Related Experiment Video
Updated: Aug 13, 2026
09:21
Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy
Published on: July 20, 2019
ONE microscopy
Ali H Shaib1, Mohamad Mahdi Alawieh2, Silvio O Rizzoli3,4,5
1Department of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany. ali.shaib@med.uni-goettingen.de.
Nature Protocols
|August 11, 2026
Summary
We developed one-step nanoscale expansion microscopy (ExM) to visualize individual protein shapes. This technique combines ExM with fluctuation analysis for super-resolution imaging on conventional microscopes.
Area of Science:
- Biophysics
- Microscopy
- Molecular Biology
Background:
- Expansion microscopy (ExM) revolutionized super-resolution imaging by physically expanding samples.
- Previous ExM methods could not achieve molecular-scale resolution for individual proteins.
- Cryo-electron microscopy and averaging techniques were previously required for single-protein shape analysis.
Purpose of the Study:
- To develop a method for visualizing individual protein shapes at molecular resolution using conventional fluorescence microscopes.
- To enable accessible and reproducible single-protein shape analysis.
- To provide a practical framework for advanced protein imaging.
Main Methods:
- Developed one-step nanoscale ExM by integrating ExM with fluctuation-based super-resolution analysis.
- Established detailed procedures for sample preparation (gel embedding, labeling), expansion, image acquisition, and data analysis.
- Introduced user-friendly software for efficient fluctuation analysis.
Main Results:
- Achieved direct visualization of individual protein shapes at molecular resolution.
- Demonstrated the capability of one-step nanoscale ExM on purified proteins, cells, and tissues.
- Made single-protein shape analysis accessible and reproducible using conventional fluorescence microscopes.
Conclusions:
- One-step nanoscale ExM overcomes previous resolution limitations in ExM.
- This technique democratizes single-protein shape analysis, previously requiring specialized equipment.
- Provides a practical and reproducible framework for protein imaging on standard fluorescence microscopes.
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