title: Combining microfluidics, optogenetics and calcium imaging to study neuronal communication in vitro.
abstract: In this paper we report the combination of microfluidics, optogenetics and calcium imaging as a cheap and convenient platform to study synaptic communication between neuronal populations in vitro. We first show that Calcium Orange indicator is compatible in vitro with a commonly used Channelrhodopsine-2 (ChR2) variant, as standard calcium imaging conditions did not alter significantly the activity of transduced cultures of rodent primary neurons. A fast, robust and scalable process for micro-chip fabrication was developed in parallel to build micro-compartmented cultures. Coupling optical fibers to each micro-compartment allowed for the independent control of ChR2 activation in the different populations without crosstalk. By analyzing the post-stimuli activity across the different populations, we finally show how this platform can be used to evaluate quantitatively the effective connectivity between connected neuronal populations.
authors.affiliation: MSC (Université Paris-Diderot, CNRS-UMR 7057), 5 Rue Thomas Mann, 75013 Paris, France; Physicochimie Curie (Institut Curie, CNRS-UMR 168, UPMC), Institut Curie, Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France; Department of Complex Systems, Weizmann Institute, Rehovot, Israel.
authors.forename: Renaud
authors.initials: R
authors.lastname: Renault
authors.affiliation: Department of Complex Systems, Weizmann Institute, Rehovot, Israel.
authors.forename: Nirit
authors.initials: N
authors.lastname: Sukenik
authors.affiliation: Physicochimie Curie (Institut Curie, CNRS-UMR 168, UPMC), Institut Curie, Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Descroix
authors.affiliation: Physicochimie Curie (Institut Curie, CNRS-UMR 168, UPMC), Institut Curie, Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Malaquin
authors.affiliation: Physicochimie Curie (Institut Curie, CNRS-UMR 168, UPMC), Institut Curie, Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Viovy
authors.affiliation: Biological Adaptation and Ageing (CNRS, UMR 8256), F-75005, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR 8256, B2A, Institut de Biologie Paris Seine, F-75005, Paris, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Peyrin
authors.affiliation: MSC (Université Paris-Diderot, CNRS-UMR 7057), 5 Rue Thomas Mann, 75013 Paris, France.
authors.forename: Samuel
authors.initials: S
authors.lastname: Bottani
authors.affiliation: MSC (Université Paris-Diderot, CNRS-UMR 7057), 5 Rue Thomas Mann, 75013 Paris, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Monceau
authors.affiliation: Department of Complex Systems, Weizmann Institute, Rehovot, Israel.
authors.forename: Elisha
authors.initials: E
authors.lastname: Moses
authors.affiliation: Physicochimie Curie (Institut Curie, CNRS-UMR 168, UPMC), Institut Curie, Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France; Biological Adaptation and Ageing (CNRS, UMR 8256), F-75005, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR 8256, B2A, Institut de Biologie Paris Seine, F-75005, Paris, France.
authors.forename: Maéva
authors.initials: M
authors.lastname: Vignes
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D002450:Cell Communication / Q000502:physiology*; D002478:Cells, Cultured; D000075402:Channelrhodopsins; D003198:Computer Simulation; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D066298:In Vitro Techniques; D044085:Microfluidics / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D062308:Optogenetics / Q000379:methods*; D051381:Rats
chemical_list: D000075402:Channelrhodopsins; D002118:Calcium