Ryan Hickey asks about the historical use of fixatives, and stains derived from
natural sources.
A good place to start is with the book by Brian Bracegirdle A History of
Microtechnique (second edition) Published by Scientific Heritage 1987.
There are many articles out there and you will
Thank you Dr Margraf, Dr Cope and now Dr Sengupta for giving up your time and
hosting the Histonet. It is an ideal way for histotechnologists and others to
be kept informed.
And Merry Christmas to everyone from the heart of Dixie!
Michael Titford
Pathology
USA Mobile
AL USA
Joyce Fortin asks about placenta's:
I work in a teaching hospital with a busy LD department and we examine every
placenta from every birth. Depending on their history the placenta may be
gross in (with four blocks) or gross only. The physicians are happy to have
a record of the weight and
Any one who has seen the CBS evening news with Scott Pelley lately (at least in
my area) will have seen an ad for a product named Salonpas. In the background
is a microscope. It is interesting that when you see a microscope in an ad on
TV, it implies that real, dedicated research has been
Victor Tobias asks about telepathology-
I seem to remember there is a rule somewhere that primary diagnosis can only be
made using a glass slide. As to chapter and verse, I don't know.
However there are reports in the literarature that it has been tried out for
F/S in Alaska and Finland.
Carol Tanck asks about material for a Middle school science day.
Good items to get hold of are plastinated organs that the young people can pick
up and handle. If you have a medical school or community college near by,
their cellular biology or anatomy departments (Titles vary in these
Leila asks about removing melanin pigment.
We run slides down to water and place in aquous 0.25% potassium Permanganate.
Wash well in tap water.Rinse in distilled water. Place in aquous 5% oxalic acid
for 5 minutes. Rinse in distilled water.Wash in tap water. continue with method.
We run
Thats's 5 minutes in the potassium permanaganate. Some stubborn melanin may
need a longer. Some authors recommend as long as 24 hours.
(It's Friday - how can you tell?)
Michael Titford
Pathology USA
Mobila AL
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And the stuff we poured down the sink without a second thought
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Ruth Yaskovich asks about staining Nissl:
Years ago, when I started in histology and there were still dinosaurs on earth,
we used cresyl fast violet for staining Nissl in celloidin sections, using a
fancy differentiation with cresote in alcohol, or something.
In paraffin sections you can
Merry Christmas to everyone from the Heart of Dixie! And thank you Dr Margraf
and Marvin Hanna for hosting the Histonet. It is a real asset to
histotechnologists.
Michael Titford
Pathology USA
Mobile AL
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Arjun Ruhella asked about fiber typing of animal soleus muscle:
No one answered him, at least in public. I guess all the hotshots are in
Vancouver!
Years ago I was involved in fiber typing and measuring of human muscle biopsy
fibers. In those human muscle biopsies, all the bundles were mixed,
Am I hearing right? The CAP has done away with the requirement to sign out
autopsies within 60 days? (But you still have to get the PAD out in two days)?
How will I ever get pathologists motivated any more?!
Michael Titford
Pathology
USA Mobile AL
Sheila asks about cut-resistant gloves:
We use the cut resistant gloves from Surgipath Medical Industries. They are of
course, cut resistant, but not cut proof or puncture proof. They fit nice and
snugly and don't impede work. They work well if you just touch a finger lightly
with a scalpel
In our frozen section room, we use plastic dropper bottles (holding about 130ml
each) to pour the solutions on the slide on a rack to stain the F/S. We
figure you don't risk getting floaters and the solutions are fresh.
However, these bottles are home made and do not look professional. We
Hazel Horn asks about the ATPase reaction:
Years ago we had problem with our ATPase reaction on F/S of muscle biopsies. We
discovered that in keeping the substrate at minus 20oC in a freezer, and
getting it out and letting it warm up to room temperature before weighing a
small amount out for
You can make your own holy water. You put regular water on a stove and boil the
hell out of it!
Mike Titford
USA - Pathology
Mobile AL
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Those of you who mourn the passing of the AFIP may be interested that a book
has been published about its history, entitled, Legacy of Excellance: The
Armed Forces Institute of Pathology 1862 - 2011 by Paul Stone, published by
The Borden Institute, Fort Detrick, Maryland. It details the AFIP
Ann Angelo asks about releasing tissues in legal cases.
I guess how you do it depends on your state and its laws, and what your own
facility lawyers require.
At our facility in the State of Alabama, to release tissues and/or medical
records we require either a release signed by the patient, a
I was a little distressed to read the message from Amy in Camp Hill,
Pennsylvania declaring she dumps everything (and I mean everything) from her
histology lab down the drain. There are a bunch of Federal Laws governing
handling and disposal of chemicals used in the histology laboratory and
Those of you that like freebies (like Dako mousepads, Thermo pens and Sigma
coffee mugs) will be amused to learn that Cell Marque are giving away packs of
playing cards. Each card shows a section stained with a different antobody (one
of theirs of course, together with the catalogue number).
Candice Camille asks about Ehrlich's hematoxylin-
The sodium iodate to ripen the hematoxylin should be added last of all, after
all the other ingredients, to ripen the stain. (So you will not see if it has
dissolved, or not!).
Better to ripen the Ehrlich's hematoxylin naturally. Do not add
Some years ago, our deionized water system became contaminated by a gram
negative organism. (I don't know what the bacteria live on, in the dark, with
only deionized water to eat). We have a contract with a deionized water
company, but they said it would be impossible to sterilize the whole
Candice Camille asks about polyvinyl alcohol.
Some time ago when I was in the Army Reserve I completed a correspondence
course for parasitology and it discussed PVA. I still have the manual. It
states: PVA is a mixture of fixative and water soluble resin that is
specifically used to fix and
An Eerdekens overseas somewhere asks about thionin staining:
An, you may have a bad batch of thionin stain. Did you just open a new jar?
Here is the USA we use stains certified for use in histology by the Biological
Stain Commission. Each lot has been tested by them for use in histology.
Nicole Cosenza asks about acetylcholinesterase methods.
Years and years ago in London we used Koelle's method to demonstrate
cholinesterase in muscle and mouse diaphragms. The method we followed was in
Pearse E, Histochemistry - Theoretical and Applied. Volume 2. Churchill
Livingstone. London
Jorge Tornero asks about Gilson's fluid.
The original Gilson's fluid was made up of Mercuric chloride 2 grams, glacial
acetic acid 0.4ml, nitric acid 1.8 ml, 95% ethyl alcohol 10 ml, and distilled
water 88ml.
I don't know about usining zinc chloride in the formula
Michael Titford
USA
Mark Elliot asks about processing FFPE tissue for EM. Hayat published a method
in his book we use. Briefly you dig out 1mm cube pieces of tissue from the
block and rotate them in xylene for one hour, followed by absolute, 95%,
70%,50% alcohols for 15 minutes each and then into your EM buffer
I am curious!! In a Histonet message last week, Tony Henwood mentioned the
formalin substitutes UMfix, Kryofix (now called Neo-fix I think) and
Spuitfix.
1) How many surgical pathology labs use formalin substitutes for routine work?
2) Which ones do they use? (which is the most popular?)
3)
Dr Cartun sent an email to the Histonet about what his institution bills for
the technical component of 88342.
Dr Cartun - Don't feel so guilty! In reality no one much pays top dollar for
any service in the hospital. Some patients are treated as DRG patients
(Hospital gets paid set amount
Thank you everyone who responded to my problem with a Leica CM 1850 cryostat
turning itself off. About 11 people responded with tips. Thank you!! The
Histonet is great!!
In answer to some enquirys:
1) The cryostat is set to defrost at 2 a.m.. The manual says it defrosts for an
hour. Jackie O'
We have a Leica CM 1850 cryostat that about once every three weeks to a month,
turnes itself off! Has anyone else experienced this? How do I fix it?
Initially, we thought someone after hours was turning it off, maybe someone in
housekeeping or a passing med tech. That was not the case.
Later
Gudrun asks about how the Gomori retic procedure works:
It is my understanding that the iron alum (ferric ammonium sulphate) acts as a
sensitizer. The iron combines with the aldehyde formed in the reticulum from
the previous oxidation step to create an aldehyde-metal complex. In the third
Thank you everyone for all the information about microarray equipment. As usual
on the Histonet, everyone was quick with the information, generous with their
time, and informative.
Michael Titford
USA Pathology
Mobile AL USA
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We are thinking of making our own microarray blocks instead of purchasing
someone elses slides. Can someone:
1) Recommend a punch to use?
2) Which size diameter tip is best? (for general use)
3) are the tips re-usable, or are they single use?
I attended April Watanabes's workshop on microarray
Before the days of automatic coverslippers, I have seen coverslipping done in
large quantities from above and from below. I guess it depends what you are
comfortable with. In either event, you want to get a squeegee effect so no
air bubbles are trapped in the mountant. You need more control
Seasons Greetings to everyone from the Heart of Dixie, and thank you Dr Linda
Margraf and colleagues for hosting the Histonet. I get to keep in touch and
not have to leave my office!
Michael Titford
USA Pathology
Mobile AL USA
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Debbie Boyd in Virginia asks about slide crushers:
At the National Society meeting in Birmingham recently, a company called
Creative Waste Solutions displayed a slide crusher. It grinds them up and the
slide labels too until it is just like very roughly ground sugar. The
instrument costs
Maria asks about fetal demise placenta:
We gross in all fetal demise placenta (usually four blocks).If the clinicians
desire genetics, they take?samples from the fetus and the placenta up on the
floor. (There is only a three day window for growing fibroblasts).
The fetus gets a gross only
One year, everyone brought in photographs of their pets. There was a
competition to guess which pets belonged to which lab employees. They say
people look like their pets
Mike Titford
Pathology
USA
Mobile AL
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I would like to add to the cooments of others concerning distilled water and
deionised water:
1) I think most histology labs use deionised water as it is cheaper and easier?
to obtain.
2) However, even deionised water will turn red with Schiff reagent.
3) For silver stain solutions, our lab
im Wheelock asks about brain containers:
We use the 5.7 liter polyethylene containers from Richard Allen (Cat # 5357).
When fixing whole brains we suspend the brain in the formalin in a hair net. We
keep the brain slices in these containers too.
Michael Titford
USA Pathology
Mobile AL USA
Kimberly Tuttle in Baltimore asks about lab inspections.
I attended a workshop at the NSH S/C in Toronto where Diane Sterchi gave a
really good workshop entitled, Cracking the code: regulatory issues in the
laboratory.? She spoke about the ISO 9000 certification requirements?that some
research
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