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Amplified Expansion Stimulated Emission Depletion Microscopy
Author(s) -
Kim Doyeon,
Kim Taeyeon,
Lee Jooyong,
Shim SangHee
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800775
Subject(s) - sted microscopy , fluorophore , microscopy , photobleaching , stimulated emission , resolution (logic) , super resolution microscopy , fluorescence , fluorescence microscope , biotinylation , avidin , chemistry , streptavidin , microscope , optics , materials science , analytical chemistry (journal) , biotin , physics , laser , biochemistry , chromatography , artificial intelligence , computer science
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.