Localization Microscopy of Actin Cytoskeleton in Human Platelets

Int J Mol Sci. 2018 Apr 11;19(4):1150. doi: 10.3390/ijms19041150.

Abstract

Here, we measure the actin cytoskeleton arrangement of different morphological states of human platelets using a new protocol for photo-switching of rhodamine class fluorophores. A new medium composition was established for imaging the cytoskeleton using Alexa Fluor 488 conjugated to phalloidin. Morphological states of platelets bound to a glass substrate are visualized and quantified by two-dimensional localization microscopy at nanoscopic resolution. Marker-less drift correction yields localization of individual Alexa 488 conjugated to phalloidin with a positional accuracy of 12 nm.

Keywords: Alexa Fluor 488; actin cytoskeleton; dSTORM; drift correction; localization microscopy; photo-switching; platelet shape change; rhodamine.

MeSH terms

  • Actins / metabolism*
  • Actins / ultrastructure
  • Blood Platelets / metabolism
  • Blood Platelets / ultrastructure*
  • Cells, Cultured
  • Fluorescent Dyes / chemistry
  • Humans
  • Microscopy, Fluorescence / methods
  • Sensitivity and Specificity

Substances

  • Actins
  • Fluorescent Dyes