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Overview
This chapter describes how to create a compensation experiment and automatically calculate
compensation values after acquiring the data. It also explains how to use these calculations for
other experiments.
Compensation involves correction for fluorescence spillover emitted by the primary fluorochrome
that is detected by the secondary fluorescent channels. For example, the excitation and the
resulting fluorescence emission for the PE fluorochrome leads to the spillover fluorescence
detected in the ECD, PC5.5, and PC7 channel. Compensation reduces the spillover fluorescence of
the PE-positive population to match the background of the PE-negative population in the secondary
channels. Compensation requires a single positive and a negative population for every single color
sample.
Properly configured compensation minimizes false data interpretation caused by spillover
fluorescence from another fluorochrome. Refer to
before and after compensation. Compensation adjustments can be completed during the data
acquisition process or after the data acquisition process is complete.
Figure 7.1 Before Compensation
Figure 7.2 After Compensation
NOTE
CytExpert compensation allows full matrix compensation, manual, and automatic.
CytExpert compensation also includes a novel Compensation Library for storage of spillover values of
dyes to easily determine the correct compensation matrix with new gain settings.
B49006AS
Compensation
Figure 7.1
and
Figure 7.2
for an example of plots
CHAPTER 7
7-1
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