Hi - we have breast cancer biopsies which have been stained for T cells, B cells etc using DAB, and we are wanting a method to determine how many immune cells there are. The problem is there are areas where there are lots of immune cells, and a scarcity in others, and some areas are mostly fat - so how best to compare between the various biopsies, which can vary in the amount of tumour/fat tissue? I feel we need some sort of measurement of percentage of immune cells compared to tumour cells. We did a basic scoring system of high/med/low, which is showing some differences between our groups. but we would like a more robust and quantitative method. Any suggestions? Thanks
I think the best method to sort between immune and tumor cells in the selected region is immunostaining followed by evaluating the immunoreactive score according to Remmele and Stegner. When you are interested in the whole tumor tissue you can use PCR or something more complicated the flow cytometer with cell sorter to sort the immune and tumor cells. Good luck
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