[Histonet] Re: PAMS Jones Basement Membrane

[gayle callis]

A stain I have never had a problem with.  However I did have the good
fortune to listen to Culling many years ago on the use of 1% periodic acid
for PAS staining and he insisted periodic acid, no matter what the
concentration, should be made fresh every time you do these stains e.g.  PAS
and PAMS.    Some people use 0.5% but we used 1% for 15 minutes, and then
did microwave staining for the methenamine silver.    A 56C - 60C water bath
also works plus monitoring the color of the sections -  the color of dark
tea.  

 

This is NOT a stable oxidizer once made up, and never should be reused under
any circumstances.   On the chance you have not used this solution in over 1
1/2 years, you may not be getting proper oxidation of the basement membranes
since the periodic acid and the methanamine silver may be bad.   Also, the
Methenamine silver should be no more than 6 months old.  We also kept our
silver nitrate solution fresh when we made up the methenamine silver
solution, and do store our silver nitrate salts in the refrigerator since
this is hygroscopic.  Check on storage of this salt in MSDS      I have
never been one to use kits for either of these stains, particularly with
periodic acid as a ready to use solution.   This goes into solution very
rapidly and be sure to oxidize for 10 minutes before going to the
methenamine silver.  None of these solutions is difficult to make up in
house.    

 

If you want, I can send my method and also a protocol pdf from HistoLogic by
Stanley Shapiro, where he used freshly made 0.5% periodic acid via private
email.   The nuclei will pick up some silver, but one should be able to
discern nuclei from basement membranes on 1 to 2 µm sections.  

 

Good luck

 

Gayle Callis

HTL/HT/MT(ASCP)

Bozeman MT  

 

  

 

 

 

You wrote: Just wanted to give a quick update on this.  I had a suggestion
for using thiosemicarbazide for 10 minutes after the periodic acid, and of
all the things we tried, this was the only thing that worked.  It eliminated
the nuclear staining and the capillaries are now picking up the silver (as
they should be!).  Unfortunately, I accidently deleted the email that
suggested this so I don't know who to thank, but it was a great suggestion!

Liz

 

From: Elizabeth Cameron

Sent: Thursday, February 16, 2012 2:17 PM

To: histonet <@t> lists.utsouthwestern.edu

Subject: Jones/PAMS

 

Hi,

I was wondering if anyone has any suggestions for a Jones/PAMS stain that is
not working properly. This is something we don't do often.  The last time we
did it was a year and a half ago, and it seemed fine at the time.

We have tried 3 or 4 protocols, including an ammoniacal silver, and it is
still not working properly.  In some protocols, our red cells are staining
but the capillaries in the glomeruli do not seem to be picking up the
silver.  In other protocols, there are nuclei of some cells that should not
be staining that are, but again, the capillaries are not.  We are working on
mouse tissue that is fixed in NBF.  The strange thing is the stain seems to
be working well on Bouins and Telly's fixed tissue.  I even tried mordanting
in Bouins!  We have tried multiple kidneys with the same results.  We are on
new bottles of silver and periodic acid, although our methenamine has been
around a while.  Any suggestions would be greatly appreciated.

Thanks!

Liz