Analysis of flow cytometry results.

[Bijou]

Hi All,

I am doing flow for the first time and need someone to tell me if the way I am analysing my results is fine:
I have a test antibody (CD34) and an isotype control. I have run each in a different tube but on the same cell line.
Now, in the results, I had to gate that particular population of course and have got mean flouresence values, which are higher for isotype controls.
Do I substract the mean of test from the isotype control mean? What's the best way to analyse flow results?

Help much appreciated.

BIJ

[CrowSan]

Sorry I am a bit confused. Your isotype control cells were more fluorescent than your samples cells? What flurophore are you using? Is it directly linked to your CD34 antibody (conjugated) or do you add a secondary?

If your samples have higher values than your controls then yes you could subtact the controls from samples to give a mean fuorescent value (if all of your CD34+ samples were positive for CD34+). Alternatively you could gate out non-fluorescent cell population from your samples and give the results as a % cells expressing CD34+ with the mean fluorescent values of the cells (hope that made sense).

CrSn