2 replies to this topic

[cardosopedro]

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.

[gradbiotech]

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

[gradbiotech]

sorry table messed up:

please find attached file for data "for forum"

Attached Files

•  for_forum.doc   61.5K   113 downloads