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Updated: Aug 30, 2025

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals
Published on: August 22, 2019
Design and Validation of a Custom-Made Laboratory Hyperspectral Imaging System for Biomedical Applications Using a
Jošt Stergar1,2, Rok Hren2, Matija Milanič1,2
1Jozef Stefan Institute, Jamova Cesta 39, SI-1000 Ljubljana, Slovenia.
Abstract:
Hyperspectral imaging (HSI) is a promising optical modality that is already being used in numerous applications. Further expansion of the capabilities of HSI depends on the modularity and versatility of the systems, which would, inter alia, incorporate profilometry, fluorescence imaging, and Raman spectroscopy while following a rigorous calibration and verification protocols, thus offering new insights into the studied samples as well as verifiable, quantitative measurement results applicable to the development of quantitative metrics. Considering these objectives, we developed a custom-made laboratory HSI system geared toward biomedical applications. In this report, we describe the design, along with calibration, characterization, and verification protocols needed to establish such systems, with the overall goal of standardization. As an additional novelty, our HSI system uses a custom-built broadband LED-based light source for reflectance imaging, which is particularly important for biomedical applications due to the elimination of sample heating. Three examples illustrating the utility and advantages of the integrated system in biomedical applications are shown. Our attempt presents both the development of a custom-based laboratory HSI system with novel LED light source as well as a framework which may improve technological standards in HSI system design.

