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Prelabeling Expansion Single‐Molecule Localization Microscopy with Minimal Linkage Error
Author(s) -
Kang Minsu,
Lee Jooyong,
Ko Sangyoon,
Shim SangHee
Publication year - 2021
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000772
Subject(s) - fluorophore , microscopy , super resolution microscopy , fluorescence microscope , streptavidin , biophysics , denaturation (fissile materials) , chemistry , resolution (logic) , fluorescence , molecule , materials science , biochemistry , biology , optics , physics , computer science , biotin , organic chemistry , artificial intelligence , nuclear chemistry
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.