Hello,
I carried out a multicolour immunophenotyping flow cytometry and now I am analyzing the data got. Basically, it's not clear to me how to analyze the data got from flowJo in order to get the graphs.
For example: n° of cells is 40304 and 0,7 ml blood volume -> 40304/0,7 to get cells/ml. Now should I 'normalize' this number to the volume of sample that I used in the experiment? I mean: if I resuspended the pellet of cells in 5 ml and I took only 2 ml for the experiment, will I do 40304/0,7* 2ml?
Thanks for your help! :)
Dear Rosa,
It will be easier if you may elaborate your experiment (marker used) and query on my email ([email protected])
You may not take percentage of cells from the dilution factor in flowcytometry.
If you need to get te absolute numbers of cells, you must have run counting beads I'm the sample while aquisition.
As Jitendra Kumar Shandilya rightly pointed out you will have to use liquid counting beads (spiked in each sample tube) to get an absolute count in flow cytometry data. Without the beads you can get a relative population count of each cell type compared to a control sample or between different treatments which may still be quite helpful.
Hi!
The great plus of flow cytometry is that normally normalization is not needed since you collect a certain number of events per sample (e.g., 10,000 events = cells). You may consider starting you data analysis with the help of FlowJo manual, which is very straightforward.