BD FACSCanto II, Reference Manual, Page: 117 / 158

Appendix A: Technical Overview 117
Spillover
The arrangement of filters and mirrors in the BD FACSCanto II enables each
PMT to receive signals predominantly from a specific fluorochrome.
However, fluorochromes emit light over a range of wavelengths. As a result, a
signal from one fluorochrome can appear in a detector used for another
fluorochrome.
Figure A-11 Spillover of FITC into PE detector
For example, FITC appears primarily in the FITC detector, but some of its
fluorescence spills over into the PE detector. PE appears primarily in the PE
detector, but some of its fluorescence spills over into the FITC detector. This
spillover must be corrected (or compensated for).
Figure A-11 shows that a portion of the FITC emission will be detected by both
the FITC and PE channels. This can be seen in an x, y plot of FITC vs PE
(Figure A-12 on page 118).
emission spectrum of FITC
emission spectrum of PE
FITC spillover into PE
BP filters
nor
malized inte
nsity