Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Minsu Kang; Jooyong Lee; Sangyoon Ko; Sang-Hee Shim

doi:10.1002/cbic.202000772

Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Chembiochem. 2021 Apr 16;22(8):1396-1399. doi: 10.1002/cbic.202000772. Epub 2021 Jan 29.

Authors

Minsu Kang^{1

2}, Jooyong Lee³, Sangyoon Ko^{1

2}, Sang-Hee Shim^{1

2}

Affiliations

¹ Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science (IBS), Seoul, 02841, Republic of Korea.
² Department of Chemistry, Korea University, Seoul, 02841, Republic of Korea.
³ Department of Biomedical Engineering, Ulsan National Institute of Science and Technology(UNIST), Ulsan, 44919, Republic of Korea.

PMID: 33325115
DOI: 10.1002/cbic.202000772

Abstract

Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ∼5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution.

Keywords: expansion microscopy; fluorescence; single-molecule localization microscopy; super-resolution microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Fluorescent Dyes / chemistry*
Microscopy, Fluorescence
Optical Imaging
Peptides / chemistry
Single Molecule Imaging*

Substances

Fluorescent Dyes
Peptides