Azido push-pull fluorogens photoactivate to produce bright fluorescent labels

J Phys Chem B. 2010 Nov 18;114(45):14157-67. doi: 10.1021/jp907080r. Epub 2009 Oct 27.

Abstract

Dark azido push-pull chromophores have the ability to be photoactivated to produce bright fluorescent labels suitable for single-molecule imaging. Upon illumination, the aryl azide functionality in the fluorogens participates in a photochemical conversion to an aryl amine, thus restoring charge-transfer absorption and fluorescence. Previously, we reported that one compound, DCDHF-V-P-azide, was photoactivatable. Here, we demonstrate that the azide-to-amine photoactivation process is generally applicable to a variety of push-pull chromophores, and we characterize the photophysical parameters including photoconversion quantum yield, photostability, and turn-on ratio. Azido push-pull fluorogens provide a new class of photoactivatable single-molecule probes for fluorescent labeling and super-resolution microscopy. Lastly, we demonstrate that photoactivated push-pull dyes can insert into bonds of nearby biomolecules, simultaneously forming a covalent bond and becoming fluorescent (fluorogenic photoaffinity labeling).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amines / chemistry
  • Animals
  • Azides / chemistry*
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry*
  • Fluorescent Dyes / metabolism
  • Photoaffinity Labels / analysis
  • Photoaffinity Labels / chemical synthesis
  • Photoaffinity Labels / chemistry
  • Photoaffinity Labels / metabolism
  • Photochemical Processes*

Substances

  • Amines
  • Azides
  • Fluorescent Dyes
  • Photoaffinity Labels