[Cytometry] Ghost population?
rvw24 at cam.ac.uk
Wed Feb 4 11:28:36 EST 2009
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?
Dr Rachael Walker
Flow Cytometry Core Facility Manager
Wellcome Trust Centre for Stem Cell Research
University of Cambridge
Tennis Court Road
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