title: Generation of living cell arrays for atomic force microscopy studies.
abstract: Atomic force microscopy (AFM) is a useful tool for studying the morphology or the nanomechanical and adhesive properties of live microorganisms under physiological conditions. However, to perform AFM imaging, living cells must be immobilized firmly enough to withstand the lateral forces exerted by the scanning tip, but without denaturing them. This protocol describes how to immobilize living cells, ranging from spores of bacteria to yeast cells, into polydimethylsiloxane (PDMS) stamps, with no chemical or physical denaturation. This protocol generates arrays of living cells, allowing statistically relevant measurements to be obtained from AFM measurements, which can increase the relevance of results. The first step of the protocol is to generate a microstructured silicon master, from which many microstructured PDMS stamps can be replicated. Living cells are finally assembled into the microstructures of these PDMS stamps using a convective and capillary assembly. The complete procedure can be performed in 1 week, although the first step is done only once, and thus repeats can be completed within 1 d.
authors.affiliation: 1] Centre Nationale de la Recherche Scientifique (CNRS), Laboratoire d'Analyse et d'Architecture des Systèmes (LAAS), Toulouse, France. [2] Université de Toulouse; LAAS, Institut des Technologies Avancées en Sciences du Vivant (ITAV), Institute de Pharmacologie et de Biologie Structurale (IPBS), Toulouse, France. [3] CNRS, Unité Mixte de Recherche (UMR) 7565, Laboratoire Structure et Réactivité des Systèmes Moléculaires Complexes (SRSMC), Vandœuvre-lès-Nancy, France. [4] Université de Lorraine, UMR 7565, Faculté de Pharmacie, Nancy, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Formosa
authors.affiliation: 1] Université de Toulouse; LAAS, Institut des Technologies Avancées en Sciences du Vivant (ITAV), Institute de Pharmacologie et de Biologie Structurale (IPBS), Toulouse, France. [2] CNRS, IPBS, UMR 5089, Toulouse, France.
authors.forename: Flavien
authors.initials: F
authors.lastname: Pillet
authors.affiliation: 1] Centre Nationale de la Recherche Scientifique (CNRS), Laboratoire d'Analyse et d'Architecture des Systèmes (LAAS), Toulouse, France. [2] Université de Toulouse; LAAS, Institut des Technologies Avancées en Sciences du Vivant (ITAV), Institute de Pharmacologie et de Biologie Structurale (IPBS), Toulouse, France. [3] L'Institut National de la Recherche Agronomie (INRA), UMR 972 Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Toulouse, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Schiavone
authors.affiliation: 1] CNRS, Unité Mixte de Recherche (UMR) 7565, Laboratoire Structure et Réactivité des Systèmes Moléculaires Complexes (SRSMC), Vandœuvre-lès-Nancy, France. [2] Université de Lorraine, UMR 7565, Faculté de Pharmacie, Nancy, France. [3] ABC Platform, Nancy, France.
authors.forename: Raphaël E
authors.initials: RE
authors.lastname: Duval
authors.affiliation: 1] Université de Toulouse; LAAS, Institut des Technologies Avancées en Sciences du Vivant (ITAV), Institute de Pharmacologie et de Biologie Structurale (IPBS), Toulouse, France. [2] Université de Toulouse, Laboratoire de Physique et Chimie de Nano-Objets (LPCNO), L'Institut National des Sciences Appliquées (INSA)-CNRS-Université Toulouse III-Paul Sabatier, Toulouse, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Ressier
authors.affiliation: 1] Centre Nationale de la Recherche Scientifique (CNRS), Laboratoire d'Analyse et d'Architecture des Systèmes (LAAS), Toulouse, France. [2] Université de Toulouse; LAAS, Institut des Technologies Avancées en Sciences du Vivant (ITAV), Institute de Pharmacologie et de Biologie Structurale (IPBS), Toulouse, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Dague
mesh_terms: D018914:Cells, Immobilized / Q000378:metabolism*; D004129:Dimethylpolysiloxanes; D018625:Microscopy, Atomic Force / Q000379:methods*; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon