Immunology Lab 542
FACScan Fluorescence Intensities from Histograms
Disk file name  Krissy Forloney, Flow Cytometry Spl 2003
Student name Krissy Forloney 
Cell type Splenocytes, TCR and CD4  Cells in scatter gate as % of total events 64% 

SAMPLE PREPARATION

Cycle No. Ab Specificity (Marker) Fluorochrome on Ab (if any)
 1  TCR alpha-beta  FITC
 1 CD4  PE 
     



RESULTS

FL1
or
FL2*		 If >1 peak Fluorescence Intensity (FI)
Control (Blank)
 Corrected Fluorescence Intensity (CFI)†
Peak No.§ Peak: % of (Events in Gate)‡ Disk File number Fluorescence Intensity (FI)
FL1  M1   57%  4  026 2.5   1.5
FL1  M2  42%   74  026  2.5  72
FL2  M1   65%  3  026  2.2  .8
FL2  M2  32%   37  026 2.2   35
             
*Specify whether FL1 or FL2 for each line in the table.
§Which peak, the first (from the left), second, ...?
‡ The peak as a % of (events in gate).
Note that (events in gate) as % of (total events) is entered at the top. If you have two equal-sized peaks, the peak % of (events in gate) should be 50% and 50%, regardless of what % of the (total events) were in the scatter gate.
†Select the appropriate control, and subtract it from your sample's FI to get the CFI.