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Super-resolution Fluorescence Microscopy

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Related Experiment Video

Updated: Dec 15, 2025

Label-Retention Expansion Microscopy LR-ExM Enables Super-Resolution Imaging and High-Efficiency Labeling

Label-Retention Expansion Microscopy LR-ExM Enables Super-Resolution Imaging and High-Efficiency Labeling

Published on: October 11, 2022

Molecular resolution imaging by post-labeling expansion single-molecule localization microscopy (Ex-SMLM).

Fabian U Zwettler¹, Sebastian Reinhard¹, Davide Gambarotto²

¹Department of Biotechnology and Biophysics, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.

Nature Communications

|July 9, 2020

Summary

Expansion microscopy combined with super-resolution techniques achieves electron microscopy resolution. This new method, post-labeling ExM, overcomes previous limitations for detailed biological imaging.

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Last Updated: Dec 15, 2025

Label-Retention Expansion Microscopy LR-ExM Enables Super-Resolution Imaging and High-Efficiency Labeling

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Published on: October 11, 2022

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Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

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Published on: December 9, 2013

Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Expansion microscopy (ExM) offers super-resolution imaging of expanded biological samples using conventional microscopes.
Combining ExM with single-molecule localization microscopy (SMLM) promises electron microscopy-level resolution.
Existing ExM-SMLM methods face challenges like protein/fluorophore loss and hydrogel incompatibility.

Purpose of the Study:

To develop a robust method combining ExM and SMLM for ultrastructural imaging.
To overcome limitations of current ExM-SMLM approaches.
To enable super-resolution imaging of immunolabeled endogenous proteins at molecular resolution.

Main Methods:

Re-embedding of expanded hydrogels for dSTORM imaging.
Post-labeling strategy for ExM-SMLM.
Utilizing microtubules and centrioles as reference structures.

Main Results:

Re-embedding enables dSTORM imaging of expanded samples.
Post-labeling ExM resolves limitations of previous super-resolution methods.
Demonstrated preservation of ultrastructural details, improved labeling efficiency, and reduced positional error.

Conclusions:

Post-labeling ExM-SMLM is a viable technique for high-resolution imaging.
This method paves the way for imaging endogenous proteins with true molecular resolution.
Overcomes key challenges in combining ExM and SMLM.