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Multicolor Caged dSTORM Resolves the Ultrastructure of Synaptic Vesicles in the Brain
Author(s) -
Lehmann Martin,
Gottschalk Benjamin,
Puchkov Dmytro,
Schmieder Peter,
Schwagerus Sergej,
Hackenberger Christian P. R.,
Haucke Volker,
Schmoranzer Jan
Publication year - 2015
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201505138
Subject(s) - ultrastructure , vesicle , synaptic vesicle , biophysics , neuroscience , chemistry , biology , anatomy , membrane , biochemistry
The precision of single‐molecule localization‐based super‐resolution microscopy, including dSTORM, critically depends on the number of detected photons per localization. Recently, reductive caging of fluorescent dyes followed by UV‐induced recovery in oxidative buffer systems was used to increase the photon yield and thereby the localization precision in single‐color dSTORM. By screening 39 dyes for their fluorescence caging and recovery kinetics, we identify novel dyes that are suitable for multicolor caged dSTORM. Using a dye pair suited for registration error‐free multicolor dSTORM based on spectral demixing (SD), a multicolor localization precision below 15 nm was achieved. Caged SD‐dSTORM can resolve the ultrastructure of single 40 nm synaptic vesicles in brain sections similar to images obtained by immuno‐electron microscopy, yet with much improved label density in two independent channels.
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