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Free keywords:
single-molecule localization microscopy, super-resolution microscopy, photoswitchable fluorophores, photokinetic analysis, kinetic models for fluorophores, membrane protein stoichiometry
Abstract:
We report on quantitative single-molecule localization microscopy, a method that next to super-resolved images of cellular structures provides information on protein copy numbers in protein clusters. This approach is based on the analysis of blinking cycles of single fluorophores, and on a model-free description of the distribution of the number of blinking events. We describe the experimental and analytical procedures, present cellular data of plasma membrane proteins and discuss the applicability of this method