pixl Real-Time PCR Platform
Instructions for Use Manual
Page 31 of 37
pixl Real-Time PCR Instructions for Use Manual
Rev. 01-28JUL2021
Appendix B: Channel Cross talk Suppression Techniques
The emission spectra of fluorophore materials used in multiplex qPCR assays can overlap
into adjacent optical channels. The pixl platform has incorporated measures to suppress
such cross talk both in hardware and software. In this section, we explain the software
measures to suppress optical channel cross talk, using experiment data as illustration.
The pixl software cross talk suppression parameters and default values are chosen
according to empirical data from most common multiplex qPCR reagent products used in
factory testing. These values can be adjusted by the user for any specific assay used (
Figure
B1
).
Example analysis of a two-channel assay utilizing HEX and ROX fluorophores is presented
(
Figure B2
) and the ROX channel demonstrates strong amplification while the HEX channel
yields negative results. This agrees with the expectation of the design of the experiment.
The fluorescence curve of the HEX channel shows an exceptionally clean flat line, this is
made possible by the crosstalk suppression.
Figure B1
. Cross talk suppression parameters.