Correlative fluorescence and electron microscopy of quantum dot labeled proteins on whole cells in liquid

Methods Mol Biol. 2014:1117:527-40. doi: 10.1007/978-1-62703-776-1_23.

Abstract

Correlative fluorescence microscopy and scanning transmission electron microscopy (STEM) of cells fully immersed in liquid is a new methodology with many application areas. Proteins, in live cells immobilized on microchips, are labeled with fluorescent quantum dot (QD) nanoparticles. In this protocol, the epidermal growth factor receptor (EGFR) is labeled. The cells are fixed after a selected labeling time, for example, 5 min as needed to form EGFR dimers. The microchip with cells is then imaged with fluorescence microscopy. Thereafter, the microchip with the labeled cells and one with a spacer are assembled in a special microfluidic device and imaged with STEM.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Microfluidic Analytical Techniques
  • Microscopy, Electron / methods*
  • Microscopy, Electron, Scanning Transmission / methods
  • Microscopy, Fluorescence / methods*
  • Proteins / chemistry*
  • Quantum Dots / chemistry*
  • Staining and Labeling

Substances

  • Proteins