2. Academic Validation
  3. Multicolor Caged dSTORM Resolves the Ultrastructure of Synaptic Vesicles in the Brain

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

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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