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The page below is a sample from the LabCE course Introduction to Flow Cytometry: Blood Cell Identification. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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T Cell Markers

Until now, the interpretation has used single color analysis. Ruling out a monoclonal B cell population does not guarantee that this is a normal peripheral blood. The T-cell lymphocyte population must also be evaluated. In order to do so, it is important to not only determine how many T cells are present, but also what subsets they represent. Dual color analysis can be used to distinguish CD4 from CD8 subsets in the CD3 population.

CD2= 91%
CD5= 90%
CD7= 91%

Image #1
CD3 is plotted with CD4. CD3/CD4 dual positive (+) cells (Q2) = 55%
Add Q1 (CD3 positive, CD4 negative cells) and Q2 percentages together to get total CD3 = 88%

Image #2
CD8 plotted with CD3. CD8/CD3 dual + cells (Q2) = 33%
Add Q1 (CD3 positive, CD8 negative cells) and Q2 percentages together to get total CD3 = 88%

Since this is the same sample and the same gated population, the total CD3 values should be very close from image to image. In the two images on the right, the total CD3 values are within one-tenth of each other (both round to 88%). The final results for the T cell markers are:
  • CD2= 91%
  • CD5= 90%
  • CD7= 91%
  • CD3= 88%
    • CD4= 55%
    • CD8= 33%