Multispectral Imaging Flow Cytometry with Spatially and Spectrally Resolving Snapshot-Mosaic Cameras for the Characterization and Classification of Bioparticles

Dittrich, Paul-Gerald; Kraus, Daniel; Ehrhardt, Enrico; Henkel, Thomas; Notni, Gunther

doi:10.3390/mi13020238

Cited by 3 publications

(1 citation statement)

References 33 publications

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“…In the meanwhile, hyperspectral imaging (HS) would incorporate features of a regular flow cytometer and a fluorescent microscope, addressing the limitations of brightfield imaging in distinguishing fluorophores spatial distribution. Implementation of hyperspectral imaging has been suggested in flow cytometrical field in prior papers [4], [5] , but the amount of data collected on utilization of lasers for excitation in tandem with hyperspectral imaging remains limited.…”

Section: Imaging In Flow Cytometrymentioning

confidence: 99%

Hyperspectral imaging for flow cytometry

Filipchuk,

Perttilä,

Colucci

et al. 2024

Multimodal Biomedical Imaging XIX

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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.

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Section: Imaging In Flow Cytometrymentioning

confidence: 99%

Hyperspectral imaging for flow cytometry

Filipchuk,

Perttilä,

Colucci

et al. 2024

Multimodal Biomedical Imaging XIX

View full text Add to dashboard Cite

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Classification of melanocytic lesions using direct illumination multispectral imaging

Goessinger,

Dittrich,

Nöcker

et al. 2024

Sci Rep

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Metasurface-enabled barcoding for compact flow cytometry

Hong,

Li,

Valentine

2024

Optica

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Flow cytometers are a vital tool for cellular phenotyping but are primarily limited to centralized laboratories due to their bulkiness and cost. Significant efforts have been made to construct on-chip flow cytometers for point-of-care applications, and a promising approach is filter-on-chip flow cytometers utilizing the conventional Bayer RGB filter on imaging cameras to miniaturize key optoelectronic components. However, conventional RGB filters fail to provide spectral channels of sufficient diversity and specificity for accurate identification of fast-moving fluorescence signals. Here, we present an optofluidic system with integrated metasurfaces that serve to increase the number and diversity of the spectral channels. Inverse design of spatially coded metasurfaces is used to maximize the classification accuracy of spectral barcodes generated along the particle trajectory obtained from single-shot imaging. The accuracy of this system is shown to be superior to generic RGB filter approaches while also realizing classification of up to 13 unique combinations of fluorophores, significantly enhancing the capability of portable flow cytometers.

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Multispectral Imaging Flow Cytometry with Spatially and Spectrally Resolving Snapshot-Mosaic Cameras for the Characterization and Classification of Bioparticles

Cited by 3 publications

References 33 publications

Hyperspectral imaging for flow cytometry

Hyperspectral imaging for flow cytometry

Classification of melanocytic lesions using direct illumination multispectral imaging

Metasurface-enabled barcoding for compact flow cytometry

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