Multicolor Caged dSTORM Resolves the Ultrastructure of Synaptic Vesicles in the Brain

  • Angew Chem Int Ed Engl. 2015 Nov 2;54(45):13230-5. doi: 10.1002/anie.201505138.
Martin Lehmann  1  2 ,  Benjamin Gottschalk  3 ,  Dmytro Puchkov  3 ,  Peter Schmieder  3 ,  Sergej Schwagerus  3 ,  Christian P R Hackenberger  3  4 ,  Volker Haucke  3  5 ,  Jan Schmoranzer  6  7  8
Affiliations
  • 1. Leibniz Institut für Molekulare Pharmakologie (FMP), Robert-Roessle-Strasse 10, 13125 Berlin (Germany). [email protected].
  • 2. Freie Universität Berlin, Department of Biochemistry, Takustrasse 6, 14195 Berlin (Germany). [email protected].
  • 3. Leibniz Institut für Molekulare Pharmakologie (FMP), Robert-Roessle-Strasse 10, 13125 Berlin (Germany).
  • 4. Humboldt Universität zu Berlin, Department of Chemistry, Brook-Taylor-Strasse. 2, 12489 Berlin (Germany).
  • 5. Freie Universität Berlin, Department of Biochemistry, Takustrasse 6, 14195 Berlin (Germany).
  • 6. Leibniz Institut für Molekulare Pharmakologie (FMP), Robert-Roessle-Strasse 10, 13125 Berlin (Germany). [email protected].
  • 7. Freie Universität Berlin, Department of Biochemistry, Takustrasse 6, 14195 Berlin (Germany). [email protected].
  • 8. Charité - Universitätsmedizin Berlin, Charité Campus Mitte, Virchowweg 6, 10117 Berlin (Germany). [email protected].
PMID: 26346505 DOI: 10.1002/anie.201505138
Abstract

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.

Keywords
NMR spectroscopy; electron microscopy; fluorescent probes; immunochemistry; single-molecule studies.
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