i am working on monitoring sebocyte lipogensis and recently started using
the ratio of FL2/FL3 fluorescence of nile red to monitor the extent of
staining of cytoplasmic lipid droplets and membranes/other irrelevant
cell material.
the interesting thing is when i plot this ratio as a function of forward
scattering, i get a nice horizontal linear ridge around FL2/FL3 of 48 with
peaks at different FSC points. what do you think may be causing this? i am
willing to fax a copy of the plot if somebody thinks that they may be able
to offer ideas.
thanks very much,
alex.