Re: [Histonet] Disposal of Bouin's Solution

[Monson, Frederick via Histonet]

https://oag.ca.gov/sites/all/files/agweb/pdfs/cci/safety/picric.pdf

Two hazards:  formaldehyde and picric acid (see attached).
While I agree with Dr. Richmond, changing protocols is not easily done.
So.  You may search for appropriate oxidizers of aldehydes.  OR!!!  You may 
take the reasonable path by sending it off periodically to a company that will 
cost enough to raise the use of the stuff to serious conversation.  

Picric acid - 
  since I used it from the late 1950's - the end of our 
"Dark Ages"[???] - 
into the mid 1970's -the age when 
students became consumers  and knowing what was on the test was paramount -  
  IS really 
hazardous - when it is dry.  [So, I always kept it covered with water 
(saturated in local conditions.]
Picric acid is the reason that you should either stop using Bouin's or waste it 
(wet!!) to a competent waste-handling company.

Cheers,

Fred Monson (5 weeks to the oblivion called 'retirement.'

-Original Message-
From: Bob Richmond via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, May 11, 2017 6:39 PM
To: Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Disposal of Bouin's Solution

Lori Jones, CT(ASCP), Pathology Supervisor, Ingalls Memorial Hospital asks:

>>We use Bouin's Solution in our pathology department and are currently
disposing of it by neutralizing it with Vytac for formalin. I can't find 
supporting documentation that this is the proper way to dispose of it. I'd like 
to know if and how other institutions are neutralizing Bouin's solution.<<

The best way to dispose of Bouin's fixative is by not buying it at all.
What do you use it for? There are substitutes for it for most purposes.

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] Happy Histotechnology Professionals Day!

[Monson, Frederick via Histonet]

And I, "O>>>>>" scary one, is following your class by leaving 38 days BEFORE I 
will be 78.
You really are old!?""@
In my family, retirement has appeared to lead directly - one way or another - 
to demise, and I have yet to discover anyone who has turned the inevitability 
of that ending.  So, I have contrived to take advantage of all that time by 
doing things that previous family have avoided.
  1.  I have mapped out a morning walk that is 10 miles from my house 
but the required exact 1 mile - thus, I will return to my car when it is 
finished.
  2.  I will return to our home and ride my bike around the block that 
is NOT a mile and has no traffic.
  3.  I am purchasing a digital camera to capture images from all of my 
sections - kept from 1965 to the present.  Since this activity is considered a 
punishment for putting it off for so long, it is clear that the effort will be 
life prolonging.
  4.  I have been practicing my aiming at 100 yards for several years 
now, and I will have time and energy to allot more time so that I can reach my  
goal of hitting the bull's eye with 5 rounds in 1 minute.  I have already made 
it clear that I will not leave this effort undone for any reason.  To make this 
effort fun, I will load my own rounds to a tenth (0.1) of a grain.  {You would 
think that we would have left weighing by seeds behind us now that we have 
grams; but I guess we will go to any length to avoid using metrics.
 4.a.  I have never been punished for joining the USMC when I needed a 
vacation from from the rigors of Lehigh University.  I enjoyed that hiatus so 
much that I have always called Paris Island, "Pleasure Island."  My older son 
joined as well, but he calls it:  "The bad place."  He has placed his 5 rounds 
in the bull's eye many times, and that explains my goal to achieve it just once.
 5.  Finally, I have signed a witnessed paper, under duress by my wife, 
that I will not go first, and I do try not to break promises to her.  

I hope that you have a similar set of activities in your plan.  However, if 
not, then would you like one of my ICS microtomes (at 2 micron intervals) to 
keep your hand in  rotating (otherwise known as 'cranking')?  BTW, has anyone 
EVER made a microtome for a lefty?  Ah!  If you were a lefty Bob, you would be 
a pathologist, because you were never comfortable cutting sections!  Wow!  I 
never thought of that before.

Cheers and reciprocating best wishes,

Fred Monson 

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmon...@wcupa.edu

From: Bob Richmond via Histonet <histonet@lists.utsouthwestern.edu>
Sent: Saturday, March 11, 2017 1:20 PM
To: Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Happy Histotechnology Professionals Day!

The old Samurai Pathologist - now retiring at 78 - thanks the many
histotechnologists who've kept him out of hot water the past 52 years.

Too bad we can't have more kids in search of a career reading Histonet - or
at least, the Help Desperately Needed notices!

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] Metabisulfite rinse for PAS reaction (potassium vs sodium)

[Monson, Frederick via Histonet]

And,  I always rinsed the Schiff with 'absolute' HOH before the tap water rinse 
- which rinses were taken from my protocol for Ehrlich's 'Acid' Hematoxylin 
that was mixed from scratch and aged in the second story deep window sills on 
the West wall of the Biology/Geology building at Lehigh University - usually 
left, after mixing, with a cotton plug, for around 6 months [with periodic 
compensatory 'absolute' water additions to the bottle].

A.  Ageless NOTE:  One of the most important phrases in organic chemistry 
is 'tautomeric shift.'  Especially for biologists - most of whom never heard 
the phrase.  Follow-up after GOOGLE:  Why is tritium so dangerous...?

B.  Current NOTE:  98% of science folk - after decades of programming - 
don't know how many digital cameras they carry with them every day.

C.  HISTO_EXPERIMENT:  next time sections are taking fresh, UN-fixed 
tissues, in the cryostat, and when time permits, do the following.
1.  set 5 or more sections on glass slides - or coverslips - 
and set them aside IN the cryostat
2.  immediately remove one and after thawing - very little time 
- and NOT treating with PA, immerse the specimen directly in Schiff's solution 
for the standard time and follow with the usual rinses and counter dye(s)-BUT, 
I left them out!
3.  at one hour intervals repeat (2) with one of the remaining 
sections.
4.  when you want a counter dye, please help yourself.
5.  if the outcome resembles mine in 1968, explain what you 
have observed.

Cheers to all,

Fred Monson
On the 16th of June, 16 days from my 78th, I will retire to do other things.  
That, at least, is the plan!!

Frederick  C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of PA
Geology-Astronomy
750 South Church St.
West Chester, PA, 19383
fmon...@wcupa.edu
610-738-0437(Work)



-Original Message-
From: Bryan Llewellyn via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Monday, March 06, 2017 9:28 PM
To: Histonet 
Subject: Re: [Histonet] Metabisulfite rinse for PAS reaction (potassium vs 
sodium)

You can use either the sodium or potassium salt. Both can also be used to make 
Schiff's reagent as well. In fact, many histotechs leave out the sulphite rinse 
step and simply rinse off in water and wash well with tap water. It seems to 
work just as well.

Bryan Llewellyn


Angela Lamberth via Histonet wrote:
> In Carson’s 3rd edition, the reducing rinse following Schiff for PAS 
> is 0.55% potassium metabisulfite (pg 137-138). For PAS/AB the reducing 
> rinse is given as sodium metabisulfite (Carson pg 149).
>
>
>
> My other texts (Sheehan/Hrapchak as well as Vacca) list the reducing 
> rinse as sodium metabisulfite which makes sense to me since the Schiff 
> is made with sodium metabisulfite.
>
>
>
> Is this a printing error in the Carson book? I appreciate any light 
> anybody can shed on this.
>


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Re: [Histonet] query

[Monson, Frederick via Histonet]

Have no expertise other with 'library.'
So, here is some new stuff that I found.

http://www.autoqbiosciences.com/products/3d-laser-nanodissection-systems/tissue-surgeon/
 

http://www.autoqbiosciences.com/_webedit/uploaded-files/All%20Files/Nano/LLS-Brosch%25C3%25BCre_TissueSurgeon_HardTissue_2014-03-18_web.pdf
 

There are lots of methods, but few that can be followed with microdissection.

Cheers,

Fred Monson


Frederick  C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of PA
Geology-Astronomy
750 South Church St.
West Chester, PA, 19383
fmon...@wcupa.edu
610-738-0437(Work)




-Original Message-
From: Nair, Indu via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Friday, March 03, 2017 1:26 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] query

Is there a protocol to section un-decalcified bone? Or is there a laboratory 
that does this routinely?
thank you
indu


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[Histonet] Old news about cryo sections

[Monson, Frederick via Histonet]

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 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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Re: [Histonet] Certification - Part I

[Monson, Frederick via Histonet]

Halloween story of a 'tech' who lacked a certificate.
T'was a Sunday, and he, a student at the age of 24,  was "ON" for the weekend 
covering blood bank, chemistry, and hematology.  A call came in for a unit of 
blood for a Friday surgical - just to bring the patient up to speed.  He, for 
reasons never given, decided to practice  'crossing' by doing up 10 units - all 
of which passed.  One he sent to the patient.  Two hours later he received a 
call for another unit, because, as he was informed, his hematocrit was 
unchanged.  What did he do?

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop:  Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmon...@wcupa.edu

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Re: [Histonet] Certification - II

[Monson, Frederick via Histonet]

He refused to send a unit without an order for a new cross match, which he got 
with some argument.  Only 4 of the 9 previously good units passed as he 
re-crossed all ten, and the one he sent did not pass the second crossing.  He 
called the pathologist on duty, who told him to give up going to medical school 
if he screwed up.  After the pathologist repeated the two experiments, he said 
a prayer and offered a compliment.  When asked how he had known, his response 
was, "I remembered the immunology I learned when I took microbiology 5 years 
earlier.

He could not get that job now, I suspect, though the conversation of 
certification remind me of what I learned about the trades and 'certification' 
in ancient Egypt.

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop:  Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmon...@wcupa.edu

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Re: [Histonet] AFB contaminant - OR - Monkish Preparation

[Monson, Frederick via Histonet]

I slways filtered my air (5um) and my water (0.22um).  Only tap water rinses 
for Hemnatoxylin, Schiff's and other rinses were not rinsed.  All of my MolBio 
and Immuno- stuff was done with filtered (clean?) water at least.

The rule is:  be Monkish at the bench and as messy as you like in your bedroom 
(if you are a male and only until you are married),

Cheers,

Fred 

Inside your water supply, lurking behind the water fawcet, is a pipe who's 
surface is lined with a BIO-film.
Further horrors await the scientist who fixes and studies the washer in the 
fawcet whose seal is the better for the 'FILM-m.'
Evolution provides variation, and every new flower that grows provides a new 
isolated environment for speciation.
My alimentary biome is unique as we are all unique.  Yukky, but true.

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmon...@wcupa.edu

From: Abbott, Tanya tanyaabb...@catholichealth.net
Sent: Thursday, July 9, 2015 3:09 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] AFB contaminant

Has anyone ever had a problem with a possible contaminant in their AFB hand 
stain? If so, how did you deal with it? Any suggestions?
Ours actually looked like an AFB like organism, which we later determined to be 
negative.
Thanks!

Tanya G. Abbott
Manager Technologist
Histology/Cytology
St Joseph Medical Center
(phone) 610-378-2635

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Re: [Histonet] Eosin

[Monson, Frederick via Histonet]

Long ago, in the foggy, foggy past,  we were admonished to insure that our 
Eosin in the bottle had a slight 'green' tinge in good daylight.

When we finally got fluorescence, we would check an HE-stained section with 
the fluorescein filters to insure the fluorescence of the Eosin. 

Very scientific, I know, but when the 'green' was gone, the Eosin was replaced.

Cheers,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop:  Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmon...@wcupa.edu

-Original Message-
From: Kienitz, Kari [mailto:kkien...@orclinic.com] 
Sent: Monday, July 13, 2015 11:21 AM
To: Hannen, Valerie; Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Eosin

It's always kind of a stab in the dark to diagnose HE problems without a 
little more information.  One thing I would try if you don't feel its an actual 
Eosin problem is your alcohol prior to the eosin.  Slide volume and humidity 
can dilute your alcohol and cause lighter cytoplasmic staining.  The alcohol 
prior to eosin should be 95% and changed daily if need be.


Kari Kienitz HT, (ASCP)
Histology Laboratory
Gastroenterology-EAST
The Oregon Clinic
 NE 99th Ave
Portland, OR  97220
503.935.8311
kkien...@orclinic.com




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missive. If you have received this in error, please notify the sender 
immediately by reply e-mail and delete this message and its attachments from 
your computer system. Thank you 
From: Hannen, Valerie [valerie.han...@parrishmed.com]
Sent: Monday, July 13, 2015 8:05 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Eosin

Good morning,

I once again am dealing with a my picky Pathologist!!  About a year ago, he 
started to complain about not having enough Eosin on his sections( first 
morning rack on Monday).. I went from changing the Eosin on Friday and stirring 
it on Monday to totally changing it  on Monday...it has all been good until the 
end of last week. The problem has started up again... little Eosin in the first 
rack of Monday morning.  Any suggestions??

Valerie Hannen,MLT(ASCP),HTL,SU (FL)
Section Chief, Histology
Parrish Medical Center
951 N. Washington Ave.
Titusville,Florida 32796
T: (321)268-6333 ext. 7506
F: (321) 268-6149
valerie.han...@parrishmed.commailto:valerie.han...@parrishmed.com
www.parrishmed.com

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Re: [Histonet] Digital Imaging Analyst Position at the University of Buffalo

[Monson, Frederick via Histonet]

A strongly recommended first interview question:  How many digital cameras do 
you have with you today?

Cheers,

FCM

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop:  Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmon...@wcupa.edu


-Original Message-
From: Featherstone, Annette via Histonet 
[mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, July 14, 2015 1:12 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] Digital Imaging Analyst Position at the University of 
Buffalo

Digital Imaging Analyst Position at The University of Buffalo:

Quicklink for Posting:  
https://www.ubjobs.buffalo.edu/applicants/Central?quickFind+58162



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[Histonet] Histonet links

[Monson, Frederick]

Histonet links appear to be functioning at a less-than-optimal level, but here 
they are anyway.
I can almost 'feel the pain' afflicting Linda and Anita, but, of course, that's 
a pompous, maladjusted, insensitive, and politically correct lie.

http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Histonethttp://lists.utsouthwestern.edu/mailman/listinfo/histonet list run by 
lindamargraf at gmail.com, anita.sengupta at 
childrens.commailto:histonet-ow...@lists.utsouthwestern.edu

The above work, but I have NOT tried the emails.  Just couldn't do it.

If the first doesn't work, then, perhaps the 'owners' are having  a problem or 
two.

Cheers,

Fred Monson

P.S.  The one thing that Darwinian Evolution must have is MAXIMUM variation, so 
get illuminated, and B_E_E_E_EG_O_O_O_D_D_D!

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop:  Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmon...@wcupa.edumailto:fmon...@wcupa.edu

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RE: [Histonet] Rolling of my ribbon ? Another paraffin question.

[Monson, Frederick]

Dear Stella,

Prior to buildings with locked windows, histologists who embedded with paraffin 
would fabricate, from just paraffin waxes of different hardness to match the 
climate of their geographic locations.  Thus, 

http://www.microscopy-uk.org.uk/mag/artjan07/yl-para1w.html

you can fabricate your own embedment with/or without the PEG (or whatever 
hygroscopic agent is added to provide 'stick') to match your needs.  Most large 
labs will not want to do this, so perhaps adding a dehumidifier to the 
microtome room to adjust the humidity to the embedment.

An accompanying message will describe one of my favorite mountants that 
requires an extended prep.

Cheers,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop:  Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmon...@wcupa.edu

HomePage:  http://cmirt.wcupa.edu (with link to instrument Scheduler)


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stella Mireles
Sent: Wednesday, November 13, 2013 3:18 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Rolling of my ribbon ? Another paraffin question.

*I am presently using a paraffin designated as an IM product.* *We are a 
facility that cuts only autopsies and have been experiencing alot of rolling of 
our sections.* *We did recently switch to this product, because of cost.*

*Question :  Is the paraffin you are using working well on autopsy tissue and 
producing ribbons right away ? Do you use it for infiltration as well as 
embedding ?*

*Thank you for your assistant.*


*Stella Walters*
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[Histonet] RE: Nitroblue Tetrazolium Chloride (NBTC)

[Monson, Frederick]

Just Google:  nitroblue tetraqzonium chloride.  Lots of sources.

Cheers,

Fred Monson

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert
Sent: Tuesday, October 12, 2010 6:41 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Nitroblue Tetrazolium Chloride (NBTC)

Does anyone know where I can get this dye?  It is to be used by a
research doc - he plans on putting the tissue in this dye and then into
10% formalin.

 

Laurie Colbert

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RE: [Histonet] Re: Responses to micron thickness for Rhodanine copper

[Monson, Frederick]

Once upon a time there was an investigator who used sections of testis of 
unspecified thickness to stain with iron hemtoxylin.  From his observations of 
the results, he concluded that there were two distinct populations of Sertoli 
cells based on his observation of two distinct populations of Sertoli cell 
nuclei - one darkly stained and one lightly stained.  The paper was published 
in Scandanavia in the late 1950's.  My unsupported conclusion was - I was very 
young at the time - that intact nuclei will hold more stain/dye than sectioned 
nuclei (not sealed to the slide) in a procedure that requires a 
'differentiation' step with the mordant.

In this case, do thicker sections hold more intact cells?

Just thought I'd ask the question.

Cheers, and I miss you folks more that you miss me, I bet!

Fred Monson

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
New Web Site:       http://cmirt.wcupa.edu/index.html 

New Scheduler:      http://cmirt.wcupa.edu/cgi-bin/ureserve_gold.pl 

Reads of the Month:  
1.  The Liver as a Lymphoid Organ, Ian Nicholas Crispe, Annual Review of 
Immunology, Apr 2009, Vol. 27: 147-163. (Access Depends on subscription.)
2. A Personal Journey of Discovery: Developing Technology and Changing 
Biology, Lee Hood, Annual Review of Analytic Chemistry, Vol. 1:  1-43.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carrie Disbrow
Sent: Monday, September 28, 2009 4:09 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: Responses to micron thickness for Rhodanine copper


Hi and thank you all for your responses!
Everyone agreed that the thicker sections allow the copper pigments to be 
better visualized. However the responders were split 50/50 in the thickness. 
There were labs that did cut the sections at 3 microns with continuous good 
results and the others at 6 microns. One lab cut the sections at 10 microns. I 
appreciate all your help and now we will test on three microns too.
Thanks again,
Carrie


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RE: [Histonet] Help With Hemo Fading

[Monson, Frederick]

Hi All,

Mordanted hematoxylin is quite colorfast when carefully 'blued' after binding 
to acid moieties in the tissues.  If one analyzes the entire process of HE 
dyeing, one can 'see' several steps in which the 'bluing' process can be 
'undone' or 'temporized'.  Note that I have always used personally prepared Gum 
Damar in Xylene to mount my sections.  It has always behaved better than 
expected.

Please recall that all of the currently used histologic procedures originated 
using water glasses from the dining room or kitchen and chemicals that were 
generated by magic.  When I was learning the procedure using the now-antiquated 
Coplin jar, I was overwhelmed by the inability of distilled water to 'blue' 
hematoxylin after acid differentiation.  For we new histologists at the time, 
the only thing that really 'set' the blue color was running tap water for 5 
minutes AFTER 5 minutes in 35% EtOH, 2% in NaHCO3.  NB, the same magic rinse 
was required to stabilize the magenta of the PAS stain.  In those days, there 
was the complaint that even some tap waters in other parts of the country left 
some lack of color fastness with mordanted hematoxylin (it must be remembered 
that the active component of the HE stain is NOT hematoxylin (those who are 
surprised must go the library!!)

In any case, I had a colleague who noticed that the slides in his collection 
were fading slowly in the closet in which he stored them (stored pretty much as 
we all do).  It was suggested that he perform a test of the storage environment 
in the following way.
  
Using an aquarium aerator placed in the closet to bubble air thru 1.5 
liters of distilled water with an appropriate indicator in a two-liter 
Ehrlenmyer with a two-hole stopper, the outlet taking the bubbled air out of 
the closet via an appropriate length of tubing.  Modify the above aerator and 
water volume to insure that the water will not be significantly evaporated 
during the overnight process (temperature, volume, and pressure are the 
appropriate variables).

This test should be run for several to many nights and days (or weeks) to 
determine whether at any time the environment 'goes' acidic.  [[The upside is 
that the experiment/test is neither labor intensive nor expensive (no grant 
proposal is required), though if you can wangle some stimulus money, you must 
let me know, because they will want to be able to count me as one of those who 
was put to work - albeit briefly.  If you run the above test for several years, 
you may be able to count me as being employed every time you refer to this 
suggestion.  [Perhaps not ethical, but we must do anything to keep unemployment 
under 10% lest there be declared another economic crisis whose fix we know by 
now will be the expected printing of more money to fund more studies of more 
fish in a pond or bugs on a wall.]]

Back to point, if at any time one observed the water to go acid, then the 
location of the storage is a likely candidate.  

If you use a machine to stain your sections, as many to most do,  then perhaps 
another environmental characteristic (such as humidity) is causing either 
continuous or seasonal ethanol dilution and thus leaving too much water in the 
sections, even though they do not appear milky when mounted.  In such a case, 
one must expect faster diffusion of acidic materials into the preparation.  NB, 
I only consider those airborne substances that are hydrogen rich - they can be 
inorganic or organic - simply anything that gets sprayed, mixed, dumped in the 
sink - anything that continuously gets aerosoled in the lab or is blown in with 
the 'conditioned' air.

Another test you might perform involves sealing slide boxes in plastic bags - 
see your local molecular biologist or check the kitchen area at KMart, Penny's, 
or Walmart for systems that pull a vacuum around food and seal it by creating a 
heat seal.  [If still unfamiliar, call Mom OR Mom-in-law!].  You must have a 
control box to compare over time.  One assumes that NOTHING can get in or out 
of such a sealed container.

The ultimate downside is that one does not or cannot control a process that one 
trusts to be automatic and credible.  

Hematologists may take note:  At the age of 40 I was laid up with a pulmonary 
'problem' that included a few days with 104-105 degree spikes, dry coughing, 
and visits from the Savior who I sent away screaming, Hell No!  I won't go! - 
and in the process scarring the b'Jesus out of my kids.  In any case after 
passing the hospitalization test and ahaving received, i.v., every known broad 
spectrum antibiotic in the pharmacy, I got my doctors to give me erythromysin 
i.v.(we don't give enythromysin orally or i.v. - it bites!), the temp went 
down, and I felt better.  [If legionella is sensitive to erythromysin, why did 
so many die from that bug in Philadelphia? (This will not be on the test, but 
it should be.)]  Thus, came the day when I was told that my blood differential 
was so