Label-retention expansion microscopy

Xiaoyu Shi; Qi Li; Zhipeng Dai; Arthur A Tran; Siyu Feng; Alejandro D Ramirez; Zixi Lin; Xiaomeng Wang; Tracy T Chow; Jiapei Chen; Dhivya Kumar; Andrew R McColloch; Jeremy F Reiter; Eric J Huang; Ian B Seiple; Bo Huang

doi:10.1083/jcb.202105067

Label-retention expansion microscopy

J Cell Biol. 2021 Sep 6;220(9):e202105067. doi: 10.1083/jcb.202105067. Epub 2021 Jul 6.

Authors

Xiaoyu Shi^#^{1

2}, Qi Li^#^{1

3}, Zhipeng Dai⁴, Arthur A Tran⁵, Siyu Feng⁶, Alejandro D Ramirez¹, Zixi Lin¹, Xiaomeng Wang¹, Tracy T Chow⁷, Jiapei Chen⁸, Dhivya Kumar⁷, Andrew R McColloch², Jeremy F Reiter^{3

7

9}, Eric J Huang⁸, Ian B Seiple^{1

3}, Bo Huang^{1

7

9}

Affiliations

¹ Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA.
² Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA.
³ Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA.
⁴ Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA.
⁵ Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, San Francisco, CA.
⁶ University of California, Berkeley-University of California, San Francisco Joint Graduate Program in Bioengineering, University of California, San Francisco, San Francisco, CA.
⁷ Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA.
⁸ Department of Pathology, University of California, San Francisco, San Francisco, CA.
⁹ Chan Zuckerberg Biohub, San Francisco, CA.

^# Contributed equally.

Abstract

Expansion microscopy (ExM) increases the effective resolving power of any microscope by expanding the sample with swellable hydrogel. Since its invention, ExM has been successfully applied to a wide range of cell, tissue, and animal samples. Still, fluorescence signal loss during polymerization and digestion limits molecular-scale imaging using ExM. Here, we report the development of label-retention ExM (LR-ExM) with a set of trifunctional anchors that not only prevent signal loss but also enable high-efficiency labeling using SNAP and CLIP tags. We have demonstrated multicolor LR-ExM for a variety of subcellular structures. Combining LR-ExM with superresolution stochastic optical reconstruction microscopy (STORM), we have achieved molecular resolution in the visualization of polyhedral lattice of clathrin-coated pits in situ.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Antibodies / chemistry
Biotin / chemistry
Cell Line, Tumor
Fluorescent Dyes / chemistry
Fluorescent Dyes / metabolism
HEK293 Cells
HeLa Cells
Humans
Mice
Microscopy, Fluorescence / methods*
Microtubules / metabolism
Microtubules / ultrastructure*
Mouse Embryonic Stem Cells / metabolism
Mouse Embryonic Stem Cells / ultrastructure*
Osteoblasts / metabolism
Osteoblasts / ultrastructure*
Staining and Labeling / methods*
Streptavidin / chemistry
Succinimides / chemistry

Substances

Antibodies
Fluorescent Dyes
Succinimides
Biotin
Streptavidin
N-hydroxysuccinimide

Abstract

Publication types

MeSH terms

Substances

Grants and funding