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-----Original Message----- From: histonet-requ...@lists.utsouthwestern.edu [mailto:histonet-requ...@lists.utsouthwestern.edu] Sent: Thursday, January 03, 2019 10:00 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 182, Issue 3 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. IHC Negative reagent controls (Cayman Fleck) 2. Re: IHC Negative reagent controls (Tony Henwood (SCHN)) 3. AM to statlab and recommendations (Heather Marlatt) 4. Re: IHC Negative reagent controls (John Garratt) 5. IHC-H&E-IHC HELP... (Curt) ---------------------------------------------------------------------- Message: 1 Date: Thu, 3 Jan 2019 02:48:10 +0000 From: Cayman Fleck <caymanfl...@hotmail.com> To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] IHC Negative reagent controls Message-ID: <bn3pr13mb06734b9c15cc4ecbe24d41e9b1...@bn3pr13mb0673.namprd13.prod.outlook. com> Content-Type: text/plain; charset="iso-8859-1" A question that came up regarding negative reagent controls for IHC...currently using Ventana i-View. Our regular negative control goes through the standard antigen retrieval steps, like 99% of our antibodies. However there are a small number of antibodies that require enzyme as well (Protease 1). I've seen a number of suggestions regarding this for the negative reagent control...some say use an additional negative control protocol that includes the protease, some say to use a single negative control protocol and just include the harshest cell conditioning that any of your protocols use (so basically use the cell conditioning + protease negative control for all antibodies)...i-View is not polymer-based so we need to continue using negative controls. Any thoughts or advice? Frank Sent from Outlook<http://aka.ms/weboutlook> ------------------------------ Message: 2 Date: Thu, 3 Jan 2019 04:06:57 +0000 From: "Tony Henwood (SCHN)" <tony.henw...@health.nsw.gov.au> To: Cayman Fleck <caymanfl...@hotmail.com> Cc: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] IHC Negative reagent controls Message-ID: <0e5c3ac509b7424e942160149bb36...@svdcmbx-mex024.nswhealth.net> Content-Type: text/plain; charset="us-ascii" Hi Cayman, Unfortunately, applying HIER to a negative control for an antibody that requires enzyme retrieval (or no retrieval at all) is not appropriate. The pre-treatment processes are different and could unmask different epitopes. If you are using a negative control then the whole procedure needs to be same with the exception or replacing the localisation antibody with an Isotypic antibody solution. (Isotype controls are primary antibodies that lack specificity to the target, but match the class and type of the primary antibody used in the application.) For example, applying citrate or EDTA HIER to sections prior to using the CD99 antibody (clone 12E7) can reveal perinuclear (golgi-like) staining of some tumours (eg some colonic carcinomas) but this is not seen if enzyme retrieval is used. Hope this is useful Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: Cayman Fleck via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Thursday, 3 January 2019 1:48 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Negative reagent controls A question that came up regarding negative reagent controls for IHC...currently using Ventana i-View. Our regular negative control goes through the standard antigen retrieval steps, like 99% of our antibodies. However there are a small number of antibodies that require enzyme as well (Protease 1). I've seen a number of suggestions regarding this for the negative reagent control...some say use an additional negative control protocol that includes the protease, some say to use a single negative control protocol and just include the harshest cell conditioning that any of your protocols use (so basically use the cell conditioning + protease negative control for all antibodies)...i-View is not polymer-based so we need to continue using negative controls. Any thoughts or advice? Frank Sent from Outlook<http://aka.ms/weboutlook> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities. ------------------------------ Message: 3 Date: Thu, 3 Jan 2019 08:19:15 -0700 From: Heather Marlatt <hmarlat...@gmail.com> To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] AM to statlab and recommendations Message-ID: <CALaVVk4g1mm33v2qi=15UM4OqBkTg1wUA0P=t3p+bp6dpwl...@mail.gmail.com> Content-Type: text/plain; charset="UTF-8" Did anyone else use American Mastertech and is now super frustrated with Stat Lab??? I?ve used AM for years and loved them, great prices and fast shipping, now cost is through the roof and they?re shipping all my stuff separately costing a fortune! Looking for recommendations on xylene alternatives and safe mounting medium. I?ve been using transcend and cover safe two American Mastertech signature products but I?m running for my life. I?ll be contacting a few of the other vendors I use as well but I?d like some recommendations on xylene alternatives we?ve been xylene free for 4 years and I?m not going back. Thank you Heather ------------------------------ Message: 4 Date: Thu, 03 Jan 2019 16:41:03 +0000 From: John Garratt <john.garr...@ciqc.ca> To: Cayman Fleck <caymanfl...@hotmail.com> Cc: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] IHC Negative reagent controls Message-ID: <roDLdKVu1mgL2MG3gSdX9mEy3xKCrHdcA_Pdd2wm98b9jpgQrtaoHaAXgaNkvRgbfG6KujZU7ly CsZYaD2omb18Ivy0R15jWj3aL5Ws1jlg=@ciqc.ca> Content-Type: text/plain; charset=UTF-8 The article referenced below may be of interest. Standardization of Negative Controls in Diagnostic Immunohistochemistry: Recommendations From the International Ad Hoc Expert Panel https://www.researchgate.net/publication/261516067_Standardization_of_Negati ve_Controls_in_Diagnostic_Immunohistochemistry_Recommendations_From_the_Inte rnational_Ad_Hoc_Expert_Panel John www.ciqc.ca ??????? Original Message ??????? On Wednesday, January 2, 2019 6:48 PM, Cayman Fleck via Histonet <histonet@lists.utsouthwestern.edu> wrote: > A question that came up regarding negative reagent controls for IHC...currently using Ventana i-View. Our regular negative control goes through the standard antigen retrieval steps, like 99% of our antibodies. However there are a small number of antibodies that require enzyme as well (Protease 1). I've seen a number of suggestions regarding this for the negative reagent control...some say use an additional negative control protocol that includes the protease, some say to use a single negative control protocol and just include the harshest cell conditioning that any of your protocols use (so basically use the cell conditioning + protease negative control for all antibodies)...i-View is not polymer-based so we need to continue using negative controls. Any thoughts or advice? > > Frank > > Sent from Outlookhttp://aka.ms/weboutlook > > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Thu, 3 Jan 2019 17:05:21 +0000 From: Curt <c.ta...@pathologyarts.com> To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] IHC-H&E-IHC HELP... Message-ID: <9c8f910f72893643b3c3793c3d67132b94d03...@pathologyserver.pathologyarts.loca l> Content-Type: text/plain; charset="us-ascii" Need help before I turn a mistake into an irreparable mistake... We have some unstained slides that were supposed to get stained with H&E but my guy stained them with IHC. It's complicated, we received slides and a block, the block was for IHC, the unstained slides were for H&E, he inverted the process) The point is, now the unstained slides are stained with IHC... I know we cannot destain the IHC but we can simply run and H&E over them... the real question I have is subsequent to the H&E... this pathologist generally likes to see the H&E then order IHC on them based on what he sees (we only have these few unstained slides, don't have blocks to recut)... So the question is... if we've already run IHC, then followed that with and H&E, can we return to run IHC on the slides again? would you want to skip any pre-treatment, antigen retrieval???? I don't see this working too well myself, if they're already stained with DAB, that would be present on the second stain... Thoughts? Thanks for your help. Curt CONFIDENTIALITY NOTE: The information transmitted, including attachments, is intended only for the person(s) or entity to which it is addressed and may contain confidential and/or privileged material. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon this information by persons or entities other than the intended recipient is prohibited. If you received this in error, please contact the sender and destroy any copies of this information. ------------------------------ Subject: Digest Footer _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ End of Histonet Digest, Vol 182, Issue 3 **************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet