Two-Color 810 nm STED Nanoscopy of Living Cells with Endogenous SNAP-Tagged Fusion Proteins

ACS Chem Biol. 2018 Feb 16;13(2):475-480. doi: 10.1021/acschembio.7b00616. Epub 2017 Oct 2.

Abstract

A 810 nm STED nanoscopy setup and an appropriate combination of two fluorescent dyes (Si-rhodamine 680SiR and carbopyronine 610CP) have been developed for near-IR live-cell super-resolution imaging. Vimentin endogenously tagged using the CRISPR/Cas9 approach with the SNAP tag, together with a noncovalent tubulin label, provided reliable and cell-to-cell reproducible dual-color confocal and STED imaging of the cytoskeleton in living cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems / genetics
  • Cell Line, Tumor
  • Color
  • Cytoskeleton / metabolism*
  • Fluorescence
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / pharmacology*
  • Fluorescent Dyes / radiation effects
  • Gene Editing
  • Humans
  • Infrared Rays
  • Microscopy, Confocal
  • Microscopy, Fluorescence / methods*
  • Organosilicon Compounds / chemical synthesis
  • Organosilicon Compounds / chemistry
  • Organosilicon Compounds / pharmacology*
  • Organosilicon Compounds / radiation effects
  • Quinolizines / chemical synthesis
  • Quinolizines / chemistry
  • Quinolizines / pharmacology*
  • Quinolizines / radiation effects
  • Recombinant Fusion Proteins / genetics
  • Vimentin / genetics

Substances

  • Fluorescent Dyes
  • Organosilicon Compounds
  • Quinolizines
  • Recombinant Fusion Proteins
  • Vimentin