Chapter 5: Cytometer QC and Setup 83
Optimizing Cytometer Settings
When you performed cytometer QC, voltage settings were adjusted to set each
parameter at a target value. These settings might not be appropriate for the
stained sample(s) you plan to analyze. Before recording data, adjust FSC, SSC,
and threshold settings; gate on the population of interest (such as lymphocytes);
and adjust voltages to optimize fluorescence signal.
For these adjustments, you will need an unstained control sample. It is important
to perform these steps in order, as some adjustments influence others.
NOTICE If the software fails to respond during acquisition and you need to
restart the software, perform a fluidics startup prior to resuming acquisition. See
BD FACSDiva Software General Issues on page 215 for more information.
1 Install the unstained control tube on the cytometer.
• Push the aspirator arm to the left.
• Place the tube onto the SIT, ensure the tube is straight, and firmly push
up until the tube comes to a complete stop and is fully seated.
• Center the aspirator arm under the tube. There are three sensor pins on
the aspirator arm. The bottom of the tube should sit within the center
of the pins.
2 Verify that the green current tube pointer is set to the unstained control
tube in the Browser. Click .
Always wear gloves when manually loading samples. A fluid flush of the
exterior of the sample injection tube (SIT) occurs between samples that
might contain biohazardous waste.
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