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Self-Interference Fluorescent Emission Microscopy for Biological Imaging

Published online by Cambridge University Press:  02 July 2020

A.K. Swan ,
Y. Tong ,
L. Moiseev ,
M. S. Ünlü  and
B.B. Goldberg
A.K. Swan
Affiliation:
Electrical and Computer Engineering, Boston University, MA, 02215Department of Physics, Boston University, Boston, MA, 02215Department of Biomedical Engineering, Boston, MA, 02215
Y. Tong
Affiliation:
Electrical and Computer Engineering, Boston University, MA, 02215Department of Physics, Boston University, Boston, MA, 02215Department of Biomedical Engineering, Boston, MA, 02215
L. Moiseev
Affiliation:
Electrical and Computer Engineering, Boston University, MA, 02215Department of Physics, Boston University, Boston, MA, 02215Department of Biomedical Engineering, Boston, MA, 02215
M. S. Ünlü
Affiliation:
Electrical and Computer Engineering, Boston University, MA, 02215Department of Physics, Boston University, Boston, MA, 02215Department of Biomedical Engineering, Boston, MA, 02215
B.B. Goldberg
Affiliation:
Electrical and Computer Engineering, Boston University, MA, 02215Department of Physics, Boston University, Boston, MA, 02215Department of Biomedical Engineering, Boston, MA, 02215
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Abstract

We present a new method of fluorescence imaging, which yields an unprecedented nm scale vertical resolution. The method uses the unique spectral signature of the fluorescent emission intensity above a reflecting surface to determine vertical position unambiguously. Emission from several heights could be resolved by deconvoluting the spectrum, so that three dimensional imaging is possible. The related technique of standing wave microscopy uses the spatial intensity variation for increased resolution. Applications of this technique include intracellular imaging and screening for specific bacteria, virus or proteins, where discrimination between raised fluorescently labeled specifically bound markers from non-specific binding is crucial.

The emission spectrum of fluorescent markers is modified in the presence of a reflecting surface. Within the wavelength range of the fluorescent emission, the selfinterference of the emitted photons from the direct and reflected path results in enhanced or suppressed emission (constructive or destructive interference) depending on the height and wavelength.

Type
Bridging the Gap Between Structural and Molecular Biology (Organized by B. Herman)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 36 - 37
Copyright
Copyright © Microscopy Society of America 2001

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References

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