Azido push-pull fluorogens photoactivate to produce bright fluorescent labels.
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).
Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
Samuel J
SJ
Lord
Hsiao-Lu D
HL
Lee
Reichel
R
Samuel
Ryan
R
Weber
Na
N
Liu
Nicholas R
NR
Conley
Michael A
MA
Thompson
Robert J
RJ
Twieg
W E
WE
Moerner
D000588:Amines / Q000737:chemistry; D000818:Animals; D001386:Azides / Q000737:chemistry*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000032:analysis / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D019920:Photoaffinity Labels / Q000032:analysis / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D055668:Photochemical Processes
D000588:Amines; D001386:Azides; D005456:Fluorescent Dyes; D019920:Photoaffinity Labels