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Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues
Tay Won Shin1,2,3, Hao Wang1,4, Chi Zhang1
1McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Nature Communications
|February 12, 2025
Summary
Ultrastructural membrane expansion microscopy (umExM) enables dense, nanoscale visualization of lipid membranes in intact tissues. This new method improves imaging resolution for studying cellular structures like neuronal axons.
Area of Science:
- Cell Biology
- Microscopy
- Neuroscience
Background:
- Lipid membranes are crucial for biological compartmentalization.
- Visualizing membranes at the nanoscale in intact tissues with high labeling density is challenging.
Purpose of the Study:
- To develop a method for high-density, nanoscale visualization of lipid membranes in intact tissues.
- To enable detailed imaging of cellular structures such as neuronal processes.
Main Methods:
- Developed ultrastructural membrane expansion microscopy (umExM).
- Combined an innovative membrane label with an optimized expansion microscopy protocol.
- Validated labeling in mouse brain slices using confocal microscopy.
Main Results:
- umExM achieved high signal-to-background ratio, uniformity, and continuity of membrane labeling.
- Imaging resolution of approximately 60 nm for membranes and proteins was achieved.
- Demonstrated utility for segmenting and tracing neuronal processes like axons.
- Achieved ~35 nm resolution by combining umExM with optical fluctuation imaging or iterative expansion.
Conclusions:
- umExM provides a powerful tool for nanoscale membrane visualization in biological tissues.
- The technique offers potential for electron microscopy-level resolution imaging of brain membranes using light microscopes.
