Amorphous Region H is drawn to include all Stem-Count Fluorospheres. Region H should be located at the top
right corner of the Histogram including the last channel of the FL1 Log (on the right) and FL2 Log scales (on the
top).
Set Region K (named “ listgate - ”) to exclude most of the double negative events from the acquisition.
Histogram 6: displays events from region E.
Verify whether the Forward and Side Scatter gain parameters of the flow cytometer are optimally set for the
processed sample. If necessary, adjust the Forward Scatter voltage/gain so that the smallest lymphocytes scatter
in the middle-left part of the histogram. Adjust the Forward Scatter to ensure that even the smallest lymphocytes
scatter above the discriminator.
Histogram 7: displays events from region H.
Region G encloses the fluorosphere singlet population only. Check that the fluorosphere singlets accumulate
homogeneously and constantly over time.
Label Region G as “CAL” to allow automatic calculation of absolute numbers of CD34
+
Stem-Trol Control Cells.
Type the correct assayed concentration of the current batch of Stem-Count Fluorospheres (refer to section 9.7 of
this document or to the instrument manual for further details).
Histogram 8: displays events from region A.
For Stem-Trol Control Cells analysis in the presence of 7-AAD Viability Dye, Histogram 8 allows you to visually
check the 7-AAD Viability Dye positive staining. Region J is not used for Histograms 1, 2, 3, and 4.
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