%0 Generic
%A Emde, Henrik von der
%C Heidelberg
%D 2024
%F heidok:35738
%R 10.11588/heidok.00035738
%T Molecule Localization with STED
%U https://archiv.ub.uni-heidelberg.de/volltextserver/35738/
%X I present a re-implementation of the super-resolution method MINSTED, which enhances the stimulated emission interaction probability of the STED (stimulated emission depletion) process and therefore reduces the optical power required by approximately one order of magnitude. This results in an enhanced localization performance, achieving uncertainties in single-molecule localization below 1nm. This molecular-scale localization precision is applied to biological specimens upon the use of the DNA-PAINT labelling scheme. The imaging of the nuclear pore complex in fixed HeLa cells is demonstrated and the colocalization of synaptic vesicle proteins in a multi-colour experiment in fixed rat hippocampal neurons was investigated. The MINSTED localization approach is additionally shown to be used for single molecule tracking at nanometre-millisecond spatio-temporal precision. The biological relevance is highlighted by tracking of the motor protein kinesin-1, clearly resolving its 16nm steps with a temporal precision of <2ms.