[Cytometry] Ghost population?

Rachael Walker rvw24 at cam.ac.uk
Wed Feb 4 11:28:36 EST 2009


Dear All

I am now the proud owner of a 50mW 561nm laser for my MoFlo (I change =

over my 561 and red diode depending on sort) and have been using it a =

lot for RFP and it seems to be working well. However, I see a population =

of cells against the axis by my negatives. I have have heard of a ghost =

population appearing on a red channel, am I seeing the same thing on my =

RFP channel? Is there anyway of getting rid of it and also what is a =

ghost population? I know I will be asked about it by my punters and I =

need to explain it in a scientific way and not by the explanation I am =

using a the moment which is, its a ghost population, it doesn't really =

matter, they really are negatives and its there because there isn't =

enough light getting through.

I see it using  590/20, 610/20, 613/20  filters, on negative cells as =

well as positive populations.  I do also see it with my red diode =

laser.  When gating cells do I include the ghost population?  Should my =

gates go all the way to the axis?

Many thanks

Rachael

-- =



Dr Rachael Walker
Flow Cytometry Core Facility Manager
Wellcome Trust Centre for Stem Cell Research
University of Cambridge
Tennis Court Road
Cambridge
CB2 1QR

01223 760227

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