Hi all:
O.K. So my samples are ready and I sit at the FACS machine. I setup the FL1,2 and 3
channel voltages so that the none stained cell population is in the lower left hand
corner. The control stained cells are nice with only a very small shift if any in the
major population in some of the channels and sporadic events with moderate staining. I
setup compensation properly for the three channels with cells stained with only single
color; the FL3 channel displays a negative population at the left hand corner and a
positive population down the axis. I put on my first sample that is highly staining
with FITC the cells being dendritic cells and the staining being for MHCII and the
staining in the FL3 channel spreads out and the negative population goes below the
origin of the axis.
To reiterate, why is it that high FITC staining induces a downward shift in the FL3
channel although the voltage for FL3 is maintained at a value set using a non-stained
population so that cells are just above the origin of the axis? How can this be solved?
Thanks,
Ashraf Abdelhafez, M.B.B.S.
Dr. Ashraf Abdelhafez
Universitätshautklinik Würzburg, AG Kämpgen
Josef-Schneider-Str. 2
97080 Würzburg, Germany
Tel. W: +49-931-201-26031
H: +49-931-2038-8318
Handy (Cellular): +49-931-8090554
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