Advances in two-photon fluorescence microscopy for high-resolution anatomical and functional imaging of cell populations in the intact brain [Elektronische Ressource] / presented by Axel Nimmerjahn

Dissertationsubmitted to theCombined Faculties for the Natural Sciences and for Mathematicsof the Ruperto-Carola University of Heidelberg, Germanyfor the degree ofDoctor of Natural Sciencespresented byDiplom-PhysicistAxel Nimmerjahnborn inKulmbach, GermanyOral Examination: July 4th, 2005Advances in Two-Photon Fluorescence Microscopyfor High-Resolution Anatomical and Functional Imagingof Cell Populations in the Intact BrainRefereesProf. Dr. Bert SakmannProf. Dr. Karlheinz MeierAdvances in Two-Photon Fluorescence Microscopy for High-ResolutionAnatomical and Functional Imaging of Cell Populations in the IntactBrainTwo-photon microscopy has enabled high-resolution imaging of single cells in the brain of anaes-thetizedanimals.Herewedevelopedtwo-photonmicroscopytowardsimagingofcellpopulationsintheneocortexofawakebehavingrodents.Forthispurpose,wedevelopedtwominiaturetwo-photonmicroscopes based on fluorescence excitation through a hollow-core photonic crystal fiber and acoherentfiber-bundle,respectively.Inaddition,wedemonstratetheirapplicabilitytoinvivoimag-ing. Furthermore, as meaningful biological application critically depends on fluorescence labeling,we developed staining methods for three different cell populations. In particular, we used Sindbis-and Lentiviral gene transfer into neurons for targeted expression of fluorescent indicators.