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Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues

delete2025-02-12
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OA
AI
T
Tay Shin
H
Hao Wang
C
Chi Zhang
B
Bobae An
Y
Yangning Lu
Z
Zhang, Elizabeth
K
Karagiannis, Emmanouil D.
K
Kang, Jeong Seuk
E
Emenari, Amauche
S
Symvoulidis, Panagiotis
A
Asano, Shoh
L
Lin, Leanne
C
Costa, Emma K.
M
Marblestone, Adam H.
K
Kasthuri, Narayanan
T
Tsai, Li-Huei
E
Edward S. Boyden *
DOI：10.1038/s41467-025-56641-zdelete
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Abstract

Abstract

En 中文
Lipid membranes are key to the nanoscale compartmentalization of biological systems, but fluorescent visualization of them in intact tissues, with nanoscale precision, is challenging to do with high labeling density. Here, we report ultrastructural membrane expansion microscopy (umExM), which combines an innovative membrane label and optimized expansion microscopy protocol, to support dense labeling of membranes in tissues for nanoscale visualization. We validate the high signal-to-background ratio, and uniformity and continuity, of umExM membrane labeling in brain slices, which supports the imaging of membranes and proteins at a resolution of similar to 60 nm on a confocal microscope. We demonstrate the utility of umExM for the segmentation and tracing of neuronal processes, such as axons, in mouse brain tissue. Combining umExM with optical fluctuation imaging, or iterating the expansion process, yields similar to 35 nm resolution imaging, pointing towards the potential for electron microscopy resolution visualization of brain membranes on ordinary light microscopes.
Keywords:
ELECTRON-MICROSCOPY
BRAIN
SEGMENTATION
FIXATION
FLUID

Journal

Nature Communications cover
Nature Communications
IF:
15.7
Papers:
9.2W
Citations:
91.2W

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No organization information available