on 2/5/01 9:29 AM, David Grehan at david.grehan@olhsc.ie wrote:

> Does anyone have any neat suggestions concerning the demonstration of
> Reed Sternberg cells when they are embedded in an intense inflammatory
> background . We get many cases of Hodgkins disease in which the
> infalmmatory background renders it difficult to be sure of CD15 positve
> RS cells . We also stain for  RS  cells using CD30 which will not light
> up inflammatory cells but is also nonspecific for this cell type. Does
> anyone double immunostain for example?
> Just curious.
> David Grehan
> 
> 
I have tried double staining for CD20/CD30 with the chromogens being DAB &
BCIP/NBT. It is still extremely difficult to see the RS cells, as there are
so many CD20 positive cells. I have found the best way to be sure if the
positive cells are RS cells, is to look for perinuclear positive staining
which is over the golgi apparatus. If the titers on your primary antibodies
are correct you should see this with CD30, and usually with CD15. Find a
good positive Hodgkins for a control & titer your antibodies on it. Hope
this helps. 

Patti Loykasek
Phenopath Lab