Abstract
Studying the formation and function of neuronal circuitry is complicated by the heterogeneity and high density of neuronal processes and synapses. Compartmentalized cell culture systems offer a simple yet powerful solution for isolation of axons from dendrites and cell somas. This chapter describes how to manufacture and use a microfluidic chip with a modular design for highly defined isolation of axons, asymmetric genetic manipulation, and whole-cell patch clamp recording. The microfluidic chip consists of detachable and resealable layers that allow multiple modes of operation during cell culture, fluidic isolation for limited transfection, and recording with low-angle electrode access. This versatile technique is useful for functional studies that require specific expression of for example optogenetic tools in presynaptic neurons or for studying the entry of pathogenic particles, such as viruses or oligomers of misfolded proteins, into presynaptic structures.
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Acknowledgements
This research was supported by the Academy of Finland (grant #253223 and grant #266820).
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Sakha, P., Brunello, C., Heikkinen, J., Jokinen, V., Huttunen, H.J. (2015). Asymmetric Genetic Manipulation and Patch Clamp Recording of Neurons in a Microfluidic Chip. In: Biffi, E. (eds) Microfluidic and Compartmentalized Platforms for Neurobiological Research. Neuromethods, vol 103. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2510-0_4
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DOI: https://doi.org/10.1007/978-1-4939-2510-0_4
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