PE and PE-CF594 will spill over slightly, but I've used them together without significant issue.
I like to use Fluorish Panel Design tool (http://www.fluorish.com/) to ensure compatibility. BD Biosciences Spectrum Viewer (see attached picture with reference to PE and PE-CF594) is also a great tool.
PE and PE-CF594 will spill over slightly, but I've used them together without significant issue.
I like to use Fluorish Panel Design tool (http://www.fluorish.com/) to ensure compatibility. BD Biosciences Spectrum Viewer (see attached picture with reference to PE and PE-CF594) is also a great tool.
I haven't used it, but useful for you, I have used PE with PE-Texas Red, which has a much greater spillover, and could distinguish both signals reliably (with compensation plus FMOs), so this should work better.
I set companstion (PE-cf594 and PE ) using BD compbead but it gives a warning. PE gives a spectral averlap in PE-cf594 channel.
How much % overlapp is indicated? You can work with big overlapps, but depends on how big. You should use FMO controls.
Dear Enrique,
Overlap was more than %100. So I could not set the compensation
Over 100% compensation does not mean that the data are wrong and can be perfectly fine. In this combination the % just seems a bit high. Check the filter settings at your instrument and compare them to the recommendation of BD. Furthermore, by balancing the voltages on the two channels you can alleviate the compensation. E.g. when you have PE on your y-axis and PE-CF594 on the x-axis and you see strong spillover (i.e. your PE-CF+ cells are in the upper right quadrant) you can increase the voltage for your PE-CF594 channel (or decrease the one for the PE channel) and your PE-CF+ population will move down to the x-axis (i.e. less compensation required). Just some thoughts. Good luck.
Agree with Gerhard. In my experience, up to 200% spill over is not the end of the study.
In any case, the use of FMOs should make you more confident. I hadn't answered due to lack of references. But I insist that PE and PE-Texas Red can indeed be used together. Go ahead!
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