%0 Generic
%A Giske, Arnold
%D 2007
%F heidok:7969
%K STED MikroskopieSTED microscopy , low temperature
%R 10.11588/heidok.00007969
%T CryoSTED microscopy - a new spectroscopic approach for improving the resolution of STED microscopy using low temperature
%U https://archiv.ub.uni-heidelberg.de/volltextserver/7969/
%X This work presents a new approach for the further improving of the resolution in the sub-diffraction fluorescence STED microscopy. The reduction of the sample temperature down to 76K eliminates the disadvantages of anti-Stokes fluorescence excitation and allows the use of wavelengths with larger STED efficiencies. On the studied dyes, this approach enhances the resolving power by a factor of 1.6 compared to room temperature and suggests further technical improvements for such resolution increase. Besides resolution, the present work solves the issue of the high triplet population in fluorescence imaging at liquid nitrogen temperature. A technique for the triplet depopulation has been developed. It provides a manifold brightness increase in the fluorescence imaging at low temperatures. The utilization of the low temperature also increases the photo-stability of the fluorescent markers. The combination of a complete triplet depopulation and higher fatigue resistance allows an enhanced signal to noise ratio in STED and fluorescence microscopy in general.