Chapter 3: Running Samples 59
Compensation controls are added to the experiment, along with a single
stained control tube for each parameter in the experiment, and an
unstained control tube (Figure 3-5). Worksheets containing appropriate
plots and gates are added for each compensation tube.
Figure 3-5 Compensation tubes
Adjusting the Voltages and Threshold
In this section, you use the unstained control tube to adjust FSC and SSC voltages
and FSC threshold, to gate the population of interest (bead singlets, in this case),
and to adjust fluorescence PMT voltages.
1 Install the unstained control tube on the cytometer.
2 Expand the Compensation Controls specimen in the Browser.
3 Click to set the current tube pointer next to the unstained control tube (it
becomes green) and click Acquire Data.
4 Adjust the FSC and SSC voltages to appropriately position the singlet bead
population for gating (Figure 3-6 on page 60).
• Click the Parameters tab in the Instrument window.
• Use the up and down arrows or drag the voltage sliders to adjust the
voltage settings.
5 Click the Threshold tab and adjust the FSC threshold, if needed.
Adjust the FSC Threshold to remove most of the debris without clipping
the singlet population (Figure 3-6 on page 60).
LSR2.book Page 59 Tuesday, April 25, 2006 3:34 PM
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