I wanted to know which marker I should use to sort the splenic macrophages from mice by flow cytometry? Actually I checked the literature and observed that people use CD11b or F4/80 as the marker for sorting of splenic macrophages. I was going to use F4/80+ CD11c- strategy but was curious to know if that is fine to proceed. I need these sorted macrophages for antigen presentation experiments. Also, can anybody give me the estimate how many macrophages (F4/80 or CD11b+) usually can be obtained by sorting one mouse spleen?
Hello Tabasum,
I recommend you to read the paper bellow. There is the possibility to identify macrophages and monocytes in spleen. This gating strategy could serve to assure that you are sorting the correct population. I'm not sure about macrophage percentage in spleen, but if you have a 5% from total events, you can recovery 1-2 million of macrophages by sorting one spleen
Hope this help,
Virginia
Hi, Is there any good way to isolate red pulp macrophages from spleen? I tried sorting the splenic macrophages by FACS using CD11b and F4/80 markers. I used 4-color staining: FITC CD11c, gated them and sorted them separately in one tube, APC F4/80 and CD11b double positive population gated and sorted them separately in the different tube simultaneously. I used PE Gr-1 also to gate out neutrophils. However, the problem is I am getting very few F4/80/CD11b double positive cells with the purity of about only 60%. The wiered thing is that I am getting more sorted DCs with high purity compared to macrophages from the same sample. Please suggest if there is any other better way to sort the splenic macrophages in good number with high purity. I have attached the sort pictures.
Thanks,
Tabasum
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