Microsonde optique et électrique pour l'enregistrement de neurones unitaires in vivo
|Advisor:||De Koninck, Yves; Vallée, Réal|
|Abstract:||The central nervous system is composed of heterogeneous populations of neurons. Studying their functional properties in the intact central nervous system (CNS) is key to be able to understand their respective role in signal processing within entire networks. To achieve this, it is essential to be able to record electrophysiologically from identified neurons in the intact tissue. Recording from identified cells types in vivo has remained a challenge, especially for local circuit neurons. Novel fluorescent labeling techniques open new possibilities on that front. To take full advantage of these recent developments, the ability to record electrophysiological signals from single neurons must be combined with optical detection of individual cells deep into CNS tissue. Here it describe the development of a novel microprobe based on a dual core optical fiber: an optical core that excites locally fluorescent labeled cells and collects back the fluorescence, and an electrolyte filled hollow core that performs classical extracellular single unit electrophysiological measurements. In contrast to previous solutions, this novel design allows production of microprobes with sufficient optical resolution for single cell detection: the microprobes could be pulled down to tips sizes of 6 µm, which is smaller than the cell body diameter of most neuron populations. It is presented the evolution of the microprobe design and the experimental setup. To characterize the optical properties of the probes, it is showed a series of in vitro experiments and numerical simulations. Then, it is presented in vivo experiment to identify and record single spinal neurons labeled retrogradely with fluorescent dyes as well as single GABAergic interneurons expressing GFP in the brain of transgenic mice. It's also established a spatial criterion to correlate optical and electrophysiological signals, confirming co-detection of single cells. This novel microprobe vastly expands possibilities for in vivo electrophysiological recording by providing parallel access to single cell optical monitoring.|
|Document Type:||Thèse de doctorat|
|Open Access Date:||18 April 2018|
|Collection:||Thèses et mémoires|
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