RE: Paraffin Block preparation from Cultured Cells

From:"Greer, Patricia" <>

When we prepare blocks from cultured cells, we mix the cells with minced
"normal" tissue and mix the tissue and cells very well - sometimes I just
keep mincing after I add the cells to the tissue.  We use pieces of lung,
kidney, liver mostly - this gives a good indication of non-specific staining
in these tissues when doing immunohistichemistry.  
When preparing the cells - to back up to the beginning - we spin the cells
down after fixation, resuspend in 70% ethanol and spin again and take off
all the alcohol except for a couple of drops.  This is then easily added to
the minced tissue.

Pat Greer
Infectious Disease Pathology 

-----Original Message-----
From: Carrie Eberhardt []
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

Any tips out there?


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