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A Green-to-Red Photoconvertible Dronpa Mutant for Multimodal Super-resolution Fluorescence Microscopy

Tijdschriftbijdrage - Tijdschriftartikel

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.
Tijdschrift: ACS Nano
ISSN: 1936-0851
Issue: 2
Volume: 8
Pagina's: 1664 - 1673
Jaar van publicatie:2014
BOF-keylabel:ja
IOF-keylabel:ja
BOF-publication weight:10
CSS-citation score:2
Authors from:Higher Education
Toegankelijkheid:Open