%0 Generic
%A Bergermann, Fabian Manuel
%D 2016
%F heidok:22323
%K Superresolution
%R 10.11588/heidok.00022323
%T Massively parallelized STED nanoscopy
%U https://archiv.ub.uni-heidelberg.de/volltextserver/22323/
%X Fluorescence microscopy constitutes a key method for the virtually  non-invasive study of biological structures and processes on a subcellular  scale. Stimulated Emission Depletion (STED) nanoscopy extends  fluorescence imaging to nanometer resolutions. However, this  method usually acquires an image by scanning small pixels in a sequential  manner, which can lead to long acquisition times of several  minutes. This limits the feasibility of many large-scale superresolution  experiments, because faster acquisition times imply sacrificing  either highest resolution, a good signal-to-noise ratio or a large image  size. To unleash the full spatio-temporal resolving power potential of  STED nanoscopy on a large imaging region, massively parallelized  acquisition is therefore inevitable.  In this thesis, I develop a comprehensive and quantitative description  of parallelized STED and validate the derived findings by means  of two experimental implementations. Investigating the key technical  parameters and their interplay reveals the possibility of reducing the  laser power by up to three orders of magnitude below the value required  for serially acquiring STED systems. This eliminates a major  bottleneck of previously reported attempts to parallelize STED nanoscopy,  limiting them to tiny image sizes or inferior resolutions. Moreover,  using a rather simple optical arrangement, I was able to enlarge  the superresolved image area to 33 μm edge length (roughly the dimensions  of a small cell), featuring a resolution down to 30 nm and a  13000-fold degree of parallelization. While the implementations presented  here should be viewed as prototypes, they prove the technical  feasibility of massively parallelized STED nanoscopy. The methods  developed in this thesis in combination with suitable high-speed detectors  bring video-rate STED nanoscopy of whole cells within reach.