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dc.contributor.authorDeNotta, Sally Anne Lenore
dc.date.accessioned2019-04-02T14:00:54Z
dc.date.available2019-04-02T14:00:54Z
dc.date.issued2018-12-30
dc.identifier.otherDeNotta_cornellgrad_0058F_11181
dc.identifier.otherhttp://dissertations.umi.com/cornellgrad:11181
dc.identifier.otherbibid: 10758090
dc.identifier.urihttps://hdl.handle.net/1813/64950
dc.description.abstractSpinal cord disorders are frequent causes of morbidity and mortality in human and veterinary medicine alike. Despite spinal cord disease affecting nearly every vertebrate species on earth, the pathophysiological mechanisms of central nervous system (CNS) disease remain poorly understood, largely due to failure of traditional ex-vivo imaging methods to capture the dynamic nature of complex neurophysiologic interactions. To address this, we have developed novel optical techniques for high-resolution, in-vivo imaging of the spinal cord in rodent models. This dissertation details novel methods for two-photon excited fluorescence (2PEF) microscopy of the spinal cord that enable direct observation of the cellular interactions occurring during health and disease. Specifically, we have developed surgical procedures that allow long term optical access in multiple regions of the mouse spinal cord, optimized labeling strategies for multicolor fluorescent in vivo imaging of cellular interactions, demonstrated successful recording of real-time calcium transients and neural activity from populations of sensory and motor neurons in the rodent spinal cord, explored and refined established models of SCI and neuropathic pain to make them suitable for serial imaging of axonal dynamics, inflammatory responses, and alterations in neural circuitry, designed and custom built a treadmill system on which a mouse can walk and run while spine-fixed under a multiphoton microscope, and implemented a recording system for capturing video of spine-fixed, running mice from which 3D spatiotemporal positioning of limbs can be measured and ultimately integrated into a comprehensive kinematic gait analysis system. Finally, we discuss the current capabilities and limitations of 2PEF microscopy for spinal cord imaging, and introduce ongoing initiatives and the inevitable transition to the next generation of multiphoton microscopy, three-photon excited fluorescence (3PEF), as a way to overcome the current challenges to 2PEF and further drive spinal cord research into the future.
dc.language.isoen_US
dc.subjectin vivo imaging
dc.subjectmicroglia
dc.subjectspinal cord
dc.subjectspinal cord injury
dc.subjectOptics
dc.subjectNeurosciences
dc.subjectMultiphoton
dc.titleIn-vivo Multiphoton Excited Fluorescence Microscopy of the Spinal Cord
dc.typedissertation or thesis
thesis.degree.disciplineMolecular and Integrative Physiology
thesis.degree.grantorCornell University
thesis.degree.levelDoctor of Philosophy
thesis.degree.namePh. D., Molecular and Integrative Physiology
dc.contributor.chairSchaffer, Chris
dc.contributor.committeeMemberDando, Robin
dc.contributor.committeeMemberLibert, Sergiy
dc.contributor.committeeMemberMiller, Andrew David
dcterms.licensehttps://hdl.handle.net/1813/59810
dc.identifier.doihttps://doi.org/10.7298/a1tm-z178


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