I have had many problems with FL4 settings on the BDCalibur concerning time delay and FL3/FL4 compensation. In addition PerCB and APC labeled antibodies are very expensive. Not only for that, most people doing three colors use Cy-chrome, or other Cy-5 tandems; when moving to four colors one have to buy again the same antibodies labelled with PerCP to replace Cy-5 ! Wouldn't be possible to do four colors using only the 488 laser ? FITC, PE, PE-Cy5 and PE-Cy-7 ? There is little overlaping with PE-Cy5 and PE-Cy-7 emission. The PMT filters could be modified accordingly, I think, and the acquisition software also. It seems feasible to me. I would be very glad to obtains suggestions on how to do that: where to buy filters, etc. Alberto. Federal University Of Rio de Janeiro BRAZIL email:alberton@acd.ufrj.br
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