Cytometer configuration question

Roberts, Daniel Daniel.Roberts at covance.com
Fri Feb 8 08:00:00 EST 2008


I am relatively new to the Canto instrumentation as well, and have just
returned from the vendor training course. I wouldn't go around changing
filters unless you are doing analysis with an antibody that you can only get
with an exotic fluorochrome. I would recommend switching the tandem dye to
something that can be compensated more efficiently. PE is great for a dim
marker, as it has one of the most efficient stokes shifts, so I would leave
that the way it is. If available, see if there is a FITC or APC tag for the
antibody you're using with the tandem fluorochromes. If you're stuck with
the PE-TR, then I yield to the expertise of others who have had to deal with
tandem compensation. I fortunately, have not :)

Dan Roberts
Covance Laboratories
Vienna VA

 -----Original Message-----
From: Grammenoudi Sofia [mailto:grammenoudi at fleming.gr] 
Sent: Wednesday, February 06, 2008 9:17 AM
To: cyto-inbox
Subject: Cytmeter configuration question

Dear all,


Since I am new to the flow cytometry field I would like to ask some
basic questions concerning instrument configuration. In our BD FACS
CANTO II the PE detector has a 556LP-585/42 configuration which follows
the 610LP mirror of RE-TR. By searching through the internet I came to
realize that (i) many fluoro's contribute error to PE and (ii) most of
you use the narrower 575/25 filter. In practice, we get high spectral
overlaps when we use PE and PE-TR markers and especially when PE
represents a dim marker. Avoiding using PE with dim markers is one
solution, although not always feasible, but does it worth changing to
the 575/25 filter to reduce this overlap? Would it help with other
fluoro's? In addition, I have attached our complete instrument
configuration and I would be grateful for any comments and improvement
tips. 


Thanks for your time. Looking forward to hear from you. 


Sofia Grammenoudi 


Flow Cytometry Lab

Institute of Immunology

Biomedical Sciences Research Center 'Al. Fleming'

34 Al. Fleming Street

166 72 Vari, 

Greece





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