MBTP 15: Immunophenotyping of neutrophils and eosinophils from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 15 (MBTP 15)

This application protocol describes the flow cytometric analysis of neutrophils and eosinophils 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.

Protocol

Gating strategy showing the analysis of neutrophils and eosinophils from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described panel to identify neutrophils and eosinophils. 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+ neutrophils and eosinophils (A) express CD11b and lack expression of T, B, and NK cell lineage markers CD3, CD19, and CD335 (B). Within CD11b+ cells, neutrophils can be distinguished by the expression of Ly-6G while eosinophils are Ly-6G- (C) and co-express Siglec-F and the chemokine receptor CD193 (D). 

Materials

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