Carrie,

First, I would immediately fix the cells in formalin.  After they are fixed,
I would follow your protocol.  I, however, use 2% agar and let it solidify
at room temperature.  This allows the cells to settle somewhat towards the
bottom.  Since the cells are already fixed, you have the luxury of working
at room temperature.  I then remove the agar, trisect it and process as
usual.

Jim

____________________________
James E. Staruk, HT(ASCP)
Mass Histology Service
http://www.masshistology.com

> -----Original Message-----
> From: Carrie Eberhardt [mailto:carrie.eberhardt@impath.com]
> Sent: Wednesday, November 15, 2000 12:53 PM
> To: 'HistoNet Server'
> Subject: Paraffin Block preparation from Cultured Cells
>
>
> Hello!  I am attempting to make paraffin blocks of cultured cells and our
> pathologist has noted that the cell density is not good enough, so I am
> asking for tips/ protocols that anyone may be using.
>
> I start with at least 6 million cells, pellet them in a 15 ml centrifuge
> tube, pour off media and add a couple drops of warm 3% Eosin/Agar
> and vortex
> to mix.  The cells are immediately chilled on ice and the agar cell pellet
> is fixed in Formalin and processed on a short cycle on an automated
> processor.  At this time, we embed the entire processed pellet, but we are
> trying to core a paraffin block and insert a cored agar pellet with no
> avail.
>
> Any tips out there?
>
> Carrie
> IMPATH-LA
>
>