Combining Microfluidics, Optogenetics and Calcium Imaging to Study Neuronal Communication In Vitro

Renaud Renault (1, 2, 3) , Nirit Sukenik (3) , Stéphanie Descroix (2) , Laurent Malaquin (2) , Jean-Louis Viovy (2) , Jean-Michel Peyrin (4) , Samuel Bottani (1) , Pascal Monceau (1) , Elisha Moses (3) , Maéva Vignes (2, 4)

1 MSC - Matière et Systèmes Complexes
2 PCC - Physico-Chimie-Curie
3 Weizmann Institute of Science [Rehovot, Israël]
4 B2A - Adaptation Biologique et Vieillissement = Biological Adaptation and Ageing

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.

Domaines: Neurosciences

Renaud Renault, Nirit Sukenik, Stéphanie Descroix, Laurent Malaquin, Jean-Louis Viovy, et al.. Combining Microfluidics, Optogenetics and Calcium Imaging to Study Neuronal Communication In Vitro. PLoS ONE, 2015, 10 (4), pp.e0120680. ⟨10.1371/journal.pone.0120680⟩. ⟨hal-01227804⟩