76 BD FACSCalibur Instructions for Use
Sensitivity Tests
After compensation is set, the Sensitivity view appears and acquisition of the
mixed-bead tube begins automatically.
The software automatically proceeds through the following Sensitivity Tests. A
minimum separation of signal-to-noise, labeled-to-unlabeled, and APC-to-PerCP
(or PerCP-Cy5.5) is encoded in each bead lot ID suffix for a particular parameter.
If a parameter’s channel separation is greater than or equal to the minimum
separation, the parameter passes the Sensitivity Test.
When complete, calibration files are saved in the BD Files > Instrument Settings
Files folder.
• Fluorescence Sensitivity Test
The software acquires 10,000 events and calculates fluorescence separation
between the means of unlabeled and labeled beads in the FL1, FL2, and
FL3 channels. FL4 sensitivity is measured by calculating the separation
between the means of the PerCP (or PerCP-Cy5.5) and APC bead signals in
the FL4 channel.
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