A peptide tag-specific nanobody enables high-quality labeling for dSTORM imaging

Nat Commun. 2018 Mar 2;9(1):930. doi: 10.1038/s41467-018-03191-2.

Abstract

Dense fluorophore labeling without compromising the biological target is crucial for genuine super-resolution microscopy. Here we introduce a broadly applicable labeling strategy for fixed and living cells utilizing a short peptide tag-specific nanobody (BC2-tag/bivBC2-Nb). BC2-tagging of ectopically introduced or endogenous proteins does not interfere with the examined structures and bivBC2-Nb staining results in a close-grained fluorophore labeling with minimal linkage errors. This allowed us to perform high-quality dSTORM imaging of various targets in mammalian and yeast cells. We expect that this versatile strategy will render many more demanding cellular targets amenable to dSTORM imaging.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • HeLa Cells
  • Humans
  • Schizosaccharomyces
  • Single Molecule Imaging / methods*
  • Single-Domain Antibodies*
  • Staining and Labeling / methods*

Substances

  • Single-Domain Antibodies