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

6 Citations (Scopus)


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
Pages (from-to)1396-1399
Number of pages4
Issue number8
Publication statusPublished - 2021 Apr 16

Bibliographical note

Funding Information:
We thank the Hyunwoo Rhee Lab (Seoul National University) for providing biotin ligase plasmids and the Hak Joong Kim Lab (Korea University) for providing the Avi plasmids. This work is supported by IBS-R023-D1.

Publisher Copyright:
© 2020 Wiley-VCH GmbH


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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry


Dive into the research topics of 'Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error'. Together they form a unique fingerprint.

Cite this