Computational Hyperspectral Microflow Cytometry
- PMID: 38770741
- DOI: 10.1002/smll.202400019
Computational Hyperspectral Microflow Cytometry
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.
Keywords: T lymphocyte analysis; computational hyperspectral fluorescence analysis; microflow cytometry; sheathless focusing; spectral reconstruction.
© 2024 The Author(s). Small published by Wiley‐VCH GmbH.
References
-
- D. A. Ateya, J. S. Erickson, P. B. Howell Jr, L. R. Hilliard, J. P. Golden, F. S. Ligler, Anal. Bioanal. Chem. 2008, 391, 1485.
-
- R.‐J. Yang, L.‐M. Fu, H.‐H. Hou, Sens. Actuators, B 2018, 266, 26.
-
- S. Stavrakis, G. Holzner, J. Choo, A. deMello, Curr. Opin. Biotechnol. 2019, 55, 36.
-
- Z. Li, S. Zhang, Q. Wei, in Smartphone Based Medical Diagnostics, (Ed: J.‐Y. Yoon), Academic Press, 2020, pp. 67–88.
-
- A. Sun, T. Li, T. Jin, Y. Li, K. Li, C. Song, L. Xi, Anal. Chem. 2021, 93, 14820.