From: Dave Coder (dave@nucleus.immunol.washington.edu)
Date: Fri Feb 04 1994 - 11:19:10 EST
Begin forwarded message: Date: 03 Feb 94 16:38:51 EST From: Marc.Langweiler@Dartmouth.EDU (Marc Langweiler) Subject: Re: Sci Fi (ve) mystery To: dcoder@u.washington.edu X-Status: ====== --- You wrote: Glad it helped. I'd look at the transmission properties of the 660LPDC. Depending how sharp the rise is around 660nm, you may reflecting some Cy5 emission since the emission peak of Cy5 isn't much beyond 660 (ML - 667 ??). --- end of quoted material --- And them Marc wrote: Dave, Thanks for your repost. In fact, I have gotten Coulter to send me an entirely new filter set. You were absolutely right about the 660LPDC - it looked really different than another of the identical filter (though we didn't see any difference upon substitution). I also contacted a "local" filter vendor (Omega Optical in Brattleboro, VT) who suggested we might be better served using a 670 BP in lieu of our 645 LPA, an opinion echoed by other respondents. ..... ============ Marc, When we first started using Cy5 somewhere in 1988 or 1989 (when Alan Waggoner had just gotten it out of the lab) I had Omega make a 670DF35 filter which worked very well. To get as much of the Cy5 signal as possible, we picked the red emission off first by using a 650SP DC (also from Omega). We could excite Cy5 (direct conjugates or Cy5-StrAv) with a 35mW HeNe laser (632nm) on an EPIC 753 with jet-in-air flow cell and get very good results. (FITC or PE sensitivity can suffer since they are now pickup farther down the optical path several other filters later. You play toward your dimmest probes and try to maximize their detection.) Omega should still offer the filters. Another source is Chroma Technology formed by people who left Omega: Chroma Technology RR 6 Box 9E Brattleboro, VT 05301 (802) 257-1800 (802) 257-9400 fax Paul Millman is with Chroma and he's the one who got me the filters originally. Dave
This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:27:05 EST
