[Histonet] RE: Color blind chart request
Bartlett, Jeanine (CDC/OID/NCEZID)
jqb7 <@t> cdc.gov
Tue Mar 13 10:38:40 CDT 2012
You probably have to have someone administer it and verify it to make it acceptable for CAP, though.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Stephen G. Ruby
Sent: Tuesday, March 13, 2012 11:37 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Color blind chart request
Google "Color blind chart".
Then select "images" from the google menu.
You will get what you want.
Select the image and print on a color printer.
Viola! Done.
Stephen G. Ruby
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Tuesday, March 13, 2012 10:32 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 100, Issue 15
Send Histonet mailing list submissions to
histonet <@t> lists.utsouthwestern.edu
To subscribe or unsubscribe via the World Wide Web, visit
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
or, via email, send a message with subject or body 'help' to
histonet-request <@t> lists.utsouthwestern.edu
You can reach the person managing the list at
histonet-owner <@t> lists.utsouthwestern.edu
When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..."
Today's Topics:
1. Re: undecalcified bone IHC (gayle callis)
2. Re: Re: undecalcified bone IHC (Victoria Baker)
3. Unsubscribe (Konop, Nicole)
4. Gram Stain (Giroux, Stacy)
5. DAKO Autostainer Issues (Friedrich_Hahn <@t> bd.com)
6. Color Blindness Testing (Laurie <@t> blufrogpath.com)
7. Re: DAKO Autostainer Issues (Drew Meyer)
8. IHC for decalcified bone protocol (Jeffery Howery)
9. Re: IHC for decalcified bone protocol (Mehmet Fatih BOZKURT)
10. RE: Gram Stain (gayle callis)
11. IHC Slides (Courtney Pierce)
12. Re: IHC Slides (Rene J Buesa)
13. RE: Gram Stain (Tony Henwood (SCHN))
14. CD34 (Chakib Boussahmain)
15. (no subject) (enrriq88 <@t> yahoo.com)
16. Liver Biopsy Processing Schedule (ADESUPO ADESUYI)
17. CD45.1 & CD45.2 IF on parrafin sections. (Thotakura, Anil Kumar)
18. IHC Processor Validation & MUM1 (Bliven, Laura)
19. Ihc validation (Sabrina Townsend)
20. RE: Re: undecalcified bone IHC (Jack Ratliff)
21. Re: CD34 (Rene J Buesa)
22. Re: IHC Processor Validation & MUM1 (Rene J Buesa)
----------------------------------------------------------------------
Message: 1
Date: Mon, 12 Mar 2012 11:04:20 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: [Histonet] Re: undecalcified bone IHC
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000001cd0072$2cbb8150$863283f0$@bresnan.net>
Content-Type: text/plain; charset="iso-8859-1"
Jeff,
It is most certainly possible to do IHC on undecalcifed bone sections
embedded in PMMA although not the easiest task. Sectioning is done on a
microtome that is powerful enough to cut the plastic and using tungsten
carbide knives. The key is total removal of the plastic from MMA embedded
bone sections to allow antibody/ immunoglobulins to access antigenic sites.
Neil Hand has done IHC successfully on PMMA embedded tissues including undecalcified bone on 2 to 3 ?m thick sections. I think one could cut thicker sections at 4 to 5 ?m and still be successful. I do not recall what Troiano et al used.
The following publications will help you and should include protocols, although conventional protocols will work according to Hand.
Blythe D. Hand N et al 1997 J Clin Path 50:45-49. The use of methyl
methacrylate resin for embedding bone marrow trephine biopsies.
Hand NM et al 1996 Antigen unmasking using microwave heating on formalin fixed tissue embedded in methyl methacrylate J Cellular Path 1:31-37
Jackson P et al. 1996 Amplification of immunocytochemical reactions by
the catalytic deposition of biotin on tissue sections. J Path
170(suppl):23A. This was about tyramide amplification when one gets a weak signal from "conventional" methods.
Hand NM, Church RJ 1998 Superheating using pressure cooking: its use and application in unmasking antigens embedded in methyl methacrylate. J Histotechnology 2`:231-236
Hand NM et al 1989 Immunohistochemistry on resin embedded tissue for light
microscopy: a novel post embedding procedure. Proceeding Royal Microscopical Society 24(1):A54-55.
Hand NM Plastic Embedding media and techniques, Ch.30, p 663-677. Theory
and Practice of Histological Technique, 5th edition by Gamble and Bancroft.
The 6th edition is updated under same title.
Use Google Scholar to find Troiano N et al from Yale on doing IHC on PMMA embedded bone sections with publications in J Histotechnology.
Hand mentioned several HIER methods, using citrate buffer. Optimizing
retrieval will depend on the antigen and you may end up doing this with some form of HIER, including microwave or other heat producing methods and with
different buffers. Enzyme digestion is also a possibility.
Hand removed MMA with xylene, warm my speed up the removal, also more than
one change for 10 - 20 minutes or longer. When I talked to him personally,
he said he had used warm xylene although temperature was not mentioned in
his chapter. After MMA removal, rehydrate section through alcohol gradient
as one does paraffin sections. He was emphatic about never allowing the
sections dry out.
Hopefully Jack Ratliff and Damien Laudier will provide more insight on this topic.
Good luck
Gayle M. Callis
HTL/HT/MT(ASCP)
******************************************
Hi Jeff,
If is it possible a few more specifics of how the tissue has been received,
processed and evaluated would help. Undecalcified bone sectioning
procedures vary and also what specific markers are you looking to do is
important.
Vikki
On Mon, Mar 12, 2012 at 11:06 AM, Rene J Buesa <rjbuesa <@t> yahoo.com>
wrote:
> Undecalcified? How are you going to section it?
> If you can section it, just use any IHC protocol for regular sections.
> Good luck!
> Ren? J.
>
> --- On Mon, 3/12/12, Jeffery Howery <Jeffery.Howery <@t> jcl.com> wrote:
>
>
> From: Jeffery Howery <Jeffery.Howery <@t> jcl.com>
> Subject: [Histonet] Undecalcified bone IHC
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Monday, March 12, 2012, 10:59 AM
>
>
> Does anyone have a protocol for Undecalcified bone for IHC?
>
------------------------------
Message: 2
Date: Mon, 12 Mar 2012 13:09:39 -0400
From: Victoria Baker <bakevictoria <@t> gmail.com>
Subject: Re: [Histonet] Re: undecalcified bone IHC
To: gayle callis <gayle.callis <@t> bresnan.net>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<CALYZRmQLbJtK4wPFEoTzW6ZmFjwMDWuTe0b5xzmjsLQgfGO0Ow <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Thank you Gayle. Vikki
On Mar 12, 2012 1:04 PM, "gayle callis" <gayle.callis <@t> bresnan.net> wrote:
> Jeff,
>
>
>
> It is most certainly possible to do IHC on undecalcifed bone sections
> embedded in PMMA although not the easiest task. Sectioning is done on a
> microtome that is powerful enough to cut the plastic and using tungsten
> carbide knives. The key is total removal of the plastic from MMA embedded
> bone sections to allow antibody/ immunoglobulins to access antigenic sites.
> Neil Hand has done IHC successfully on PMMA embedded tissues including
> undecalcified bone on 2 to 3 ?m thick sections. I think one could cut
> thicker sections at 4 to 5 ?m and still be successful. I do not
> recall what Troiano et al used.
>
>
>
> The following publications will help you and should include protocols,
> although conventional protocols will work according to Hand.
>
>
>
> Blythe D. Hand N et al 1997 J Clin Path 50:45-49. The use of methyl
> methacrylate resin for embedding bone marrow trephine biopsies.
>
> Hand NM et al 1996 Antigen unmasking using microwave heating on
> formalin fixed tissue embedded in methyl methacrylate J Cellular Path
> 1:31-37
>
> Jackson P et al. 1996 Amplification of immunocytochemical reactions by
> the catalytic deposition of biotin on tissue sections. J Path
> 170(suppl):23A. This was about tyramide amplification when one gets a
> weak signal from "conventional" methods.
>
> Hand NM, Church RJ 1998 Superheating using pressure cooking: its use
> and application in unmasking antigens embedded in methyl methacrylate.
> J Histotechnology 2`:231-236
>
> Hand NM et al 1989 Immunohistochemistry on resin embedded tissue for
> light
> microscopy: a novel post embedding procedure. Proceeding Royal
> Microscopical Society 24(1):A54-55.
>
> Hand NM Plastic Embedding media and techniques, Ch.30, p 663-677. Theory
> and Practice of Histological Technique, 5th edition by Gamble and
> Bancroft.
> The 6th edition is updated under same title.
>
>
>
> Use Google Scholar to find Troiano N et al from Yale on doing IHC on
> PMMA embedded bone sections with publications in J Histotechnology.
>
>
>
>
>
> Hand mentioned several HIER methods, using citrate buffer. Optimizing
> retrieval will depend on the antigen and you may end up doing this
> with some form of HIER, including microwave or other heat producing
> methods and with
> different buffers. Enzyme digestion is also a possibility.
>
>
>
> Hand removed MMA with xylene, warm my speed up the removal, also more than
> one change for 10 - 20 minutes or longer. When I talked to him
> personally,
> he said he had used warm xylene although temperature was not mentioned in
> his chapter. After MMA removal, rehydrate section through alcohol
> gradient
> as one does paraffin sections. He was emphatic about never allowing the
> sections dry out.
>
>
>
> Hopefully Jack Ratliff and Damien Laudier will provide more insight on
> this topic.
>
>
>
> Good luck
>
>
>
> Gayle M. Callis
>
> HTL/HT/MT(ASCP)
>
>
>
>
>
>
>
>
>
>
>
>
>
> ******************************************
>
>
>
> Hi Jeff,
>
>
>
> If is it possible a few more specifics of how the tissue has been
> received,
>
> processed and evaluated would help. Undecalcified bone sectioning
>
> procedures vary and also what specific markers are you looking to do
> is
>
> important.
>
>
>
> Vikki
>
> On Mon, Mar 12, 2012 at 11:06 AM, Rene J Buesa <rjbuesa <@t>
> yahoo.com>
> wrote:
>
>
>
> > Undecalcified? How are you going to section it?
>
> > If you can section it, just use any IHC protocol for regular sections.
>
> > Good luck!
>
> > Ren? J.
>
> >
>
> > --- On Mon, 3/12/12, Jeffery Howery <Jeffery.Howery <@t> jcl.com> wrote:
>
> >
>
> >
>
> > From: Jeffery Howery <Jeffery.Howery <@t> jcl.com>
>
> > Subject: [Histonet] Undecalcified bone IHC
>
> > To: histonet <@t> lists.utsouthwestern.edu
>
> > Date: Monday, March 12, 2012, 10:59 AM
>
> >
>
> >
>
> > Does anyone have a protocol for Undecalcified bone for IHC?
>
> >
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 3
Date: Mon, 12 Mar 2012 12:35:07 -0500
From: "Konop, Nicole" <NKonop <@t> chw.org>
Subject: [Histonet] Unsubscribe
To: "'histonet-request <@t> lists.utsouthwestern.edu'"
<histonet-request <@t> lists.utsouthwestern.edu>,
"'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<A733875C1AA92D4EBD9A35A9506DFE3A5DAF600DF6 <@t> C1EXCPWV02.chwi.chswi.org>
Content-Type: text/plain; charset="us-ascii"
Nicole Anne Konop BS, HTL(ASCP)
Histology Team Lead
Children's Hospital of Wisconsin
(414)266-6580 Direct Line
(414)907-0366 Pager
(414)266-2524 Histology Department
------------------------------
Message: 4
Date: Mon, 12 Mar 2012 12:35:47 -0500
From: "Giroux, Stacy" <Stacy.Giroux <@t> stjohn.org>
Subject: [Histonet] Gram Stain
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<29CCF3EC44815745864D9F84A1ED4B51B3E0709B0E <@t> AUSP03VMBX11.apptixhealth.net>
Content-Type: text/plain; charset="iso-8859-1"
Hi,
Our lab is currently transitioning from the Brown & Brenn gram stain due to no longer wanting to store picric acid due to its potential hazards. Our pathologists have requested a gram stain for paraffin embedded tissue that looks similar but does not use picric acid or ether. Does anyone have any suggestions on stains that could be used or gram stain kits that are available for purchase that are good?
Thank you for your help,
Stacy
CONFIDENTIALITY NOTICE:
This email message and any accompanying data or files is confidential and may contain privileged information intended only for the named recipient(s). If you are not the intended recipient(s), you are hereby notified that the dissemination, distribution, and or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at the email address above, delete this email from your computer, and destroy any copies in any form immediately. Receipt by anyone other than the named recipient(s) is not a waiver of any attorney-client, work product, or other applicable privilege.
------------------------------
Message: 5
Date: Mon, 12 Mar 2012 14:31:44 -0400
From: Friedrich_Hahn <@t> bd.com
Subject: [Histonet] DAKO Autostainer Issues
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF3E9D799B.1E276F03-ON852579BF.0064FB8C-852579BF.0065C7D9 <@t> bd.com>
Content-Type: text/plain; charset="US-ASCII"
We have been having issues with our Autostainer since bringing it out
of storage. All parts are in excellent shape, according to the
technician that performed the PM. It worked fine for a month of so,
but then began malfunctioning after only a couple of staining runs.
At some point in the staining process, the machine appears to lose its
XY homing and jams itself against the front right corner, dispensing
buffer onto the floor of the chamber and occasionally making grinding
noises. We've had a technician take a look at it, 'clean' the
software, and we thought we had it resolved, but the problem appears
to be back.
Has anyone experienced this issue? If so, was it hardware- or
software- based?
Thanks in advance!
-----------------------------------------
*******************************************************************
IMPORTANT MESSAGE FOR RECIPIENTS IN THE U.S.A.: This message may
constitute an advertisement of a BD group's products or services or a
solicitation of interest in them. If this is such a message and you
would like to opt out of receiving future advertisements or
solicitations from this BD group, please forward this e-mail to
optoutbygroup <@t> bd.com.
*******************************************************************
This message (which includes any attachments) is intended only for the
designated recipient(s). It may contain confidential or proprietary
information and may be subject to the attorney-client privilege or
other confidentiality protections. If you are not a designated
recipient, you may not review, use, copy or distribute this message.
If you received this in error, please notify the sender by reply
e-mail and delete this message. Thank you.
*******************************************************************
Corporate Headquarters Mailing Address: BD (Becton, Dickinson and
Company) 1 Becton Drive Franklin Lakes, NJ 07417 U.S.A.
*******************************************************************
------------------------------
Message: 6
Date: Mon, 12 Mar 2012 11:34:20 -0700
From: <Laurie <@t> blufrogpath.com>
Subject: [Histonet] Color Blindness Testing
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<20120312113420.295dc6182df7e5cbb4f32bc101c30dcc.f82e5fb846.wbe <@t> email15.secureserver.net>
Content-Type: text/plain; charset="utf-8"
Does anyone know of any (free) online testing for color blindness Does anyone have an alternate method that they have used to satisfy CAP requirements?
Laurie
------------------------------
Message: 7
Date: Mon, 12 Mar 2012 14:42:44 -0400
From: Drew Meyer <41dmb41 <@t> gmail.com>
Subject: Re: [Histonet] DAKO Autostainer Issues
To: "Friedrich_Hahn <@t> bd.com" <Friedrich_Hahn <@t> bd.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <5FAEEDF0-63D7-4EBC-9A3E-AEF9D5297130 <@t> gmail.com>
Content-Type: text/plain; charset=us-ascii
There's a circuit board on the arm near the barcode scanner that controls the movement and orientation of the arm. If that circuitboard malfunctions, then you can see some of the erratic behavior that you are describing. That's one possibility I can think of that might cause that kind of problem. However, the technician who looked at it should be able to troubleshoot and replace that board if it is a problem.
Drew
Sent from my iPhone
On Mar 12, 2012, at 2:31 PM, Friedrich_Hahn <@t> bd.com wrote:
>
> We have been having issues with our Autostainer since bringing it out
> of storage. All parts are in excellent shape, according to the
> technician that performed the PM. It worked fine for a month of so,
> but then began malfunctioning after only a couple of staining runs.
> At some point in the staining process, the machine appears to lose its
> XY homing and jams itself against the front right corner, dispensing
> buffer onto the floor of the chamber and occasionally making grinding
> noises. We've had a technician take a look at it, 'clean' the
> software, and we thought we had it resolved, but the problem appears
> to be back.
> Has anyone experienced this issue? If so, was it hardware- or
> software- based?
> Thanks in advance!
>
> -----------------------------------------
> *******************************************************************
> IMPORTANT MESSAGE FOR RECIPIENTS IN THE U.S.A.: This message may
> constitute an advertisement of a BD group's products or services or a
> solicitation of interest in them. If this is such a message and you
> would like to opt out of receiving future advertisements or
> solicitations from this BD group, please forward this e-mail to
> optoutbygroup <@t> bd.com.
> *******************************************************************
> This message (which includes any attachments) is intended only for the
> designated recipient(s). It may contain confidential or proprietary
> information and may be subject to the attorney-client privilege or
> other confidentiality protections. If you are not a designated
> recipient, you may not review, use, copy or distribute this message.
> If you received this in error, please notify the sender by reply
> e-mail and delete this message. Thank you.
> *******************************************************************
> Corporate Headquarters Mailing Address: BD (Becton, Dickinson and
> Company) 1 Becton Drive Franklin Lakes, NJ 07417 U.S.A.
> *******************************************************************
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 8
Date: Mon, 12 Mar 2012 11:49:10 -0700
From: "Jeffery Howery" <Jeffery.Howery <@t> jcl.com>
Subject: [Histonet] IHC for decalcified bone protocol
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<DB6A4527C2995D4288E5224B16012B6181E53A <@t> JCLNMMAIL12.jclnt.jcl.com>
Content-Type: text/plain; charset="us-ascii"
Anyone have a Protocol for Decalcified bone for IHC?
------------------------------
Message: 9
Date: Mon, 12 Mar 2012 20:51:26 +0200
From: Mehmet Fatih BOZKURT <fbozkurt <@t> gmail.com>
Subject: Re: [Histonet] IHC for decalcified bone protocol
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<CA+pSMGT6Uiy+xx08H=iT6kJTZcXu9VzqHo=1vcKSDZSztr7N4A <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
cool !!
On Mon, Mar 12, 2012 at 8:49 PM, Jeffery Howery <Jeffery.Howery <@t> jcl.com>wrote:
> Anyone have a Protocol for Decalcified bone for IHC?
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
--
Mehmet Fatih BOZKURT, DVM, PhD
Afyon Kocatepe University
Faculty of Veterinary Medicine
Department of Pathology
03030, ANS Campus
Afyonkarahisar-TURKEY
Tel: +902722281312-109
------------------------------
Message: 10
Date: Mon, 12 Mar 2012 13:08:55 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: RE: [Histonet] Gram Stain
To: "'Giroux, Stacy'" <Stacy.Giroux <@t> stjohn.org>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000001cd0083$93bf5c80$bb3e1580$@bresnan.net>
Content-Type: text/plain; charset="us-ascii"
Stacy,
You can buy a complete Brown and Brenn staining kit from Newcomer Supply
which is very good, and not change your method. The picric acid/acetone
mixture is included and you wouldn't have to store stock picric acid nor ether in the lab but just this small amount of Picric acid/acetone that
comes in the kit. Disposal should not be that much of a problem. They
also have a Hucker Twort Gram stain that uses acetone for destaining and you supply the acetone. Poly Scientific has Gram Stain kit (Hucker
modification) which uses Grams decolorizer, a mixture of acetone and alcohol. They have excellent staining kits too.
Gayle Callis
HTL/HT/MT(ASCP)
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Giroux, Stacy
Sent: Monday, March 12, 2012 11:36 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Gram Stain
Hi,
Our lab is currently transitioning from the Brown & Brenn gram stain due to no longer wanting to store picric acid due to its potential hazards. Our pathologists have requested a gram stain for paraffin embedded tissue that looks similar but does not use picric acid or ether. Does anyone have any suggestions on stains that could be used or gram stain kits that are available for purchase that are good?
Thank you for your help,
Stacy
CONFIDENTIALITY NOTICE:
This email message and any accompanying data or files is confidential and may contain privileged information intended only for the named recipient(s).
If you are not the intended recipient(s), you are hereby notified that the dissemination, distribution, and or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at the email address above, delete this email from your computer, and destroy any copies in any form immediately. Receipt by anyone other than the named recipient(s) is not a waiver of any attorney-client, work product, or other applicable privilege._______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 11
Date: Mon, 12 Mar 2012 15:35:48 -0400
From: Courtney Pierce <Courtney.Pierce <@t> quintiles.com>
Subject: [Histonet] IHC Slides
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<E596AA0B8A45794A8EF828F5158C44E24B31614F41 <@t> USADC-AMBXD00.quintiles.net>
Content-Type: text/plain; charset="us-ascii"
Having problems with IHC slides not looking the same between runs. Does it have to do with how long and at what temp they are in the oven for?
Courtney Pierce
IHC Specialist
Quintiles
Translational R&D - Oncology
Innovation
Navigating the new health
610 Oakmont Lane
Westmont, IL 60559
Office: + 630-203-6234
courtney.pierce <@t> quintiles.com
clinical | commercial | consulting | capital
********************** IMPORTANT--PLEASE READ ************************ This electronic message, including its attachments, is COMPANY CONFIDENTIAL and may contain PROPRIETARY or LEGALLY PRIVILEGED information. If you are not the intended recipient, you are hereby notified that any use, disclosure, copying, or distribution of this message or any of the information included in it is unauthorized and strictly prohibited. If you have received this message in error, please immediately notify the sender by reply e-mail and permanently delete this message and its attachments, along with any copies thereof. Thank you.
************************************************************************
------------------------------
Message: 12
Date: Mon, 12 Mar 2012 12:52:12 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] IHC Slides
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>, Courtney Pierce
<Courtney.Pierce <@t> quintiles.com>
Message-ID:
<1331581932.14770.YahooMailClassic <@t> web162101.mail.bf1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Not really. Time in the oven and temp is usually not an issue as long as the tissue is well fixed/processed/infiltrated.
Ren? J.
--- On Mon, 3/12/12, Courtney Pierce <Courtney.Pierce <@t> quintiles.com> wrote:
From: Courtney Pierce <Courtney.Pierce <@t> quintiles.com>
Subject: [Histonet] IHC Slides
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Date: Monday, March 12, 2012, 3:35 PM
Having problems with IHC slides not looking the same between runs. Does it have to do with how long and at what temp they are in the oven for?
Courtney Pierce
IHC Specialist
Quintiles
Translational R&D - Oncology
Innovation
Navigating the new health
610 Oakmont Lane
Westmont, IL 60559
Office: + 630-203-6234
courtney.pierce <@t> quintiles.com
clinical | commercial | consulting | capital
**********************? IMPORTANT--PLEASE READ? ************************ This electronic message, including its attachments, is COMPANY CONFIDENTIAL and may contain PROPRIETARY or LEGALLY PRIVILEGED information.? If you are not the intended recipient, you are hereby notified that any use, disclosure, copying, or distribution of this message or any of the information included in it is unauthorized and strictly prohibited.? If you have received this message in error, please immediately notify the sender by reply e-mail and permanently delete this message and its attachments, along with any copies thereof. Thank you.
************************************************************************
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 13
Date: Mon, 12 Mar 2012 23:38:11 +0000
From: "Tony Henwood (SCHN)" <tony.henwood <@t> health.nsw.gov.au>
Subject: [Histonet] RE: Gram Stain
To: "'Giroux, Stacy'" <Stacy.Giroux <@t> stjohn.org>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6D6BD1DE8A5571489398B392A38A715760A20038 <@t> xmdb02.nch.kids>
Content-Type: text/plain; charset="us-ascii"
Stacy,
There are several Gram stain variants that do not use Picric acid.
Tworts variant uses fast green and neutral red after the crystal violet-Iodine steps.
Brown-Hopps method uses Basic Fuchsin, Gallego's differentiator and 1.5% Tartrazine after the crystal-violet steps, to stain the gram negative bugs.
A simple red counterstain is to stain sections with 1% aqueous neutral red for 3 min after the crystal violet-Iodine steps
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Giroux, Stacy
Sent: Tuesday, 13 March 2012 4:36 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Gram Stain
Hi,
Our lab is currently transitioning from the Brown & Brenn gram stain due to no longer wanting to store picric acid due to its potential hazards. Our pathologists have requested a gram stain for paraffin embedded tissue that looks similar but does not use picric acid or ether. Does anyone have any suggestions on stains that could be used or gram stain kits that are available for purchase that are good?
Thank you for your help,
Stacy
CONFIDENTIALITY NOTICE:
This email message and any accompanying data or files is confidential and may contain privileged information intended only for the named recipient(s). If you are not the intended recipient(s), you are hereby notified that the dissemination, distribution, and or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at the email address above, delete this email from your computer, and destroy any copies in any form immediately. Receipt by anyone other than the named recipient(s) is not a waiver of any attorney-client, work product, or other applicable privilege._______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
*********************************************************************************
This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.
Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead
This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
*********************************************************************************
------------------------------
Message: 14
Date: Mon, 12 Mar 2012 18:19:14 -0700 (PDT)
From: Chakib Boussahmain <chak_bou <@t> yahoo.com>
Subject: [Histonet] CD34
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<1331601554.30627.YahooMailClassic <@t> web161804.mail.bf1.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Hello histonet,
Does anyone using CD34? if so, can you share the staining protocol with us? and what are you using for antigen retrieval?
Thank you so much.
Chakib Boussahmain
HTL(ASCP)
------------------------------
Message: 15
Date: Mon, 12 Mar 2012 18:25:44 -0700 (PDT)
From: enrriq88 <@t> yahoo.com
Subject: [Histonet] (no subject)
To: "Histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<1331601944.56861.YahooMailNeo <@t> web113503.mail.gq1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Please?unsubscribe.
thanks
------------------------------
Message: 16
Date: Mon, 12 Mar 2012 21:26:20 -0400
From: ADESUPO ADESUYI <adesupo2002 <@t> hotmail.com>
Subject: [Histonet] Liver Biopsy Processing Schedule
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <SNT136-W37FC7903FD34A8ECB5992EB6580 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
Hi,
I was wondering if anyone can share their LIVER Biopsy Processing Schedule with me?
Thanks,
Ade
------------------------------
Message: 17
Date: Tue, 13 Mar 2012 11:06:01 +0000
From: "Thotakura, Anil Kumar" <a.thotakura <@t> imperial.ac.uk>
Subject: [Histonet] CD45.1 & CD45.2 IF on parrafin sections.
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <CB84DA98.13C80%a.thotakura <@t> imperial.ac.uk>
Content-Type: text/plain; charset="us-ascii"
Dear All,
I want to do immunofluorescence on formalin fixed paraffin sections, Can you guys help me sending protocol how to do it ?
Thank you very much for your help.
BW
Anil
------------------------------
Message: 18
Date: Tue, 13 Mar 2012 08:29:17 -0500
From: "Bliven, Laura" <bliven.laura <@t> marshfieldclinic.org>
Subject: [Histonet] IHC Processor Validation & MUM1
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <201203131329.q2DDTXa8031218 <@t> mailhost3.mfldclin.edu>
Content-Type: text/plain; charset="iso-8859-1"
1. Any feedback on the process for validating a new tissue processor? Finding 80 to 100 breast specimens that are large enough to divide up and retest Estrogen and Progesterone would take a long time to complete. Also, how many antibodies and specimens should be tested? As we all know, some tumors are very rare and it's only after they are processed that we really know their diagnosis. We will be keeping some current processors, just adding a new one.
2. Also, any feedback on clones for MUM1? The antibody we are currently validating is staining some follicular lymphomas which should generally be negative. I do not feel the antigen retrieval is too harsh and believe it may be the clone.
Thanks,
Laura
______________________________________________________________________
The contents of this message may contain private, protected and/or privileged information. If you received this message in error, you should destroy the e-mail message and any attachments or copies, and you are prohibited from retaining, distributing, disclosing or using any information contained within. Please contact the sender and advise of the erroneous delivery by return e-mail or telephone. Thank you for your cooperation.
------------------------------
Message: 19
Date: Tue, 13 Mar 2012 08:54:27 -0500
From: Sabrina Townsend <histogirl4 <@t> gmail.com>
Subject: [Histonet] Ihc validation
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <27E73F93-AB37-4260-8BEA-35F23AF4F022 <@t> gmail.com>
Content-Type: text/plain; charset=us-ascii
I am having to validate antibodies and was wondering if you have to do a parallel study or just use known positive and negative tissue?
~Sabrina~
------------------------------
Message: 20
Date: Tue, 13 Mar 2012 10:50:23 -0400
From: Jack Ratliff <ratliffjack <@t> hotmail.com>
Subject: RE: [Histonet] Re: undecalcified bone IHC
To: Gayle Callis <gayle.callis <@t> bresnan.net>, Histonet
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BLU167-W13B54FA2D93AE7D10CB5E8AE580 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
I might also add that Neil Hand is co-speaking with myself and Philip Seifert this year at the annual National Society for Histotechnology - Symposium/Convention in Vancouver B.C. Our workshop is titled:
Resin Applications Forum: Methods for Processing, Special Staining, Immunohistochemical and In Situ Hybridization of Soft and Hard Tissue Including Medical Device Implants
During the last 50 years, numerous histological procedures have been described on resin embedded tissue. While different types of resins are available for different purposes, the acrylics provide the widest range of techniques, especially for light microscopy applications. However, as demand from H&E to more sophisticated techniques increases, so too have the problems, and nowhere is this more apparent and controversial than in the application of immunohistochemistry on resin sections. This workshop will provide a review and discussion for those individuals that currently work with and/or are just getting started working with soft and hard tissue specimens and specifically the various resins (i.e. MMA, GMA, Technovit, Acrylosin, etc.) associated with their specific tissue interests. The workshop will also detail the preparation and staining of sections of soft and hard tissue, including implants (e.g. undemineralized bone and cardiovascular stents), for immunohistochemical and in situ hybridization staining using different acrylic and epoxy resin embedding media. Specific problems and pitfalls, either technical or operational associated with certain resin embedding procedures, will be illustrated and examined. Particular emphasis will be given to procedures which have been used extensively for routine diagnostic, and research purposes, i.e. those that WORK! Individuals with a current or future intent to process and cut undemineralized tissue or tissue containing foreign implant materials using acrylic or epoxy resins are strongly encouraged to attend this workshop!
Please feel free to contact me if you would like more information about the workshop as information relevant to the exact date and time becomes available. All I know at this time is that the NSH meeting is September 29th - October 3rd, 2012.
Best Regards,
Jack
Jack Ratliff
Hard Tissue Histologist
Chairman, Hard Tissue Committee - National Society for Histotechnology
> From: gayle.callis <@t> bresnan.net
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Mon, 12 Mar 2012 11:04:20 -0600
> Subject: [Histonet] Re: undecalcified bone IHC
>
> Jeff,
>
>
>
> It is most certainly possible to do IHC on undecalcifed bone sections
> embedded in PMMA although not the easiest task. Sectioning is done on
> a microtome that is powerful enough to cut the plastic and using
> tungsten carbide knives. The key is total removal of the plastic from
> MMA embedded bone sections to allow antibody/ immunoglobulins to access antigenic sites.
> Neil Hand has done IHC successfully on PMMA embedded tissues including
> undecalcified bone on 2 to 3 ?m thick sections. I think one could cut
> thicker sections at 4 to 5 ?m and still be successful. I do not recall
> what Troiano et al used.
>
>
>
> The following publications will help you and should include protocols,
> although conventional protocols will work according to Hand.
>
>
>
> Blythe D. Hand N et al 1997 J Clin Path 50:45-49. The use of methyl
> methacrylate resin for embedding bone marrow trephine biopsies.
>
> Hand NM et al 1996 Antigen unmasking using microwave heating on
> formalin fixed tissue embedded in methyl methacrylate J Cellular Path
> 1:31-37
>
> Jackson P et al. 1996 Amplification of immunocytochemical reactions by
> the catalytic deposition of biotin on tissue sections. J Path
> 170(suppl):23A. This was about tyramide amplification when one gets a
> weak signal from "conventional" methods.
>
> Hand NM, Church RJ 1998 Superheating using pressure cooking: its use
> and application in unmasking antigens embedded in methyl methacrylate.
> J Histotechnology 2`:231-236
>
> Hand NM et al 1989 Immunohistochemistry on resin embedded tissue for
> light
> microscopy: a novel post embedding procedure. Proceeding Royal
> Microscopical Society 24(1):A54-55.
>
> Hand NM Plastic Embedding media and techniques, Ch.30, p 663-677.
> Theory and Practice of Histological Technique, 5th edition by Gamble and Bancroft.
> The 6th edition is updated under same title.
>
>
>
> Use Google Scholar to find Troiano N et al from Yale on doing IHC on
> PMMA embedded bone sections with publications in J Histotechnology.
>
>
>
>
>
> Hand mentioned several HIER methods, using citrate buffer. Optimizing
> retrieval will depend on the antigen and you may end up doing this
> with some form of HIER, including microwave or other heat producing
> methods and with different buffers. Enzyme digestion is also a possibility.
>
>
>
> Hand removed MMA with xylene, warm my speed up the removal, also more
> than one change for 10 - 20 minutes or longer. When I talked to him
> personally, he said he had used warm xylene although temperature was
> not mentioned in his chapter. After MMA removal, rehydrate section
> through alcohol gradient as one does paraffin sections. He was
> emphatic about never allowing the sections dry out.
>
>
>
> Hopefully Jack Ratliff and Damien Laudier will provide more insight on
> this topic.
>
>
>
> Good luck
>
>
>
> Gayle M. Callis
>
> HTL/HT/MT(ASCP)
>
>
>
>
>
>
>
>
>
>
>
>
>
> ******************************************
>
>
>
> Hi Jeff,
>
>
>
> If is it possible a few more specifics of how the tissue has been
> received,
>
> processed and evaluated would help. Undecalcified bone sectioning
>
> procedures vary and also what specific markers are you looking to do
> is
>
> important.
>
>
>
> Vikki
>
> On Mon, Mar 12, 2012 at 11:06 AM, Rene J Buesa <rjbuesa <@t>
> yahoo.com>
> wrote:
>
>
>
> > Undecalcified? How are you going to section it?
>
> > If you can section it, just use any IHC protocol for regular sections.
>
> > Good luck!
>
> > Ren? J.
>
> >
>
> > --- On Mon, 3/12/12, Jeffery Howery <Jeffery.Howery <@t> jcl.com> wrote:
>
> >
>
> >
>
> > From: Jeffery Howery <Jeffery.Howery <@t> jcl.com>
>
> > Subject: [Histonet] Undecalcified bone IHC
>
> > To: histonet <@t> lists.utsouthwestern.edu
>
> > Date: Monday, March 12, 2012, 10:59 AM
>
> >
>
> >
>
> > Does anyone have a protocol for Undecalcified bone for IHC?
>
> >
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 21
Date: Tue, 13 Mar 2012 08:06:35 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] CD34
To: histonet <@t> lists.utsouthwestern.edu, Chakib Boussahmain
<chak_bou <@t> yahoo.com>
Message-ID:
<1331651195.30522.YahooMailClassic <@t> web162106.mail.bf1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Becton-Dickinson Monoclonal antibody at 1:100 for 30 minutes after HIER (citrate at pH6) with placenta as control.
Ren? J.
--- On Mon, 3/12/12, Chakib Boussahmain <chak_bou <@t> yahoo.com> wrote:
From: Chakib Boussahmain <chak_bou <@t> yahoo.com>
Subject: [Histonet] CD34
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, March 12, 2012, 9:19 PM
Hello histonet,
Does anyone using CD34? if so, can you share the staining protocol with us? and what are you using for antigen retrieval?
Thank you so much.
Chakib Boussahmain
HTL(ASCP)
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 22
Date: Tue, 13 Mar 2012 08:25:50 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] IHC Processor Validation & MUM1
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>, LauraBliven
<bliven.laura <@t> marshfieldclinic.org>
Message-ID:
<1331652350.10883.YahooMailClassic <@t> web162101.mail.bf1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
CAP has some guidelines as to the number of specimens and antibodies to test. I think to remember that the lowest amount was 25 specimens, but I do not remember about the antibodies but it makes sense to use only those your use to work with.
Ren? J.
--- On Tue, 3/13/12, Bliven, Laura <bliven.laura <@t> marshfieldclinic.org> wrote:
From: Bliven, Laura <bliven.laura <@t> marshfieldclinic.org>
Subject: [Histonet] IHC Processor Validation & MUM1
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Date: Tuesday, March 13, 2012, 9:29 AM
1. Any feedback on the process for validating a new tissue processor? Finding 80 to 100 breast specimens that are large enough to divide up and retest Estrogen and Progesterone would take a long time to complete. Also, how many antibodies and specimens should be tested? As we all know, some tumors are very rare and it's only after they are processed that we really know their diagnosis. We will be keeping some current processors, just adding a new one.
2. Also, any feedback on clones for MUM1? The antibody we are currently validating is staining some follicular lymphomas which should generally be negative. I do not feel the antigen retrieval is too harsh and believe it may be the clone.
Thanks,
Laura
______________________________________________________________________
The contents of this message may contain private, protected and/or privileged information.? If you received this message in error, you should destroy the e-mail message and any attachments or copies, and you are prohibited from retaining, distributing, disclosing or using any information contained within.? Please contact the sender and advise of the erroneous delivery by return e-mail or telephone.? Thank you for your cooperation.
-----Inline Attachment Follows-----
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
End of Histonet Digest, Vol 100, Issue 15
*****************************************
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
More information about the Histonet
mailing list