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Title: Improved methods for functional neuronal imaging with genetically encoded voltage indicators
Author: Quicke, Peter Ernest Hosking
ISNI:       0000 0004 8499 5104
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2019
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Voltage imaging has the potential to revolutionise neuronal physiology, enabling high temporal and spatial resolution monitoring of sub- and supra-threshold activity in genetically defined cell classes. Before this goal is reached a number of challenges must be overcome: novel optical, genetic, and experimental techniques must be combined to deal with voltage imaging's unique difficulties. In this thesis three techniques are applied to genetically encoded voltage indicator (GEVI) imaging. First, I describe a multifocal two-photon microscope and present a novel source localisation control and reconstruction algorithm to increase scattering resistance in functional imaging. I apply this microscope to image population and single-cell voltage signals from voltage sensitive fluorescent proteins in the first demonstration of multifocal GEVI imaging. Second, I show that a recently described genetic technique that sparsely labels cortical pyramidal cells enables single-cell resolution imaging in a one-photon widefield imaging configuration. This genetic technique allows simple, high signal-to-noise optical access to the primary excitatory cells in the cerebral cortex. Third, I present the first application of lightfield microscopy to single cell resolution neuronal voltage imaging. This technique enables single-shot capture of dendritic arbours and resolves 3D localised somatic and dendritic voltage signals. These approaches are finally evaluated for their contribution to the improvement of voltage imaging for physiology.
Supervisor: Schultz, Simon ; Neil, Mark ; Knöpfel, Thomas Sponsor: Engineering and Physical Sciences Research Council
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral