[Histonet] PCNA on methanol fixed cell smears along with Ki67..

[Choi Ul Soo]

Dear histonetters!
 
Hello, I am a graduate student, DVM, studying on
immunocytochemical evaluation of some proliferation markers on methanol 
fixed 
cell smears from canine mammary tumors. Markers are PCNA, Ki67 antibodies, 
and some others. Smears have been stained with Diff Quik, a modified Wright 
stain using methanol fixation. 
As it turned out, I have only a few cases PCNA positive i.e. 7 / 47 cases. 
Correponding tissue sections have also been immunostained and all are 
positive with PCNA with variable positivities. I have used Oncogene Cat # 
NA03 PCNA antibody. 

The reagents, and immunostaining procedures are same both for smears and 
tissue sections except that the smears are stored ones and transferred to 
adhesive slides using liquid mounting medium. 
 
I have successfully retrieved Ki67 antigens on methanol fixed smears after 
trasferring to adhesive slides using antigen retrieval. 
 
My question is that 'Is there anybody who has ever tried PCNA on methanol 
fixed smears, and got similar experiences
Or, who has ever heard of similar issues of PCNA like this?' 

Any information of any kind regarding this issue would be appreciated. I 
would gratefully receive any guidances or advices on this problem, because 
I should write up a 'discussion' section of the article. 
 
Lots of thanks in advance!
 
Ul Soo Choi
DVM, graduate student, 
VMTH of CVM, Seoul National Univ., 
Seoul, Korea

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