RE: CMV Ag
Edoardo (coledoco@tin.it)
Thu, 14 Aug 1997 08:33:02 +0200
Dear Haywood
we developed an improved method for detecting
CMV Ag by flow cytometry by using the Caltag
Fix&Perm kit (remeber that fixation IS the crucial
turning point for this analysis either on slide or
by flow) and the DAKO CMV Ag antibody (diluited 1:5);
after 30' incubation at 4=B0C in dark , a secondary
FITC-conjugated anti-mouse Ab is added (DAKO diluted 1:100).
All samples were run together with a control for
autofluorescence (that's higher for PMN) by using
the DAKO secondary antibody alone.
In our hands , after severals samples analyzed with
flow and standard immunophluorescence with optical
microscopy, we found a perfect correlation of data.
I've heard that now a new Ab for CMV (with better
specificity) is available from Serotec.
Gating a bitmap on the PMN region you must analyze
a very large number of cells since sometimes the
parasitemia is very low : if you have hardware troubles
(low RAM) you can avoid to save your data as listmode files.
If you need some further informations feel free to
contact me .
Edoardo Colombo MD
Director Flow Lab
Dept. of Pathology
Valduce Hospital
COMO , Italy
coledoco@tin.it