Ronnie,
I have compared a number of methods for preparing cell blocks for IHC and
have the following comment;
Overall, I much prefer the plasma/thrombin method.
Primarily because freshly harvested cells can be collected in plasma,
transported, aliquoted and the cell concentration easily adjusted, prior to
addition of thrombin.
Furthermore, unfixed cells are now completely surrounded by the same
proteinaceous intercellular matrix found in all solid tissues.
This proteinaceous matrix is a necessary and crucial component of all
fixation reactions in tissue and actively participates in the fixation
process.
To my way of thinking, cell blocks produced this way more closely mimic
solid tissues for IHC control purposes!
Once clotted (15 mins including retraction) the cell blocks can be optimally
fixed and processed.
We have used this method for preparing numerous cell blocks for IHC.
The only background staining seen is a specific background, due to secondary
reagents being insufficiently absorbed against human Ig's, this is easily
corrected as necessary.
One caveat, this type of specific background will interfere with
interpretation of staining for Human Ig's, Kappa/Lambda light chains and
albumen,
particularly if agonal imbibition by dying cells occurs.
Bryan
----- Original Message -----
From: "Houston, Ronald"
To:
Sent: Tuesday, May 29, 2007 4:15 PM
Subject: [Histonet] Cell Block preparation
Would anyone care to comment on the pros and cons of preparing cell
blocks using the Plamsa/thrombin technique and the Agar method which is
more prominent in the European field?
I am particularly interested in the prevalence of background staining in
ICC. Does the use of plasma interfere with interpretation of the
staining results? I know there have been reports of extraneous tissue
being found in cell blocks coming from a commercially prepared clotting
agent.
Thanks
Ronnie
Ronnie Houston, MS, HT(ASCP)QIHC
Anatomic Pathology Manager
Columbus Children's Hospital
700 Children's Drive
Columbus, OH 43205
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