MBTP 14: Immunophenotyping of macrophages from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 14 (MBTP 14)

This application protocol describes the flow cytometric analysis of macrophages after spleen dissociation from healthy C57BL/6 mice. Mouse spleens are easily dissociated using the gentleMACS™ Dissociators to quickly obtain viable single cell suspensions ready for flow cytometric analysis.


Gating strategy showing the analysis of red pulp macrophages from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described panel. Samples were initially gated on single cells using FSC-H/FSC-A gating and on live cells using Viobility-negative gating (data not shown). CD45+ red pulp macrophages (A) lack expression of T, B, and NK cell lineage markers CD3, CD19, and CD335 (B) and express CD11b, F4/80, and CD68 (C). In addition, red pulp macrophages also characteristically express Mer and CD64 (D). 


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