Hi everyone! I'm new to the world of flow cytometry, I want to count the T cells in the rat liver, After getting the monocytes from the liver, I stained the CD3, then first I gate the monocytes and let the machine count 10,000 cells then stop, see the percentage of CD3+, is it OK? Using 10,000 multiply CD3+ percentage？
Mingjie,
I am not a flow expert, however when I am using flow I typically am only looking for the percentages of each cell type and not the actual number as the number really depends on where your sample came from and how it was processed. I usually am looking at the different liver cell types and different hepatocyte zone types found. So, unless you have a really good method for normalization and to account for loss of cells during sample processing, I would think the better way to report your data would be to discuss the percentage of all cells types that your T cells make up instead of quantifying the cell number itself. But, again, I am not a flow expert and others may have better ideas for how you can actually quantify more accurately. (I just know from my experience that depending on your methods your numbers/percentages can change and are not always consistent).
Good luck!
Hi
I use to count the number of the total cells using a cell counter and then detect and calculate the percentage of the target in the total using flow cytometry.
Hello,
I agree with previous answer. FACS will only give you a percentage of your population of interest. You can assume that a event correspond to one cell passing through but that can also correspond to debris or doublet cells.
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