Computational Hyperspectral Microflow Cytometry Articles uri icon

published in

publication date

  • July 2024

start page

  • 1

end page

  • 12

issue

  • 30

volume

  • 20

International Standard Serial Number (ISSN)

  • 1613-6810

Electronic International Standard Serial Number (EISSN)

  • 1613-6829

abstract

  • Miniaturized flow cytometry has significant potential for portable applications, such as cell-based diagnostics and the monitoring of therapeutic cell manufacturing, however, the performance of current techniques is often limited by the inability to resolve spectrally-overlapping fluorescence labels. Here, the study presents a computational hyperspectral microflow cytometer (CHC) that enables accurate discrimination of spectrally-overlapping fluorophores labeling single cells. CHC employs a dispersive optical element and an optimization algorithm to detect the full fluorescence emission spectrum from flowing cells, with a high spectral resolution of ≈3 nm in the range from 450 to 650 nm. CHC also includes a dedicated microfluidic device that ensures in-focus imaging through viscoelastic sheathless focusing, thereby enhancing the accuracy and reliability of microflow cytometry analysis. The potential of CHC for analyzing T lymphocyte subpopulations and monitoring changes in cell composition during T cell expansion is demonstrated. Overall, CHC represents a major breakthrough in microflow cytometry and can facilitate its use for immune cell monitoring.

subjects

  • Biology and Biomedicine
  • Electronics

keywords

  • computational hyperspectral fuorescence analysis; microflow cytometry; spectral reconstruction; sheathless focusing; t lymphocyte analysis