Back in the 1960's when I was learning all about the PAS method, I got my hands
on a new CRYO-Stat (-30_!!!!!!!!!!!!!!!) and tried my 'stuff' on fresh frozen
sections. Having been in the habit of controlling every variable I could name,
I included a 'Schiff' control with each PAS.
1. Immediate reactions yielded negative control and positive
PAS.
It was late in the day, and I left the rest of my sections in the bottom of the
cryostat and went home to the family.
2. Next morning reactions showed similar PAS and visible, but
light, 'pink' in the control.
3. By 4:00pm I had very positive controls and similar PAS's.
Since I had just finished getting A's in organic, I was ready for
'auto-aldehydes' via atmosphere, BUT my total funding for my thesis came to a
little past $350.00, so I could not purchase or connive oxidation inhibitors or
N2 to run thru the cryostat.
4. That experience, and things I kept hearing from all
directions, led me to begin my histology and electron microscopy classes with
biologists that "...everything you will 'see' this semester will be artifact!"
5. The last extension of that occurred when I had close to
$900k of grant money, and I tested the dogma of permeability in an attempt to
discover why 'God' would lay such a rich capillary bed under the epithelium of
the urinary bladder of mammals when those epithelia were proved by "Ussing's
Chamber" to be impermeable to almost everything!
6. My rule has been. "All experiments performed on dead or
even freshly exposed tissue and cells are very likely artefactual."
7. Knowing that before I started, I still could only do what
was possible, and almost all of that was not kind to the subject beyond
anesthetics.
8. Yet; I am always aware of the truth of my rule, and that
many times we perform experiments of color about which we are not as careful as
we might be about the variables that we haven't attended to.
9. The most frightening experience I had when I was 'doing'
science was when the last paper I wrote was accepted without comment. I have
since determined that the explanation was the temporal irrelevance of the
effort - i.e., it was almost 30 years too late to be of any real help.
10. I may at this moment be one of the very few US-university
trained histologists, NOT a pathologist, that remains alive and working. Our
$900k NIH grant may have been the last time they gave money to anatomist
histologists. We did anatomy/histology and undergraduate physiology, and were
published - except for the non-Ussing experiments.
11. OK here's the summary. 33 rabbits. Osmostic measurements
on urine taken at catheterizing and that collected over the subsequent 1 hour.
1250 mosm +_ 50(?) for the urine in the bladder at the start. 350+_25(?) mosm
after the hour collection that did not spend (much) time in the bladder. In
human bladder mucosa (harvested from brain-dead subjects) I subsequently found
aquaporins and a urea transporter. Then, time ran out, and the anatomists were
unmasked.
I must recommend to those of you who wonder where we are going with biology
these days- and years ahead. Well, I strongly recommend that you search for a
copy of the Hershey&Chase paper of 1952 and the "Structure of the T4
baseplate...." with movies taken from 3D-Tomographic and Structural Biological
crystallographic data [
http://www.nature.com/nature/journal/v533/n7603/full/nature17971.html ] . One
wonders if the T4 phage is always the same - what is its structural and
'functional range.'
One more that I received from a non-scientist this morning:
https://www.wired.com/2016/09/gorgeous-unsettling-video-evolution-action/#slide-1
Cheers to you all who labor among the rainbows of our tissues,
Fred Monson who is being call 'dead wood' by few. I fear soon there will be
many.
Still, cheers!!
-----Original Message-----
From: HistoQuenie via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Wednesday, September 14, 2016 1:07 PM
To: Tyrone Genade via Histonet < >
Subject: Re: [Histonet] head kidney issues
Good morning Mr. Genade
Try using acetone/H202 in your blocking step. It may solve your problem with
endogenous peroxide.
Use the same amounts you would use for your blocking solution. I hope this
helps.
Cynthia James
Sent from Mail for Windows 10
From: Tyrone Genade via Histonet
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