Bright photoactivatable fluorophores for single-molecule imaging

Nat Methods. 2016 Dec;13(12):985-988. doi: 10.1038/nmeth.4034. Epub 2016 Oct 24.

Abstract

Small-molecule fluorophores are important tools for advanced imaging experiments. We previously reported a general method to improve small, cell-permeable fluorophores which resulted in the azetidine-containing 'Janelia Fluor' (JF) dyes. Here, we refine and extend the utility of these dyes by synthesizing photoactivatable derivatives that are compatible with live-cell labeling strategies. Once activated, these derived compounds retain the superior brightness and photostability of the JF dyes, enabling improved single-particle tracking and facile localization microscopy experiments.

MeSH terms

  • Animals
  • COS Cells
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cell Membrane Permeability
  • Embryonic Stem Cells
  • Fluorescent Dyes / chemical synthesis*
  • Fluorescent Dyes / chemistry*
  • Fluorescent Dyes / metabolism
  • Fluorescent Dyes / radiation effects
  • Humans
  • Ligands
  • Light
  • Mice
  • Microscopy, Fluorescence
  • Molecular Structure
  • Photochemical Processes*
  • Photochemistry / methods
  • Recombinant Fusion Proteins / metabolism
  • Single Molecule Imaging / methods*
  • Small Molecule Libraries / chemical synthesis*
  • Small Molecule Libraries / chemistry*
  • Small Molecule Libraries / metabolism
  • Small Molecule Libraries / radiation effects
  • Spectrometry, Fluorescence
  • Spectrophotometry, Ultraviolet
  • Staining and Labeling

Substances

  • Fluorescent Dyes
  • Ligands
  • Recombinant Fusion Proteins
  • Small Molecule Libraries