Re: Spin on IHC buffers
Thanks,
Part of the senseless babbling I was referring to involves osmotic
balance but it didn't make much sense the way I described it. I'm glad that
I was on the same wavelength though.
The tissue is formalin fixed and paraffin embedded. So does that prevent
the deleterious effects? If it does then people are wasting a lot of money
on buffer. I for one am not quite ready to take such a chance but it bears
consideration.
Amos
----- Original Message -----
From: "Monson, Frederick C."
To: "'Amos Brooks'"
Cc: "'List-HistoPath'"
Sent: Thursday, February 21, 2002 4:26 PM
Subject: RE: Spin on IHC buffers
> Hi Amos,
> One of my favorite sub-subjects. Colligative properties of
> solutions. I think the words you want are "osmotic pressure". If
sections
> are fresh-frozen and thick enough to house unsectioned cells, then just as
> in a suspension of RBC's, diHOH will 'fill them to bursting', because of
> "osmosis". Same thing to the unfixed "ground substance" of connective
> tissue. Maybe I should issue forth on "Chromatin" and "Metachromasia".
Ah
> yes! My alcohol of choice for dehydrating tissues stained with
> metachromatic dyes is Tertiary ButAnal ("TBA" - Now why would anyone think
> to give the suffix of that word so much prominence?) In any case, with
> TBA, the metachromasia assuredly survives. I've had some in Azure B (DNA
> vs. RNA) mounted in Damar for 20 years and still see azure and purple.
> Get those people free tickets to see "Osmosis Jones".
> By the way, if the tissue is fixed, diHOH probably won't have any
> deleterious effect on the results. I mean, after we use the magic tap
water
> on the Schiff's Reagent, don't we rinse in diHOH before we enter the
> alcohols. No? For Heaven's sake, why not!!!???!!! Wouldn't we
contaminate
> the alcohol dilutions we made with diHOH?
> Has anyone ever determined why the tap water in Duluth works better
> on Schiff's than that in Minneapolis? I never did hear how that debate
was
> resolved. I missed most of it by becoming electron microscopic.
> Well, Amos, I am taking tomorrow off, because I need to replace a
> windshield wiper motor. I'm starting at the exhaust pipe and I'm going to
> work my way forward until I find that little bugger. Then, after lunch,
> while I revel in my mechanical skills, I'm going to watch the US team beat
> the Russians in ice hockey and, hopefully, the Canadians beat the
> Liliputians.
>
> My best wishes for all to have a peaceful weekend, especially our
> neighbors who are sailing with the Navy, flying with the Air Force
> (robotically with the Army) and sand flea hunting with the Marines.
Smack!
>
> Fred Monson
>
> Frederick C. Monson, PhD
> The best research
> Center for Advanced Scientific Imaging
> occurs before work
> West Chester University
> at the bench.
> West Chester, Pennsylvania, USA, 19383
> 610-738-0437
> fmonson@wcupa.edu
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