Re: Spin on IHC buffers
I quit using PBS w ith tween 20 approximately 4 years ago. I have had
great success with using just DI H2O as my rinse. If I am going to store
my slides for a while before I can continue with my staining, then I use
PBS for a holding solution.
I am talking about FFPE tissue of course.
Sorry if I am alittle late responding, I was away on vacation last week.
On Thu, 21 Feb 2002 17:56:47 -0500 Amos Brooks
> Part of the senseless babbling I was referring to involves
> 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
> the deleterious effects? If it does then people are wasting a lot of
> on buffer. I for one am not quite ready to take such a chance but it
> ----- 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
> > 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
> > tissue. Maybe I should issue forth on "Chromatin" and
> > 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
> > 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
> > on the Schiff's Reagent, don't we rinse in diHOH before we enter
> > alcohols. No? For Heaven's sake, why not!!!???!!! Wouldn't we
> > the alcohol dilutions we made with diHOH?
> > Has anyone ever determined why the tap water in Duluth works
> > on Schiff's than that in Minneapolis? I never did hear how that
> > 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
> > 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
> > 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
> > email@example.com
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