Re: 10%/alcohol formalin; also VIPs
<< Previous Message | Next Message >>
From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> |
To: | Elizabeth Wenig <se59aw@yahoo.com> |
Reply-To: | |
Date: | Sun, 25 Jul 1999 01:28:46 -0400 (EDT) |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
On Sat, 24 Jul 1999, Elizabeth Wenig wrote:
> Thanks for your response! Could you try to address some other issues on
> the subject,eg: the effect/wear of the salts on the VIPs, the
> processing quality of certain tissues,etc.
> In appreciation,
> Elizabeth
25/7/1999
Dear Elizabeth,
It's nice to be appreciated, but:-
I'm not sure what you mean. An alcoholic formaldehyde fixative
usually does not contain any salts: typically it's alcohol,
formalin, acetic acid and water, with many variations in the
concentrations. I would advise against buying a commercial
mixture unless its exact composition is declared and you
intend to use it all in the near future. These mixtures
change with time: ethyl acetate is slowly formed by reaction
of ethanol and acetic acid. (I know at least one paper that
provides evidence of improved fixation because of chemical
changes in older solutions. In that case the fixative also
contained picric acid.) If you mix a formal-acetic-alcohol
(FAA) combination shortly before using, you know it will be
The same every time. If you are not satisfied with the result
you can vary concentrations one at a time.
As for "VIP," to me this is vasoactive intestinal polypeptide.
I have immunostained for it in material fixed in neutral
buffered formaldehyde and in neutral formaldehyde with picrate,
and so have many others. I do not know if it is preserved by FAA.
I cannot think of a reason why its antigenicity should not be
well preserved, but the only way to know is by trying it out with
a known positive control such as a sensory ganglion or (more
difficult) small intestine or skin. VIP is present in the smaller
neurons of sensory ganglia.
The easiest sensory ganglion to find is the trigeminal ganglion
of the rat, which is conspicuous after removing the brain, lying
lateral and posterior to the pituitary gland. To get it out, you
need a sharp-pointed scalpel blade (#11) to cut through the
overlying dura. This is a much easier job than finding and
removing a spinal ganglion or any of the other cranial nerve
ganglia.
This presumption about VIP may be wrong; you actually said
"VIPs" in your message. If you really mean very important
persons, an alcoholic cocktail is certainly the one
of choice, if they do not arrive already pickled. Yeltsin's
fixative, said to be about 45% ethanol, is well known to
be very effective if a large volume is provided.
You replied to my reply after a 10 day delay, so if you have a
further question please be more prompt, because for 4 weeks
starting 28th July I will be globe trotting and completely
unsubscribed, unscribed, unsusribed and utherwize unobtainable
on HistoNet (if I can get the spellings right).
J.A.K.
> --- "J. A. Kiernan" <jkiernan@julian.uwo.ca> wrote:
> > On Thu, 15 Jul 1999, Elizabeth Wenig wrote:
> >
> > > Can I please have some pros and cons concerning
> > the use of
> > > comercialized, pre-made 10% formalin verses a
> > scratch measured recipe
> > > of 37% formaldahyde, absolute alcohol and h2o? I
> > have a few of my own,
> > > but need some more!
> >
> > If you have the recipe, the ingredients, a
> > graduated
> > cylinder and a bottle, why would you even think of
> >
> > paying extra to buy the stuff pre-mixed?
> > John Kiernan
> > London, Canada
> >
-------------------------------------------------------------------
<< Previous Message | Next Message >>