Dear Joseph, I've used APC-Cy7, excited by Spectra-Physics He-Ne 35mW 635nm red laser and filters: 3-way beam splitter (mirror) -> 710 DMLP -> 780/60 BP. Anti-mouse CD8 alpha APC-Cy7 gave only 1-log difference, but nice separation from background (which is lower on red excitation). This was with red-sensitive Hamamatsu R-3896 PMT and to obtain same mean value on positive population with R-928 PMT, voltage had to go from 750 to 800! This change would probably give more difference with something brighter, if possible - same CD8 was quite bright coupled with PE-Cy5. Sasha. ************ Dr Sasha Sreckovic Dept Path & Micro University of Bristol University Walk Bristol, BS8 1TD, UK Sasa.Sreckovic@bristol.ac.uk +44-(0)117-928-8606 ************ ---------- >From: Joseph Webster <J.Webster@centenary.usyd.edu.AU> >To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >Subject: PMT for Cy7? >Date: Wed, Mar 29, 2000, 3:22 AM > > > We want to try out APC/Cy7 on our FACStar Plus, some clues > would be helpful from anyone who has used it on a StarPlus > or Vantage. > > We have a red HeNe laser that gives 35-40mW. > We have ordered Omega filters 710DRLP and 740EFLP to read > APC and APC/Cy7 in FL4 & FL5. > > I believe the standard PMT will probably not be red-sensitive > enough for Cy7. > If it won't work, what PMT should I buy? > Also what's the cheapest source for us in Australia? > > What have I missed or forgotten? > > Thanks, Joseph. > > Joseph Webster, Flow Cytometry Facility, > Centenary Institute > Locked Bag No.6, Newtown, NSW 2042, AUSTRALIA. > Ph: 61-2-9565-6110 Fax: 61-2-9565-6101 > e-mail: J.Webster@centenary.usyd.edu.au >
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