A user's guide to multicolor flow cytometry panels for comprehensive immune profiling
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A user's guide to multicolor flow cytometry panels for comprehensive immune profiling. / Holmberg-Thyden, Staffan; Grønbæk, Kirsten; Gang, Anne Ortved; El Fassi, Daniel; Hadrup, Sine Reker.
I: Analytical Biochemistry, Bind 627, 114210, 2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - A user's guide to multicolor flow cytometry panels for comprehensive immune profiling
AU - Holmberg-Thyden, Staffan
AU - Grønbæk, Kirsten
AU - Gang, Anne Ortved
AU - El Fassi, Daniel
AU - Hadrup, Sine Reker
N1 - Publisher Copyright: © 2021 The Author(s)
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - Flow cytometry
KW - Immune monitoring
KW - Immunology
KW - Myelodysplastic syndrome
KW - Unsupervised clustering
U2 - 10.1016/j.ab.2021.114210
DO - 10.1016/j.ab.2021.114210
M3 - Journal article
C2 - 34033799
AN - SCOPUS:85106548951
VL - 627
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
M1 - 114210
ER -
ID: 272028953