Large field-of-view nanometer-sectioning microscopy by using metal-induced energy transfer and biexponential lifetime analysis
Open Access
- 19 January 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Communications Biology
- Vol. 4 (1), 1-7
- https://doi.org/10.1038/s42003-020-01628-3
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.Keywords
Funding Information
- National Research Foundation of Korea (2016M3A7B6908929, 2020R1A2C2011942, NRF-2017R1A2B4003950)
- Korea Institute for Advancement of Technology (P0011925)
This publication has 39 references indexed in Scilit:
- Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thalianaPlant Methods, 2011
- Axial Nanometer Distances Measured by Fluorescence Lifetime Imaging MicroscopyNano Letters, 2010
- Differential Evanescence Nanometry: Live-Cell Fluorescence Measurements with 10-nm Axial Resolution on the Plasma MembraneBiophysical Journal, 2008
- High-throughput screening of cellular features using high-resolution light-microscopy; Application for profiling drug effects on cell adhesionJournal of Structural Biology, 2007
- Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization MicroscopyBiophysical Journal, 2006
- Imaging Intracellular Fluorescent Proteins at Nanometer ResolutionScience, 2006
- Quantitating fluorescence intensity from fluorophores: Practical use of MESF valuesJournal of Research of the National Institute of Standards and Technology, 2002
- Fluorescence near interfaces: The role of photonic mode densityJournal of Modern Optics, 1998
- Enhancement and inhibition of electromagnetic radiation in plane-layered media IPlane-wave spectrum approach to modeling classical effectsJournal of the Optical Society of America B, 1997
- Sellmeier coefficients and dispersion of thermo-optic coefficients for some optical glassesApplied Optics, 1997