Hi everyone, In our quest to run more colors on the sorter, we have been using Coulter's ECD (PE-TxRed) conjugates on our Vantage SE. The vantage is equipped with Enterprise and HeNe lasers. We do not have any means to cross-beam compensate. So, plugging the Bandpass for RED613 (610bp20) into FL3 has made it possible to sort four colors on our system along with the traditional, FITC, PE and APC. We have also chosen ECD because PerCP quenches about 1.5logs on our Vantage and PE-Cy5 crosses too much into APC. BUT... I have users here who want it to work on the Calibur, because they don't want to buy or can't buy PerCP conjugates or PE-Cy5 conjugates and because they want to directly compare emissions from both machines. But that is another problem of mine - I digress... So, since we cannot change out the optics for FL3 on the Calibur, they are collecting only the far red tail of the emission with the standard 675LPfilter; they are completely missing the emission peak. I've told them that the conjugate wasn't designed to work on a Calibur, but the user "saw signals" and now I'm dealing with their "it works fine for me" syndrome, when I ask them to refrain from promoting this antibody combination to other users who are beginning 4 color work. Could anyone provide me references that would help get my point across? If there is someone out there who does this combination successfully on their Calibur and really trusts their data, could you share with me the typical instrument settings values you're coming up with? I understand that there are instrument to instrument differences but would still like to know... Thanks, Lora W. Barsky, Research Specialist CHILDRENS HOSPITAL LOS ANGELES Research Immunology/Bone Marrow Transplant Flow Cytometry Core 4650 Sunset Blvd. Mail Stop #62 Los Angeles, CA 90027 (323) 669-5935
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