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Fast reversibly photoswitching red fluorescent proteins for live-cell RESOLFT nanoscopy.
Pennacchietti, Francesca; Serebrovskaya, Ekaterina O; Faro, Aline R; Shemyakina, Irina I; Bozhanova, Nina G; Kotlobay, Alexey A; Gurskaya, Nadya G; Bodén, Andreas; Dreier, Jes; Chudakov, Dmitry M; Lukyanov, Konstantin A; Verkhusha, Vladislav V; Mishin, Alexander S; Testa, Ilaria.
Nat Methods ; 15(8): 601-604, 2018 08.
Article in English | MEDLINE | ID: mdl-29988095

ABSTRACT

Reversibly photoswitchable fluorescent proteins (rsFPs) are gaining popularity as tags for optical nanoscopy because they make it possible to image with lower light doses. However, green rsFPs need violet-blue light for photoswitching, which is potentially phototoxic and highly scattering. We developed new rsFPs based on FusionRed that are reversibly photoswitchable with green-orange light. The rsFusionReds are bright and exhibit rapid photoswitching, thereby enabling nanoscale imaging of living cells.


Subject(s)
Luminescent Proteins/chemistry , Luminescent Proteins/metabolism , Cell Line , Humans , Intravital Microscopy/methods , Kinetics , Light , Microscopy, Fluorescence/methods , Nanotechnology , Photochemical Processes , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/metabolism , Spectrophotometry , Red Fluorescent Protein
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