Re: PMT for Cy7?

From: Dr Sasha Sreckovic (sasa.sreckovic@bris.ac.uk)
Date: Sun Apr 02 2000 - 11:00:24 EST


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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