Quantifying histogram data - Flow Cytometry analysis (Jan/24/2010 )

Hi.

What is the best method to quantify your data from flow cytometry histograms?

As far as I looked, you can use the mean fluorescence intensity to compare two experimental conditions. However, this is not always a good strategy. For instance, in my ROS measurements I get small (unespecific) peaks of fluorescence behind the main peak, and this affects the mean fluorescence.

Another possibility is to create and quantify regions at the right and left of a certain point. However, I never know how to decide where should be this reference point...


Thanks for any reply.

Hi there,

I am willing to compare GFP amounts in different samples analyzed through FACS:

sample 1: histogram analysis

Marker Left, Right Events % Gated % Total Mean Geo Mean CV Median Peak Ch
All 1, 9910 5000 100.00 100.00 177.79 8.01 639.27 7.17 1
M1 11, 9910 1500 33.33 33.33 525.62 33.24 344.98 16.70 9910

sample 2: histogram analysis

All 1, 9910 5000 100.00 100.00 1620.37 51.29 203.12 16.85 9910
M1 11, 9910 2570 50.21 50.00 2813.57 343.81 139.85 230.82 9910

Is there anyway to plot RFU plot between them. It is evident that sample 2 has far more cells expressing GFP than sample 1

sorry table messed up:

please find attached file for data "for forum"