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OMIP-057

Mouse γδ T‐Cell Development Characterized by a 14 Color Flow Cytometry Panel

Terkild Brink Buus, Mia Hamilton Jee, Niels Ødum Cytometry PART A, Volume 95, Issue 7, 726-729 (2010)

PURPOSE: This panel was designed to quantify the distribution of developing γδ T cells within seven development stages related to the programming of distinct effector phenotypes in the murine thymus. Furthermore, the panel was designed to assess the expression of additional surface markers at each development stage within two of the major γδ T‐cell subsets identified by the usage of different V‐segments in their T‐cell receptor (TCR): TCRVγ1.1+ and TCRVγ2+. The panel was developed using thymus from adult C57Bl/6 mice magnetically depleted of CD4 and CD8 expressing cells in order to enrich for the γδ T‐cell population.

CELL TYPE: Thymocytes (thymus lymphocytes)

MACHINE: BD LSR Fortessa

Marker Target Clone Fluorochrome Purpose
Dead cells Mouse PI n/a
CD8 Mouse 53-6.7 BUV395 Depletion of aß T cells and progenitor
CD4 Mouse Gk1.5 BUV737 Depletion of aß T cells and progenitor
CD3 Mouse 145-2C11 Brilliant Violet 786 T cell
CLEC12A Mouse 5D3/CLEC12A Brilliant Violet 421 Population A, B
CD24 Mouse M1/69 Brilliant Violet 510 Population A-F
CD25 Mouse PC61 APC-Cy7 Population A
CD73 Mouse TY/11.8 Brilliant Violet 605 Population E, F, G
CD117 Mouse 2B8 APC-R700 Population E
CD200 Mouse OX-90 eFluor 660 Population D, E, F
TCR delta Mouse GL-3 PE-CF594 ?d T
TCR V gamma 1.1 Mouse 2.11 FITC ?d T-cell subset
TCR V gamma 2 Mouse UC3-10A6 PE-Cy7 ?d T-cell subset