Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Minsu Kang, Jooyong Lee, Sangyoon Ko, Sang Hee Shim

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
JournalChemBioChem
DOIs
Publication statusAccepted/In press - 2020

Keywords

  • expansion microscopy
  • fluorescence
  • single-molecule localization microscopy
  • super-resolution microscopy

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

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