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Proceedings Paper

Spectral illumination system utilizing spherical reflection optics
Author(s): Samantha Gunn Mayes; Craig Browning; Samuel A. Mayes; Marina Parker; Thomas C. Rich; Silas J. Leavesley
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Paper Abstract

Fluorescence imaging microscopy has traditionally been used because of the high specificity that is achievable through fluorescence labeling techniques and optical filtering. When combined with spectral imaging technologies, fluorescence microscopy can allow for quantitative identification of multiple fluorescent labels. We are working to develop a new approach for spectral imaging that samples the fluorescence excitation spectrum and may provide increased signal strength. The enhanced signal strength may be used to provide increased spectral sensitivity and spectral, spatial, and temporal sampling capabilities. A proof of concept excitation scanning system has shown over 10-fold increase in signal to noise ratio compared to emission scanning hyperspectral imaging. Traditional hyperspectral imaging fluorescence microscopy methods often require minutes of acquisition time. We are developing a new configuration that utilizes solid state LEDs to combine multiple illumination wavelengths in a 2-mirror assembly to overcome the temporal limitations of traditional hyperspectral imaging. We have previously reported on the theoretical performance of some of the aspects of this system by using optical ray trace modeling. Here, we present results from prototyping and benchtop testing of the system, including assembly, optical characterization, and data collection. This work required the assembly and characterization of a novel excitation scanning hyperspectral microscopy system, containing 12 LEDs ranging from 365- 425 nm, 12 lenses, a spherical mirror, and a flat mirror. This unique approach may reduce the long image acquisition times seen in traditional hyperspectral imaging while maintaining high specificity and sensitivity for multilabel identification and autofluorescence imaging in real time.

Paper Details

Date Published: 17 February 2020
PDF: 7 pages
Proc. SPIE 11243, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII, 112430L (17 February 2020); doi: 10.1117/12.2546395
Show Author Affiliations
Samantha Gunn Mayes, Univ. of South Alabama (United States)
Craig Browning, Univ. of South Alabama (United States)
Samuel A. Mayes, Univ. of South Alabama (United States)
Marina Parker, Univ. of South Alabama (United States)
Thomas C. Rich, Univ. of South Alabama (United States)
Silas J. Leavesley, Univ. of South Alabama (United States)


Published in SPIE Proceedings Vol. 11243:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII
Daniel L. Farkas; Attila Tarnok, Editor(s)

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