Analysis of Lung Immune Cells by Flow Cytometry

Analysis of cell populations in BALF or single cell suspension of lung homogenate was conducted as before^{46 (link)}. Cells were labeled with a combination of phycoerythrin (PE)-labeled anti-NK 1.1 (PK136; eBioscience), fluorescein (FITC)-labeled anti-Ly6G (RB6-8C5; eBioscience), allophycocyanin (APC)-labeled anti-MHCII (M5/114.15.2; eBioscience), PerCP-Cy5.5-labelled anti-CD11c (N418; Biolegend), or phycoerythrin (PE)-labeled CD4 (RM4-5; Biolegend) alone. Neutrophils (PMNs) were defined as Ly6G⁺/MHCII⁻ , Macrophages as CD11c⁺/MHCII^low-mid and Dendritic cells (DCs) as CD11c⁺/MHCII^high. Fc block (anti-mouse CD16/32) was also added to each sample (93; eBioscience). Uniform dyed microspheres (Bangs Laboratories, Inc) were added to calculate the concentration of cellular components. Samples were run on FACS Calibur and analyzed on FlowJo (ver 10.0.6).

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Ahn D., Parker D., Planet P.J., Nieto P.A., Bueno S.M, & Prince A. (2014). Secretion of IL-16 through TNFR1 and calpain-caspase signaling contributes to MRSA pneumonia. Mucosal immunology, 7(6), 1366-1374.

Publication 2014

labeled anti-NK 1.1 (PK136; FITC)-labeled anti-Ly6G (RB6-8C5; PerCP-Cy5.5-labelled anti-CD11c (N418; RM4-5; Fc block (anti-mouse CD16/32) Uniform dyed microspheres

Allophycocyanin Cells Cellular components Cy5 5 Dendritic cells Fitc Fluorescein Lung Macrophages Microspheres Mouse Neutrophils Phycoerythrin Populations

Corresponding Organization :

Other organizations : Columbia University, Pontificia Universidad Católica de Chile

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