Agilent Technologies NovoCyte Quanteon Operator's Manual page 35

Flow cytometer
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Introduction
Figure 7.
Fluorescence Compensation Theory
Fluorochromes emit light over a range of wavelengths (Figure 7). Optical filters are
used to limit the range of wavelength measured by a given detector. However,
when two or more fluorochromes are used in the same test, the overlap in
wavelength ranges often makes it impossible to isolate light from a given
fluorochrome from another fluorochrome using optical filters. As a result, light
emitted from one fluorochrome will appear in a detector intended for detecting the
other fluorochrome (Figure 8). This is known as fluorescence spillover. Spillover
can be corrected mathematically by using a method called compensation.
Figure 8.
NovoCyte Quanteon Flow Cytometer Operator's Guide
Emission Spectra of Commonly Used Fluorochromes
Spillover from the FITC Fluorochrome to the PE Detector
35

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