hybridization, General Chemistry, DNA, 021001 nanoscience & nanotechnology, Acceptor, Fluorescence, 0104 chemical sciences, Microsecond, Förster resonance energy transfer, Quantum dot, Optoelectronics, 0210 nano-technology, business, Luminescence, Peptides, Biosensor
Abstract: The unique photophysical properties of semiconductor quantum dot (QD) bioconjugates offer many advantages for active sensing, imaging, and optical diagnostics. In particular, QDs have been widely adopted as either donors or acceptors in Forster resonance energy transfer (FRET)-based assays and biosensors. Here, we expand their utility by demonstrating that QDs can function in a simultaneous role as acceptors and donors within time-gated FRET relays. To achieve this configuration, the QD was used as a central nanoplatform and coassembled with peptides or oligonucleotides that were labeled with either a long lifetime luminescent terbium(III) complex (Tb) or a fluorescent dye, Alexa Fluor 647 (A647). Within the FRET relay, the QD served as a critical intermediary where (1) an excited-state Tb donor transferred energy to the ground-state QD following a suitable microsecond delay and (2) the QD subsequently transferred that energy to an A647 acceptor. A detailed photophysical analysis was undertaken for each step of the FRET relay. The assembly of increasing ratios of Tb/QD was found to linearly increase the magnitude of the FRET-sensitized time-gated QD photoluminescence intensity. Importantly, the Tb was found to sensitize the subsequent QD-A647 donor-acceptor FRET pair without significantly affecting the intrinsic energy transfer efficiency within the second step in the relay. The utility of incorporating QDs into this type of time-gated energy transfer configuration was demonstrated in prototypical bioassays for monitoring protease activity and nucleic acid hybridization; the latter included a dual target format where each orthogonal FRET step transduced a separate binding event. Potential benefits of this time-gated FRET approach include: eliminating background fluorescence, accessing two approximately independent FRET mechanisms in a single QD-bioconjugate, and multiplexed biosensing based on spectrotemporal resolution of QD-FRET without requiring multiple colors of QD.
Published: 2012
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49. Multiplexed Tracking of Protease Activity Using a Single Color of Quantum Dot Vector and a Time-Gated Förster Resonance Energy Transfer Relay

Author: Kimihiro Susumu, Igor L. Medintz, Niko Hildebrandt, Anthony P. Malanoski, W. Russ Algar, and Michael H. Stewart
Subjects: Time Factors, Molecular Sequence Data, chemistry.chemical_element, Color, Nanotechnology, Terbium, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Multiplexing, Analytical Chemistry, law.invention, Relay, law, Quantum Dots, Fluorescence Resonance Energy Transfer, Chymotrypsin, Trypsin, Amino Acid Sequence, Enzyme Assays, Chemistry, business.industry, technology, industry, and agriculture, equipment and supplies, 021001 nanoscience & nanotechnology, Fluorescence, 0104 chemical sciences, Förster resonance energy transfer, Quantum dot, Optoelectronics, 0210 nano-technology, Luminescence, business, Peptides, Biosensor
Abstract: Semiconductor quantum dots (QDs) are attractive probes for optical sensing and imaging due to their unique photophysical attributes and nanoscale size. In particular, the development of assays and biosensors based on QDs and Förster resonance energy transfer (FRET) continues to be a prominent focus of research. Here, we demonstrate the application of QDs as simultaneous donors and acceptors in a time-gated FRET relay for the multiplexed detection of protease activity. In contrast to the current state-of-the-art, which uses multiple colors of QDs, multiplexing was achieved using only a single color of QD. The other constituents of the FRET relay, a luminescent terbium complex and fluorescent dye, were assembled to QDs via peptides that were selected as substrates for the model proteases trypsin and chymotrypsin. Loss of prompt FRET between the QD and dye signaled the activity of chymotrypsin; loss of time-gated FRET between the terbium and QD signaled the activity of trypsin. We applied the FRET relay in a series of quantitative, real-time kinetic assays of increasing biochemical complexity, including multiplexed sensing, measuring inhibition in a multiplexed format, and tracking the proteolytic activation of an inactive pro-protease to its active form in a coupled, multienzyme system. These capabilities were derived from a ratiometric analysis of the two FRET pathways in the relay and permitted extraction of initial reaction rates, enzyme specificity constants, and apparent inhibition constants. This work adds to the growing body of research on multifunctional nanoparticles and introduces multiplexed sensing as a novel capability for a single nanoparticle vector. Furthermore, the ability to track both enzymes within a coupled biological system using one vector represents a significant advancement for nanoparticle-based biosensing. Prospective applications in biochemical research, applied diagnostics, and drug discovery are discussed.
Published: 2012
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50. Evaluating Quantum Dot Performance in Homogeneous FRET Immunoassays for Prostate Specific Antigen

Author: Niko Hildebrandt, Olivier Lefebvre, Karl David Wegner, Shashi Bhuckory, Xue Qiu, Institut d'électronique fondamentale (IEF), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Geometry and Probability for Motion and Action (E-MOTION), Laboratoire d'informatique GRAphique, VIsion et Robotique de Grenoble (GRAVIR - IMAG), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS)-Inria Grenoble - Rhône-Alpes, Institut National de Recherche en Informatique et en Automatique (Inria), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de Recherche en Informatique et en Automatique (Inria)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Grenoble (INPG)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de Recherche en Informatique et en Automatique (Inria)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Grenoble (INPG)-Inria Grenoble - Rhône-Alpes
Subjects: Photoluminescence, [SDV]Life Sciences [q-bio], Analytical chemistry, bioconjugation, 02 engineering and technology, lcsh:Chemical technology, 010402 general chemistry, 01 natural sciences, Biochemistry, Article, Analytical Chemistry, homogeneous, medicine, [CHIM]Chemical Sciences, antibodies, lcsh:TP1-1185, immunoassay, Electrical and Electronic Engineering, Instrumentation, ComputingMilieux_MISCELLANEOUS, clinical diagnostics, Detection limit, [PHYS]Physics [physics], Qdots, Bioconjugation, medicine.diagnostic_test, Chemistry, business.industry, terbium, time-gating, TPSA, 021001 nanoscience & nanotechnology, Atomic and Molecular Physics, and Optics, 0104 chemical sciences, Prostate-specific antigen, Förster resonance energy transfer, FRET, serum, Quantum dot, Immunoassay, Optoelectronics, 0210 nano-technology, business, Plate reader
Abstract: The integration of semiconductor quantum dots (QDs) into homogeneous Förster resonance energy transfer (FRET) immunoassay kits for clinical diagnostics can provide significant advantages concerning multiplexing and sensitivity. Here we present a facile and functional QD-antibody conjugation method using three commercially available QDs with different photoluminescence (PL) maxima (605 nm, 655 nm, and 705 nm). The QD-antibody conjugates were successfully applied for FRET immunoassays against prostate specific antigen (PSA) in 50 µL serum samples using Lumi4-Tb (Tb) antibody conjugates as FRET donors and time-gated PL detection on a KRYPTOR clinical plate reader. Förster distance and Tb donor background PL were directly related to the analytical sensitivity for PSA, ...which resulted in the lowest limits of detection for Tb-QD705 (2 ng/mL), followed by Tb-QD655 (4 ng/mL), and Tb-QD605 (23 ng/mL). Duplexed PSA detection using the Tb-QD655 and Tb-QD705 FRET-pairs demonstrated the multiplexing ability of our immunoassays. Our results show that FRET based on QD acceptors is suitable for multiplexed and sensitive biomarker detection in clinical diagnostics.
Published: 2015

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