What method do you use to measure the antibody-dependent phagocytosis of cancer cells by macrophages? Do you prefer to use microscopy or flow cytometry for this analysis?
Thanks,
Chris
The easiest,high-throughput method is to incubate fluorescent, antigen-coated beads with antibodies and then "feed" them to phagocytic cells (either primary phagocytic cells or phagocytic cell lines). Phagocytosis is measured by the increase in fluorescence intensity (MFI) of the phagocytoci cells. for example: Malar J. 2016 May 31;15:301. doi: 10.1186/s12936-016-1348-9.
let me know if you need a protocol...
To my understanding, flow cytometry is best to as it tells you both qualitative and quantitative results unlike microscopy. Here is how you can proceed :
labelled your tumour cells with a cell staining fluorescence dye known as Carboxyfluorescein succinimidyl ester( my favorite dye )and mix these cells with your effector cells and tumour cell antigen specific monoclonal antibodies followed by incubation. label your cells antibodies conjugated with any florescence dye and use cell liability marker such as FVD. Then using FC machine, count the number of killed or phagocytozed and live cells. it will tell you both qualitative and quantitative figure. good luck.
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