[Histonet] Keratin Artifact
Has anyone experienced the following artifact? When performing Keratin IHC
stains from previously frozen lymph node tissue in paraffin blocks. I am
seeing plasma cells and the occasional histiocyte showing light but positive
staining. It is only recently that we have seen this happen. The same artifact
is reproducible using Bectin -Dickinson's pre-dilute Cam 5.2 diluted to 1:20
and 1:40 among different lot #'s and unopened antibody vials to eliminate
contamination. Instrumentation utilized is the Dako autostainer and Vision
BioSystems Bond instrument. The same staining pattern is also seen when stained
on Ventana's Nexes instrument using the same Cam 5.2 and again a new
unopened antibody. It is also seen when staining on Ventana's Benchmark XT
instrument using Ventana's Keratin 5D3 antibody. This seems to happen mostly but not
always on tissue previously used for frozen section snap frozen with O.C.T.
The frozen tissue is thawed and placed in unbuffered zinc formalin and
processed. Comparative studies using NBF netted same results.
Keratin stains performed on colon tissue or similar tissue not frozen
but processed in the same processor does not show this artifact. Any ideas of
what might be the cause would be appreciated.
Could this have something to do with radiation procedures of
James M. Hart
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