Correlative stochastic optical reconstruction microscopy and electron microscopy

PLoS One. 2015 Apr 15;10(4):e0124581. doi: 10.1371/journal.pone.0124581. eCollection 2015.

Abstract

Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • Epithelial Cells / ultrastructure
  • Humans
  • Microscopy, Electron / instrumentation
  • Microscopy, Electron / methods*
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Microtomy
  • Microtubules / ultrastructure
  • Orthomyxoviridae / ultrastructure
  • Staining and Labeling / methods*
  • Stochastic Processes
  • Tissue Embedding
  • Virus Release / physiology