Amplified Expansion Stimulated Emission Depletion Microscopy

Doyeon Kim, Taeyeon Kim, Jooyong Lee, Sang-Hee Shim

Research output: Contribution to journalArticle

Abstract

Expansion microscopy (ExM) enhances spatial resolution by using a swellable polymer that expands the sample volume by a factor of ≈4 in one dimension and a factor of ≈64 in volume. Combining ExM with stimulated emission depletion (STED) microscopy, referred to as ExSTED, increases the resolution to up to 10 nm. However, photobleaching is a critical issue in ExSTED because the sample expansion lowers the fluorophore density whereas high-resolution STED requires high depletion intensity. To overcome these issues, we developed extremely bright expansion nanoscopy by using biotin–avidin signal amplification to increase the labeling density. Our method provides up to sevenfold increases in fluorescence signal intensity in expanded samples, thus enabling the use of STED imaging with maximum depletion intensities of a commercial microscope in the order of GW cm −2 . We demonstrated the method by using biotinylated antibodies and genetic incorporation approaches that allow localization of biotin in a specific molecule or organelle.

Original languageEnglish
Pages (from-to)1260-1265
Number of pages6
JournalChemBioChem
Volume20
Issue number10
DOIs
Publication statusPublished - 2019 May 15

Fingerprint

Stimulated emission
Microscopy
Microscopic examination
Photobleaching
Biotin
Organelles
Fluorophores
Polymers
Fluorescence
Labeling
Amplification
Microscopes
Antibodies
Imaging techniques
Molecules

Keywords

  • antibodies
  • expansion microscopy
  • fluorescence
  • stimulated emission depletion microscopy
  • super-resolution microscopy

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

Cite this

Amplified Expansion Stimulated Emission Depletion Microscopy. / Kim, Doyeon; Kim, Taeyeon; Lee, Jooyong; Shim, Sang-Hee.

In: ChemBioChem, Vol. 20, No. 10, 15.05.2019, p. 1260-1265.

Research output: Contribution to journalArticle

Kim, Doyeon ; Kim, Taeyeon ; Lee, Jooyong ; Shim, Sang-Hee. / Amplified Expansion Stimulated Emission Depletion Microscopy. In: ChemBioChem. 2019 ; Vol. 20, No. 10. pp. 1260-1265.
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