Abstract
The field of flow cytometry has witnessed rapid technological advancements in the last few decades. While the founding principles of fluorescent detection on cells (or particles) within a uniform fluid stream remains largely unchanged, the availability more sensitive cytometers with the ability to multiplex more and more florescent signals has resulted in very complex high-order assays. This results in the co-use of fluorophores with increased levels of emission overlap and/or spillover spreading than in years past and thus requires careful and well thought out planning for flow cytometry assay development. As an example, we present the development of a large 18-color (20 parameter) flow cytometry assay designed to take an in depth analysis of effector lymphocyte phenotypes, with careful attention to assay controls and panel design.
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Acknowledgments
This work was supported by the Moffitt Cancer Center—Innovative Core Projects (Project number 16060201), NCI–NIH (1 R01 CA148995-01; P30CA076292; P50CA168536), the V Foundation, the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation, and the Chris Sullivan Foundation.
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Nakagawa, R., Brayer, J., Restrepo, N., Mulé, J.J., Mailloux, A.W. (2021). High-Dimensional Flow Cytometry Analysis of Regulatory Receptors on Human T Cells, NK Cells, and NKT Cells. In: Markowitz, J. (eds) Translational Bioinformatics for Therapeutic Development. Methods in Molecular Biology, vol 2194. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0849-4_14
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DOI: https://doi.org/10.1007/978-1-0716-0849-4_14
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