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Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

DOI:
10.3791/53186-v

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09:16 min

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September 11, 2015

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,
1INM-Leibniz Institute for New Materials, 2Department of Physics,University of Saarland

Chapters

  • 00:05Title
  • 01:45Preparation of Silicon Nitride Membrane Microchips
  • 02:56Preparation of Cells on Microchips
  • 03:52EGFR Labeling, Fixation, and Fluorescence Microscopy
  • 06:12Wet Environmental Scanning Electron Microscopy-Scanning Transmission Electron Microscopy of Whole Cells (ESEM-STEM)
  • 07:20Results: EGFR Stoichiometry in Labeled COS7 Cells
  • 08:45Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes the labeling of epidermal growth factor receptor (EGFR) on COS7 fibroblast cells, and subsequent correlative light- and electron microscopy of whole cells in hydrated state. The label contained fluorescent quantum dots. The protocol can be used to study the stoichiometry of EGFR at the single molecule level.

Tags

Keywords: Epidermal Growth Factor ReceptorEGFRCOS7 Fibroblast CellsCorrelative MicroscopyFluorescence MicroscopyEnvironmental Scanning Electron Microscopy (ESEM)Scanning Transmission Electron Microscopy (STEM)Quantum DotsReceptor StoichiometryReceptor DistributionReceptor Clustering

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