Multicolor caged dSTORM resolves the ultrastructure of synaptic vesicles in the brain
M. Lehmann; B. Gottschalk; D. Puchkov; P. Schmieder; S. Schwagerus; C. Hackenberger; V. Haucke; J. Schmoranzer*
Angew. Chem. Int. Ed. Engl. 54, 13230 - 13235 (2015); Angew. Chem. 127, 13428 - 13433 (2015)
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.