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Adaptive optics allows STED-FCS measurements in the cytoplasm of living cells

Barbotin, Aurélien; Galiani, Silvia; Urbancic, Iztok; Eggeling, Christian; Booth, Martin J.
in: Optics Express (2019) 23378

Fluorescence correlation spectroscopy in combination with super-resolution stimulated emission depletion microscopy (STED-FCS) is a powerful tool to investigate molecular diffusion with sub-diffraction resolution. It has been of particular use for investigations of two dimensional systems like cell membranes, but has so far seen very limited applications to studies of three-dimensional diffusion. One reason for this is the extreme sensitivity of the axial (z) STED depletion pattern to optical aberrations. We present here an adaptive optics-based correction method that compensates for these aberrations and allows STED-FCS measurements in the cytoplasm of living cells.

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