[Histonet] RE: Histonet Digest, Vol 26, Issue 20

[Pixley, Sarah (pixleysk)]

Dear John Kiernan and histonetters:

I read with great interest the article you suggested on the PLP
fixative. And I searched PubMed for similar articles. But I do not see a
good analysis of what might be the alternative to PLP. The authors of
the paper, Hixson et al., suggest that paraformaldehyde and lysine might
be just as effective as PLP, with lowered cell toxicity, but they don't
do an analysis of the effects on morphology or antigenicity. Have you
tried anything like this? Does a combo of PF and lysine provide a better
fixation of carbohydrate antigens than PLP?  Do you know if these
authors or anyone else followed up on this and developed and described a
better fixative in more detail? 

The reason that I ask is that I have an antigen that is particularly
sensitive to fixation and the monoclonal antibody to it works only with
the PLP fixative. Supposedly the antigen is the carbohydrate portion of
a cell surface receptor. But the fixation is so light that I lose almost
all my cultured cells during processing for IHC (I have very lightly
attached cultured cells). I have been searching for an alternative to
PLP for years! So I thank you greatly for this article. I guess my next
step would be to try different concentrations of paraformaldehyde and
lysine. But if this has been tried before, I would appreciate any
suggestions!

Sarah Pixley