You probably have to have someone administer it and verify it to make it acceptable for CAP, though.
-----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephen G. Ruby Sent: Tuesday, March 13, 2012 11:37 AM To: histonet@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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Tuesday, March 13, 2012 10:32 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 100, Issue 15 Send Histonet mailing list submissions to histonet@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-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@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_h...@bd.com) 6. Color Blindness Testing (lau...@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) (enrri...@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.cal...@bresnan.net> Subject: [Histonet] Re: undecalcified bone IHC To: "'Histonet'" <histonet@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 <bakevicto...@gmail.com> Subject: Re: [Histonet] Re: undecalcified bone IHC To: gayle callis <gayle.cal...@bresnan.net> Cc: Histonet <histonet@lists.utsouthwestern.edu> Message-ID: <calyzrmqlbjtk4wpfeotzw6zmfjwmdwute0b5xzmjslqgfgo...@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.cal...@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@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 3 Date: Mon, 12 Mar 2012 12:35:07 -0500 From: "Konop, Nicole" <nko...@chw.org> Subject: [Histonet] Unsubscribe To: "'histonet-requ...@lists.utsouthwestern.edu'" <histonet-requ...@lists.utsouthwestern.edu>, "'histonet@lists.utsouthwestern.edu'" <histonet@lists.utsouthwestern.edu> Message-ID: <a733875c1aa92d4ebd9a35a9506dfe3a5daf600...@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.gir...@stjohn.org> Subject: [Histonet] Gram Stain To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <29ccf3ec44815745864d9f84a1ed4b51b3e0709...@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_h...@bd.com Subject: [Histonet] DAKO Autostainer Issues To: histonet@lists.utsouthwestern.edu Message-ID: <of3e9d799b.1e276f03-on852579bf.0064fb8c-852579bf.0065c...@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 optoutbygr...@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: <lau...@blufrogpath.com> Subject: [Histonet] Color Blindness Testing To: histonet@lists.utsouthwestern.edu Message-ID: <20120312113420.295dc6182df7e5cbb4f32bc101c30dcc.f82e5fb846....@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 <41dm...@gmail.com> Subject: Re: [Histonet] DAKO Autostainer Issues To: "friedrich_h...@bd.com" <friedrich_h...@bd.com> Cc: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <5faeedf0-63d7-4ebc-9a3e-aef9d5297...@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_h...@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 > optoutbygr...@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@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.how...@jcl.com> Subject: [Histonet] IHC for decalcified bone protocol To: <histonet@lists.utsouthwestern.edu> Message-ID: <db6a4527c2995d4288e5224b16012b6181e...@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 <fbozk...@gmail.com> Subject: Re: [Histonet] IHC for decalcified bone protocol To: Histonet@lists.utsouthwestern.edu Message-ID: <CA+pSMGT6Uiy+xx08H=iT6kJTZcXu9VzqHo=1vcksdzsztr7...@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 cool !! On Mon, Mar 12, 2012 at 8:49 PM, Jeffery Howery <jeffery.how...@jcl.com>wrote: > Anyone have a Protocol for Decalcified bone for IHC? > > _______________________________________________ > Histonet mailing list > Histonet@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.cal...@bresnan.net> Subject: RE: [Histonet] Gram Stain To: "'Giroux, Stacy'" <stacy.gir...@stjohn.org>, <histonet@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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Giroux, Stacy Sent: Monday, March 12, 2012 11:36 AM To: histonet@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@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.pie...@quintiles.com> Subject: [Histonet] IHC Slides To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <e596aa0b8a45794a8ef828f5158c44e24b31614...@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.pie...@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 <rjbu...@yahoo.com> Subject: Re: [Histonet] IHC Slides To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu>, Courtney Pierce <courtney.pie...@quintiles.com> Message-ID: <1331581932.14770.yahoomailclas...@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.pie...@quintiles.com> wrote: From: Courtney Pierce <courtney.pie...@quintiles.com> Subject: [Histonet] IHC Slides To: "histonet@lists.utsouthwestern.edu" <histonet@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.pie...@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@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.henw...@health.nsw.gov.au> Subject: [Histonet] RE: Gram Stain To: "'Giroux, Stacy'" <stacy.gir...@stjohn.org>, "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <6d6bd1de8a5571489398b392a38a715760a20...@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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Giroux, Stacy Sent: Tuesday, 13 March 2012 4:36 AM To: histonet@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@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_...@yahoo.com> Subject: [Histonet] CD34 To: histonet@lists.utsouthwestern.edu Message-ID: <1331601554.30627.yahoomailclas...@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: enrri...@yahoo.com Subject: [Histonet] (no subject) To: "Histonet@lists.utsouthwestern.edu" <Histonet@lists.utsouthwestern.edu> Message-ID: <1331601944.56861.yahoomail...@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 <adesupo2...@hotmail.com> Subject: [Histonet] Liver Biopsy Processing Schedule To: <histonet@lists.utsouthwestern.edu> Message-ID: <snt136-w37fc7903fd34a8ecb5992eb6...@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.thotak...@imperial.ac.uk> Subject: [Histonet] CD45.1 & CD45.2 IF on parrafin sections. To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <cb84da98.13c80%a.thotak...@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.la...@marshfieldclinic.org> Subject: [Histonet] IHC Processor Validation & MUM1 To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <201203131329.q2ddtxa8031...@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 <histogi...@gmail.com> Subject: [Histonet] Ihc validation To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <27e73f93-ab37-4260-8bea-35f23af4f...@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 <ratliffj...@hotmail.com> Subject: RE: [Histonet] Re: undecalcified bone IHC To: Gayle Callis <gayle.cal...@bresnan.net>, Histonet <histonet@lists.utsouthwestern.edu> Message-ID: <blu167-w13b54fa2d93ae7d10cb5e8ae...@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.cal...@bresnan.net > To: histonet@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@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 <rjbu...@yahoo.com> Subject: Re: [Histonet] CD34 To: histonet@lists.utsouthwestern.edu, Chakib Boussahmain <chak_...@yahoo.com> Message-ID: <1331651195.30522.yahoomailclas...@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_...@yahoo.com> wrote: From: Chakib Boussahmain <chak_...@yahoo.com> Subject: [Histonet] CD34 To: histonet@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@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 <rjbu...@yahoo.com> Subject: Re: [Histonet] IHC Processor Validation & MUM1 To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu>, LauraBliven <bliven.la...@marshfieldclinic.org> Message-ID: <1331652350.10883.yahoomailclas...@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.la...@marshfieldclinic.org> wrote: From: Bliven, Laura <bliven.la...@marshfieldclinic.org> Subject: [Histonet] IHC Processor Validation & MUM1 To: "histonet@lists.utsouthwestern.edu" <histonet@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.? 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