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

Opt Express. 2019 Aug 5;27(16):23378-23395. doi: 10.1364/OE.27.023378.

Abstract

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.

MeSH terms

  • Cell Survival
  • Cytoplasm / metabolism*
  • Diffusion
  • Humans
  • Microscopy, Fluorescence / methods*
  • Optics and Photonics*
  • Solutions
  • Spectrometry, Fluorescence / methods*

Substances

  • Solutions