Journal article
A user's guide to multicolor flow cytometry panels for comprehensive immune profiling
University of Copenhagen1
T-Cells and Cancer, Experimental & Translational Immunology, Department of Health Technology, Technical University of Denmark2
Department of Health Technology, Technical University of Denmark3
Experimental & Translational Immunology, Department of Health Technology, Technical University of Denmark4
Multicolor flow cytometry is an essential tool for studying the immune system in health and disease, allowing users to extract longitudinal multiparametric data from patient samples. The process is complicated by substantial variation in performance between each flow cytometry instrument, and analytical errors are therefore common.
Here, we present an approach to overcome such limitations by applying a systematic workflow for pairing colors to markers optimized for the equipment intended to run the experiments. The workflow is exemplified by the design of four comprehensive flow cytometry panels for patients with hematological cancer.
Methods for quality control, titration of antibodies, compensation, and staining of cells for obtaining optimal results are also addressed. Finally, to handle the large amounts of data generated by multicolor flow cytometry, unsupervised clustering techniques are used to identify significant subpopulations not detected by conventional sequential gating.
Language: | English |
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Year: | 2021 |
Pages: | 114210 |
ISSN: | 10960309 and 00032697 |
Types: | Journal article |
DOI: | 10.1016/j.ab.2021.114210 |
ORCIDs: | Hadrup, Sine Reker and 0000-0001-8226-1658 |