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dc.contributor.authorAkbari, Najva
dc.date.accessioned2022-01-24T18:08:20Z
dc.date.issued2021-12
dc.identifier.otherAkbari_cornellgrad_0058F_12871
dc.identifier.otherhttp://dissertations.umi.com/cornellgrad:12871
dc.identifier.urihttps://hdl.handle.net/1813/110919
dc.description125 pages
dc.description.abstractMultiphoton microscopy has enabled unprecedented access to biological systems in their native environment. Two-photon microscopy was invented at Cornell University in 1990 and has since been adopted all over the world for deep in vivo imaging. While three-photon microscopy was demonstrated not much later (1996), the required technology and incentive for further development of this technique was not available until much later in 2013 once again at Cornell, the original home of multiphoton imaging. Three-photon microscopy has since enabled unprecedented access deep in highly scattering biological tissue.While the advantage of three-photon microscopy for deep imaging has been demonstrated, the depth limit of this technique had not been established. In this dissertation I provide a comprehensive theoretical and experimental investigation of the depth limit of multiphoton microscopy techniques. I demonstrate experimentally that high spatial resolution diffraction-limited imaging at a depth of 10 scattering mean-free paths, which is nearly twice the transport mean free path, is possible with multiphoton microscopy. Our results indicate that the depth limit of three-photon microscopy is significantly beyond what has been achieved in biological tissues so far, and further technological development is required to reach the full potential of three-photon microscopy. Additionally, I demonstrate unique possibilities that multiphoton microscopy enables in neuroscience research by applying three-photon microscopy and third harmonic generation microscopy to a miniature adult vertebrate, Danionella dracula. My work demonstrates that two- and three-photon microscopy can be used to access the entire depth of the adult wild type Danionella dracula brain without any modifications to the animal other than mechanical stabilization. These results show that multiphoton microscopy is ideal for readily penetrating the entire adult brain within the geometry of these miniature animals’ head structures, without the need for pigment removal. With multiphoton microscopy enabling optical access to the adult brain and a repertoire of methods that allow observation of the larval brain, Danionella provides a model system for readily studying the entire brain over the lifetime of a vertebrate for the first time.
dc.language.isoen
dc.subjectDanionella
dc.subjectMultiphoton Microscopy
dc.subjectNeuroscience
dc.subjectZebrafish
dc.titleTHE DEPTH LIMIT OF MULTIPHOTON MICROSCOPY AND APPLICATIONS TO IMAGING MINIATURE ADULT VERTEBRATE BRAINS
dc.typedissertation or thesis
dc.description.embargo2023-01-05
thesis.degree.disciplineApplied Physics
thesis.degree.grantorCornell University
thesis.degree.levelDoctor of Philosophy
thesis.degree.namePh. D., Applied Physics
dc.contributor.chairXu, Chris
dc.contributor.committeeMemberSchaffer, Chris
dc.contributor.committeeMemberFetcho, Joseph R.
dcterms.licensehttps://hdl.handle.net/1813/59810.2
dc.identifier.doihttps://doi.org/10.7298/459n-qk89


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