Dear FACS Vantage Operators/Users, We have discovered recently that we had been running our Vantage with a different filter set on FL2 than what is issued on both the FACScan and the FACSCalibur. We had been running with a 575BP26 for quite a while to analyze PE. All of our work "seemed" OK; bright markers were only slightly dimmer than what was seen on the analysis machines. But when we needed to do some dim marker sorts and were coming up with negative expression, we had a problem. So, we ran some tests by carefully placing the isotype control in the same channels on all machines. The Vantage (with the 575BP26 installed) gave us results that were dimmer than the Calibur by 192 channels! Then when I installed a 585BP42, which is the same filter used in the Calibur, I was only 62 channels dimmer. The 62 channel difference was also seen on FL1, so I felt the drop was due to laser alignment and stream in air issues. What a relief!! I kick myself for not checking the filter sets earlier. I discussed these issues with my field engineers and we agreed that a 575BP26 should work for PE, but really, it doesn't. I was loosing a lot of sensitivity!!! Hoping others may learn from my experiences, I'm giving the following advice: be sure to run PE with 585BP42 filter set in FL2 on the FACSVantage, when using an Enterprise Argon/UV laser. Thanks for your time, Lora Barsky Children's Hospital Los Angeles Research Immunology/BMT 323 669-5935
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:53 EST