Localization STED (LocSTED) microscopy with 15 nm resolution


Puthukodan S., Murtezi E., Jacak J., Klar T. A.

NANOPHOTONICS, vol.9, no.4, pp.783-792, 2020 (Journal Indexed in SCI) identifier identifier

  • Publication Type: Article / Article
  • Volume: 9 Issue: 4
  • Publication Date: 2020
  • Doi Number: 10.1515/nanoph-2019-0398
  • Title of Journal : NANOPHOTONICS
  • Page Numbers: pp.783-792

Abstract

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 similar 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.