Hi Antonia,

I perfuse with PLP for 10 minutes and put the tissue in PLP for overnight
immersion. I freeze the tissue for frozen sectioning the next day. I kept
some tissue in PLP for about 2 weeks and compared it with the overnight
fixed. The retrieval I used on the overnight fixed tissue did not work for
the 2 weeks fixed tissue. Desmin stained only very faint. So my first
thought was that I was using underfixed tissue.

Veronique Andriessen BAS
Lab. Molecular Liver Cell Biology,
Free University Brussels (VUB)
Belgium

-----Original Message-----
From: Antonia Abeyta [mailto:ALAbeyta@salud.unm.edu]
Sent: donderdag 24 juni 2004 15:37
To: vandries@vub.ac.be
Subject: Re: [Histonet] PLP fixative


What are you perfusing with?  We perfuse with saline and 20 minutes in
PLP does the job.

Antonia


>>> "Veronique Andriessen"  6/24/2004 2:49:12 AM
>>>
Hi Histonetters,

It has been some time, but I would like to thank everyone for their
kind
replies on my last post on doublestaining for somatostatin receptor
and
desmin.
I have tried the PLP fixative and it gives the best staining for the
somatostatin receptor until now. Desmin works pretty well with a bit
of
retrieval.
I've seen some inconsistancy in retrieval time so I'm wondering if my
tissue
is completely fixed. Does anybody know how long tissue must stay in
PLP
fixative (after perfusion) to be completely fixed?

Thanks

Veronique Andriessen BAS
Lab. Molecular Liver Cell Biology,
Free University Brussels (VUB)
Belgium




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