MBTP 11: Immunophenotyping of T cells from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 11 (MBTP 11)

This application protocol describes the flow cytometric analysis of major T cell subsets after spleen dissociation from healthy C57BL/6 mice. Mouse spleens can easily be dissociated using the gentleMACS™ Dissociators to quickly obtain viable single cell suspensions ready for downstream applications, such as flow cytometric analysis.

Protocol

Gating strategy showing the analysis of T cell subsets from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described T cell panel to identify major T cell subsets. CD4+ and CD8+ T cells were identified from CD3-expressing cell subsets (A, B). Different subsets of naive, central memory (CM), effector memory (EM), CD127-, and KLRG1-expressing T cells were analyzed within CD4+ (C, D, E) and CD8+ T cells (F, G, H) respectively. The differential expression of the adhesion molecules CD62L and CD44 was used to identify naive T cells from CM and EM T cells. Naive T cells expressed high levels of CD62L and showed absent or low expression of CD44 (C). Both CM and EM populations expressed CD44. CM T cells also expressed CD62L, whereas expression of CD62L was absent in EM T cells. Analysis of CD127 and KLRG1 was included in this panel to allow the study of T cell responses during infection. During the course of an infection the levels of KLRG1 increase, especially on CD8+ T cells.

Materials

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