Chapter 4: Instrument QC 79
Optimizing Settings
Because leucocytes have different optical properties than BD Calibrite beads, it is
important to optimize instrument settings for cell samples. Optimize settings for
2-color setup using a blood sample stained with any combination of reagents that
identifies separate non-overlapping cell populations and stains brightly, such as
FITC- and PE-labeled monoclonal antibodies. Optimization following 3- and 4-
color setup varies depending on the application. Refer to the appropriate
application note or reagent package insert for examples.
This section describes how to optimize settings in BD FACSComp software. For
optimization in BD CellQuest Pro software, see Chapter 5.
1 Prepare optimization samples.
For the Loader, place tubes in rack positions 5–38.
2 Click Optimization at the Summary view or click the icon in the icon bar.
The Optimization icon is available after the last assay in the run.
3 Choose an option from the optimization menu (Figure 4-1).
Plot parameters change for the chosen option, and associated instrument
settings are downloaded to the cytometer. Preset options are as follows:
; Tip Use BD FACSComp preferences to change default options (page 33).
You can also change plot parameters by clicking an axis label and choosing
a new parameter, or download a different instrument settings file (page 82).
Option
Instrument Settings
File Name
Plot 1
Parameters
Plot 2
Parameters
Plot 3
Parameters
1 MultiTEST.opt FL3 vs SSC FL1 vs FL2 FL3 vs FL4
2 TriTEST.opt FL3 vs SSC FL1 vs FL2 FL3 vs FL2
3 Simultest.opt FSC vs SSC FL1 vs FL2 FSC vs SSC
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