Chapter 5: Optimizing cytometer settings 83
Since the application settings have been optimized for your
sample, the cytometer settings should not require adjustment
other than the changing of FSC and SSC voltages to place the
beads on scale.
6. Adjust the P1 gate to surround only the singlets.
7. Right-click the P1 gate and select Apply to All Compensation
Controls.
The P1 gate on each stained control worksheet is updated with
your changes.
8. Click Record Data.
9. When recording is finished, remove the unstained control tube
from the cytometer.
10. Click Next Tube.
11. Install the next tube onto the cytometer and repeat steps 8
through 10 until data for all stained control tubes has been
recorded.
12. Double-click the first stained control tube to display the
corresponding worksheet.
Caution! Do not change the PMT voltages after the
first compensation control has been recorded. In
order to calculate compensation, all controls must
be recorded with the same PMT voltage settings. If
you need to adjust the PMT voltage for a
subsequent compensation control, you must record
all compensation controls again.
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