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Article Dans Une Revue ACS Nano Année : 2014

Green-to-red photoconvertible Dronpa mutant for multimodal super-resolution fluorescence microscopy

Résumé

Advanced imaging techniques crucially depend on the labels used. In this work, we present the structure-guided design of a fluorescent protein that displays both reversibly photochromic and green-to-red photoconversion behavior. We first designed ffDronpa, a mutant of the photochromic fluorescent protein Dronpa that matures up to three times faster while retaining its interesting photochromic features. Using a combined evolutionary and structure-driven rational design strategy, we developed a green-to-red photoconvertible ffDronpa mutant, called pcDronpa, and explored different optimization strategies that resulted in its improved version, pcDronpa2. This fluorescent probe combines a high brightness with low photobleaching and photoblinking. We herein show that, despite its tetrameric nature, pcDronpa2 allows for multimodal subdiffraction imaging by sequentially imaging a given sample using both super-resolution fluctuation imaging and localization microscopy.

Dates et versions

hal-03513242 , version 1 (05-01-2022)

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Citer

Benjamien Moeyaert, Ngan Nguyen Bich, Elke de Zitter, Susana Rocha, Koen Clays, et al.. Green-to-red photoconvertible Dronpa mutant for multimodal super-resolution fluorescence microscopy. ACS Nano, 2014, 8 (2), pp.1664-1673. ⟨10.1021/nn4060144⟩. ⟨hal-03513242⟩
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