[Histonet] Re: Histonet Digest, Vol 99, Issue 19
Madeleine Huey
madeleinehuey <@t> gmail.com
Wed Feb 15 23:04:34 CST 2012
From: "del phillips" <dphillips <@t> vetmed.lsu.edu>
Subject: [Histonet] IHC and bone implants
Does anyone have any suggestions for staining bone implants? I am having
trouble with wash offs. I use charged slides and they are still washing off.
Thanks
Del
Del,
Since you did not mention what AR buffer & time for your IHC and
here's my suggestion;
1) HIER with Diva (Biocare) in a Pascal (or similar) pressure cooker for 3 min
2) Rinse slide immediately after pressure cooker is done (NO cooling
in room temperature for any length of time)
3) Proceed your IHC protocol
Note: If you still have problem after following the above
instruction, then try Fisher Scientific "Ultra Stick slides - Cat. #
???". It's relatively same price as regular charge slides, cheaper
than Dako IHC slides.
In the past, I have done IHC/IFC with 100 um Frozen section with cheap
pressure cooker & the thick section still stick to the slide with
excellent morphology.
Good Luck!
Madeleine Huey BS, HTL/QIHC
Supervisor - Pathology (IPOX & Histology)
madeleine_h <@t> elcaminohospital.org
On Wed, Feb 15, 2012 at 10:01 AM,
<histonet-request <@t> lists.utsouthwestern.edu> wrote:
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> Today's Topics:
>
> 1. Re: good source for APES to make silane treated slides?
> (Geoff McAuliffe)
> 2. wrinkles (wassan alkadhumi)
> 3. (no subject) (Ingles Claire )
> 4. Re: H. pylori (Richard Cartun)
> 5. RE: IHC and bone implants (Jack Ratliff)
> 6. Re: (no subject) (Rene J Buesa)
> 7. Re: wrinkles (Rene J Buesa)
> 8. QA by pathologists (Bob Richmond)
> 9. New DNA Probes for Kappa and Lambda (Vickroy, Jim)
> 10. RE: (no subject) (Tony Henwood (SCHN))
> 11. endogeneous peroxidase blocking (Marilyn Tyler)
> 12. (no subject) (Sheree H)
> 13. Meditech users (Sheila Adey)
> 14. Independent Service Technician (Stella Mireles)
> 15. Re: bone implants (Reynolds,Donna M)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Tue, 14 Feb 2012 13:44:55 -0500
> From: Geoff McAuliffe <mcauliff <@t> umdnj.edu>
> Subject: Re: [Histonet] good source for APES to make silane treated
> slides?
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <4F3AABA7.40709 <@t> umdnj.edu>
> Content-Type: text/plain; CHARSET=US-ASCII; format=flowed
>
> Sigma Chemical, catalog number A3648
>
> Geoff
>
>
> On 2/14/2012 11:18 AM, Joseph Madary wrote:
>>
>>
>>
>> Nick Madary, HT/HTL(ASCP)QIHC
>> George Washington University
>> Pathology Core Laboratory
>> Ross Hall, Room 706
>> 23rd and I Street NW
>> Washington D.C. 20037
>> 202.994.8916
>> patjnm <@t> gwumc.edu
>>
>>
>> _______________________________________________
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>
> --
> --
> **********************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane, Piscataway, NJ 08854
> voice: (732)-235-4583
> mcauliff <@t> umdnj.edu
> **********************************************
>
>
>
> ------------------------------
>
> Message: 2
> Date: Tue, 14 Feb 2012 10:44:33 -0800 (PST)
> From: wassan alkadhumi <w_alkadhumi <@t> yahoo.com>
> Subject: [Histonet] wrinkles
> To: h n <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <1329245073.26066.YahooMailNeo <@t> web45203.mail.sp1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Dear histonet members
> First I wish every one a happy valentine day!!!
> At our hospital the lab temperature is not controlled. In summer it is very hot and in winter it is cold. When I trim the paraffin embedded tissue my sections are faultless . Now because it is winter I have a problem having wrinkles in the sectionsafter collecting them from the water bath. Does anyone recommend me to buy slide drying hotplate for the slides?especially for the IHC slides, if you recommend me to buy it at what temperature should I use it and how it would effect the baking time afterward?
>
> Thanks
> Wassan/Histotechnican
> Shorsh General Hospital
> North of Iraq
>
> ------------------------------
>
> Message: 3
> Date: Tue, 14 Feb 2012 13:22:16 -0600
> From: "Ingles Claire " <CIngles <@t> uwhealth.org>
> Subject: [Histonet] (no subject)
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <064F1ACBAE8A78469AE2E41D533D87E505A7DC <@t> UWHC-MAIL2.uwhis.hosp.wisc.edu>
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hey histo gurus:
> Just a little twist for you. We are planning on doing GMS and PASD on frozen skin sections. Is there any change in staining protocol for paraffin vs frozen. We plan on fixing in ETOH. I have a co-worker saying that we don't need to use diastase on the PASD because there are already active "enzymes" in the tissue as it is fresh. He also said that we don't need to heat the silver for the GMS. Not sure why. These changes don't seem right to me. But I have been wrong before (Yes, I admit it). Thanks for all the wisdom out there floating in the electrons. We are staining for fungus BTW.
> Claire
>
>
>
> ------------------------------
>
> Message: 4
> Date: Tue, 14 Feb 2012 14:48:38 -0500
> From: "Richard Cartun" <Rcartun <@t> harthosp.org>
> Subject: Re: [Histonet] H. pylori
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>, "Toni Rathborne"
> <trathborne <@t> somerset-healthcare.com>
> Message-ID: <4F3A7445.7400.0077.1 <@t> harthosp.org>
> Content-Type: text/plain; charset=US-ASCII
>
> Keep in mind that many of the commercially-available monoclonal antibodies to H. pylori do not label Helicobacter helilmannii, often seen in children, especially those with exposure to animals.
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & Immunopathology
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT 06102
> (860) 545-1596 Office
> (860) 545-2204 Fax
>
>
>>>> "Rathborne, Toni" <trathborne <@t> somerset-healthcare.com> 2/14/2012 10:21 AM >>>
> Happy Valentines Day!
>
> Our lab is looking into ordering H. pylori now and I was wondering what clone, or whose antibody everyone is using.
> As always, thanks for your responses. I know I can always count on this group to offer opinions and suggestions :)
>
> Toni
>
>
>
>
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> ------------------------------
>
> Message: 5
> Date: Tue, 14 Feb 2012 15:36:39 -0500
> From: Jack Ratliff <ratliffjack <@t> hotmail.com>
> Subject: RE: [Histonet] IHC and bone implants
> To: <dphillips <@t> vetmed.lsu.edu>, Histonet
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <BLU167-W1480233215BC1A532B4323AE7C0 <@t> phx.gbl>
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> Del,
>
> I have a few questions for you to better understand your problem.
>
> 1) Are you talking about decalcified paraffin sections or undecalcified resin embedded specimens?
>
> 2) Is the implant washing off or both the bone and implant?
>
> 3) What type of implant/material is it?
>
> 4) If decalcified and paraffin embedded, what temperature and how long do you melt your specimens to the slide on the slide warmer?
>
> 5) If undecalcified and resin embedded, what type of resin and how thick are your sections?
>
> 6) Lastly, what step do you notice that the section or implant becomes detached from the slide?
>
> Please forgive me for the list of questions, but a little more information will help to pinpoint the problem and help get straight to the best corrective action.
>
> Best Regards,
>
> Jack
>
>
> Jack Ratliff
> Chairman, Hard Tissue Committee - National Society for Histotechnology
> Senior Histologist, Biomimetic Therapeutics, Inc.
>
>
>
>
>> From: dphillips <@t> vetmed.lsu.edu
>> To: Histonet <@t> lists.utsouthwestern.edu
>> Date: Mon, 13 Feb 2012 09:53:15 -0600
>> CC:
>> Subject: [Histonet] IHC and bone implants
>>
>> Does anyone have any suggestions for staining bone implants? I am having
>> trouble with wash offs. I use charged slides and they are still washing off.
>>
>>
>>
>> Thanks
>>
>> Del
>>
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 6
> Date: Tue, 14 Feb 2012 12:55:30 -0800 (PST)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] (no subject)
> To: histonet <@t> lists.utsouthwestern.edu, Ingles Claire
> <CIngles <@t> uwhealth.org>
> Message-ID:
> <1329252930.27538.YahooMailClassic <@t> web162102.mail.bf1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> About not using diastase for PASD because "there are active enzymes in the tissue": that is wrong because those are the same enzymes active in any tissue just before it is fixed. Also remember that you are going to fix with EthOL. The whole issue behind the PASD (as you well know) is to compare the results of the PAS with and without diastase, so your co-worker is wrong.
> As to not heating the silver for GMS I do see any reason to not doing that. If you do not heat it you will get something different to GMS. Perhaps your co-worker is fearful that the section may peel-off because of the heat but, other than that, there is no reason.
> René J.
>
>
>
>
> --- On Tue, 2/14/12, Ingles Claire <CIngles <@t> uwhealth.org> wrote:
>
>
> From: Ingles Claire <CIngles <@t> uwhealth.org>
> Subject: [Histonet] (no subject)
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Tuesday, February 14, 2012, 2:22 PM
>
>
> Hey histo gurus:
> Just a little twist for you. We are planning on doing GMS and PASD on frozen skin sections. Is there any change in staining protocol for paraffin vs frozen. We plan on fixing in ETOH. I have a co-worker saying that we don't need to use diastase on the PASD because there are already active "enzymes" in the tissue as it is fresh. He also said that we don't need to heat the silver for the GMS. Not sure why. These changes don't seem right to me. But I have been wrong before (Yes, I admit it). Thanks for all the wisdom out there floating in the electrons. We are staining for fungus BTW.
> Claire
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 7
> Date: Tue, 14 Feb 2012 12:58:24 -0800 (PST)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] wrinkles
> To: h n <histonet <@t> lists.utsouthwestern.edu>, wassan alkadhumi
> <w_alkadhumi <@t> yahoo.com>
> Message-ID:
> <1329253104.25837.YahooMailClassic <@t> web162102.mail.bf1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> If you consider that it\\your lab is cold now in winter, have you checked out the temperature in the water bath? Perhaps it is too cold to allow the sections to expand. If the water bath keeps its temperature (about 45ºC) the room temperature is irrelevant.
> René J.
>
> --- On Tue, 2/14/12, wassan alkadhumi <w_alkadhumi <@t> yahoo.com> wrote:
>
>
> From: wassan alkadhumi <w_alkadhumi <@t> yahoo.com>
> Subject: [Histonet] wrinkles
> To: "h n" <histonet <@t> lists.utsouthwestern.edu>
> Date: Tuesday, February 14, 2012, 1:44 PM
>
>
> Dear histonet members
> First I wish every one a happy valentine day!!!
> At our hospital the lab temperature is not controlled. In summer it is very hot and in winter it is cold. When I trim the paraffin embedded tissue my sections are faultless . Now because it is winter I have a problem having wrinkles in the sectionsafter collecting them from the water bath. Does anyone recommend me to buy slide drying hotplate for the slides?especially for the IHC slides, if you recommend me to buy it at what temperature should I use it and how it would effect the baking time afterward?
>
> Thanks
> Wassan/Histotechnican
> Shorsh General Hospital
> North of Iraq
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 8
> Date: Tue, 14 Feb 2012 16:33:36 -0500
> From: Bob Richmond <rsrichmond <@t> gmail.com>
> Subject: [Histonet] QA by pathologists
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> <CAOKsRH7UME8Tzq65sA_tEWjozDt+t_PKiU90LiFxfuTQiGL5pw <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Akemi Allison asks:
>
>>>What % QA do most pathologists do internally? I'm trying to update my AP Manual. My last CLIA inspection, they told me to put in my AP Manual that it is done and get a binder labeled "discrepancies". If there are any...<<
>
> As far as I know, there is no percentage requirement like the
> burdensome and useless 10% rescreen on Pap smears. In my travels I've
> seen:
>
> One three-pathologist group did 100% review before sign-out.
>
> At least two services I've seen did a 10% review, cases chosen at
> random. This seemed really pointless.
>
> One service did a 10% review, with the originating pathologist
> choosing the cases to be reviewed. This fairly productive.
>
> One five-pathologist service meticulously documented informal
> consultation, both in the report and on a separate piece of paper.
> They aimed at, and got, a 2.8% review. In a multiple pathologist
> practice, I think this was the most productive method I ever saw.
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
>
>
>
> ------------------------------
>
> Message: 9
> Date: Tue, 14 Feb 2012 16:31:29 -0600
> From: "Vickroy, Jim" <Vickroy.Jim <@t> mhsil.com>
> Subject: [Histonet] New DNA Probes for Kappa and Lambda
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <24A4826E8EF0964D86BC5317306F58A55FDC5DE697 <@t> mmc-mail.ad.mhsil.com>
> Content-Type: text/plain; charset="us-ascii"
>
> In the past we have used a cocktail DNA probe (ISH) for Kappa and Lambda. Has anybody had any experience validating and setting up the new probes from Ventana which we have to mix the four probes together and form our own cocktail? Please share with me your thoughts. I am seriously considering sending this test out because of the infrequency, time, ASR hassle, costs of separate detection kits, etc.
>
> Jim
>
> James Vickroy BS, HT(ASCP)
>
> Surgical and Autopsy Pathology Technical Supervisor
> Memorial Medical Center
> 217-788-4046
>
>
> ________________________________
> This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited.
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>
> ------------------------------
>
> Message: 10
> Date: Tue, 14 Feb 2012 23:04:54 +0000
> From: "Tony Henwood (SCHN)" <tony.henwood <@t> health.nsw.gov.au>
> Subject: RE: [Histonet] (no subject)
> To: "'Ingles Claire '" <CIngles <@t> uwhealth.org>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <6D6BD1DE8A5571489398B392A38A715760A1AAFF <@t> xmdb02.nch.kids>
> Content-Type: text/plain; charset="us-ascii"
>
> The reason for using diastase is to remove the glycogen from the tissues. AND if you do not heat the methenamine silver solution for the GMS it will take forever to stain, in fact all you will get is background spots. This I can verify, have you ever forgot to turn on the water bath before staining? - several hours later - nuthin!!
>
> Who ever is giving you the advice I would wonder about their histotechnology education and training (in fact have they had any training at all?).
>
> Do not listen to them. Extrapolate from cytology. GMS and DiPAS have been done on cytology smears fixed in ethanol as well as air-dried for years see:
>
> Loughman NT. "Pneumocystis carinii: rapid diagnosis with the microwave oven" Acta Cytologica. 33(3):416-7, 1989 May-Jun.
>
> PINTOZZI, R. L. "Modified Grocott's methenamine silver nitrate method for quick staining of Pneumocystis carinii" J Clin Pathol 1978 31: 803-805
>
> 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 Ingles Claire
> Sent: Wednesday, 15 February 2012 6:22 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] (no subject)
>
> Hey histo gurus:
> Just a little twist for you. We are planning on doing GMS and PASD on frozen skin sections. Is there any change in staining protocol for paraffin vs frozen. We plan on fixing in ETOH. I have a co-worker saying that we don't need to use diastase on the PASD because there are already active "enzymes" in the tissue as it is fresh. He also said that we don't need to heat the silver for the GMS. Not sure why. These changes don't seem right to me. But I have been wrong before (Yes, I admit it). Thanks for all the wisdom out there floating in the electrons. We are staining for fungus BTW.
> Claire
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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>
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>
>
> ------------------------------
>
> Message: 11
> Date: Wed, 15 Feb 2012 08:14:10 +0200
> From: "Marilyn Tyler" <Marilyn.Tyler <@t> uct.ac.za>
> Subject: [Histonet] endogeneous peroxidase blocking
> To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <4F3B6952020000900012F35B <@t> gwiasmtp.uct.ac.za>
> Content-Type: text/plain; charset="windows-1252"
>
> Morning All
>
> Posing this question for a colleague. Suggestions for
> immunohistochemistry blocking for endogeneous peroxidase probably caused
> by red blood cells in mouse lung tissue. Using 1% hydrogen
> peroxide/dist.water for 15mins and still getting non specific staining.
>
>
> Thank you
>
> Marilyn
>
>
> Medical School UCT
> Dept of Surgery J50/26 or 30
> Surgical research. OMB
> Groote Schuur Hospital
> Observatory
>
> 021-4066227
>
>
>
>
>
> ###
>
> UNIVERSITY OF CAPE TOWN
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> ------------------------------
>
> Message: 12
> Date: Wed, 15 Feb 2012 01:27:14 -0800 (PST)
> From: Sheree H <equineshowmom01 <@t> yahoo.com>
> Subject: [Histonet] (no subject)
> To: dennis.barry <@t> thrivent.com, lmphillips <@t> charter.net,
> melissa.zak-walser <@t> jacobs.com, shiloy <@t> charter.net,
> histonet <@t> lists.utsouthwestern.edu, skiptokatblack <@t> bellsouth.net,
> agianakas <@t> bellsouth.net
> Message-ID:
> <1329298034.80174.YahooMailMobile <@t> web125806.mail.ne1.yahoo.com>
> Content-Type: text/plain; charset=us-ascii
>
> http://www.kingtigertkdwilson.com/modules/mod_wdbanners/myfriends.php?central164.gif
>
> ------------------------------
>
> Message: 13
> Date: Wed, 15 Feb 2012 05:52:19 -0500
> From: Sheila Adey <sadey <@t> hotmail.ca>
> Subject: [Histonet] Meditech users
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <BAY154-W170EA5670179EC364AA7B5C67D0 <@t> phx.gbl>
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> Hello:Are any of the Meditech users having the pathologists order their levels in the computer?We would like to implement this procedure. Any suggestions. :)ThanksSheila
>
> ------------------------------
>
> Message: 14
> Date: Wed, 15 Feb 2012 09:43:50 -0600
> From: Stella Mireles <estellamireles <@t> gmail.com>
> Subject: [Histonet] Independent Service Technician
> To: Histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> <CABm4S9PMnoaVZThYMTPJMc4k3LQ2bAvJ2+VfyQnX83Rh50jM1Q <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Is there anyone out there that does repairs on Sakura equipment and works
> for themselves
> in the Texas area.
>
>
> ------------------------------
>
> Message: 15
> Date: Wed, 15 Feb 2012 10:49:27 -0600
> From: "Reynolds,Donna M" <dreynold <@t> mdanderson.org>
> Subject: [Histonet] Re: bone implants
> To: "'histonet <@t> lists.utsouthwestern.edu'"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <785BBF0C5F49CE41BA74460A43A08F023055EE4325 <@t> DCPWVMBXC0VS3.mdanderson.edu>
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> I don't know about implants but we were having a terrible time with bone marrows coming off. Usually they came off during the HEIR using the steamer.
> At that point we were placing our slides lying flat in a 55-60º oven overnight which usually helps tremendously with difficult tissue. This was followed with routine deparaffinization and hydration. It was suggested to us to try a 70º water bath for 2-4 hours for HEIR. I was doubtful that my antibodies would work but they did. In fact the staining was better with 2 hr HEIR than with 4.
>
> Donna Reynolds Chief Histology laboratory, Research
> Department Cancer Biology
> U.T. M.D. Anderson Cancer Center, Houston, TX.
> 713-792-8106
>
> Message: 4
> Date: Mon, 13 Feb 2012 09:53:15 -0600
> From: "del phillips" <dphillips <@t> vetmed.lsu.edu>
> Subject: [Histonet] IHC and bone implants
> To: <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000a01ccea67$9958da90$cc0a8fb0$@vetmed.lsu.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Does anyone have any suggestions for staining bone implants? I am having
> trouble with wash offs. I use charged slides and they are still washing off.
>
>
>
> Thanks
>
> Del
>
>
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>
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>
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> End of Histonet Digest, Vol 99, Issue 19
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