RE: Spin on IHC buffers
|From:||"Monson, Frederick C." |
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
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!
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
> From: Amos Brooks
> Sent: Wednesday, February 20, 2002 7:26 PM
> To: histonet
> Subject: Spin on IHC buffers
> I have heard of a lab recently that does not use buffer for their IHC
> staining anymore. They have switched to deionized water. Now, of course I
> know not to do this but I was hard pressed to put into words why. I would
> thrilled to hear a more eloquent explanation than what I am babbling on
> Amos Brooks
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