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1. Mechanical stress shapes the cancer cell response to neddylation inhibition

Author

Frédérique Mittler, Patricia Obeïd, Vincent Haguet, Cédric Allier, Sophie Gerbaud, Anastasia V. Rulina, Xavier Gidrol, and Maxim Y. Balakirev

Subjects
MLN4924, Rho GTPases, Mechanical stress, Tight junctions, Metastasis, Prostate cancer, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Abstract Background The inhibition of neddylation by the preclinical drug MLN4924 represents a new strategy to combat cancer. However, despite being effective against hematologic malignancies, its success in solid tumors, where cell–cell and cell-ECM interactions play essential roles, remains elusive. Methods Here, we studied the effects of MLN4924 on cell growth, migration and invasion in cultured prostate cancer cells and in disease-relevant prostate tumoroids. Using focused protein profiling, drug and RNAi screening, we analyzed cellular pathways activated by neddylation inhibition. Results We show that mechanical stress induced by MLN4924 in prostate cancer cells significantly affects the therapeutic outcome. The latter depends on the cell type and involves distinct Rho isoforms. In LNCaP and VCaP cells, the stimulation of RhoA and RhoB by MLN4924 markedly upregulates the level of tight junction proteins at cell–cell contacts, which augments the mechanical strain induced by Rho signaling. This “tight junction stress response” (TJSR) causes the collapse of cell monolayers and a characteristic rupture of cancer spheroids. Notably, TJSR is a major cause of drug-induced apoptosis in these cells. On the other hand, in PC3 cells that underwent partial epithelial-to-mesenchymal transition (EMT), the stimulation of RhoC induces an adverse effect by promoting amoeboid cell scattering and invasion. We identified complementary targets and drugs that allow for the induction of TJSR without stimulating RhoC. Conclusions Our finding that MLN4924 acts as a mechanotherapeutic opens new ways to improve the efficacy of neddylation inhibition as an anticancer approach. Graphical Abstract

Published
2022
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3. High-Content Monitoring of Drug Effects in a 3D Spheroid Model

Author

Frédérique Mittler, Patricia Obeïd, Anastasia V. Rulina, Vincent Haguet, Xavier Gidrol, and Maxim Y. Balakirev

Subjects
spheroids, 3D model, prostate cancer, MLN4924, HCS screening, drug discovery, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

A recent decline in the discovery of novel medications challenges the widespread use of 2D monolayer cell assays in the drug discovery process. As a result, the need for more appropriate cellular models of human physiology and disease has renewed the interest in spheroid 3D culture as a pertinent model for drug screening. However, despite technological progress that has significantly simplified spheroid production and analysis, the seeming complexity of the 3D approach has delayed its adoption in many laboratories. The present report demonstrates that the use of a spheroid model may be straightforward and can provide information that is not directly available with a standard 2D approach. We describe a cost-efficient method that allows for the production of an array of uniform spheroids, their staining with vital dyes, real-time monitoring of drug effects, and an ATP-endpoint assay, all in the same 96-well U-bottom plate. To demonstrate the method performance, we analyzed the effect of the preclinical anticancer drug MLN4924 on spheroids formed by VCaP and LNCaP prostate cancer cells. The drug has different outcomes in these cell lines, varying from cell cycle arrest and protective dormancy to senescence and apoptosis. We demonstrate that by using high-content analysis of spheroid arrays, the effect of the drug can be described as a series of EC50 values that clearly dissect the cytostatic and cytotoxic drug actions. The method was further evaluated using four standard cancer chemotherapeutics with different mechanisms of action, and the effect of each drug is described as a unique multi-EC50 diagram. Once fully validated in a wider range of conditions, this method could be particularly valuable for phenotype-based drug discovery.

Published
2017
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4. Polarization Induced Electro-Functionalization of Pore Walls: A Contactless Technology

Author

Aurélie Bouchet-Spinelli, Emeline Descamps, Jie Liu, Abdulghani Ismail, Pascale Pham, François Chatelain, Thierry Leïchlé, Loïc Leroy, Patrice Noël Marche, Camille Raillon, André Roget, Yoann Roupioz, Neso Sojic, Arnaud Buhot, Vincent Haguet, Thierry Livache, and Pascal Mailley

Subjects
nanopore, micropore, clef, biosensing, electro-functionalization, contactless, Biotechnology, TP248.13-248.65
Abstract

This review summarizes recent advances in micro- and nanopore technologies with a focus on the functionalization of pores using a promising method named contactless electro-functionalization (CLEF). CLEF enables the localized grafting of electroactive entities onto the inner wall of a micro- or nano-sized pore in a solid-state silicon/silicon oxide membrane. A voltage or electrical current applied across the pore induces the surface functionalization by electroactive entities exclusively on the inside pore wall, which is a significant improvement over existing methods. CLEF’s mechanism is based on the polarization of a sandwich-like silicon/silicon oxide membrane, creating electronic pathways between the core silicon and the electrolyte. Correlation between numerical simulations and experiments have validated this hypothesis. CLEF-induced micro- and nanopores functionalized with antibodies or oligonucleotides were successfully used for the detection and identification of cells and are promising sensitive biosensors. This technology could soon be successfully applied to planar configurations of pores, such as restrictions in microfluidic channels.

Published
2019
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5. Selective individual primary cell capture using locally bio-functionalized micropores.

Author

Jie Liu, Radoslaw Bombera, Loïc Leroy, Yoann Roupioz, Dieudonné R Baganizi, Patrice N Marche, Vincent Haguet, Pascal Mailley, and Thierry Livache

Subjects
Medicine, Science
Abstract

BACKGROUND: Solid-state micropores have been widely employed for 6 decades to recognize and size flowing unlabeled cells. However, the resistive-pulse technique presents limitations when the cells to be differentiated have overlapping dimension ranges such as B and T lymphocytes. An alternative approach would be to specifically capture cells by solid-state micropores. Here, the inner wall of 15-µm pores made in 10 µm-thick silicon membranes was covered with antibodies specific to cell surface proteins of B or T lymphocytes. The selective trapping of individual unlabeled cells in a bio-functionalized micropore makes them recognizable just using optical microscopy. METHODOLOGY/PRINCIPAL FINDINGS: We locally deposited oligodeoxynucleotide (ODN) and ODN-conjugated antibody probes on the inner wall of the micropores by forming thin films of polypyrrole-ODN copolymers using contactless electro-functionalization. The trapping capabilities of the bio-functionalized micropores were validated using optical microscopy and the resistive-pulse technique by selectively capturing polystyrene microbeads coated with complementary ODN. B or T lymphocytes from a mouse splenocyte suspension were specifically immobilized on micropore walls functionalized with complementary ODN-conjugated antibodies targeting cell surface proteins. CONCLUSIONS/SIGNIFICANCE: The results showed that locally bio-functionalized micropores can isolate target cells from a suspension during their translocation throughout the pore, including among cells of similar dimensions in complex mixtures.

Published
2013
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6. Polarization Induced Electro-Functionalization of Pore Walls: A Contactless Technology

Author

Patrice N. Marche, Jie Liu, Yoann Roupioz, Thierry Livache, Emeline Descamps, Loïc Leroy, Camille Raillon, André Roget, Thierry Leïchlé, Francois Chatelain, Aurélie Bouchet-Spinelli, Vincent Haguet, Arnaud Buhot, Neso Sojic, Pascale Pham, Abdulghani Ismail, Pascal Mailley, Chimie pour la Reconnaissance et l’Etude d’Assemblages Biologiques (CREAB), 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), Équipe MICrosystèmes d'Analyse (LAAS-MICA), Laboratoire d'analyse et d'architecture des systèmes (LAAS), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Université Fédérale Toulouse Midi-Pyrénées, Département Microtechnologies pour la Biologie et la Santé (DTBS), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Alloimmunité-Autoimmunité-Transplantation (A2T), Institut Universitaire d'Hématologie (IUH), Université Paris Diderot - Paris 7 (UPD7)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Équipe Microsystèmes électromécaniques (LAAS-MEMS), Georgia Tech Lorraine [Metz], Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Ecole Supérieure d'Electricité - SUPELEC (FRANCE)-Georgia Institute of Technology [Atlanta]-CentraleSupélec-Ecole Nationale Supérieure des Arts et Metiers Metz-Centre National de la Recherche Scientifique (CNRS), 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]), Institut des Sciences Moléculaires (ISM), Université Montesquieu - Bordeaux 4-Université Sciences et Technologies - Bordeaux 1-École Nationale Supérieure de Chimie et de Physique de Bordeaux (ENSCPB)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Biomicrotechnologie et génomique fonctionnelle (BIOMICS), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), 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 National de la Santé et de la Recherche Médicale (INSERM)-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 National de la Santé et de la Recherche Médicale (INSERM)-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), Université Toulouse Capitole (UT Capitole), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université Toulouse Capitole (UT Capitole), Université de Toulouse (UT), Ecole Nationale Supérieure des Arts et Metiers Metz-Georgia Institute of Technology [Atlanta]-Ecole Supérieure d'Electricité - SUPELEC (FRANCE)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Université Montesquieu - Bordeaux 4-Université Sciences et Technologies - Bordeaux 1 (UB)-École Nationale Supérieure de Chimie et de Physique de Bordeaux (ENSCPB)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), ANR-17-EURE-0003,CBH-EUR-GS,CBH-EUR-GS(2017), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse Capitole (UT Capitole), and Descamps, Emeline

Subjects
Silicon, Materials science, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, lcsh:Biotechnology, Clinical Biochemistry, chemistry.chemical_element, Nanotechnology, Biosensing Techniques, Review, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, electro-functionalization, Nanopores, lcsh:TP248.13-248.65, Electric Impedance, micropore, nanopore, Polarization (electrochemistry), Silicon oxide, contactless, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, CLEF, [SCCO.NEUR]Cognitive science/Neuroscience, [SCCO.NEUR] Cognitive science/Neuroscience, technology, industry, and agriculture, Membranes, Artificial, Electrochemical Techniques, General Medicine, Microporous material, 021001 nanoscience & nanotechnology, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Nanopore, Membrane, chemistry, Surface modification, biosensing, 0210 nano-technology, Biosensor, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

This review summarizes recent advances in micro- and nanopore technologies with a focus on the functionalization of pores using a promising method named contactless electro-functionalization (CLEF). CLEF enables the localized grafting of electroactive entities onto the inner wall of a micro- or nano-sized pore in a solid-state silicon/silicon oxide membrane. A voltage or electrical current applied across the pore induces the surface functionalization by electroactive entities exclusively on the inside pore wall, which is a significant improvement over existing methods. CLEF’s mechanism is based on the polarization of a sandwich-like silicon/silicon oxide membrane, creating electronic pathways between the core silicon and the electrolyte. Correlation between numerical simulations and experiments have validated this hypothesis. CLEF-induced micro- and nanopores functionalized with antibodies or oligonucleotides were successfully used for the detection and identification of cells and are promising sensitive biosensors. This technology could soon be successfully applied to planar configurations of pores, such as restrictions in microfluidic channels.

Published
2019

7. Encapsulated organoids & organ-on-a-chip platform for cancer modeling

Author

Xavier Gidrol, A. Tollance, S. Porte, C. Belda-Marin, Vincent Haguet, Patricia Obeid, Nathalie Picollet-D'hahan, A. Romero-Millan, Bastien Laperrousaz, Donald K. Martin, Frédérique Kermarrec, Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-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), Laboratoire de Physique de l'ENS Lyon (Phys-ENS), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Biogeochemistry Research Centre, School of Chemistry, 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 National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), and École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL)

Subjects
010302 applied physics, Scaffold, Computer science, [SDV]Life Sciences [q-bio], Microfluidics, Cancer, Nanotechnology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, medicine.disease, 01 natural sciences, Organ-on-a-chip, 3. Good health, 0103 physical sciences, Organoid, medicine, ComputingMilieux_MISCELLANEOUS
Abstract

This review highlights our major developments in the fields of organoids and organ-on-chip to address issues in fundamental and biomedical research by modeling development and cancer. We illustrate how contemporary miniaturized technologies (microfluidics, 3D scaffolding, 3D imaging) combined with RNAi-based organoids HTS (High-Throughput Screening), would help forming and analyzing consistent, efficient and reproducible organoids. Moreover, we illustrate the potential of engineered organ-on-chip devices for creating novel human organ and disease models, with particular focus on “prostate-on-a-chip”.

Published
2017

8. High-Content Monitoring of Drug Effects in a 3D Spheroid Model

Author

Patricia Obeid, Xavier Gidrol, Vincent Haguet, Anastasiia V. Rulina, Maxim Y Balakirev, Frédérique Mittler, Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-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), Inflammasome NLRP3 – NLRP3 Inflammasome, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Biomicrotechnologie et génomique fonctionnelle (BIOMICS), 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 National de la Santé et de la Recherche Médicale (INSERM)-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 National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Drug, 3D model, Cancer Research, Cell cycle checkpoint, [SDV]Life Sciences [q-bio], media_common.quotation_subject, Cell, MLN4924, spheroids, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Computational biology, Biology, lcsh:RC254-282, drug discovery, 03 medical and health sciences, LNCaP, ubiquitin, medicine, Methods, ComputingMilieux_MISCELLANEOUS, media_common, Drug discovery, Spheroid, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, prostate cancer, Anticancer drug, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 3. Good health, Nedd8, 030104 developmental biology, medicine.anatomical_structure, Oncology, Cell culture, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.IMM]Life Sciences [q-bio]/Immunology, HCS screening
Abstract

International audience; A recent decline in the discovery of novel medications challenges the widespread use of 2D monolayer cell assays in the drug discovery process. As a result, the need for more appropriate cellular models of human physiology and disease has renewed the interest in spheroid 3D culture as a pertinent model for drug screening. However, despite technological progress that has significantly simplified spheroid production and analysis, the seeming complexity of the 3D approach has delayed its adoption in many laboratories. The present report demonstrates that the use of a spheroid model may be straightforward and can provide information that is not directly available with a standard 2D approach. We describe a cost-efficient method that allows for the production of an array of uniform spheroids, their staining with vital dyes, real-time monitoring of drug effects, and an ATP-endpoint assay, all in the same 96-well U-bottom plate. To demonstrate the method performance, we analyzed the effect of the preclinical anticancer drug MLN4924 on spheroids formed by VCaP and LNCaP prostate cancer cells. The drug has different outcomes in these cell lines, varying from cell cycle arrest and protective dormancy to senescence and apoptosis. We demonstrate that by using high-content analysis of spheroid arrays, the effect of the drug can be described as a series of EC50 values that clearly dissect the cytostatic and cytotoxic drug actions. The method was further evaluated using four standard cancer chemotherapeutics with different mechanisms of action, and the effect of each drug is described as a unique multi-EC50 diagram. Once fully validated in a wider range of conditions, this method could be particularly valuable for phenotype-based drug discovery.

Published
2017

9. Contactless Electrofunctionalization of a Single Pore

Author

Thierry Livache, Emeline Descamps, Vincent Haguet, Francois Chatelain, Pascal Mailley, and Aurélie Bouchet

Subjects
Membranes, Materials science, Nanotechnology, General Chemistry, Models, Theoretical, Polypyrrole, Biomaterials, chemistry.chemical_compound, Membrane, chemistry, Electric field, Electrode, Deposition (phase transition), General Materials Science, Porous medium, Porosity, Biosensor, Biotechnology, Voltage
Abstract

Customized pores are smart components that find challenging applications in a variety of fields including purification membranes and biosensing systems. The incorporation of recognition probes within pores is therefore a challenge, due to the technical difficulty of delimiting the area functionalized and obtaining the localized, specific chemical modification of pore walls. An innovative approach, named contactless electrofunctionalization (CLEF), is presented to overcome this problem. CLEF allows easy, one-step modification of the inner surface of a pore etched in a dielectric membrane. The pore wall is coated under the influence of an electric field created by the application of a voltage between two electrodes, located near but not in contact with the pore openings. This specific localization of the deposited material within the pore is extremely rapid. Coatings were reliably and reproducibly obtained using polypyrrole co-polymers bearing oligonucleotides, demonstrating that this technology has a promising future in the design of biosensors. Moreover, the versatility of this process allows the deposition of various electroactive entities such as iridium oxide and therefore indicates a strong potential for diverse applications involving porous materials.

Published
2009

10. 2D and 3D cell microarrays in pharmacology

Author

Xavier Gidrol, Nathalie Picollet-D'hahan, Lamya Ghenim, Vincent Haguet, Brigitte Fouque, and Béatrice Schaack

Subjects
Automation, Laboratory, Pharmacology, Miniaturization, Microarray, Drug discovery, Microarray analysis techniques, Cell, Cell Culture Techniques, Nanotechnology, Biology, Microarray Analysis, High-Throughput Screening Assays, medicine.anatomical_structure, Cell culture, Drug Discovery, medicine, Animals, Humans, Biological Assay, DNA microarray
Abstract

To analyze the phenotypic consequences of perturbing mammalian cells with drugs, there is an increasing need for systematic cell-based assays in an HTS format. Cell microarrays provide an attractive solution as they offer more than a simple miniaturization and mechanization of conventional microtiter plates. While standard monolayer two-dimensional culture conditions are poor mimics of the cellular environment in situ, microfabricated systems enable three-dimensional organotypic cell cultures and have the potential to provide biological insight not achievable before. This article compares different cell microarray formats and evaluates their potential use in the drug discovery process.

Published
2009

11. Time-lapse contact microscopy of cell cultures based on non-coherent illumination

Author

François Perraut, Nathalie Picollet-D'hahan, Francois Chatelain, Marion Gabriel, Vincent Haguet, Cyrille Pornin, Marc R. Block, Stéphane Gétin, Dorothée Balle, Stéphanie Bigault, Xavier Gidrol, Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-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), Département d'Optronique (DOPT), Laboratoire d'Electronique et des Technologies de l'Information (CEA-LETI), Université Grenoble Alpes (UGA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Département Microtechnologies pour la Biologie et la Santé (DTBS), 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 (UGA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Dynamique des systèmes d'adhérence et différenciation (DySAD), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), 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)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Materials science, [SDV]Life Sciences [q-bio], Population, Cell Culture Techniques, Holography, Video microscopy, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Time-Lapse Imaging, Article, law.invention, Optics, Cell Movement, law, Microscopy, Cell Adhesion, Humans, Image sensor, education, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, Cell Proliferation, education.field_of_study, Microscopy, Video, Multidisciplinary, business.industry, Ray, Pinhole (optics), Photonics, business
Abstract

Video microscopy offers outstanding capabilities to investigate the dynamics of biological and pathological mechanisms in optimal culture conditions. Contact imaging is one of the simplest imaging architectures to digitally record images of cells due to the absence of any objective between the sample and the image sensor. However, in the framework of in-line holography, other optical components, e.g., an optical filter or a pinhole, are placed underneath the light source in order to illuminate the cells with a coherent or quasi-coherent incident light. In this study, we demonstrate that contact imaging with an incident light of both limited temporal and spatial coherences can be achieved with sufficiently high quality for most applications in cell biology, including monitoring of cell sedimentation, rolling, adhesion, spreading, proliferation, motility, death and detachment. Patterns of cells were recorded at various distances between 0 and 1000 μm from the pixel array of the image sensors. Cells in suspension, just deposited or at mitosis focalise light into photonic nanojets which can be visualised by contact imaging. Light refraction by cells significantly varies during the adhesion process, the cell cycle and among the cell population in connection with every modification in the tridimensional morphology of a cell.

Published
2015

12. A statistically inferred microRNA network identifies breast cancer target miR-940 as an actin cytoskeleton regulator

Author

Vincent Haguet, Stéphanie Combe, Ricky Bhajun, Christian Lajaunie, Xavier Gidrol, Amandine Pitaval, Eric Sulpice, Patricia Obeid, Laurent Guyon, Itebeddine Ghorbel, Biomicrotechnologie et génomique fonctionnelle (BIOMICS), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-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)-Institut National de la Santé et de la Recherche Médicale (INSERM)-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 de Bioinformatique (CBIO), MINES ParisTech - École nationale supérieure des mines de Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Cancer et génome: Bioinformatique, biostatistiques et épidémiologie d'un système complexe, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), ANR et CEA, Mines Paris - PSL (École nationale supérieure des mines de Paris), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Centre de Géosciences (GEOSCIENCES), MINES ParisTech - École nationale supérieure des mines de Paris-PSL Research University (PSL), and MINES ParisTech, Bibliothèque

Subjects
Kinase, Mesenchymal Transition, [SDV]Life Sciences [q-bio], Cells, Gene regulatory network, Gene-Expression, Breast Neoplasms, Biology, Discovery, Article, Cell Movement, Myosin, microRNA, Gene expression, Humans, Gene Regulatory Networks, Disease, ComputingMilieux_MISCELLANEOUS, Cellular Senescence, Cell Proliferation, Monomeric GTP-Binding Proteins, Genetics, Regulation of gene expression, [SDV.BIBS] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Multidisciplinary, Gene Expression Profiling, Computational Biology, Molecular Sequence Annotation, Animal Development, Actin cytoskeleton, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Cell biology, Gene Expression Regulation, Neoplastic, Gene expression profiling, Actin Cytoskeleton, MicroRNAs, Gene Ontology, Female, Databases, Nucleic Acid, Prediction, Signal Transduction
Abstract

International audience; MiRNAs are key regulators of gene expression. By binding to many genes, they create a complex network of gene co-regulation. Here, using a network-based approach, we identified miRNA hub groups by their close connections and common targets. In one cluster containing three miRNAs, miR-612, miR-661 and miR-940, the annotated functions of the co-regulated genes suggested a role in small GTPase signalling. Although the three members of this cluster targeted the same subset of predicted genes, we showed that their overexpression impacted cell fates differently. miR-661 demonstrated enhanced phosphorylation of myosin II and an increase in cell invasion, indicating a possible oncogenic miRNA. On the contrary, miR-612 and miR-940 inhibit phosphorylation of myosin II and cell invasion. Finally, expression profiling in human breast tissues showed that miR-940 was consistently downregulated in breast cancer tissues M icroRNAs are a class of endogenous, small (19–25 nucleotides), single-stranded non-coding RNAs that regulate gene expression in all eukaryotic organisms. In metazoans, microRNAs most commonly bind to the 39 untranslated region (39UTR) of their mRNA target transcript and cause translational repression and/or mRNA degradation. Every microRNA is predicted to regulate from a dozen to thousands of genes, including transcription factors. This fine-tuning of protein expression is known to be involved in many physiological processes, such as development, apoptosis, signal transduction and even cancer progression 1,2. More than 2,000 mature human microRNAs are listed in the 20 th release of miRBase: http://www.mirbase.org (2014) (Date of access:19/08/2013), and some authors hypothesise that the majority of human genes are regulated by microRNAs 3. Since their discovery in 1993 4 , a fair understanding of their role in animal development and in the onset and progression of diseases 2 , as well as of their potential use in therapies 5 , has been gathered. However, the cooperative behaviour of microRNAs is still under investigation. A growing body of experimental evidence suggests that microRNAs can regulate genes through complementarity, meaning that microRNAs can act together to regulate individual genes or groups of genes involved in similar processes 6. For example, Hu and co-workers demonstrated that transducing a cocktail of precursor microRNAs (miR-21, miR-24 and miR-221) can result in more effective engraftment of transplanted cardiac progenitor cells 7. Consistent with these discoveries, Zhu et al. demonstrated that miR-21 and miR-221 coregulate 56 gene ontology (GO) processes 8. In the same study, the authors also showed that cotransfection of miR-1 and miR-21 increases H 2 O 2-induced myocardial apoptosis and oxidative stress. These recent findings support the idea of microRNA-mediated cooperative regulation but also argue for the use of systemic approaches, notably based on graph theory, to decipher individual and complementary roles of microRNAs. Some work has been conducted to use recent high-throughput experiment-derived data sets to infer microRNA synergistic relationships 9–12. Herein, we present a microRNA network based on target similarities among microRNAs to infer clusters of microRNAs. Clusters are defined as groups of microRNAs sharing a set of common targets, predicted by either DIANA-microT v3 13 or TargetScan v6.2 14. Some authors have used GO enrichment analysis as a confirmatory tool for their clustering approach 11. In our case, GO enrichment is not used to infer networks but as a way to estimate the probable metabolic pathway(s) a cluster of microRNAs could co-regulate. Moreover, the novelty of our approach is to consider not only clusters of microRNAs but also OPEN

Published
2015

13. Selective Individual Primary Cell Capture Using Locally Bio-Functionalized Micropores

Author

Radoslaw Bombera, Vincent Haguet, Dieudonné R. Baganizi, Yoann Roupioz, Patrice N. Marche, Pascal Mailley, Loïc Leroy, Thierry Livache, Jie Liu, ROUPIOZ, Yoann, Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), 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), Groupe matière condensée et matériaux (GMCM), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), 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), 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), Contrôle moléculaire de la réponse immune specifique, Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Biopuces (BIOPUCES), 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])-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Institut de Chimie du CNRS (INC)-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)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Fluorescence-lifetime imaging microscopy, Immunoconjugates, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Polymers, T-Lymphocytes, Cell, lcsh:Medicine, Cell Separation, Physical Chemistry, 01 natural sciences, Mice, chemistry.chemical_compound, Engineering, Fluorescence microscope, Biomacromolecule-Ligand Interactions, lcsh:Science, ComputingMilieux_MISCELLANEOUS, B-Lymphocytes, 0303 health sciences, Multidisciplinary, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Porous Materials, Silicon Dioxide, Microspheres, medicine.anatomical_structure, Membrane, Oligodeoxyribonucleotides, Microtechnology, Material Films, Material by Structure, Research Article, Surface Chemistry, Silicon dioxide, Materials Science, Biophysics, Bioengineering, Cell Surface Proteins, Antibodies, Material by Attribute, 03 medical and health sciences, medicine, Animals, Pyrroles, Biology, 030304 developmental biology, 010401 analytical chemistry, lcsh:R, Microporous material, [CHIM.ORGA] Chemical Sciences/Organic chemistry, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Semiconductors, Transmission microscopy, Immunology, lcsh:Q, Filtration, Spleen
Abstract

BACKGROUND: Solid-state micropores have been widely employed for 6 decades to recognize and size flowing unlabeled cells. However, the resistive-pulse technique presents limitations when the cells to be differentiated have overlapping dimension ranges such as B and T lymphocytes. An alternative approach would be to specifically capture cells by solid-state micropores. Here, the inner wall of 15-µm pores made in 10 µm-thick silicon membranes was covered with antibodies specific to cell surface proteins of B or T lymphocytes. The selective trapping of individual unlabeled cells in a bio-functionalized micropore makes them recognizable just using optical microscopy. METHODOLOGY/PRINCIPAL FINDINGS: We locally deposited oligodeoxynucleotide (ODN) and ODN-conjugated antibody probes on the inner wall of the micropores by forming thin films of polypyrrole-ODN copolymers using contactless electro-functionalization. The trapping capabilities of the bio-functionalized micropores were validated using optical microscopy and the resistive-pulse technique by selectively capturing polystyrene microbeads coated with complementary ODN. B or T lymphocytes from a mouse splenocyte suspension were specifically immobilized on micropore walls functionalized with complementary ODN-conjugated antibodies targeting cell surface proteins. CONCLUSIONS/SIGNIFICANCE: The results showed that locally bio-functionalized micropores can isolate target cells from a suspension during their translocation throughout the pore, including among cells of similar dimensions in complex mixtures.

Published
2013

14. Polarization-Induced Local Pore-Wall Functionalization for Biosensing: From Micropore to Nanopore

Author

André Roget, Emeline Descamps, Loïc Leroy, Pascale Pham, Arnaud Buhot, Pascal Mailley, Fabien Sauter-Starace, Vincent Haguet, Aurélie Bouchet, Thierry Livache, Jie Liu, Buhot, Arnaud, Chimie pour la Reconnaissance et l’Etude d’Assemblages Biologiques (CREAB), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-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)-Université Grenoble Alpes (UGA)-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), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Biomicrotechnologie et génomique fonctionnelle (BIOMICS), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-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 (UGA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-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 (UGA), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Équipe MICrosystèmes d'Analyse (LAAS-MICA), Laboratoire d'analyse et d'architecture des systèmes (LAAS), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J), 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 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 National de la Santé et de la Recherche Médicale (INSERM)-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 National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Laboratoire Biopuces (BIOPUCES), 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), 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), and 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)

Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology, Silicon, chemistry.chemical_element, Nanotechnology, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, Analytical Chemistry, Nanopores, Electricity, Electric field, Polarization (electrochemistry), ComputingMilieux_MISCELLANEOUS, Membranes, Artificial, Microporous material, 021001 nanoscience & nanotechnology, Silicon Dioxide, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Nanopore, Membrane, chemistry, Surface modification, Microtechnology, 0210 nano-technology, Biosensor
Abstract

The use of biological-probe-modified solid-state pores in biosensing is currently hindered by difficulties in pore-wall functionalization. The surface to be functionalized is small and difficult to target and is usually chemically similar to the bulk membrane. Herein, we demonstrate the contactless electrofunctionalization (CLEF) approach and its mechanism. This technique enables the one-step local functionalization of the single pore wall fabricated in a silica-covered silicon membrane. CLEF is induced by polarization of the pore membrane in an electric field and requires a sandwich-like composition and a conducting or semiconducting core for the pore membrane. The defects in the silica layer of the micropore wall enable the creation of an electric pathway through the silica layer, which allows electrochemical reactions to take place locally on the pore wall. The pore diameter is not a limiting factor for local wall modification using CLEF. Nanopores with a diameter of 200 nm fabricated in a silicon membrane and covered with native silica layer have been successfully functionalized with this method, and localized pore-wall modification was obtained. Furthermore, through proof-of-concept experiments using ODN-modified nanopores, we show that functionalized nanopores are suitable for translocation-based biosensing.

Published
2012

15. Electrochemically induced maskless metal deposition on micropore wall

Author

Jie Liu, Vincent Haguet, Pascal Mailley, Pascale Pham, Fabien Sauter-Starace, Thierry Livache, and Clément Hébert

Subjects
Biomaterials, Metal deposition, Materials science, Fabrication, Foundation (engineering), General Materials Science, Nanotechnology, General Chemistry, Microporous material, Biotechnology
Abstract

By applying an external electric field across a micropore via an electrolyte, metal ions in the electrolyte can be reduced locally onto the inner wall of the micropore, which was fabricated in a silica-covered silicon membrane. This maskless metal deposition on the silica surface is a result of the pore membrane polarization in the electric field.

Published
2011

16. Self-arraying of charged levitating droplets

Author

Christian Jeandey, Henri Grateau, Jérémie Nussbaumer, Paul Kauffmann, Pascale Pham, Vincent Haguet, Gilbert Reyne, and Alain Masse

Subjects
Interaction forces, business.industry, Chemistry, Diamagnetic levitation, Centripetal force, Molecular physics, Analytical Chemistry, Magnetic field, Physics::Fluid Dynamics, Optics, Physics::Plasma Physics, Condensed Matter::Superconductivity, Magnet, Levitation, Diamagnetism, business, Droplet evaporation
Abstract

Diamagnetic levitation of water droplets in air is a promising phenomenon to achieve contactless manipulation of chemical or biochemical samples. This noncontact handling technique prevents contaminations of samples as well as provides measurements of interaction forces between levitating reactors. Under a nonuniform magnetic field, diamagnetic bodies such as water droplets experience a repulsive force which may lead to diamagnetic levitation of a single or few micro-objects. The levitation of several repulsively charged picoliter droplets was successfully performed in a ~1 mm(2) adjustable flat magnetic well provided by a centimeter-sized cylindrical permanent magnet structure. Each droplet position results from the balance between the centripetal diamagnetic force and the repulsive Coulombian forces. Levitating water droplets self-organize into satellite patterns or thin clouds, according to their charge and size. Small triangular lattices of identical droplets reproduce magneto-Wigner crystals. Repulsive forces and inner charges can be measured in the piconewton and the femtocoulomb ranges, respectively. Evolution of interaction forces is accurately followed up over time during droplet evaporation.

Published
2011

17. Diamagnetically trapped arrays of living cells above micromagnets

Author

Ammara Ith, Vincent Haguet, Franz Bruckert, Victor Gaude, Paul Kauffmann, Daniel O'Brien, Stéphanie Combe, Nora Dempsey, Béatrice Schaack, Florian Boué, Gilbert Reyne, Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire des matériaux et du génie physique (LMGP ), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National Polytechnique de Grenoble (INPG)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Micro et NanoMagnétisme (NEEL - MNM), Institut Néel (NEEL), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS), Institut National Polytechnique de Grenoble (INPG)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Micro et NanoMagnétisme (MNM)

Subjects
Materials science, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Cell Survival, Biomedical Engineering, Cell Culture Techniques, Contrast Media, Bioengineering, Nanotechnology, Gadolinium, 02 engineering and technology, Trapping, Cell Growth Processes, 01 natural sciences, Biochemistry, Jurkat cells, Suspension (chemistry), Jurkat Cells, Planar, Meglumine, Heterocyclic Compounds, 0103 physical sciences, Organometallic Compounds, Humans, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Microscale chemistry, 010302 applied physics, General Chemistry, Equipment Design, [CHIM.MATE]Chemical Sciences/Material chemistry, 021001 nanoscience & nanotechnology, Remanence, Tissue Array Analysis, Magnet, Magnets, Diamagnetism, 0210 nano-technology
Abstract

International audience; Cell arrays are of foremost importance for many applications in pharmaceutical research or fundamental biology. Although arraying techniques have been widely investigated for adherent cells, organization of cells in suspension has been rarely considered. The arraying of non-adherent cells using the diamagnetic repulsive force is presented. A planar arrangement of Jurkat cells is achieved at the microscale above high quality microfabricated permanent magnets with remanent magnetization of J(r) approximate to 1 T, in the presence of a paramagnetic contrast agent. The cytotoxicity of three Gd based contrast agents, Gd-DOTA, Gd-BOPTA and Gd-HP-DO3A, is studied. Among them, Gd-HP-DO3A appears to be the most biocompatible toward Jurkat cells. In close agreement with analytical simulations, diamagnetically 'suspended' cells have been successfully arrayed above square and honeycomb-like micromagnet arrays, which act as a "diamagnetophobic" surface. Living cell trapping is achieved in a simple manner using concentrations of Gd-HP-DO3A as low as 1.5 mM.

Published
2011

18. Contactless dielectrophoretic handling of diamagnetic levitating water droplets in air

Author

Paul Kauffmann, Vincent Haguet, Pascale Pham, Thibault Honegger, A Masse, Gilbert Reyne, M Kustov, David Peyrade, Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire des technologies de la microélectronique (LTM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), and Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
010302 applied physics, Materials science, 010401 analytical chemistry, Microfluidics, [SPI.NRJ]Engineering Sciences [physics]/Electric power, Mechanics, Maxwell stress tensor, Dielectrophoresis, 01 natural sciences, 0104 chemical sciences, Electronic, Optical and Magnetic Materials, Magnetic field, Physics::Fluid Dynamics, Nuclear magnetic resonance, 0103 physical sciences, Levitation, Diamagnetism, Digital microfluidics, Electrical and Electronic Engineering, Magnetic levitation, ComputingMilieux_MISCELLANEOUS
Abstract

Micromagnets have been reported to successfully levitate picoliter water droplets in air. Manipulation of levitating water droplets is a contamination-free alternative to digital-microfluidic labs-on-chip where droplets are handled in channels or on a substrate. An integrated electromagnetic hybrid device combining dielectrophoretic (DEP) forces to control the position of levitating droplets along a magnetic groove, is proposed. Diamagnetic forces are analytically computed with CADES while Comsol Multiphysics™ (FEM) is used for the DEP forces. Approximations of the `point dipole model' are compared to the Maxwell Stress Tensor method applied on the 3-D model. Based on these results, an electric sequence for polarizing planar parallel Indium Tin Oxide (ITO) electrodes is proposed in order to provide a stable and accurate control of the droplet microposition along the gap. The use of these electrodes as micro-conductors to produce variable magnetic fields for magnetophoretic droplet actuation is considered and compared to DEP actuation.

Published
2010

19. Electrofunctionalization: Small 20/2009

Author

Vincent Haguet, Aurélie Bouchet, Emeline Descamps, Thierry Livache, Francois Chatelain, and Pascal Mailley

Subjects
Biomaterials, General Materials Science, General Chemistry, Biotechnology
Published
2009

20. Magnetophoretic and Dielectrophoretic Actuations Coupled with Diamagnetic Trapping in Air and Liquids

Author

Vincent Haguet, H. Chetouani, Gilbert Reyne, Paul Kauffmann, Pascale Pham, Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Garcia, Sylvie, Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
010302 applied physics, Materials science, [SPI.NRJ]Engineering Sciences [physics]/Electric power, 02 engineering and technology, Trapping, 021001 nanoscience & nanotechnology, 01 natural sciences, Atomic and Molecular Physics, and Optics, Chemical physics, 0103 physical sciences, Diamagnetism, Electrical and Electronic Engineering, 0210 nano-technology, ComputingMilieux_MISCELLANEOUS, [SPI.NRJ] Engineering Sciences [physics]/Electric power
Abstract

International audience; We have previously developed microsystems for contactless handling of diamagnetic bodies in air and liquids [1]. Magnetic levitation of droplets, particles and cells is achieved by specific configurations of microscale permanent magnet devices. In this work, two new actuation technologies are presented, which could both guide particles and solve some of the current limitations of dielectrophoresis (DEP). Dielectrophoretic and magnetophoretic actuations are combined with diamagnetic levitation to move and position micro-particles along linear magnetostatic traps. Interestingly, magnetic levitation combined with magnetophoresis appears relevant for the actuation of particles in air. Dielectrophoresis in liquids is also enhanced by this technique, including in conductive solvents.

Published
2009

21. Principle and Analysis of a Two-dimensional On-Chip Magnetophoresis of Bioparticules for Contamination-free Biochemical Reactors

Author

J.-F. Jacquot, Vincent Haguet, H. Chetouani, Gilbert Reyne, Christian Jeandey, H. Rostaing, C. Dieppedale, T. Kerlin, Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Garcia, Sylvie

Subjects
010302 applied physics, Materials science, business.industry, [SPI.NRJ]Engineering Sciences [physics]/Electric power, 02 engineering and technology, 021001 nanoscience & nanotechnology, 01 natural sciences, Biomagnetism, Electronic, Optical and Magnetic Materials, Paramagnetism, Nuclear magnetic resonance, Magnet, 0103 physical sciences, Levitation, Optoelectronics, Diamagnetism, Magnetic nanoparticles, Wafer, Electrical and Electronic Engineering, 0210 nano-technology, business, Magnetic levitation, ComputingMilieux_MISCELLANEOUS, [SPI.NRJ] Engineering Sciences [physics]/Electric power
Abstract

A new on chip magnetic device was designed, fabricated and tested. This system enables precise contactless arrangement, arraying and trapping of diamagnetic microbeads. The device is composed of holes matrix throughout thick CoPt magnetic layers electroplated on silicon wafer. The device was experienced with diamagnetic polystyrene-latex beads of mean diameter 3 urn in a paramagnetic buffered solution. Using this system, diamagnetic micron-sized beads were successfully controlled and trapped in spatial stable levitation within magnetic wells. Contactless passive levitation brings amazing possibilities to bio-chips. Compared with existing approaches, it allows precise passive contactless arraying of microbeads or unlabelled cells and enables contamination-free biochemical micro-reactors

Published
2007

22. Magnetic levitation of microdroplets in air

Author

Vincent Haguet, Jeandey, C., Chetouani, H., Reyne, G., Chatelain, F., Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), and Garcia, Sylvie

Subjects
[SPI.NRJ]Engineering Sciences [physics]/Electric power, ComputingMilieux_MISCELLANEOUS, [SPI.NRJ] Engineering Sciences [physics]/Electric power
Abstract

International audience

Published
2006

23. Diamagnetic levitation with permanent magnets for contactless guiding and trapping of microdroplets and particles in air and liquids

Author

H. Rostaing, H. Chetouani, Christian Jeandey, Vincent Haguet, Gilbert Reyne, C. Dieppedale, Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Garcia, Sylvie, Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Riado, Elise, and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology

Subjects
010302 applied physics, Materials science, Condensed matter physics, Magnetism, [SPI.NRJ]Engineering Sciences [physics]/Electric power, 02 engineering and technology, Trapping, 021001 nanoscience & nanotechnology, 01 natural sciences, Spin-stabilized magnetic levitation, Electronic, Optical and Magnetic Materials, Paramagnetism, Magnet, 0103 physical sciences, Levitation, Diamagnetism, Electrical and Electronic Engineering, 0210 nano-technology, Magnetic levitation, ComputingMilieux_MISCELLANEOUS, [SPI.NRJ] Engineering Sciences [physics]/Electric power
Abstract

Diamagnetic levitation was applied to micro-positioning and trapping of diamagnetic micron-sized particles and micro-droplets. Scale reduction laws for magnetism indicate that diamagnetism is highly efficient for magnetic MEMS. We demonstrate here that diamagnetic bodies can be contactless guided along magnetic grooves or trapped in magnetic wells both in air and in liquids. Levitation of water, ethanol and oil microdroplets was achieved within a 1.6 mm bore of a cylindrical magnet and in micromachined bulk magnets. Thick permanent micromagnets were also electroplated above silicon wafers in various matrix configurations. Diamagnetic levitation on micromagnet arrays was tested with 3 mum-large diamagnetic latex beads in a paramagnetic buffered solution. Trapping and arraying of microparticles in stable spatial levitation within magnetic wells were successfully achieved

Published
2006

24. Theoretical characterization of the electronic properties of extended thienylenevinylene oligomers

Author

Vincent Haguet, Didier Stiévenard, Christophe Delerue, Guy Allan, Jean Roncali, Pierre Frère, Christophe Krzeminski, Eric Levillain, Institut d’Électronique, de Microélectronique et de Nanotechnologie - UMR 8520 (IEMN), Centrale Lille-Institut supérieur de l'électronique et du numérique (ISEN)-Université de Valenciennes et du Hainaut-Cambrésis (UVHC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Université Polytechnique Hauts-de-France (UPHF), Chimie, Ingénierie Moléculaire et Matériaux d'Angers (CIMMA), and Université d'Angers (UA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Chemical Physics (physics.chem-ph), Materials science, General Physics and Astronomy, FOS: Physical sciences, 02 engineering and technology, Dihedral angle, Computational Physics (physics.comp-ph), 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 0104 chemical sciences, [PHYS.PHYS.PHYS-COMP-PH]Physics [physics]/Physics [physics]/Computational Physics [physics.comp-ph], Molecular wire, Delocalized electron, Tight binding, Chemical physics, Ab initio quantum chemistry methods, Physics - Chemical Physics, Density functional theory, Physical and Theoretical Chemistry, Local-density approximation, 0210 nano-technology, Electronic band structure, Physics - Computational Physics
Abstract

International audience; We present semiempirical tight binding calculations on thienylenevinylene oligomers up to the hexadecamer stage (n=16) and ab initio calculations based on the local density approximation up to n=8. The results correctly describe the experimental variations of the gap versus size, the optical spectra, and the electrochemical redox potentials. We propose a simple model to deduce from the band structure of the polymer chain the electronic states of the oligomers close to the gap. We analyze the evolution of the gap as a function of the torsion angle between consecutive cells: the modifications are found to be small up to a ~30° angle. We show that these oligomers possess extensive pi-electron delocalization along the molecular backbone which makes them interesting for future electronic applications such as molecular wires.

Published
1999

25. Monitoring impedance changes associated with motility and mitosis of a single cell

Author

Xavier Gidrol, Takeshi Ishibashi, Yu Hoshino, Lamya Ghenim, Vincent Haguet, Matsuhiko Nishizawa, and Hirokazu Kaji

Subjects
Conductometry, Cell division, Cell Culture Techniques, Biomedical Engineering, Mitosis, Motility, Bioengineering, Equipment Design, General Chemistry, Adhesion, Microfluidic Analytical Techniques, Biology, Biochemistry, Cell biology, Equipment Failure Analysis, Prophase, Cell Movement, Humans, Plethysmography, Impedance, Cell adhesion, Metaphase, HeLa Cells, Anaphase
Abstract

We present a device enabling impedance measurements that probe the motility and mitosis of a single adherent cell in a controlled way. The micrometre-sized electrodes are designed for adhesion of an isolated cell and enhanced sensitivity to cell motion. The electrode surface is switched electro-chemically to favour cell adhesion, and single cells are attracted to the electrode using positive dielectrophoresis. Periods of linear variation in impedance with time correspond to the motility of a single cell adherent to the surface estimated at 0.6 μm h(-1). In the course of our study we observed the impedance changes associated with mitosis of a single cell. Electrical measurements, carried out concomitantly with optical observations, revealed three phases, prophase, metaphase and anaphase in the time variation of the impedance during cell division. Maximal impedance was observed at metaphase with a 20% increase of the impedance. We argue that at mitosis, the changes detected were due to the charge density distribution at the cell surface. Our data demonstrate subtle electrical changes associated with cell motility and for the first time with division at the single-cell level. We speculate that this could open up new avenues for characterizing healthy and pathological cells.

Published
2010

27. Monitoring impedance changes associated with motility and mitosis of a single cell.

Author

Lamya Ghenim, Yu Hoshino, Takeshi Ishibashi, Vincent Haguet, and Xavier Gidrol

Subjects
MITOSIS, CELL adhesion, CELL motility, MICROMETERS, DIELECTROPHORESIS, IMPEDANCE spectroscopy
Abstract

We present a device enabling impedance measurements that probe the motility and mitosis of a single adherent cell in a controlled way. The micrometre-sized electrodes are designed for adhesion of an isolated cell and enhanced sensitivity to cell motion. The electrode surface is switched electro-chemically to favour cell adhesion, and single cells are attracted to the electrode using positive dielectrophoresis. Periods of linear variation in impedance with time correspond to the motility of a single cell adherent to the surface estimated at 0.6 μm h−1. In the course of our study we observed the impedance changes associated with mitosis of a single cell. Electrical measurements, carried out concomitantly with optical observations, revealed three phases, prophase, metaphase and anaphase in the time variation of the impedance during cell division. Maximal impedance was observed at metaphase with a 20% increase of the impedance. We argue that at mitosis, the changes detected were due to the charge density distribution at the cell surface. Our data demonstrate subtle electrical changes associated with cell motility and for the first time with division at the single-cell level. We speculate that this could open up new avenues for characterizing healthy and pathological cells. [ABSTRACT FROM AUTHOR]

Published
2010
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28. Organoids-based High Content Screening in Oncology

Author

Xavier Gidrol, Laperrousaz, B., Patricia Obeid, Porte, S., Gerbeau, S., Kermarrec, F., Vincent Haguet, Dolega, M., Christophe Le Priol, cedric allier, Nathalie Picollet-d'Hahan, Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut National de la Santé et de la Recherche Médicale (INSERM)-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), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), 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 National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), and Bertacchi Griffi, Nathalie

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio]
Abstract

International audience; To analyze the phenotypic consequences of genetic perturbation in mammalian cells with drugs, RNAi or expression vectors, there are increasing needs for systematic cell-based high content screening (HCS) approaches, particularly in oncology. Although several groups are performing HCS in human cell lines, the real challenge in translational research remains screening on a reduced number of primary cells directly obtained from patients in a microenvironment that would resemble the original tissues. While standard monolayer two-dimensional culture conditions or even spheroids resulting form the forced aggregation of cells are poor mimics of the tumour natural history, microfabricated systems enable three-dimensional clonal organoids cultures and have the potential to provide biological insight not achievable before. We use microfluidics and MEMS (MicroElectroMechanical Systems) to analyze the phenotypic consequences of genetic perturbations (RNAi-based HCS) in 3D organoids. Specifically, we present a novel approach based on a flowfocusing microfluidic system that encapsulates either single prostatic or mammary cell in Matrigel beads and assay for development of organoids. Furthermore, we developed new imaging technology to monitor live organoids self-assembly and inter organoids cell trafficking.

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