Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells.

ABSTRACT: We present new fluorophore-conjugates for dual-color photoactivation and super-resolution imaging inside live mammalian cells. These custom-designed, photo-caged Q-rhodamines and fluoresceins are cell-permeable, bright and localize specifically to intracellular targets. We utilized established orthogonal protein labeling strategies to precisely attach the photoactivatable fluorophores to proteins with subsequent activation of fluorescence by irradiation with UV light. That way, diffusive cytosolic proteins, histone proteins as well as filigree mitochondrial networks and focal adhesion proteins were visualized inside living cells. We applied the new photoactivatable probes in inverse fluorescence recovery after photo-bleaching (iFRAP) experiments, gaining real-time access to protein dynamic

SUBMITTER: Hauke S 

PROVIDER: S-EPMC5351804 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature