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분자 분광학 및 동력학 연구단
분자 분광학 및 동력학 연구단
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Amplified Expansion Stimulated Emission Depletion Microscopy

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Title
Amplified Expansion Stimulated Emission Depletion Microscopy
Author(s)
Doyeon Kim; Kim, Taeyeon; Lee, Jooyong; Sang-Hee Shim
Publication Date
2019-05
Journal
CHEMBIOCHEM, v.20, no.10, pp.1260 - 1265
Publisher
WILEY-V C H VERLAG GMBH
Abstract
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim 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
URI
https://pr.ibs.re.kr/handle/8788114/6311
ISSN
1439-4227
Appears in Collections:
Center for Molecular Spectroscopy and Dynamics(분자 분광학 및 동력학 연구단) > Journal Papers (저널논문)
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