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Two‐Color RESOLFT Nanoscopy with Green and Red Fluorescent Photochromic Proteins
Author(s) -
LavoieCardinal Flavie,
Jensen Nickels A.,
Westphal Volker,
Stiel Andre C.,
Chmyrov Andriy,
Bierwagen Jakob,
Testa Ilaria,
Jakobs Stefan,
Hell Stefan W.
Publication year - 2014
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201301016
Subject(s) - fluorescence , photochromism , microscopy , superresolution , super resolution microscopy , optics , materials science , fluorescence microscope , resolution (logic) , chemistry , nanotechnology , physics , image (mathematics) , computer science , artificial intelligence
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.