I will sort some cells using FACS sorting and I need to compare them to unsorted cells. Do I have to make the unsorted cells pass through the same steps like the sorted one but without staining to be compared? (I mean multiple centrifugations and pass through the FACS machine) because i know these steps may be hard on the cells and may affect their response and growth rate
I know that but what about the unsorted cells ? should they pass through the sorting machine too ?
Hi Osama, if you are fortunate enough to have additional FACS sorting time, then go for it. I agree with you that the stresses imposed by sorting could affect the cells and that running your cells through the FACS sorter would be the best control, however I doubt whether many researchers would bother. It would be worth trying at least once to see whether it makes a difference to your cells. Regards Rosemary
I suppose the negative population will be the best control for your positive sorted cells. You can collect both at the same time (just one sorting). However if you need the original population as a control for your experimental purpose you can sort and collect the unstained or isotype control sample which should be treated the same as your stained sample.
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