Chapter 5: Cytometer QC and Setup 67
6 Optimize settings, as needed.
Tip Threshold and side scatter are the most frequently optimized
parameters for TBNK assays.
• To adjust the optimization gate, click —the acquisition
display stops updating. Move the gate by dragging the gate border, or
resize the gate by selecting a corner and dragging. When you are ready
to proceed, click .
• To adjust detectors, thresholds, or spectral overlap settings, click on the
corresponding tab. Use controls in the tab to adjust the settings. For
instructions, see Using Cytometer Controls on page 71.
Tip Spectral overlap values are automatically recalculated when you adjust
voltages.
For clinical applications that use tandem conjugates such as APC-Cy7 or
PE-Cy7, spectral overlap varies from lot to lot. Because BD FACSCanto
clinical software setup targets the average lot, you might need to adjust
spectral overlap settings for these reagents.
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