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.
Highlights
Behind the apparent simplicity of dissociated neuronal cultures lies an incredibly rich catalogue of behaviors present in vivo, including growth and differentiation [1, 2], plasticity [3, 4], electrical activity [5,6,7,8,9,10] and information processing [11,12,13]
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 demonstrate the possibility of combining optogenetics, calcium imaging and microfluidics to study neuronal connectivity
Summary
Behind the apparent simplicity of dissociated neuronal cultures lies an incredibly rich catalogue of behaviors present in vivo, including growth and differentiation [1, 2], plasticity [3, 4], electrical activity [5,6,7,8,9,10] and information processing [11,12,13]. The culture of primary neurons, extracted from healthy or diseased animals, has rapidly become an invaluable tool to understand the nervous system, from the molecular actors of neuronal functions, to the behaviors produced by embodied cultures. Interfacing such cultures is a necessary step to understand many of these processes. For this purpose, Microelectrodes Arrays (MEAs) have remained the favorite alternative.
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