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Updated: Dec 13, 2025
13:13
Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy
Published on: December 3, 2012
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Light microscopy of proteins in their ultrastructural context
Ons M'Saad1,2, Joerg Bewersdorf3,4,5,6
1Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.
Nature Communications
|August 2, 2020
Summary
Researchers developed a new method to visualize nanoscale protein distribution within cells. This technique uses physical expansion and bulk labeling to reveal cellular nanoarchitecture with standard light microscopy.
Area of Science:
- Cell biology
- Microscopy
- Biophysics
Background:
- Visualizing nanoscale protein distribution in cellular ultrastructure is difficult with fluorescence microscopy.
- Electron microscopy (EM) provides ultrastructural context but lacks molecular specificity.
Purpose of the Study:
- To develop a novel optical contrast method for revealing cellular nanoarchitecture.
- To enable visualization of protein distribution at the nanoscale using conventional confocal microscopy.
Main Methods:
- Physical expansion of the intracellular space by 13 to 21-fold.
- Simultaneous retention of proteins during expansion.
- Bulk (pan) labeling of the proteome for protein density visualization.
Main Results:
- Achieved an optical contrast equivalent to electron microscopy (EM).
- Revealed cellular ultrastructure and nanoarchitecture using standard light microscopy.
- Resolved local protein densities within the decrowded intracellular space.
Conclusions:
- The developed method overcomes limitations of traditional fluorescence microscopy for ultrastructural protein localization.
- This technique offers a new principle for optical contrast, bridging the gap between light and electron microscopy.
- Enables detailed visualization of cellular nanoarchitecture with accessible confocal microscopy.
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