Tumor bearing lungs were minced and digested enzymatically using collagenase type 2 (Worthington, Lakewood, NJ) and DNaseI (Roche Applied Science, Indianapolis, IN, USA) at 37°C with constant shaking. The Csf1r-YFP or F4/80 positive myeloid populations (TIMs) were sorted on the BD FACS Aria/ AriaIII. For bone marrow studies, bone marrow precursors were flushed out from the femurs and tibiae of metastatic tumor bearing mice and myeloid cells were sorted using PE conjugated anti-CD11b (BD Pharmingen, San Jose, CA, USA) and APC conjugated anti-GR1 antibodies (eBioscience, San Diego, CA, USA).
Facs aria ariaiii
Manufactured by BD
The BD FACS Aria and AriaIII are high-performance cell sorters designed for advanced flow cytometry applications. They offer precise cell sorting capabilities for a wide range of sample types and research purposes.
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2 protocols using facs aria ariaiii
1
Isolation of Tumor-Infiltrating Myeloid Cells
2
Isolation of Tumor-Infiltrating Myeloid Cells
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Tumor bearing lungs were minced and digested enzymatically using collagenase type 2 (Worthington, Lakewood, NJ) and DNaseI (Roche Applied Science, Indianapolis, IN, USA) at 37°C with constant shaking. The Csf1r-YFP or F4/80 positive myeloid populations (TIMs) were sorted on the BD FACS Aria/ AriaIII. For bone marrow studies, bone marrow precursors were flushed out from the femurs and tibiae of metastatic tumor bearing mice and myeloid cells were sorted using PE conjugated anti-CD11b (BD Pharmingen, San Jose, CA, USA) and APC conjugated anti-GR1 antibodies (eBioscience, San Diego, CA, USA).
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