From: Ingel Demedts (M.DemedtsIngelK@rug.ac.be)
Date: Fri Jul 04 2003 - 10:28:26 EST
Hi, has anyone ever tried the tandem conjugates AlexaFluor 680-APC, AlexaFluor 700-APC and Alexa Fluor 750-APC? We have a BDFACS Vantage SE, using 2 lasers (Argon and HeNe). The fluorochromes we use most of the times are FITC (FL1), PE (FL2), PE-Cy5(FL3) and APC(FL5). I recently tried a 5-colour experiment using APC-Cy7 in our FL4 channel (which has a 780DF/60 filter in front of it; FL4 and FL5 are separated by a 710 LP filter). The APC-Cy7 signal however was not quite strong and I'd like to try one of the above mentioned Alexa dyes.I suppose that with our optics, the best choice would be AlexaFluor750-APC, which, according to the manufacturer, has an absorption max at 650 nm and an emission max at 775 nm. A second problem was that I had to compensate a lot for spillover from APC in the FL4 channel (up to 80%!!). Is this what some of you have also noticed, or is this really not acceptable and possibly due to insufficient alignment? Thanks for your comments, Ingel Dr. Demedts Ingel Ghent University Hospital Department of Respiratory Diseases De Pintelaan 185 9000 Ghent Belgium tel 0032-9-2406279 M.DemedtsIngelK@rug.ac.be
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