Validation of excitation-scan hyperspectral multi-faceted mirror array prototype system advancements to hyperspectral imaging applications

Marina Parker; Sam A. Mayes; Craig M. Browning; Joshua Deal; Samantha Gunn-Mayes; Thomas C. Rich; Silas J. Leavesley

doi:10.1117/12.2607659

2 March 2022 Validation of excitation-scan hyperspectral multi-faceted mirror array prototype system advancements to hyperspectral imaging applications

Marina Parker, Sam A. Mayes, Craig M. Browning, Joshua Deal, Samantha Gunn-Mayes, Thomas C. Rich, Silas J. Leavesley

Author Affiliations +

Proceedings Volume 11966, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX; 1196606 (2022) https://doi.org/10.1117/12.2607659
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Hyperspectral imaging technologies (HSI) have undergone rapid development since their beginning stages. While original applications were in remote sensing, other uses include agriculture, food safety and medicine. HSI has shown great utility in fluorescence microscopy for detecting signatures from many fluorescent molecules; however, acquisitions speeds have been slow due to light losses associated with spectral filtering. Therefore, we designed a novel light emitting diode (LED)-based rapid excitation scanning hyperspectral imaging platform allowing users to obtain simultaneous measurements of fluorescent labels without compromising acquisition speeds. Previously, we reported our results of the optical ray trace simulations and the geometrical capability of designing a multifaceted mirror imaging system as an initial approach to combine light at many wavelengths. The design utilized LEDs and a multifaceted mirror array to combine light sources into a liquid light guide. The computational model was constructed using Monte Carlo optical ray software (TracePro, Lambda Research Corp.). Recent prototype validation results show that when compared to a commercial emission scanning spectral confocal microscope (Zeiss-LSM-980), the novel LED-based excitation scanning HSI prototype successfully detected and separated six fluorescent labels from a custom 6-label African green monkey kidney epithelial cells. We report on the prototype’s ability to overcome limitations of acquisition speeds, sensitivity, and specificity present in conventional systems. Future work will evaluate prototype’s light losses to determine latent design modifications needed to demonstrate the system’s feasibility as a promising solution for overcoming HSI acquisition speeds. This work was supported by NSF award MRI1725937.

Conference Presentation

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Marina Parker, Sam A. Mayes, Craig M. Browning, Joshua Deal, Samantha Gunn-Mayes, Thomas C. Rich, and Silas J. Leavesley "Validation of excitation-scan hyperspectral multi-faceted mirror array prototype system advancements to hyperspectral imaging applications", Proc. SPIE 11966, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX, 1196606 (2 March 2022); https://doi.org/10.1117/12.2607659

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