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Chlorophyll fluorescence is a ubiquitous tool in basic and applied plant science research. Various standard commercial instruments are available for characterization of photosynthetic material like leaves or microalgae, most of which integrate the overall fluorescence signals above a certain cut-off wavelength. However, wavelength-resolved (fluorescence signals appearing at different wavelengths having different time dependent decay) signals contain vast information required to decompose complex signals and processes into their underlying components that can untangle the photo-physiological process of photosynthesis. Hence, to address this we describe an advanced chlorophyll fluorescence spectrometer - ChloroSpec - allowing three-dimensional simultaneous detection of fluorescence intensities at different wavelengths in a time-resolved manner. We demonstrate for a variety of typical examples that most of the generally used fluorescence parameters are strongly wavelength dependent. This indicates a pronounced heterogeneity and a highly dynamic nature of the thylakoid and the photosynthetic apparatus under actinic illumination. Furthermore, we provide examples of advanced global analysis procedures integrating this three-dimensional signal and relevant information extracted from them that relate to the physiological properties of the organism. This conveniently obtained broad range of data can make ChloroSpec a new standard tool in photosynthesis research., (© 2024 The Authors. Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

Gemigliptin-Rosuvastatin single-pill combination is a promising therapeutic tool in the effective control of hyperglycemia and hypercholesterolemia. Organic sensors with high quantum yields have profoundly significant applications in the pharmaceutical industry, such as routine quality control of marketed formulations. Herein, the fluorescence sensor, 2-Morpholino-4,6-dimethyl nicotinonitrile 3, (λex; 226 nm, λem; 406 nm), was synthesized with a fluorescence quantum yield of 56.86% and fully characterized in our laboratory. This sensor showed high efficiency for the determination of Gemigliptin (GEM) and Rosuvastatin (RSV) traces through their stoichiometric interactions and simultaneously fractionated by selective solvation. The interaction between the stated analytes and sensor 3 was a quenching effect. Various experimental parameters and the turn-off mechanism were addressed. The adopted approach fulfilled the ICH validation criteria and showed linear satisfactory ranges, 0.2-2 and 0.1-1 μg/mL for GEM and RSV, respectively with nano-limits of detection less than 30 ng/mL for both analytes. The synthesized sensor has been successfully applied for GEM and RSV co-assessment in their synthetic polypill with excellent % recoveries of 98.83 ± 0.86 and 100.19 ± 0.64, respectively. No statistically significant difference between the results of the proposed and reported spectrophotometric methods in terms of the F- and t-tests. Ecological and whiteness appraisals of the proposed study were conducted via three novel approaches: the Greenness Index via Spider Diagram, the Analytical Greenness Metric, and the Red-Green-Blue 12 model. The aforementioned metrics proved the superiority of the adopted approach over the previously published one regarding eco-friendliness and sustainability. Our devised fluorimetric turn-off sensing method showed high sensitivity, selectivity, feasibility, and rapidity with minimal cost and environmental burden over other sophisticated techniques, making it reliable in quality control labs., (© 2024. The Author(s).)

In this work, a flow-based spectrofluorimetric method for iodine determination was developed. The system consisted of a miniaturized chip-based flow manifold for solutions handling and with integrated spectrofluorimetric detection. A multi-syringe module was used as a liquid driver. Iodide was quantified from its catalytic effect on the redox reaction between Ce(IV) and As(III), based on the Sandell-Kolthoff reaction. The method was applied for the determination of iodine in salt, pharmaceuticals, supplement pills, and seaweed samples without off-line pre-treatment. An in-line oxidation process, aided by UV radiation, was implemented to analyse some samples (supplement pills and seaweed samples) to eliminate interferences and release iodine from organo-iodine compounds. This feature, combined with the fluorometric reaction, makes this method simpler, faster, and more sensitive than the classic approach of the Sandell-Kolthoff reaction. The method allowed iodine to be determined within a range of 0.20-4.0 µmol L -1 , with or without the in-line UV digestion, with a limit of detection of 0.028 µmol L -1 and 0.025 µmol L -1 , respectively.

A sensing platform with both ratiometric fluorescence and colorimetric responses towards copper(II) ions (Cu 2+ ) and D-penicillamine (D-pen) was constructed based on carbon dots (CDs). o-Phenylenediamine (OPD) was employed as a chromogenic development reagent for reaction with Cu 2+  to generate the oxidation product 2,3-diaminophenazine (oxOPD), which not only emits green fluorescence at 555 nm, but also quenches the blue fluorescence of CDs at 443 nm via the inner filter effect (IFE) and Förster resonance energy transfer (FRET). Additionally, oxOPD exhibits obvious absorption at 420 nm. Since the intense chelation affinity of D-pen to Cu 2+ greatly inhibits the oxidation of OPD, the intensity ratio of fluorescence at 443 nm to that at 555 nm (F 443 /F 555 ) and the absorbance at 420 nm (A 420 ) were conveniently employed as spectral response signals to represent the amount of D-pen introduced into the testing system. This dual-signal sensing platform exhibits excellent selectivity and sensitivity towards both Cu 2+ and D-pen, with low detection limits of 0.019 μM and 0.092 μM, respectively. In addition, the low cytotoxicity of the testing reagents involved in the proposed sensing platform facilitates its application for live cell imaging., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

A novel spectrofluorimetric sensing platform was designed for Ractopamine measurement in aqueous and plasma samples. d-penicillamine functionalized graphene quantum dots (DPA-GQDs) was utilized as a fluorescence probe, which was synthesized through the pyrolysis of citric acid in the presence of DPA. This one-pot down-top strategy causes to high-yield controllable synthesis method. The reaction time and probe concentration were optimized. Then, the fluorescence intensity of aqueous samples containing different Ractopamine concentrations and 500 ppm DPA-GQDs were measured at 25°C with an excitation wavelength of 274 nm. The sensing platform was also applied to detect Ractopamine in untreated plasma samples. The fluorescence spectroscopy technique responses indicated a linear relationship between the peak fluorescence intensity and ractopamine concentration in the range of 0.25-15 ppm with low limit of quantification of 0.25 ppm was for aqueous and plasma samples, respectively., (© 2021 John Wiley & Sons Ltd.)

A colorimetric and fluorescent dual-channel detection method for acid phosphatase (ACP) activity has been constructed, based on the internal filtering effect between oxidized 3,3',5,5'-tetramethylbenzidine (oxTMB) and rhodamine B (RB). Au 3+ , which in situ form gold nanoparticles (AuNPs), can oxidize colorless 3,3',5,5'-tetramethylbenzidine (TMB) to oxTMB (blue color). The fluorescence of RB can be quenched by oxTMB due to the spectral overlap of emission of RB and absorption of oxTMB. By means of the above process, ACP can be determined because ACP promotes the hydrolysis of 2-phospho-L-ascorbic acid trisodium salt (AAP) to generate ascorbic acid (AA), which can inhibit the internal filtering effect between RB and oxTMB. No material preparation was needed for the determination of ACP. The colorimetric and fluorimetric methods can quantify ACP in the range 0.06-5.0 mU/mL and 0.03-5.0 mU/mL, respectively. Furthermore, a smartphone-assisted sensing platform has been constructed for on-site monitoring of ACP in the range 0.75-50 mU/mL, and the detection limit is 0.3 mU/mL. The methods developed can measure ACP in human serum successfully., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

A smartphone-based dual-wavelength digital imaging platform containing red (539-695 nm) and blue (389-511 nm) band-pass filters was developed for point-of-care (POC) testing of alkaline phosphatase (ALP) activity. The platform was based on dual-emitting fluorescent nanohybrids (AuNC@NAN), the ratiometric probe, which had a fluorescence "on-off-on-off" response. The probe comprised red-emitting gold nanoclusters (AuNCs) acting as the signal report units and blue-emitting N-(2-aminoethyl-1,8-naphthalimide) (NAN) acting as an internal reference. The different responses of the ratiometric probes resulted in a continuous color-multiplexing change from pink-red to dark-purple upon exposure to ALP. The dual-wavelength digital imaging platform was employed to acquire images of AuNC or NAN fluorescence signals without the influence of background light. Unlike the classical one-time digital imaging mode, the accurate red (R) and blue (B) channel values of the generated images can help to directly judge or eliminate the disturbance from unavoidable interfering factors. The R/B values were successfully employed for determining the ALP activity at a range 2.0 to 35.0 mU·mL -1 with the detection limit of 1.04 mU·mL -1 . Such sensing imaging platform is also successful in determining ALP activity in human serum with 94.9-105% recoveries and relative standard deviation in the range 4.2-5.6%. A novel dual-wavelength smartphone-based digital imaging platform was proposed for simultaneous readout of the reporting and internal reference signals from dual-emitting ratiometric fluorescence probes, which allowed us to the accurate, reliable, and highly sensitive assay of ALP activity in complex samples.

We developed a facile detection method of spermine based on the fluorescence (FL) quenching of the ciprofloxacin-Tb3+ complex, which shows astrong green emission. Ciprofloxacin (CP) makes efficient bondings to Tb3+ ion as a linker molecule through carboxylic and ketone groups to form a kind of lanthanide coordination polymer. The addition of spermine that competes with Tb3+ ions for the interaction with CP due to its positive charge brings about weakened coordination linkage of CP and Tb3+. The probe exhibited high sensitivity, selectivity, and good linearity in the range of 2-180 μM with a low limit of detection of 0.17 μM. Moreover, we applied this method on the paper strip test (PST), along with the integration of a smartphone and Arduino-based device. The practical reliability of the developed probe was evaluated on human serum samples with acceptable analytical results., Competing Interests: The authors have declared that no competing interests exist.

Background: Finding and preserving normal parathyroid glands or localizing and removing diseased parathyroid glands are crucial steps to successful thyroid and parathyroid operations. Using near-infrared fluorescence detection to identify parathyroid glands during thyroid and parathyroid operations has lately gained widespread recognition, with 2 Food and Drug Administration-cleared devices currently in the market. We aim to update the endocrine surgery community on how near-infrared fluorescence detection can be most optimally used for rapid intraoperative parathyroid gland identification or preservation., Methods: A literature review was performed using the key terms: "parathyroid," "near infrared," and "fluorescence" in relevant search engines. Based on the reviewed literature and expert surgeons' opinions, recommendations were formulated for applying near-infrared fluorescence detection to identify or preserve parathyroid glands during cervical endocrine surgery., Results: The scope of near-infrared fluorescence detection can be broadly categorized into (1) using near-infrared auto-fluorescence to identify or locate both healthy and diseased parathyroid glands, and (2) using contrast-enhanced near-infrared fluorescence to evaluate parathyroid gland perfusion. The benefits and pitfalls for both near-infrared-based approaches are described herein., Conclusion: Near-infrared fluorescence detection appears helpful for identification and likely preservation of parathyroid glands. We hope these recommendations will be valuable to the practicing endocrine surgeon as they consider incorporating these intraoperative adjuncts in their surgical practice., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Thrombin is an important enzyme that plays a pivotal role in the blood clotting pathways. An imbalance in the activity of this enzyme is clinically known to be associated with various diseases, such as thrombosis, inflammation, atherosclerosis, and haemophilia, suggesting the need to devise sensors for Thrombin detection. However, the majority of the fluorescence-based Thrombin assays rely on fluorescence labelling assays or Thrombin specific recognition biomolecules, such as, aptamers or antibody which requires sophisticated techniques and makes it very expensive. Herein, we report a simple, selective, sensitive and label-free fluorescence detection scheme for Thrombin which is based on the interaction between Thrombin and a fluorescent complex of Heparin with a molecular rotor dye, Thioflavin-T. The detection scheme exploits selective interaction between cationic Thrombin and anionic Heparin to modulate the monomer-aggregate equilibrium of the Thioflavin-T-Heparin system. Importantly, the present system offers a ratiometric response that has the ability for robust quantification of Thrombin concentration even in complex medium. The involvement of all commercially available components is a crucial advantage of this detection scheme. Further, the detection scheme also shows reasonable response in diluted serum matrix., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10-18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo., (© 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

A sensor system was designed for the detection of Enoxaparin (Enox), a low molecular weight heparin (LMWH) that was run over the fluorescence quenching mechanism of fluorescein (FL) dye. At nanomolar concentrations, FL probe was subjected to fluorescence quenching by Fe(III). Fluorescence quenching mechanism of FL by Fe(III) was examined using various analytical techniques such as UV-vis absorption, fluorescence, and Fourier transform Infrared spectroscopy techniques, as well as with scanning electron microscope. The results indicated that photoinduced electron transfer process occurred between FL and Fe and that FL was quenched both statically and dynamically. Thermodynamic parameters showed that the interactions between them were predominantly hydrophobic interactions. Enox caused FL to recover its lost fluorescence properties and an increase was observed in the intensity of the fluorescence. Enox was detected successfully with the turn on fluorescence sensor. The developed Enox biosensor exhibited linearity in the range of 0-1.1 μg/ml. For Enox detection, the limit of detection was measured as 255 ng/mL. Enox biosensor was presented as a practical, simple, and applicable sensor system with high sensitivity and good selectivity. Enox is a medication usually monitored indirectly over anticoagulation. This study was presented as an alternative method for monitoring Enox directly. HIGHLIGHTS: Fluorescence quenching of Fluorescein dye by Fe(III) was studied in detail. The presence of enoxaparin enhanced the fluorescence properties of the fluorescein dye. A sensitive, simple and effective sensor system for determination of Enoxaparin, a low molecular weight heparin was shaped in the aqueous media. It was presented as a new method for Enoxaparin to be followed directly.

A thiourea-based fluorescent chemosensor NADA, (E)-2-(4-(diethylamino)-2-hydroxybenzylidene)-N-(naphthalen-1-yl)hydrazine-1-carbothioamide, has been designed and synthesized. NADA could detect Ga 3+ through a fluorescent turn-on with a low detection limit (0.29 μM). Importantly, NADA could effectively discriminate Ga 3+ from Al 3+ and In 3+ . The binding mechanism of NADA with Ga 3+ was identified by ESI-mass, NMR titration, and DFT calculations.

Methanol is highly toxic for human, so methanol detection is valuable especially in water and ethanol medium without complicated and time consuming procedure. In this work, we present a new fluorescence probe for direct detection of methanol in aqueous and ethanol medium based on the ZnS:Mn 2+ quantum dot (QDs) and soluble N-methylpolypyrrole (NMPPy) hybrid. Moreover, the number of spectroscopic techniques were used to study the chemical composition and optical properties of the resultant QDs as well as investigation on the sensing mechanism toward methanol. Also, methanol can be determined by using ZnS:Mn 2+ QDs/NMPPy hybrid based switchable fluorescence sensing system, with high sensitivity, high selectivity and a very good detection limit of 1 mM with linearity in the concentration range of 25-230 mM (~0.1-0.9% v/v) in aqueous solution. Finally, the ZnS:Mn 2+ QDs/NMPPy hybrid as optical sensor was successfully utilized to determine the amount of methanol in real alcoholic beverage samples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Capillary electrophoresis (CE) is a highly efficient separation method capable of handling small sample volumes (∼pL) and low (∼yoctomole) detection limits and, as such, is ideal for applications that require high sensitivity, such as single-cell analysis (Chen et al. Anal. Chem. 1996 , 68 (4), 690-696; Cohen et al. Annu. Rev. Anal. Chem. 2008 , 1 (1), 165-190; Vickerman et al. ACS Chem. Biol. 2018 , 13 (7), 1741-1751). Low-cost CE instrumentation is quickly expanding, but low-cost, open-source fluorescence detectors with ultrasensitive detection limits are lacking (Vickerman et al. ACS Chem. Biol. 2018 , 13 (7), 1741-1751; Fang et al. Electrophoresis 2016 , 37 (17-18), 2376-2383; Casto et al. Anal. Chem. 2019 , 40 (1), 65-78). Silicon photomultipliers (SiPM) are inexpensive, low-footprint detectors with the potential to fill the role as a detector when cost, size, and customization are important. In this work, we demonstrate the use of a SiPM in CE with zeptomolar detection limits and a dynamic range spanning 5 orders of magnitude, comparable to photomultiplier detectors. The performance of these detectors was measured using a continuous wave excitation laser in an epifluorescence detection configuration. We characterize the performance of the SiPM as a highly sensitive detector by measuring enzyme activity in single cells. This simple, small footprint, and low-cost (<$130) light detection circuit will be beneficial for open-source, portable, and budget-friendly instrumentation requiring high sensitivity.

A novel molecularly imprinted ratiometric fluorescent probe was fabricated by simple sol-gel polymerization for selective and sensitive assay of C-type natriuretic peptide (CNP) in biosamples. Both the nitrobenzoxadiazole (NBD) and carbon dots (CDs) were located on the surface of silica, used as the detection signal and reference signal, respectively. For the turn-on-based probe, the fluorescence intensity of NBD could be quantitatively enhanced by CNP based on the strategy of photo-induced electron transfer (PET), while the fluorescence of CDs remained unchanged. The obtained probe exhibited excellent recognition selectivity and fast kinetics to CNP templates, and also showed good stability. The linear range of CNP determination was 5-80 pg mL -1 with a low detection limit of 2.87 pg mL -1 . Finally, the probe was successfully applied to determine CNP in human serum samples and attained high recoveries between 97.3 and 104% with precisions below 4.7%. The result indicates that the proposed method has promising potential for the assay of trace peptides in complex matrices. Schematic illustration for the formation and determination mechanism of the probe.

In this paper, a novel and facile synthetic method of 3-mercaptopropionic acid functionalized copper nanoclusters with aggregation-induced emission (AIE) induced by Cu 2+ (Cu 2+ @MPA-Cu NCs) was developed by a one-pot reaction as a fluorescent probe for the detection of sulfide ion (S 2- ). The prepared Cu 2+ @MPA-Cu NCs behaved as aggregated clusters and had strong pink fluorescence under 365 nm UV light with excellent fluorescence emission at 610 nm. The quantum yield increased from 0.56% to 4.8% before and after Cu 2+ added. The presence of S 2- would strongly bind to Cu 2+ , which caused the structure of the aggregated Cu 2+ @MPA-Cu NCs to be destroyed and then the fluorescence quenched. On the basis of this principle, a fluorescent probe was constructed for the detection of S 2- with a very good linearity in the range 0-600 μM ( R 2 = 0.9843) and a detection limit of 26.3 nM. Finally, the nanohybrids were successfully demonstrated for the application in the selective detection of S 2- in food additives. This study essentially paved a new avenue for effectively developing an easy sensor platform for S 2- measurements in food additives.

Carbon dots (CDs) have been a new class of fascinating carbon-based fluorescent nanomaterials. In the present work, new N-doped CDs with highly crystalline graphite structures are prepared from renewable precursors, chitosan and tartaric acid, and are well characterized. The prepared CDs are applied as a biocompatible fluorescent sensor for the sequential detection of Fe 3+ and AA. Among various transition metal ions, Fe 3+ can selectively quench the fluorescence of CDs. Upon the further addition of AA, the quenched fluorescence of CDs is then restored as Fe 3+ is reduced to Fe 2+ by AA, which can be utilized for the fluorescent determination of AA. A good linear relationship in the range of 0-150 μM of AA concentration is established with a low detection limit of 0.02 µM. Moreover, the practical applications of this fluorescent sensing method in measurement of AA in food samples are successfully realized with satisfactory results., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A reaction-based luminescence chemosensor was synthesized for sulfite detection based on a fluorescein derivative with thermally activated delayed fluorescence (TADF). The chemosensor exhibited a fluorescence turn-on effect on sulfite with good sensitivity and selectivity. Importantly, utilizing the long luminescence lifetime of the TADF compound, the chemosensor realized photoluminescence lifetime imaging for sulfite in living cells with the luminescence lifetime distribution mainly around 14 μs.

The estimation of yeast viability with B- and N-doped carbon dots (BN-CDs) was investigated in this paper. BN-CDs with a fluorescent quantum yield of 65.47% were prepared by a one-step hydrothermal method. The size distribution of BN-CDs was relatively narrow, with the majority falling within 7.5-8.5 nm, and they were mainly composed of carbon, oxygen, nitrogen, and boron. BN-CDs were shown to have strong and stable fluorescence. They exhibited excitation-independent photoluminescence property, which could avoid the autofluorescence and limitation of the excitation source. Dead and live yeast cells were distinguished well by BN-CD staining in a short time, and there was no strict requirement for light protection. The application of BN-CDs in beer brewing can solve the problem of estimation of yeast viability.

Fluorescence imaging technology has been extensively applied in chemical and biological research profiting from its high sensitivity and specificity. Much attention has been devoted to breaking the light diffraction-limited spatial resolution. However, it remains a great challenge to improve the axial resolution in a way that is accessible in general laboratories. Surface plasmon-coupled emission (SPCE), generated by the interactions between surface plasmons and excited fluorophores in close vicinity of the thin metal film, offers an opportunity for optical imaging with potential application in analysis of molecular and biological systems. Benefiting from the highly directional and distance-dependent properties, SPCE imaging (SPCEi) has displayed excellent performance in bioimaging with improved sensitivity and axial confinement. Herein, we give a brief overview of the development of SPCEi. We describe the unique optical characteristics and constructions of SPCEi systems and highlight recent advances in the use of SPCEi for biological applications. We hope this review provides readers with both the insights and future prospects of SPCEi as a new promising imaging platform for potentially widespread applications in biological research and medical diagnostics. Graphical abstract.

Fluorescence capillary analysis (FCA) realizes trace-level analysis of micro-volume samples; it is easy to operate, extremely low in analytical cost and can significantly lessen environmental pollution from analytical chemistry waste. FCA has the characteristics of green analytical chemistry and has been applied in clinical, biochemical, pharmaceutical, food safety and other fields. FCA basically involves a micro-volume glass capillary, a capillary holder and an ordinary fluorescence detector. The capillary is not only a container for chemical reaction and detection but also functions as a carrier to immobilize enzymes, gene probes or reagents; it can be used repeatedly or can be disposable. In analysis, the capillary which is modified with functional reagents sucks in a measured liquid for the reaction and is then inserted into the holder within the fluorescent detector for measurement. The immobilized FCA method has been successfully used in the determination of reduced coenzyme I, ethanol in liqueur, lactic acid in dairy products, pyruvic acid and glucose in serum, trace-level sulfated bile acid in urine, the ratio of pyruvic/lactic acid in serum, and pyruvic acid in cells as well as in DNA end-labeling and dyeing methods. Further, FCA can also be extended to capillary arrays to complete multipurpose simultaneous determinations and can be combined with mobile phones as fluorescence detectors for use in mobile health analytical technology. FCA will produce considerable social benefits in medicine, pharmacy, fermentation of food, environmental protection and other fields. Therefore, the relevant contents are presented in this tutorial review.

A novel quinazolinone based turn-on fluorescence probe for sensitive monitoring hypochlorite was prepared using the mild condensation reaction between 2-(2'-hydroxyphenyl)-4(3H)-quinazolinone derivative and 4-methylbenzenesulfonyl hydrazide. The probe exhibited specific selectivity to ClO - with obvious optical signal changes from weak fluorescence at 560 nm to a strong fluorescence emission at 520 nm and color changes from colorless to yellow, which could be noticed by the naked eye. The detection limit toward hypochlorite is as low as 11.4 nM. Moreover, the probe could sensitively response to ClO - in living cells with satisfying imaging effect and has been successfully applied to the determination of ClO - in practical water samples, which indicated that the probe has certain application potential for hypochlorite monitoring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A highly accurate, simple and sensitive spectrofluorimetric analytical method for dapagliflozin (DGF) quantitation was developed. The proposed method was successively applied to DGF analysis in both its pure and pharmaceutical dosage forms. This method was developed to investigate DGF stability in its degradation products, as laid out in International Council for Harmonisation (ICH) rules. Kinetics of alkaline degradation of DGF was also calculated. The half-life time (t 1/2 ) of the reaction was 75.32 min. An alkaline degradation pathway was described. The present study involved measurement of the second-derivative synchronous fluorescence intensity of DGF at Δλ = 30 nm. Peak amplitude was measured at 322 nm. Linear range of the calibration curve was 0.1-1.0 μg ml -1 . Lower detection and quantitation limits were 0.023 and 0.071 μg ml -1 , respectively, and indicated good sensitivity of the proposed method. Mean per cent recovery was 99.78 ± 1.78%. The proposed analytical approach was successfully applied to DGF in the quality control laboratory and would be suitable as a stability-indicating assay., (© 2019 John Wiley & Sons, Ltd.)

The direct determination of alogliptin benzoate (ALO) using fluorescence has not yet been accomplished because ALO cannot fluoresce naturally. Accordingly, it should be derivatized first with a fluorogenic reagent to enhance the sensitivity required for its bioanalysis. This method is the first spectrofluorimetric assay for ALO quantification exploiting the nucleophilic nature of its amino group to react with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in borate buffer at pH 8.5 to produce a strong fluorescent compound that is excited at and emits at wavelengths 470 and 527 nm, respectively. Experimental variables concerning the conditions of reaction and fluorogenic intensity were carefully investigated and optimized. Linearity was from 1-250 ng ml -1 with a lower detection limit of 0.29 ng ml -1 and a lower quantification limit of 0.88 ng ml -1 . Validation of the current study was accomplished with mean per cent recovery of 100.62 ± 1.59 in tablets and 99.86 ± 0.82 in human plasma. Furthermore, the current method has been utilized in the bioanalysis of ALO in real rat plasma after oral administration with a simple specimen preparation. The developed method has proven to be a promising alternative method for ALO analysis in bioequivalence studies., (© 2019 John Wiley & Sons, Ltd.)

Rifampicin is a common antibiotic used in human and veterinary medicine to treat tuberculosis and other diseases caused by numerous pathogenic bacteria. However, the excessive or improper use of rifampicin usually leads to a series of problems, including bacterial resistance, excessive drug-resistance and water pollution. Thus, it is of great importance to develop selective and sensitive assays for monitoring rifampicin in biological systems. In this study, we designed a fluorescence "turn-off" strategy for the trace detection of rifampicin based on a glutathione-stabilized copper nanoclusters (GSH-Cu NC) sensor. In an aqueous solution, the fluorescence of the GSH-Cu NCs at 632 nm can be quenched effectively and selectively by rifampicin due to the inner-filter effect (IFE) of fluorescence mechanism. Distinctively, this GSH-Cu NC sensor exhibited excellent fluorescence sensing capability for the trace detection of rifampicin with a very low limit of detection (LOD) of 16 pM in a wide linear range from 50 to 10 000 pM. It is not only more sensitive than the other methods previously reported for the detection of rifampicin, but also has an outstanding selectivity and strong anti-interference in complex samples. Furthermore, the as-developed GSH-Cu NCs were also successfully applied to determine rifampicin in different real samples with quantitative spike recoveries ranging from 97% to 105%.

Cellulose paper has strong potential as an analytical platform owing to its unique characteristics. In the present study, we investigated a procedure for functionalizing the surface of cellulose paper by dip-coating a mixture of a functional polymer and a perfluoroalkylated surfactant (surfactant 1). The functional polymer comprised a mixture of methyl methacrylate and poly(ethylene glycol) methacrylate monomers. The monomer ratio in the functional polymer affected the hydrophilicity and water absorbance of the cellulose paper after dip-coating. Furthermore, the presence of surfactant 1 during dip-coating promoted the surface segregation of poly(ethylene glycol) (PEG) moieties in the polymer, which enhanced the hydrophilicity, prevented nonspecific protein adsorption, and maintained the water absorbance of the dip-coated cellulose paper. Dip-coating with another functional polymer containing biotin groups produced a cellulose paper with a biotin-decorated surface in a one-step procedure. The displayed biotin groups immobilized avidin on the surface, and the PEG moieties in the polymer prevented nonspecific protein adsorption. We then immobilized a thrombin-binding DNA aptamer on the avidin-immobilized cellulose paper to prepare a paper-based analytical device. It is possible to visualize thrombin in model solutions and serum using the paper-based analytical device.

We present a dual-channel mobile lidar system based on laser-induced fluorescence (LIF) for real-time standoff detection and concentration distribution analysis of tryptophan. The system employs an ultraviolet laser excitation source and signal detectors for receiving fluorescence signals within two different wavelength bands. The performed experiments measured tryptophan aerosols at two different standoff distances. Moreover, distilled water and ethanol solutions were also detected for comparison. The results show that the system can detect LIF signals of tryptophan, give early warnings, locate the diffusion sources, and monitor the variation of the aerosol concentration distribution in real time.

Alkaline phosphatase (ALP) is a membrane-bound enzyme widely present in biological tissues and is extensively used as a biomarker in clinical diagnosis because abnormal ALP levels in organisms are often closely related to many diseases. In order to meet the urgent needs of human health and the development of precision medicine, the classic method of detecting ALP seems to be unable to meet a "tailor-made" treatment plan for patients. In order to get the best therapeutic effect and the lowest side effects, the researchers have made considerable efforts and established numerous strategies for detecting ALP. In this review, we focus on the current development and basic principles of fluorescence strategy to detect ALP, and compare the advantages and disadvantages of detecting ALP based on different fluorescent detection modes. In addition, to more intuitively understand the performance of different materials for detecting ALP, a table is presented. Finally, the future trends and prospective in this field will be discussed, it could be speculated that invent and design the detection mechanism based on the relationship between the new material and ALP is the key to effective detection of ALP, and process data with a computer analog digital platform will be a welcome and obsessive technology. It is hoped that some insights and inspiration for future research work will be provided., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A digital fluorescence detector (DFD), a handheld fluorescence detection device, can convert the fluorescence signal of samples into the corresponding fluorescer concentration. Herein, by adopting a DFD as the readout, a novel intelligent platform was developed based on a ratiometric paper-based device (RPD) for multiple aminoglycoside detection. There are five layers and four parallel channels contained in the designed RPD, functioning as reagent storage, fluidic path control and signal processing, respectively. The rationale of this design lies in the fact that aptamer/graphitic carbon nitride nanosheet (Apt/g-C 3 N 4 NS) modified layers can catalyze o -phenylenediamine to fluorescent 2,3-diaminophenazine (DAP) in the presence of H 2 O 2 . When Apt was removed from nanosheets via the Apt-target reaction, the peroxidase-like activity would be decreased, thus decreasing the production of DAP. All the changes of the fluorescence DAP signal can be read out using a portable DFD. Based on the DFD signal change related to the concentration of the target, a quantitative reaction platform was established. Furthermore, the sample flow and Apt-target reaction time can be reasonably regulated using the H 2 O 2 -cleavable hydrophobic compound modified layer placed between the target recognition region and detection region. Then, the practicality of this platform was verified through realizing sensitive analysis of streptomycin, tobramycin, and kanamycin simultaneously. Overall, with merits including portability and ease of operation, the platform shows great potential in on-site simultaneous detection of multiple targets, especially in resource-limited settings.

Timely and effective detection of bacterial pathogens is of great importance to reduce morbidity rates from bacterial infections. Recently, enzyme-activated fluorogenic probes, which invoke enzymatic catalysis to trigger fluorescence emission, have been superior sensors for bacterial infections needed for accurate diagnoses. Here, a fluorescent sensor for nitroreductase (NTR) detection is described. It is based on a cyanine fluorophore and utilizes photoinduced electron transfer to generate a rapid 10-fold fluorescence response after being catalytically reduced by NTR. It has enabled selective and sensitive visualization of NTR activity in vitro and in living bacterial pathogens. Thus, the probe has great potential to provide a rapid, noninvasive tool to diagnose infections and guide antimicrobial selection., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Single molecule detection provides detailed information about molecular structures and functions but it generally requires the presence of a fluorescent marker which can interfere with the activity of the target molecule or complicate the sample production. Detecting a single protein with its natural UV autofluorescence is an attractive approach to avoid all the issues related to fluorescence labeling. However, the UV autofluorescence signal from a single protein is generally extremely weak. Here, we use aluminum plasmonics to enhance the tryptophan autofluorescence emission of single proteins in the UV range. Zero-mode waveguide nanoapertures enable the observation of the UV fluorescence of single label-free β-galactosidase proteins with increased brightness, microsecond transit times, and operation at micromolar concentrations. We demonstrate quantitative measurements of the local concentration, diffusion coefficient, and hydrodynamic radius of the label-free protein over a broad range of zero-mode waveguide diameters. Although the plasmonic fluorescence enhancement has generated a tremendous interest in the visible and near-infrared parts of the spectrum, this work pushes further the limits of plasmonic-enhanced single molecule detection into the UV range and constitutes a major step forward in our ability to interrogate single proteins in their native state at physiological concentrations.

Non-invasive detection of urinary bladder cancer remains a significant challenge. Urinary volatile organic compounds (VOCs) are a promising alternative to cell-based biomarkers. Herein, we demonstrate a novel diagnosis system based on an optic fluorescence sensor array for detecting urinary bladder cancer VOCs biomarkers. This study describes a fluorescence-based VOCs sensor array detecting system in detail. The choice of VOCs for the initial part was based on an extensive systematic search of the literature and then followed up using urinary samples from patients with urinary bladder transitional cell carcinoma. Canonical discriminant analysis and partial least squares discriminant analysis (PLS-DA) were employed and correctly detected 31/48 urinary bladder cancer VOC biomarkers and achieved an overall 77.75% sensitivity and 93.25% specificity by PLS-DA modelling. All five urine samples from bladder cancer patients, and five healthy controls were successfully identified with the same sensor arrays. Overall, the experiments in this study describe a real-time platform for non-invasive bladder cancer diagnosis using fluorescence-based gas-sensor arrays. Pure VOCs and urine samples from the patients proved such a system to be promising; however, further research is required using a larger population sample., (© 2018 The Authors. Journal of Biophotonics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A novel ratiometric fluorescence molecularly imprinted sensor based on a dual-emission quantum dot hybrid was fabricated and used as an alternative analytical tool for the detection of tetracycline. In the synthesis process, red-emitting quantum dots (r-QDs) and green-emitting quantum dots (g-QDs) were modified by two different methods. Afterward, a stepwise precipitation polymerization imprinting reaction was performed to prepare the novel ratiometric fluorescence molecularly imprinted sensor (MIP-g/r-QD sensor). The MIP-g/r-QD sensor integrated the advantages of molecularly imprinted polymers and ratiometric fluorescence probes. The specific recognition sites in the polymer layers could adsorb tetracycline molecules, and then they caused fluorescence quenching behavior of g-QDs via an electron transfer process. Under the optimal conditions, a linear relationship was obtained covering the range from 10 to 160 μmol/L, with a correlation coefficient of 0.9976 and a high imprinting factor of about 3.3. Moreover, the novel MIP-g/r-QD sensor was successfully applied to detect tetracycline in milk samples. This work provides a new way to fabricate an efficient ratiometric fluorescence molecularly imprinted sensor based on quantum dots for convenient, fast, and highly selective and sensitive detection of organic molecules. Graphical abstract A novel ratiometric fluorescence molecularly imprinted sensor based on a dual-emission quantum dot hybrid was fabricated and used as an alternative analytical tool for the detection of tetracycline. AIBN azoisobutyronitrile, EGDMA ethylene glycol dimethacrylate, KH-570 3-(methacryloyloxy)propyltrimethoxysilane, OVDAC octadecyl-4-vinylbenzyldimethylammonium chloride, PDDA poly(diallyldimethylammonium chloride), QD quantum dot, TEOS tetraethoxysilane.

To explore the effect of signal acquisition way on screening ability, the multiplexing fiber optic laser induced fluorescence spectroscopy (LIFS) system with one-, two- and three-probe, were employed respectively to detect artificially aflatoxin B 1 (AFB 1 , 5, 10, 20, 30, and 50 ppb) contaminated 300 pistachio kernels in this study. Compared to one- and two-probe modes, highest accuracy (≥97.0%) by support vector machine (SVM) employing 390-660 nm were obtained using three-probe, which also showed the most attractive precision (root mean square error of prediction (RMSEP) < 4.5 ppb) for AFB 1 by stepwise multiple linear regression (SMLR) using 174-1100 nm. These suggested that the effective collection of spatial information could improve the performance of model, and the three-probe LIFS had a preliminary feasibility for discriminating pistachios contaminated with low concentration of AFB 1 . Further study on classifying naturally contaminated samples is needed to validate the applicability of this system., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We propose a handheld projective imaging device for orthotopic projection of near-infrared fluorescence images onto target biological tissue at visible wavelengths without any additional visual aid. The device integrates a laser diode light source module, a camera module, a projector, an ultrasonic distance sensor, a Raspberry Pi single-board computer, and a battery module in a rugged handheld unit. It is calibrated at the detected working distance for seamless coregistration between fluorescence emission and projective imaging at the target tissue site. The proposed device is able to achieve a projection resolution higher than 314  μm and a planar projection bias less than 1 mm at a projection field of view of 58  ×  108  mm2 and a working distance of 27 cm. Technical feasibility for projective imaging is verified in an ex vivo model of chicken breast tissue using indocyanine green as a fluorescence agent. Clinical utility for image-guided surgery is demonstrated in a clinical trial where sentinel lymph nodes in breast cancer patients are identified and resected under the guidance of projective imaging. Our ex vivo and in vivo experiments imply the clinical utility of deploying the proposed device for image-guided surgical interventions in resource-limited settings.

Grating-coupled surface plasmon field-enhanced fluorescence spectroscopy (GC-SPFS) with optical bound/free (B/F) separation technique was developed by employing a highly directional fluorescence with polarization of surface plasmon-coupled emission (SPCE) to realize highly sensitive immunoassay regardless of the ligand affinity. A highly sensitive immunoassay system with GC-SPFS was constructed using a plastic sensor chip reproducibly fabricated in-house by nanoimprinting and applied to the quantitative detection of an anti-lysozyme single-domain antibody (sdAb), to compare conventional washing B/F separation with optical B/F separation. Differences in the affinity of the anti-lysozyme sdAb, induced by artificial mutation of only one amino acid residue in the variable domain were attributed to higher sensitivity than that of the conventional Biacore surface plasmon resonance (SPR) system. The detection limit (LOD; means of six replicates of the zero standard plus three standard deviations) of the GC-SPFS immunoassay with optical B/F separation, was estimated to be 1.2 ng/ml with the low-affinity ligand (mutant sdAb Y52A: KD level was of the order of 10-7 ~ 10-6 M) and was clearly improved as compared to that (LOD: 9.4 ng/ml) obtained with the conventional washing B/F separation. These results indicate that GC-SPFS with the optical B/F separation technique offers opportunities to re-evaluate low-affinity biomaterials that are neither fully utilized nor widespread, and could facilitate the creation of novel and innovative methods in drug and diagnostic development., Competing Interests: We have the following interests: T.K., K.N., Y.N. are employed by Konica Minolta, Inc‥ The patent related to this research is disclosed as follow. “Surface plasmon enhanced fluorescence measurement device and surface plasmon enhanced fluorescence measurement method.” PCT/JP2014/002009, Priority date: 8 Apr. 2014. There are no further patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.

Atrazine is a common agricultural pesticide which has been reported to occur widely in surface drinking water, making it an environmental pollutant of concern. In the quest for developing sensitive detection methods for pesticides, the use of quantum dots (QDs) as sensitive fluorescence probes has gained momentum in recent years. QDs have attractive and unique optical properties whilst coupling of QDs to molecularly imprinted polymers (MIPs) has been shown to offer excellent selectivity. Thus, the development of QD@MIPs based fluorescence sensors could provide an alternative for monitoring herbicides like atrazine in water. In this work, highly fluorescent CdSeTe/ZnS QDs were fabricated using the conventional organometallic synthesis approach and were then encapsulated with MIPs. The CdSeTe/ZnS@MIP sensor was characterized and applied for selective detection of atrazine. The sensor showed a fast response time (5 min) upon interaction with atrazine and the fluorescence intensity was linearly quenched within the 2-20 mol L -1 atrazine range. The detection limit of 0.80 × 10 -7  mol L -1 is comparable to reported environmental levels. Lastly, the sensor was applied in real water samples and showed satisfactory recoveries (92-118%) in spiked samples, hence it is a promising candidate for use in water monitoring., (© 2019 John Wiley & Sons, Ltd.)

Recently, luminescent metal-organic framework (MOF) materials have attracted considerable attention in fluorescence sensing. In this essay, we prepared a new cluster-based Ca II -MOFs {[Ca 1.5 (μ 8 -HL 1 )(DMF) 2 ]·DMF} n ( 1 ) with good water dispersibility, excellent photoluminescence properties (FL quantum yield of 20.37%) and great fluorescence stability. Further, it was employed to design as an "off-on" fluorescence sensor for sensitive detection of l-cysteine. This proposed strategy was that fluorescence of Ca II -MOFs 1 was quenched for providing a low fluorescence background by the introduction of Pb 2+ forming the Ca II -MOFs 1 /Pb 2+ hybrid system. The quenching effect could be ascribed to the static quenching mechanism because of the formation of ground-state complexes and coordination interactions between the free carboxyl of H 4 L 1 ligands of Ca II -MOFs 1 and Pb 2+ . Then, with the addition of l-cysteine into the Ca II -MOFs 1 /Pb 2+ hybrid system, the fluorescence signal was immediately restored. This result was because the Pb 2+ was gradually released from the hybrid system by chelation interactions between the -SH groups of l-cysteine and Pb 2+ . This method received a relative wide linear range varying from 0.05 to 40 μM and a low detection limit of 15 nM for detection of l-cysteine. This proposed strategy was also successfully applied to detect l-cysteine in human serum samples with satisfactory recoveries from 95.9 to 101.5%.

As a marker of malignant tumors, miRNA is closely related to the occurrence and metastasis of tumors. How to achieve rapid and sensitive real-time detection is important for clinical prevention and treatment of cancer. In this study, an intelligent detection platform based on smartphone image processing technology made point-of-care testing a reality. This new smart approach could detect multiple targets simultaneously and sensitively. Hydrogel microparticles of different coding modes (shapes, numbers) were prepared by flow lithography to detect different miRNAs. After sandwich immunoassays, different shapes of hydrogels showed different fluorescence intensities depending on their targets. Images were captured by a smartphone and then analyzed by image recognition processing software installed on the smartphone. The concentration of miRNA was obtained within 10 s. The entire reaction process did not exceed 2 h. This intelligent and portable detection platform for miRNAs was reliable and the limit of detection reached the femtomole level. This work provided a demonstration of intelligent, portable, real-time detection of tumor markers.

In recent years, there has been an increase in pesticide use to improve crop production due to the growth of agricultural activities. Consequently, various pesticides have been present in the environment for an extended period of time. This review presents a general description of recent advances in the development of methods for the quantification of pesticides used in agricultural activities. Current advances focus on improving sensitivity and selectivity through the use of nanomaterials in both sensor assemblies and new biosensors. In this study, we summarize the electrochemical, optical, nano-colorimetric, piezoelectric, chemo-luminescent and fluorescent techniques related to the determination of agricultural pesticides. A brief description of each method and its applications, detection limit, purpose-which is to efficiently determine pesticides-cost and precision are considered. The main crops that are assessed in this study are bananas, although other fruits and vegetables contaminated with pesticides are also mentioned. While many studies have assessed biosensors for the determination of pesticides, the research in this area needs to be expanded to allow for a balance between agricultural activities and environmental protection.

Fluctuation correlation spectroscopy (FCS) is a single-molecule/particle detection technique based on measuring signal fluctuations in a highly focused detection volume. Multiple-parameter information can be obtained from the FCS measurement including the amplitude, characteristic diffusion time of correlation curve, and brightness of the adopted probes. The multiple-parameter change is related with physical or chemical change occurring in the probes. Meanwhile, the detection method has advantages such as short sample time in seconds, sample volume with low limit in femtoliters, and mixing to detection without any separations. These advantages make the FCS technique suitable for homogeneous analysis. In this review, we summarized recent novel applications of FCS and its variants in homogeneous analysis including nucleic acid analysis, protein analysis, enzyme activity assay, direct characterization of nanoparticles in solution, and others. Graphical abstract.

The precise cellular function of peroxynitrite (ONOO - ) in biosystems remains elusive, primarily owing to being short of ultrasensitive techniques for monitoring its intracellular distribution. In this work, a novel rhodamine B cyclic 1,2-dimethylhydrazine fluorescent chemodosimeter RDMH-PN for highly specific and ultrasensitive monitoring of basal ONOO - in biosystems was rationally designed. The fluorescence titration experiments demonstrated that RDMH-PN was capable of quantitatively detecting 0-100 nM ONOO - (limit of detection = 0.68 nM). In addition, RDMH-PN has outstanding performances of ultrafast measurement, naked-eye detection, and preeminent selectivity toward ONOO - to accurately detect intracellular basal ONOO - . Finally, it has been confirmed that RDMH-PN could not only map the intracellular basal ONOO - level by inhibition tests but also trace the fluctuations of endogenous and exogenous ONOO - levels with diverse stimulations in live cells and zebrafish.

Designing organic saccharide sensors for use in aqueous solution is a nontrivial endeavor. Incorporation of hydrogen bonding groups on a sensor's receptor unit to target saccharides is an obvious strategy but not one that is likely to ensure analyte-receptor interactions over analyte-solvent or receptor-solvent interactions. Phenylboronic acids are known to reversibly and covalently bind saccharides (diols in general) with highly selective affinity in aqueous solution. Therefore, recent work has sought to design such sensors and understand their mechanism for allowing fluorescence with bound saccharides. In past work, binding orientations of several saccharides were determined to dimethylaminomethylphenylboronic acid (DMPBA) receptors with an anthracene fluorophore; however, the binding orientation of d-fructose to such a sensor could not be determined. In this work, we investigate the potential binding modes by generating 20 possible bidentate and six possible tridentate modes between fructose and DMPBA, a simplified receptor model. Gas phase and implicit solvent geometry optimizations, with a myriad functional/basis set pairs, were carried out to identify the lowest energy bidentate and tridentate binding modes of d-fructose to DMPBA. An interesting hydrogen transfer was observed during selected bidentate gas phase optimizations; this transfer suggests a strong sharing of the hydrogen atom between the boronate hydroxyl and amine nitrogen.

Cancer-associated mRNA (mRNA) is an important biomarker for early diagnosis, prognosis, and prediction of treatment responses. Despite recent developments in fluorescence live cell imaging, reliable detection and quantification of mRNA in living cells still remain challenging due to a complicated intracellular environment. Herein, we present a fluorescent nanosensor for live-cell monitoring of cancer-related mRNAs involved in the canonical Wnt/β-catenin signaling pathway. The nanosensor enables rapid and accurate assessment of gene downregulation efficiency in a dose- and time-dependent manner by measuring quantitative fluorescence signal corresponding to β-catenin or its target mRNA levels in living cells. It is expected that the fluorescent nanosensor will be applicable to high-throughput screening for the efficient drug discovery and insightful understanding of the molecular mechanisms of potential drug candidate for cancer treatment.

Considerable interest in using lanthanide complexes in optics have been well-known persisted for a long time. But such molecular-based edifices have been excluded from practical application because of their poor thermal or photo stabilities. Here a novel europium embedded layered double hydroxide (Mg-Al LDH-Eu) has been established and such an inorganic-organic framework demonstrates improved thermal performance due to hydrolysis and poly condensation of the trimethoxysilyl-unit. In addition, the incorporation of a functional building block such as ethylenediamine triacetic acid can significantly minimize the negative effects of hydroxyl groups. In the presence of tetracycline (Tc), the nanoprobe exhibits an "off-on" change in aqueous solution, and the red luminescence can be excited in the visible light range (405 nm). It provides a very sensitive signal response to Tc with an excellent linear relation in the range of 0.1 μM to 5.0 μM, and the detection limit of this probe is measured to be 7.6 nM. This nanoplatform exhibits low cytotoxicity during in vitro experiments and can be employed for the detection of tetracycline in 293T cells.

Super-resolution microscopy techniques enable optical imaging in live cells with unprecedented spatial resolution. They unfortunately lack the temporal resolution required to directly investigate cellular dynamics at scales sufficient to measure molecular diffusion. These fast time scales are, on the other hand, routinely accessible by spectroscopic techniques such as fluorescence correlation spectroscopy (FCS). To enable the direct investigation of fast dynamics at the relevant spatial scales, FCS has been combined with super-resolution stimulated emission depletion (STED) microscopy. STED-FCS has been applied in point or scanning mode to reveal nanoscale diffusion behavior of molecules in live cells. In this protocol, we describe the technical details of performing point STED-FCS (pSTED-FCS) and scanning STED-FCS (sSTED-FCS) measurements, from calibration and sample preparation to data acquisition and analysis. We give particular emphasis to 2D diffusion dynamics in cellular membranes, using molecules tagged with organic fluorophores. These measurements can be accomplished within 4-6 h by those proficient in fluorescence imaging.

The present paper studies the applicability of a portable cost-effective spectroscopic system for the optical screening of skin tumors. in vivo studies of Raman scattering and autofluorescence (AF) of skin tumors with the 785 nm excitation laser in the near-infrared region included malignant melanoma, basal cell carcinoma and various types of benign neoplasms. The efficiency of the portable system was evaluated by comparison with a highly sensitive spectroscopic system and with the diagnosis accuracy of a human oncologist. Partial least square analysis of Raman and AF spectra was performed; specificity and sensitivity of various skin oncological pathologies detection varied from 78.9% to 100%. Hundred percent accuracy of benign and malignant skin tumors differentiation is possible only with a combined analysis of Raman and AF signals., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Cell culture assays for therapeutic drug screening today are fully automated. Vitality of the cells is monitored by different sensors. For such a system, we propose a new reader unit, which is capable of reading two different fluorescent sensors and electrical impedance in 24-well-plates. Main goals are to reduce cost, complexity and size while achieving a similar performance as the existing reader unit. To achieve this, measurement electronics and signal paths for frequency domain fluorescence and bio-impedance measurement are combined. Central component is an integrated circuit for impedance spectroscopy. A new compact and economic optical setup is developed to read two different sensor spots on the bottom of the well. Measurement errors introduced by different components like DFT leakage, and frequency dependent signal delays are evaluated and compensated. A set of commercially available fluorescence sensor spots is used to verify the read out performance. The results are usable, with noise slightly higher than commercial readers. To verify the impedance measurement accuracy, measurements of known resistances are conducted. In the relevant impedance and frequency range for biological applications a suitable accuracy is achieved. Due to the higher sampling rate of the new reader, the higher noise can be reduced through averaging. The new system is significantly smaller and cheaper to manufacture than commercially available devices., (Copyright © 2018 Elsevier B.V. All rights reserved.)