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Large field-of-view nanometer-sectioning microscopy by using metal-induced energy transfer and biexponential lifetime analysis

Authors
 Wonsang Hwang  ;  Jinwon Seo  ;  DongEun Kim  ;  Chang Jun Lee  ;  In-Hong Choi  ;  Kyung-Hwa Yoo  ;  Dug Young Kim 
Citation
 COMMUNICATIONS BIOLOGY, Vol.4(1) : 91, 2021-01 
Journal Title
 COMMUNICATIONS BIOLOGY 
Issue Date
2021-01
Abstract
Total internal reflection fluorescence (TIRF) microscopy, which has about 100-nm axial excitation depth, is the method of choice for nanometer-sectioning imaging for decades. Lately, several new imaging techniques, such as variable angle TIRF microscopy, supercritical-angle fluorescence microscopy, and metal-induced energy transfer imaging, have been proposed to enhance the axial resolution of TIRF. However, all of these methods use high numerical aperture (NA) objectives, and measured images inevitably have small field-of-views (FOVs). Small-FOV can be a serious limitation when multiple cells need to be observed. We propose large-FOV nanometer-sectioning microscopy, which breaks the complementary relations between the depth of focus and axial sectioning by using MIET. Large-FOV imaging is achieved with a low-magnification objective, while nanometer-sectioning is realized utilizing metal-induced energy transfer and biexponential fluorescence lifetime analysis. The feasibility of our proposed method was demonstrated by imaging nanometer-scale distances between the basal membrane of human aortic endothelial cells and a substrate.
Files in This Item:
T202101293.pdf Download
DOI
10.1038/s42003-020-01628-3
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Choi, In Hong(최인홍) ORCID logo https://orcid.org/0000-0001-9851-0137
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/182453
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