Ultrahigh-throughput single-molecule spectroscopy and spectrally resolved super-resolution microscopy.
By developing a wide-field scheme for spectral measurement and implementing photoswitching, we synchronously obtained the fluorescence spectra and positions of ∼10(6) single molecules in labeled cells in minutes, which consequently enabled spectrally resolved, 'true-color' super-resolution microscopy. The method, called spectrally resolved stochastic optical reconstruction microscopy (SR-STORM), achieved cross-talk-free three-dimensional (3D) imaging for four dyes 10 nm apart in emission spectrum. Excellent resolution was obtained for every channel, and 3D localizations of all molecules were automatically aligned within one imaging path.
Department of Chemistry, University of California, Berkeley, Berkeley, California, USA.|Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
Zhengyang
Z
Zhang
Department of Chemistry, University of California, Berkeley, Berkeley, California, USA.
Samuel J
SJ
Kenny
Department of Chemistry, University of California, Berkeley, Berkeley, California, USA.
Margaret
M
Hauser
Department of Chemistry, University of California, Berkeley, Berkeley, California, USA.
Wan
W
Li
Department of Chemistry, University of California, Berkeley, Berkeley, California, USA.|Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
Ke
http://orcid.org/0000-0002-2788-194X
K
Xu
D002232:Carbocyanines / Q000032:analysis; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000032:analysis; D057166:High-Throughput Screening Assays / Q000379:methods; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods*
C569686:Alexa Fluor 647; D002232:Carbocyanines; D005456:Fluorescent Dyes