STED nanoscopy of the living brain

Berning, Sebastian

dc.contributor.advisor	Hell, Stefan Prof. Dr.	de
dc.contributor.author	Berning, Sebastian	de
dc.date.accessioned	2012-05-03T18:34:32Z	de
dc.date.accessioned	2013-01-18T14:23:54Z	de
dc.date.available	2013-01-30T23:51:28Z	de
dc.date.issued	2012-05-03	de
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-000D-F0B1-E	de
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-3200
dc.description.abstract	Im Laufe des letzten Jahrzehnts ist eine neue Klasse von Fluoreszenzmikroskopen entwickelt worden, welche eine vielfach bessere r	de
dc.format.mimetype	application/pdf	de
dc.language.iso	eng	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/3.0/	de
dc.title	STED nanoscopy of the living brain	de
dc.type	doctoralThesis	de
dc.title.translated	STED-Mikroskopie des intakten Gehirns	de
dc.contributor.referee	Hell, Stefan Prof. Dr.	de
dc.date.examination	2011-12-13	de
dc.subject.dnb	500 Naturwissenschaften	de
dc.subject.gok	RPV 280	de
dc.subject.gok	RPV 720	de
dc.subject.gok	MED 531	de
dc.subject.gok	MED 272	de
dc.description.abstracteng	Over the past decade, a new class of fluorescence microscopes has evolved which thoroughly break the diffraction limit of classical far-field light microscopy and thus provide significantly improved spatial resolution. Especially so-called STED microscopes have shown to provide valuable information otherwise inaccessible when investigating molecular processes in living biological specimens. Naturally, the question arises whether the findings obtained from cultured cells or tissue samples can be directly transferred to the whole living organism. Especially in the case of the brain, where neurons process information only by linking to thousands of neighboring neurons, it is often questionable to deduce the function of the whole organ solely from observations in isolated systems. Imaging directly in the cortex of living higher animals therefore represents the gold standard in the neurosciences. As STED microscopy is becoming increasingly popular for experiments in cultured brain slices, it is important to verify the obtained results accordingly. The scope of this thesis is to establish STED microscopy directly in the living mouse brain. In order to identify potential problems arising from aberrations induced by the refractive index mismatch between the immersion system and the brain tissue, the system is first assessed numerically. Following this, a new microscope in upright configuration is constructed addressing the specific requirements for in-vivo experiments. As a result, it is shown for the first time that super-resolution light microscopy in living higher organisms is indeed possible. Using the newly established protocols for preparation and imaging, < 70nm structural features are routinely observed in transgenic mice expressing the popular fluorescent proteins EGFP and EYFP. Time-lapse recordings of single dendritic spines reveal subtle morphological changes on the nanometer scale as well as the timescales on which they take place. Furthermore, images of glial cells such as astrocytes and oligodendrocytes exhibit small features which could previously only be seen in electron micrographs. Finally, a new recording scheme is demonstrated which enables in-vivo STED microscopy with two super-resolved spectral channels in double-transgenic mice. Initial experiments show that this is a promising method for future studies of interactions at the finest processes of neurons and glial cells.	de
dc.contributor.coReferee	Schmidt, Christoph F. Prof. Dr.	de
dc.contributor.thirdReferee	Grubmüller, Helmut Prof. Dr.	de
dc.subject.topic	Göttingen Graduate School for Neurosciences and Molecular Biosciences (GGNB)	de
dc.subject.ger	STED	de
dc.subject.ger	Mikroskopie	de
dc.subject.ger	Mikroskop	de
dc.subject.ger	Dendriten	de
dc.subject.ger	Maus	de
dc.subject.ger	Gehirn	de
dc.subject.eng	STED	de
dc.subject.eng	microscopy	de
dc.subject.eng	microscope	de
dc.subject.eng	dendrites	de
dc.subject.eng	mouse	de
dc.subject.eng	brain	de
dc.subject.bk	42.12	de
dc.subject.bk	42.03	de
dc.subject.bk	33.18	de
dc.identifier.urn	urn:nbn:de:gbv:7-webdoc-3491-0	de
dc.identifier.purl	webdoc-3491	de
dc.affiliation.institute	Göttinger Graduiertenschule für Neurowissenschaften und Molekulare Biowissenschaften (GGNB)	de
dc.identifier.ppn	737899379	de

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GGNB - Göttinger Graduiertenzentrum für Neurowissenschaften, Biophysik und molekulare Biowissenschaften [1152]
GGNB - Göttingen Graduate Center for Neurosciences, Biophysics and Molecular Biosciences

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