[Flow cytometry] T cell phenotypes（T cells subsets）gating strategy

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How to design a panel for analyzing T cell subsets in peripheral blood by using Flow Cytometry?

First of all, you have to know which lasers you have in your machine. Base on the lasers you have, design a panel with suitable fluorochromes conjugated with your cell markers.

To gate in different T cell subsets, you will need to have following CD markers as below (CD Markers panel) with desired fluorochromes. By these, you can first gate CD4 and CD8 T cells and then their subsets, such as T Central Memory cells (ＴCM cells), T Effector Memory cells (ＴEM cells), T Terminal Effector cells (ＴTE cells), T stem cell memory, T regulatory cells (Treg cells) and Treg subsets (ＴregCM cells, ＴregEM cells and ＴregTE cells).

CD Markers panel（BD biosciences）
  1. CD45 KrO
  2. CD3 APC750
  3. CD4 APC
  4. CD8 APC700
  5. CD45RA FITC
  6. CCR7 PE
  7. PD-1 PC5.5
  8. CD57 PB
  9. CD27 PC7
  10. CD28 ECD
  11. CD127 BV650
  12. CD25 BV785
  13. CD14 BUV395
  14. CD16 BUV496
  15. HLA-DR BUV661
  16. CD38 BUV525
  17. CD95 BUV405
  18. Promokine 840 viability
  19. Forward Scatter
  20. Side Scatter
  21. Time
Here is an example of T cell subsets gating strategy:
Figure 1: T cell subsets gating strategy