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Updated: Jul 7, 2026

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells
11:45

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

Published on: February 27, 2020

Cryo-immunogold electron microscopy

Peter J Peters1, Erik Bos, Alexander Griekspoor

  • 1Netherlands Cancer Institute, Amsterdam, The Netherlands.

Current Protocols in Cell Biology
|January 30, 2008
PubMed

Insights

High-resolution cryo-immunogold electron microscopy offers sensitive detection of protein and antigen localization within cells. This method preserves antigenicity by avoiding harsh organic solvents, enabling ultrastructural topological biochemistry studies.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microscopy

Background:

  • Subcellular localization of proteins is crucial for understanding cellular functions.
  • Existing immunodetection methods can compromise antigenicity due to harsh processing steps.

Purpose of the Study:

  • To describe a high-resolution cryo-immunogold electron microscopy technique for sensitive subcellular protein localization.
  • To enable topological biochemistry studies at the ultrastructural level.

Main Methods:

  • Chemical fixation of cells or tissues using aldehydes.
  • Embedding fixed material in gelatin.
  • Cryosectioning and mounting ultrathin sections on Formvar-coated grids.
  • High-resolution cryo-immunogold electron microscopy for immunodetection.

Main Results:

  • Achieved sensitive immunodetection of antigens on ultrathin sections.
  • Preserved protein antigenicity by omitting harsh organic solvents.
  • Enabled ultrastructural analysis of protein localization.

Conclusions:

  • Cryo-immunogold electron microscopy is a highly sensitive method for subcellular antigen localization.
  • The technique provides superior preservation of antigenicity compared to traditional methods.
  • Facilitates detailed ultrastructural studies of protein distribution and function.

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