A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow

Methods Cell Biol. 2017:140:149-164. doi: 10.1016/bs.mcb.2017.03.008. Epub 2017 Apr 19.

Abstract

While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam-scanning electron microscopy data. Here we demonstrate the entire integrated CLEM workflow using fluorescently tagged MCF7 breast cancer cells.

Keywords: 3D FIB-SEM; CLEM; Correlative imaging technique; Electron microscopy; Fluorescence microscopy.

Publication types

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

MeSH terms

  • Green Fluorescent Proteins / metabolism
  • Humans
  • Imaging, Three-Dimensional*
  • MCF-7 Cells
  • Microfluidics
  • Microscopy, Electron / methods*
  • Microscopy, Fluorescence / methods*
  • Recombinant Fusion Proteins / metabolism

Substances

  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins