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European Biophysics Journal

, Volume 48, Issue 5, pp 485–490 | Cite as

New coumarin- and phenoxazine-based fluorescent probes for live-cell STED nanoscopy

  • Stane Pajk
  • Hana Majaron
  • Matej Novak
  • Boštjan Kokot
  • Janez ŠtrancarEmail author
Biophysics Letter

Abstract

The potential of live-cell stimulated emission depletion (STED) nanoscopy has not yet been fully exploited. Currently, the main limitation is the small number of fluorophores and probes that can sustain high light intensity/high dose employed in STED. Namely, fluorophores suitable for STED nanoscopy must be bright and highly photostable and exhibit a large Stokes shift. To expand the list of available probes, we synthesized and evaluated several new membrane probes for live-cell STED nanoscopy. Of the tested probes, probes MePyr500, ThiaCN545 and NB640 not only allow high-resolution STED images, but also partition into the intracellular membranes relatively quickly, thus lacking the selectivity of labelling solely the plasma membrane. During experiments, cytotoxicity was observed merely with the probe ThiaCN545, which blebs the plasma membrane. In comparison with commercially available CellMask Orange and STAR RED (KK114) DPPE, all our tested probes exhibited better photostability with the exception of NB640, which had the fastest bleaching rate of all tested probes. The best overall results can be assigned to the probe MePyr500, providing high-resolution STED images as well as high photostability with no noticeable cytotoxicity, making it an excellent candidate for further development.

Keywords

STED Super-resolution Fluorescence Live cell Membrane probes 

Notes

Acknowledgements

Funding was provided by Horizon 2020 Framework Programme (Grant No. 686098).

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Copyright information

© European Biophysical Societies' Association 2019

Authors and Affiliations

  • Stane Pajk
    • 1
    • 2
  • Hana Majaron
    • 2
  • Matej Novak
    • 1
  • Boštjan Kokot
    • 2
  • Janez Štrancar
    • 2
    Email author
  1. 1.Faculty of PharmacyUniversity of LjubljanaLjubljanaSlovenia
  2. 2.Jožef Stefan InstituteLjubljanaSlovenia

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