[Histonet] RE: Histonet Digest, Vol 134, Issue 28
John O'Brien
john <@t> imebinc.com
Fri Jan 23 13:22:07 CST 2015
COLD plate is way to cold.
Regards
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Today's Topics:
1. RE: rodent eye (Gowan,Christie C)
2. Cracking paraffin blocks (Wheelock, Timothy R.)
3. RE: Cheap Disposable Blades for Facing In (Bea DeBrosse-Serra)
4. Re: Cracking paraffin blocks (Hans B Snyder)
5. Amyloid by Congo Red (Jeffrey Robinson)
6. Thermo IHC (Cheri Miller)
7. Problem with cracked paraffin blocks (Wheelock, Timothy R.)
8. Re: Amyloid by Congo Red (Michael Ann Jones)
----------------------------------------------------------------------
Message: 1
Date: Wed, 21 Jan 2015 16:23:06 +0000
From: "Gowan,Christie C" <christiecgowan <@t> dermatology.med.ufl.edu>
Subject: RE: [Histonet] rodent eye
To: Casie Phillips <casie4384 <@t> gmail.com>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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Hi Casie,
Hope by now you have rec'd some good tips on rodent eye prep. The only thing
I have to offer is that we always used Davidson's fixative for 24 hours and
then transferred to 70% ETOH. This worked beautifully preserving all eye
components. Good luck and don't forget to check the Histonet archives where
I know rodent eyes have been discussed in the past.
Christie Gowan HT (ASCP)
Department of Dermatology
4037 NW 86th Terrace, 4th Fl
Mohs Laboratory
Gainesville, FL 32606
Phone: 352 594-1529
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu
[histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Casie Phillips
[casie4384 <@t> gmail.com]
Sent: Thursday, January 15, 2015 2:53 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] rodent eye
Good afternoon,
I am currently working with Lewis rats performing corneal alkali injuries at
varying strengths. Is there someone there that has prior experience working
with a rat eye and would be willing to share information on the most
effective ways to preserve, fix and cut the cornea sample.
We are interested in using the cornea without using the whole globe if
possible. For now we will be using basic H&E staining with a possibility of
immunohistochemistry at a later time. The main outcome we are looking for is
to find the presence of neutrophils in the cornea. A second objective is to
look for any damaged or newly reconstructed tissue.
I would greatly appreciate any advice relating to the type of paraffin used,
the ideal length of time to save the tissue and any assistance you can
suggest for completing this process successfully.
Thank you for your time. Any assistance will be greatly appreciated.
Sincerely,
Casie Phillips
Casie4384 <@t> gmail.com
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------------------------------
Message: 2
Date: Fri, 23 Jan 2015 14:57:15 +0000
From: "Wheelock, Timothy R." <twheelock <@t> mclean.harvard.edu>
Subject: [Histonet] Cracking paraffin blocks
To: "'Histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
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<69718C0B0B3C414D9F8E7214AD400CC9773EABC4 <@t> PHSX10MB11.partners.org>
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Hi Everyone:
Recently, I purchased the Medite Valida Embedding Center, which I demoed
previously without a problem, if I recall right.
I am having problems now with blocks developing cracks on the cold plate.
The cracks run either through the wax right next to the specimen, or more
frequently, right through the tissue itself.
I use Surgipath Embedding Media (EM-400).
The surface of the Valida's cold plate is -14C.
The company has not seen this happening before, but they are looking into
this further.
Also, I had the same problem when I demoed the Thermo-Fisher HistoStar.
I do not think that this is a problem inherent with any particular machine.
Has anyone encountered this problem before?
If so, how did you resolve it?
Is it possible that the -14C cold plate is too cold? Should I warm it up a
bit?
The surface of the Valida cold plate is, I believe, made out of smooth
stainless steel.
My old (24 years) Shandon Embedding Center's cold plate is made out of cast
aluminum and has a slightly "rougher" texture.
Could this produce a different way of conducting heat out of the paraffin
block than the Shandon?
Perhaps the stainless steel draws heat from the blocks faster than the
aluminum, and thus causes the cracking?
I have used the Shandon Embedding Center for over 24 years.
I have never had a problem with blocks that crack.
Thanks for any thoughts that you may have on this.
Tim Wheelock
Harvard Brain Bank
McLean Hospital
Belmont, MA
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------------------------------
Message: 3
Date: Fri, 23 Jan 2015 15:49:27 +0000
From: Bea DeBrosse-Serra <BDeBrosse-Serra <@t> isisph.com>
Subject: RE: [Histonet] Cheap Disposable Blades for Facing In
To: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>, Sandra Cheasty
<cheastys <@t> svm.vetmed.wisc.edu>
Cc: "Histonet \(histonet <@t> lists.utsouthwestern.edu\)"
<histonet <@t> lists.utsouthwestern.edu>
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We are doing the same.
Beatrice DeBrosse-Serra HT(ASCP)QIHC
Isis Pharmaceuticals
Antisense Drug Discovery
2855 Gazelle Ct.
Carlsbad, CA 92010
760-603-2371
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jennifer
MacDonald
Sent: Friday, January 23, 2015 2:59 AM
To: Sandra Cheasty
Cc: Histonet (histonet <@t> lists.utsouthwestern.edu)
Subject: Re: [Histonet] Cheap Disposable Blades for Facing In
We save our blade from the previous day to use for facing. We keep them in
slide mailers.
Sent from my iPad
> On Jan 22, 2015, at 4:21 PM, Sandra Cheasty
<cheastys <@t> svm.vetmed.wisc.edu> wrote:
>
> Hello everyone,
>
> Does anyone have a source for cheap, low-profile
> blades
for facing in blocks?
>
> Thanks!
> Sandy
>
>
>
> Sandra J. Cheasty, HT (ASCP)
>
> Histology & Necropsy Supervisor, President Keith Richards Fan Club
>
> UW-Madison, School of Veterinary Medicine
>
>
>
>
>
>
>
>
>
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------------------------------
Message: 4
Date: Fri, 23 Jan 2015 12:07:03 -0500
From: Hans B Snyder <hans <@t> histologistics.com>
Subject: Re: [Histonet] Cracking paraffin blocks
To: "Wheelock, Timothy R." <twheelock <@t> mclean.harvard.edu>
Cc: "Histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
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Hello Tim,
I also have had this cracking problem in the past but since resolved.
I'm not sure about the specifics of the cracking problem but the cold plate
might have something to do with it. Our cold plate is set to -5C and use
Paraplast from McCormick Scientific. Our paraffin is roughly the same
melting temp as yours, so probably not much difference.
Unfortunately trial and error is apart of our jobs since there is relatively
little uniformity in histology equipment and products. Have you tried
setting the cold plate to a warmer temperature yet? It's worth a try and if
that doesn't work maybe the paraffin has been getting too hot. I know when
the paraffin gets too hot, the wax and plastic separate or break down and
cause inconsistencies in the paraffin. To test this, take an external
thermometer and place inside the paraffin tank. Then record the temp every
hour for some time.
This will tell you if the tank itself is a consistent temperature.
Also, are the blocks taken off the cold plate and immediately jarred loose
from the mold? Sometimes when I do this the shock of pried out of the mold
can cause the paraffin to become brittle and break rather than bend.
Let me know how it goes.
Thank you
Hans B Snyder
Histologistics
60 Prescott Street
Worcester, MA 01605
508-308-7800
hans <@t> histologistics.com
On Fri, Jan 23, 2015 at 9:57 AM, Wheelock, Timothy R.
<twheelock <@t> mclean.harvard.edu> wrote:
> Hi Everyone:
>
> Recently, I purchased the Medite Valida Embedding Center, which I demoed
previously without a problem, if I recall right.
> I am having problems now with blocks developing cracks on the cold plate.
> The cracks run either through the wax right next to the specimen, or more
frequently, right through the tissue itself.
> I use Surgipath Embedding Media (EM-400).
> The surface of the Valida's cold plate is -14C.
>
> The company has not seen this happening before, but they are looking into
this further.
> Also, I had the same problem when I demoed the Thermo-Fisher HistoStar.
> I do not think that this is a problem inherent with any particular
machine.
> Has anyone encountered this problem before?
> If so, how did you resolve it?
>
> Is it possible that the -14C cold plate is too cold? Should I warm it up a
bit?
> The surface of the Valida cold plate is, I believe, made out of smooth
stainless steel.
> My old (24 years) Shandon Embedding Center's cold plate is made out of
cast aluminum and has a slightly "rougher" texture.
> Could this produce a different way of conducting heat out of the paraffin
block than the Shandon?
> Perhaps the stainless steel draws heat from the blocks faster than the
aluminum, and thus causes the cracking?
>
> I have used the Shandon Embedding Center for over 24 years.
> I have never had a problem with blocks that crack.
>
> Thanks for any thoughts that you may have on this.
>
> Tim Wheelock
> Harvard Brain Bank
> McLean Hospital
> Belmont, MA
>
>
> The information in this e-mail is intended only for the person to whom
> it is addressed. If you believe this e-mail was sent to you in error
> and the e-mail contains patient information, please contact the
> Partners Compliance HelpLine at http://www.partners.org/complianceline
> . If the e-mail was sent to you in error but does not contain patient
> information, please contact the sender and properly dispose of the e-mail.
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Fri, 23 Jan 2015 17:43:33 +0000
From: Jeffrey Robinson <JRobinson <@t> pathology-associates.com>
Subject: [Histonet] Amyloid by Congo Red
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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Greetings to all Histotechs- Here's an amyloid question for the braintrust.
We are cutting our slides and controls at 9 and staining in Congo Red for 1
hour. The control stains fine but the patient tissue is staining negative
even on cases that the pathologist assures us should be positive for
amyloid. We are using the Leica APEX charged slides with control and
patient tissue on the same slide. Does anyone have any thoughts on why the
patient tissue is not staining? Thanks!
Jeff Robinson HT, HTL, Senior Histotechnologist, Sierra Pathology Lab,
Clovis, CA.
This email and attachments may contain PHI that is privileged and
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------------------------------
Message: 6
Date: Fri, 23 Jan 2015 11:54:06 -0600
From: Cheri Miller <cmiller <@t> physlab.com>
Subject: [Histonet] Thermo IHC
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Cc: "histonet-bounces <@t> lists.utsouthwestern.edu"
<histonet-bounces <@t> lists.utsouthwestern.edu>
Message-ID: <E3C81A010935EA41B379AC765103F3BF4000D6C102 <@t> olsrv12>
Content-Type: text/plain; charset="iso-8859-1"
Hi Histonetters! I need some help. We just acquired a Thermo IHC system. Can
anyone help us cut through the *****88*888 and give me some helpful tips?
We have practical experience on the Ventana systems only. Has anyone been
successful at using only 1 HIER buffer? Any tips on how to shorten the
offline retrieval process? Currently we are at over 40 mins. Seems to me a
ph of 7.6 -7.8 should work for most all of our antibodies. Thanks, Cheri
Cheryl A. Miller HT ASCP cm
Physicians Laboratory, P.C.
4840 F St.
Omaha , NE. 68117
402 731 4145 ext. 532
Cell 402 980 2537
Fax 402 731 8653
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Message: 7
Date: Fri, 23 Jan 2015 17:55:12 +0000
From: "Wheelock, Timothy R." <twheelock <@t> mclean.harvard.edu>
Subject: [Histonet] Problem with cracked paraffin blocks
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
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Hi again everyone:
I want to thank you all for your advice.
The consensus is that I have my new embedding center's cold plate set way
too low at -14C, and that I should try raising it to around -5C to cure my
cracked block problem.
I will run some test blocks over the weekend, and then embed them at this
new temperature on Monday.
Otherwise my Valida embedding center is working very nicely (as did the
HistoStar when I demoed it).
The cassette holding tank can accommodate a large number of brain specimens
and the controls are very easy to use.
Thanks again for your advice.
Have a great weekend.
Tim
Tim Wheelock
Harvard Brain Bank
McLean Hospital
Belmont, MA
The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail contains patient information, please contact the Partners Compliance
HelpLine at http://www.partners.org/complianceline . If the e-mail was sent
to you in error but does not contain patient information, please contact the
sender and properly dispose of the e-mail.
------------------------------
Message: 8
Date: Fri, 23 Jan 2015 17:58:31 +0000
From: Michael Ann Jones <mjones <@t> metropath.com>
Subject: Re: [Histonet] Amyloid by Congo Red
To: Jeffrey Robinson <JRobinson <@t> pathology-associates.com>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <D0E7D992.65C3%mjones <@t> metropath.com>
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What protocol and reagents are you using for the stain?
Michael Ann Jones, HT (ASCP)
Histology Manager
Metropath
7444 W. Alaska Dr. #250
Lakewood, CO 80226
303.634.2511
Mjones <@t> metropath.com
On 1/23/15, 10:43 AM, "Jeffrey Robinson"
<JRobinson <@t> pathology-associates.com> wrote:
>Greetings to all Histotechs- Here's an amyloid question for the
>braintrust. We are cutting our slides and controls at 9 and staining
>in Congo Red for 1 hour. The control stains fine but the patient
>tissue is staining negative even on cases that the pathologist assures
>us should be positive for amyloid. We are using the Leica APEX charged
>slides with control and patient tissue on the same slide. Does anyone
>have any thoughts on why the patient tissue is not staining? Thanks!
>
>Jeff Robinson HT, HTL, Senior Histotechnologist, Sierra Pathology Lab,
>Clovis, CA.
>
>
>This email and attachments may contain PHI that is privileged and
>confidential and is not intended for any unauthorized person. If you,
>the reader, are not the intended recipient you are hereby notified that
>any dissemination, distribution or copying of this communication is
>strictly prohibited. Do not read the email but instead reply to the
>sender and destroy the message and any attachments. Thank you.
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
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