Determening monocyte derived macrophages in scatterplot

Dear flowers,

We have stimulated human monocytes with M-CSF and have monitored CD71,
CD14 and HLA-DR by flow cytometry as the cells progresses to macrophages.
We have not succeeded in finding a marker in which the expression profile
increases as the cells progresses to macrophages. Is there a well
expressed marker to distinguish between monocytes and monocyte dervied
macrophages? (We know that CD204 and CD206 should be compatible markers,
however, they are not competent for flow cytometry).

We do observe that there is a great change in scatter properties when
monocytes become macrophages (4-5 days after stimulation). The macrophages
are large cells and are more spread out in a scatterplot.

Is it possible to use the scatterplot as a certain evaluation of 
monocytes becoming macrophages? We may want to take advantage of the
“polluting” lymphocyte population, as a fixed location in the scatterplot
(adjust the voltage to position the lymphocytes in a lymphocyte gate) and
then estimate the movement of cells from the monocyte population /gate and
evaluate these as machrophages.

I would very much like some feedback on this issue.


Regards, Hans Christian  Dalsbotten
Flow Cytometry Core Facility, Dep. of Clinical Chemistry
Ulleval University Hospital
Oslo, Norway