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.
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37070 Göttingen (Germany), Fax: (+49) 551 201-2505; Equal contributions.
Flavie
F
Lavoie-Cardinal
Nickels A
NA
Jensen
Volker
V
Westphal
Andre C
AC
Stiel
Andriy
A
Chmyrov
Jakob
J
Bierwagen
Ilaria
I
Testa
Stefan
S
Jakobs
Stefan W
SW
Hell
D002478:Cells, Cultured; D003116:Color; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D008856:Microscopy, Fluorescence; D036103:Nanotechnology / Q000379:methods*; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
RESOLFT; fluorescent proteins; nanoscopy; photoswitching; superresolution imaging