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. Epub 2015 Sep 8.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology
  • Brain / ultrastructure*
  • Color*
  • Fluorescent Dyes / chemistry*
  • Image Processing, Computer-Assisted*
  • Mice
  • Microscopy, Fluorescence / methods*
  • Molecular Structure
  • NIH 3T3 Cells
  • Synaptic Vesicles / ultrastructure*


  • Fluorescent Dyes