Hyperspectral cytometry at the single-cell level using a 32-channel photodetector.
type: Distribution
access URL: https://hal.science/hal-00691181
format: HTML
issued: 2026-05-20T19:51:28.032388
modified: 2026-05-20T19:51:27.985868

Hyperspectral cytometry at the single-cell level using a 32-channel photodetector.
type: Dataset
contact point: Organization
  fn: CCSD
keyword: classification
keyword: system
keyword: sdvotlife-sciences-q-bioother-q-bioot
keyword: infoeu-reposemanticsarticle
keyword: next-generation-instruments
keyword: flow-cytometry
keyword: resolution-spectral-analysis
keyword: fluorescence-spectra
keyword: hyperspectral-cytometry
keyword: journal-articles
keyword: nanoparticles
landing page: the journal of the International Society for Analytical Cytology
description: Despite recent progress in cell-analysis technology, rapid classification of cells remains a very difficult task. Among the techniques available, flow cytometry (FCM) is considered especially powerful, because it is able to perform multiparametric analyses of single biological particles at a high flow rate-up to several thousand particles per second. Moreover, FCM is nondestructive, and flow cytometric analysis can be performed on live cells. The current limit for simultaneously detectable fluorescence signals in FCM is around 8-15 depending upon the instrument. Obtaining multiparametric measurements is a very complex task, and the necessity for fluorescence spectral overlap compensation creates a number of additional difficulties to solve. Further, to obtain well-separated single spectral bands a very complex set of optical filters is required. This study describes the key components and principles involved in building a next-generation flow cytometer based on a 32-channel PMT array detector, a phase-volume holographic grating, and a fast electronic board. The system is capable of full-spectral data collection and spectral analysis at the single-cell level. As demonstrated using fluorescent microspheres and lymphocytes labeled with a cocktail of antibodies (CD45/FITC, CD4/PE, CD8/ECD, and CD3/Cy5), the presented technology is able to simultaneously collect 32 narrow bands of fluorescence from single particles flowing across the laser beam in <5 μs. These 32 discrete values provide a proxy of the full fluorescence emission spectrum for each single particle (cell). Advanced statistical analysis has then been performed to separate the various clusters of lymphocytes. The average spectrum computed for each cluster has been used to characterize the corresponding combination of antibodies, and thus identify the various lymphocytes subsets. The powerful data-collection capabilities of this flow cytometer open up significant opportunities for advanced analytical approaches…
issued: 2026-05-20T19:51:28.011729
language: en
modified: 2026-05-20T19:51:28.011734

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type: Agent