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

Angewandte Chemie
Martin LehmannJan Schmoranzer

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.

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Citations

Mar 14, 2017·Chemical Reviews·Markus Sauer, Mike Heilemann
Dec 12, 2017·Frontiers in Cellular Neuroscience·Janosch P Heller, Dmitri A Rusakov
Sep 14, 2019·Chemistry : a European Journal·Joshua K G KarlssonAnthony Harriman
Sep 5, 2018·Journal of Cell Science·Volker Haucke, Michael M Kozlov
Feb 6, 2021·Journal of Neurochemistry·Callista B Harper, Karen J Smillie
Sep 18, 2018·Chemical Reviews·Honglin Li, Joshua C Vaughan
Apr 19, 2019·ACS Chemical Biology·Fadi M Jradi, Luke D Lavis
Sep 2, 2017·Journal of the American Chemical Society·Megan S MichieMartin J Schnermann
Aug 18, 2020·ACS Nano·Dominic A HelmerichMarkus Sauer

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