Localization STED (LocSTED) microscopy with 15 nm resolution

Sujitha Puthukodan, Eljesa Murtezi, Jaroslaw Jacak, Thomas A. Klar

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)


We present localization with stimulated emission depletion (LocSTED) microscopy, a combination of STED and single-molecule localization microscopy (SMLM). We use the simplest form of a STED microscope that is cost effective and synchronization free, comprising continuous wave (CW) lasers for both excitation and depletion. By utilizing the reversible blinking of fluorophores, single molecules of Alexa 555 are localized down to ~5 nm. Imaging fluorescently labeled proteins attached to nanoanchors structured by STED lithography shows that LocSTED microscopy can resolve molecules with a resolution of at least 15 nm, substantially improving the classical resolution of a CW STED microscope of about 60 nm. LocSTED microscopy also allows estimating the total number of proteins attached on a single nanoanchor.

Original languageEnglish
Pages (from-to)783-792
Number of pages10
Issue number4
Publication statusPublished - 1 Apr 2020


  • protein nanoanchors
  • single-molecule imaging
  • single-molecule localization microscopy (SMLM)
  • stimulated emission depletion (STED) microscopy


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