[Histonet] Old news about cryo sections

Monson, Frederick FMonson at wcupa.edu
Wed Sep 14 13:15:24 CDT 2016

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 at 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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