Two-color RESOLFT nanoscopy with green and red fluorescent photochromic proteins.
Up to now, all demonstrations of reversible saturable optical fluorescence transitions (RESOLFT) superresolution microscopy of living cells have relied on the use of reversibly switchable fluorescent proteins (RSFP) emitting in the green spectral range. Here we show RESOLFT imaging with rsCherryRev1.4, a new red-emitting RSFP enabling a spatial resolution up to four times higher than the diffraction barrier. By co-expressing green and red RSFPs in living cells we demonstrate two-color RESOLFT imaging both for single ("donut") beam scanning and for parallelized versions of RESOLFT nanoscopy where an array of >23,000 "donut-like" minima are scanned simultaneously.
['Lavoie-Cardinal|Flavie|F|', 'Jensen|Nickels A|NA|', 'Westphal|Volker|V|', 'Stiel|Andre C|AC|', 'Chmyrov|Andriy|A|', 'Bierwagen|Jakob|J|', 'Testa|Ilaria|I|', 'Jakobs|Stefan|S|', 'Hell|Stefan W|SW|']
D002478:Cells, Cultured D003116:Color D049452:Green Fluorescent Proteins D006367:HeLa Cells D006801:Humans D008164:Luminescent Proteins D008856:Microscopy, Fluorescence D036103:Nanotechnology
RESOLFT nanoscopy fluorescent proteins photoswitching superresolution imaging