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Novel tools for simultaneous optogenetic manipulation and calcium imaging in the zebrafish nervous system

Gheisari, Ali

Abstract (englisch):
The large number of neurons and neural interconnection makes the nervous system a highly dense and complex network. Understanding the functionality of such a network requires not only high-throughput recording of neural activities with cellular resolution but also a non-invasive and precisely defined interaction with the neurons. This work demonstrates two pivotal steps towards these aims:
(i) Developing a fluorescence microscope based on Bessel light-sheet illumination to record neural activities by means of calcium imaging. Chapter II describes the successful development and construction of an in vivo light-sheet fluorescence microscope (in vivo LSFM) based on conventional digitally laser scanning light-sheet microscopy (DSLM) with interchangeable Gaussian and Bessel illumination modalities to compare the performance of both methods. The Bessel illumination modality in comparison to the Gaussian one reveals not only a two-fold improvement in axial resolution but also a reduction by the factor of 4 in the shadowing artifact and consequently, 35 times improvement in detecting the correct neural activities from the calcium signals.
(ii) Providing a suitable transgenic zebrafish model for optogenetic manipulation; Optogenetics apply light to facilitate the interaction with a genetically-engineered cell and/or populations of cells. ... mehr

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DOI: 10.5445/IR/1000071261
Zugehörige Institution(en) am KIT Zoologisches Institut (ZOO)
Publikationstyp Hochschulschrift
Jahr 2017
Sprache Englisch
Identifikator urn:nbn:de:swb:90-712618
KITopen-ID: 1000071261
Verlag KIT, Karlsruhe
Umfang 151 S.
Abschlussart Dissertation
Fakultät Fakultät für Chemie und Biowissenschaften (CHEM-BIO)
Institut Zoologisches Institut (ZOO)
Prüfungsdatum 27.04.2017
Referent/Betreuer Prof. M. Bastmeyer (KIT); Prof. F. S. Pavone (Univ. d. Studi, Firenze)
Bemerkung zur Veröffentlichung Zugl.: Florenz, Univ. d. Studi, Diss.
Schlagworte Fluorescence, Light Microscopy, Optical sectioning, Light sheet microscopy, Bessel Light sheet, Neuronal activity recording, in vivo imaging, Optogenetics, transgenic zebrafish, Chlamydomonas channelrhodopsin1, Volvox channelrhodopsin1, C1V1
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