Gating strategy used for flow cytometry of human PBMCs.
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posted on 2019-02-27, 18:55 authored by Peter M. Mihailovic, Wai Man Lio, Romana Herscovici, Kuang-Yuh Chyu, Juliana Yano, Xiaoning Zhao, Jianchang Zhou, Bo Zhou, Michael R. Freeman, Wei Yang, Prediman K. Shah, Bojan Cercek, Paul C. DimayugaPBMCs were collected after 72 hours in culture with and without stimulation with Keratin 8 peptide. Positive control was stimulation with 0.5x T cell stimulation cocktail. Cells were collected and stained for flow cytometry. Cell singlets were selected and non-viable cells were excluded, followed by size-gating of viable cells. Cells were plotted on CD62L and CCR7 based on CD3+CD4+ cells or CD3+CD8+ cells. CD62L(-)CCR7(-) Effector cells were selected as T Effector Memory (TEM) or T Effector Memory RA+ (TEMRA) based on CD45RO/CD45RA stain.
