2010
DOI: 10.1038/nmeth.1477
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A photoactivatable marker protein for pulse-chase imaging with superresolution

Abstract: IrisFP is a photoactivatable fluorescent protein that combines irreversible photoconversion from a green- to a red-emitting form with reversible photoswitching between a fluorescent and a nonfluorescent state in both forms. Here we introduce a monomeric variant, mIrisFP, and demonstrate how its multiple photoactivation modes can be used for pulse-chase experiments combined with subdiffraction-resolution imaging in living cells by using dual-color photoactivation localization microscopy (PALM).

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Cited by 113 publications
(115 citation statements)
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“…We also applied STED-RICS to the study of focal adhesion dynamics. In previous work, we observed assembly and disassembly of individual focal adhesions by using superresolution PALM imaging combined with pulse-chase experiments, based on the photoactivatable fluorescent protein mIrisFP fused to paxillin 16 . Here we have investigated focal adhesions of Xenopus laevis tissue culture (XTC) cells using b1-integrin, immunostained with the monoclonal antibody 8C8 labelled with Atto647N (Fig.…”
Section: Probing Lipid Dynamics In a Model Membrane By Sted-ricsmentioning
confidence: 99%
“…We also applied STED-RICS to the study of focal adhesion dynamics. In previous work, we observed assembly and disassembly of individual focal adhesions by using superresolution PALM imaging combined with pulse-chase experiments, based on the photoactivatable fluorescent protein mIrisFP fused to paxillin 16 . Here we have investigated focal adhesions of Xenopus laevis tissue culture (XTC) cells using b1-integrin, immunostained with the monoclonal antibody 8C8 labelled with Atto647N (Fig.…”
Section: Probing Lipid Dynamics In a Model Membrane By Sted-ricsmentioning
confidence: 99%
“…3). Application-wise, a tunable signal detection reporter offers opportunities for advanced protein discovery strategies involving protein turnover at the cellular membrane, pulse-chase of vesicular traffic or post-cell-labeling assays that require the removal of signal (Bodor et al, 2012;Fuchs et al, 2010;Giepmans et al, 2006;Lippincott-Schwartz and Patterson, 2003;Mizukami et al, 2012). Furthermore, in order to increasingly accelerate the off-rates of signal detection using multiselective FAPs, the future focus will be placed on isolating weakaffinity fluorogens.…”
Section: Discussionmentioning
confidence: 99%
“…Zunächst kommt es ähnlich wie bei den Kaedeähnlichen PSFPs bei Bestrahlung mit violettem Licht zu einem irreversiblen Umschalten von IrisFP, [64] seiner monomeren Variante mIrisFP [57] und NijiFP [58] in gleichzeitig grüne und rote Formen. Die anschließende Bestrahlung der grünen Formen dieser FPs mit 488 nm und wenig intensiven Lasern mit 405 nm führt zu einem reversiblen An-und Ausschalten des grünen Chromophors.…”
Section: Photoaktivierbare Rfpsunclassified
“…[82] In Versuchsaufbauten zur superauflçsenden Bildgebung wurden die reversibel und irreversibel schaltbaren mIrisFP und NijiFP in einer Kopplung mit einer selektiven "Pulse-Chase"-Photomarkierung verwendet. [57,58] …”
Section: V Verkhusha Et Alunclassified