Hyperspectral imaging for flow cytometry

Flow cytometry is a widely used analysis technique in biomedical sciences. It has found extensive utilization in both clinical diagnostics and cutting-edge biological research. As the method has been gaining greater recognition, its underlying technologies have undergone rapid development to further expand its range of applications. A notable trend is the introduction of imaging modalities to flow cytometry to expand the information content of the analyzed sample. The introduction of a camera component to the already well-established detectors, such as photo multiplier tubes (PMTs) or avalanche photo diodes (APDs), adds intricacy to the arrangement of optical subsystem in flow cytometer. Moreover, it brings forth additional requirements for effectively coordinating information capture among different detector types. An appealing alternative to address this challenge is hyperspectral imaging - a technique which enables capturing of the spatial and spectral information simultaneously. Yet, there has not been much research performed to study applications of hyperspectral imaging in combination with narrow bandwidth illumination commonly used in flow cytometry. In this work, we investigate the applicability of hyperspectral imaging to flow cytometrical systems, where a multiple wavelength laser system is utilized for sample illumination. A four-wavelength laser illumination platform developed by Modulight Corporation is utilized as the light source. Our main objective is to assess the hyperspectral imaging component's ability to distinguish between the illuminating light and the fluorescence emitted by the sample. Furthermore, we carefully evaluate the quality of the obtained hyperspectral images and explore the potential to differentiate samples based on the collected spatial data.