Multicolor Caged dSTORM Resolves the Ultrastructure of Synaptic Vesicles in the Brain. Issue 45 (8th September 2015)

• Record Type: Journal Article
• Authors: Lehmann, Martin; Gottschalk, Benjamin; Puchkov, Dmytro; Schmieder, Peter; Schwagerus, Sergej; Hackenberger, Christian P. R.; Haucke, Volker; Schmoranzer, Jan
• Is Part Of: Angewandte Chemie international edition. Volume 54:Issue 45(2015)
• Page Start: 13230
• Page End: 13235
• Publication Date: 2015-09-08
• Subjects: electron microscopy -- fluorescent probes -- immunochemistry -- NMR spectroscopy -- single‐molecule studies; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201505138
• Languages: English
• ISSNs: 1433-7851

Abstract

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. Abstract : dSTORM gets colorful : A screen for fluorescence recovery following reduction revealed several dyes suitable for high‐resolution caged dSTORM imaging. Using dye pairs, multicolor localization precision could be achieved below 15 nm. This system is used to resolve the ultrastructure of single synaptic vesicles. Scale bar: 20 nm.