[Histonet] techniques for archaeological bone
Hi all... Had someone in here who is interested in sectioning old bones - any ideas as to what would be the best embedding medium? These are bones from a graveyard more than 100 years old - not fossils. Decal is out of the question - so it would have to be a resin of some sort. Tips, ideas, contacts and methods would be very welcome at this stage -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa +27 11 717 2298 (tel fax) 073 5574456 (emergencies only) Question: Are rhinos overweight unicorns? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Secondary antibody causing nuclear staining?
I understand the point about the biotin and I should have said that when using the ABC method we have taken to always using an avidin/biotin blocking kit. We are using biotinylated secondary antibodies from Vector. I have seen the same problem occur in our anti-mouse, anti-rabbit and anti-goat. In my last run I had stomach fundus as well as skin melanoma, both had pos.nuclei in the negative (no primary). In another run I had colon ca and breast ca, the breast ca had fewer pos. nuclei than the colon ca but they were still there. Some days the positive nuclei are stronger in a sample that was just weakly positive before. Just want to understand what it is and what effects it. Thank you all for your ideas. Eva Permaul Georgetown University On Mon, Jul 23, 2012 at 7:16 PM, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: I should have added that this was from the workshop notes on a Hypotheticals Workshop I ran last year at our Australian National Meeting. 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 Tony Henwood (SCHN) Sent: Tuesday, 24 July 2012 9:00 AM To: 'Eva Permaul'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Secondary antibody causing nuclear staining? It is possible that this is due to Biotin nuclei where excess biotin is found in the nuclei of some cells, see below: Optically clear nuclei have been reported in endometrial epithelium associated with first and second trimester abortions (Sickel di Sant'Agnese 1994). Optically clear nuclei have also been found in different types of tissues of diverse organs such as ovary, thyroid and lung (Nakatani et al 1994, Mount Cooper 2001). The optically clear nuclei contain excess biotin. Endogenous biotin immunoreactivity is generally not visualized in formalin fixed, paraffin-embedded tissues unless a heat-induced antigen retrieval step has been introduced (Mount Cooper 2001). In this placental section, optically clear nuclei (containing biotin) bind to the streptavidin of the ABC technique giving a reaction similar to that seen with CMV containing cells. If a polymer method (or even the original Sternberger's PAP method) is used then this anomalous staining will disappear, thus allowing confident demonstration of CMV infected nuclei. The false-positive staining pattern caused by endogenous biotin can be cytoplasmic or nuclear. A report of positive immunoreactivity of hepatocellular carcinomas for inhibin was later determined to be a false-positive finding due to cytoplasmic endogenous biotin. Steroid cell tumours of the ovary were found to demonstrate endogenous biotin cytoplasmic staining in 36% of cases. Immunoreactivity for anti-Herpes virus immunohistochemical staining in a series of endometria was also later determined to be a false-positive result due to biotin. The prominent intranuclear inclusions, resembling herpes virus cytopathic effect, were caused by intranuclear biotin and not viral particles. Similar false positive staining for CMV in products of conception has also been reported (Mount Cooper 2001). False-positive staining can be cytoplasmic or nuclear. When cytoplasmic, the appearance of the false signal is that of a dull brown granular or fluffy staining pattern. If this quality of staining is observed with several different antibodies, endogenous staining by biotin should be considered. When nuclear, a false-positive reaction may be associated with optically clear nuclei identified on HE stained sections. False-positive staining due to endogenous biotin, however, does not occur in a cell membrane pattern (Mount Cooper 2001). Mount SL Cooper K (2001) Beware of biotin: a source of false-positive immunohistochemistry Current Diagnostic Pathology 7:161-167. Nakatani et al (1994) Am J Surg Pathol 18(6):637-642. Sickel di Sant'Agnese (1994) Arch Pathol Lab Med 118:831-833 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 Eva Permaul Sent: Monday, 23 July 2012 11:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Secondary antibody causing nuclear staining? Hello, I have noticed that our biotinylated secondary antibodies on occasion cause nuclear staining in some samples.
Re: [Histonet] Secondary antibody causing nuclear staining?
Are you getting false positives and variations on the same control tissue for different days ? Sent from my iPhone On Jul 24, 2012, at 8:13 AM, Eva Permaul e...@georgetown.edu wrote: I understand the point about the biotin and I should have said that when using the ABC method we have taken to always using an avidin/biotin blocking kit. We are using biotinylated secondary antibodies from Vector. I have seen the same problem occur in our anti-mouse, anti-rabbit and anti-goat. In my last run I had stomach fundus as well as skin melanoma, both had pos.nuclei in the negative (no primary). In another run I had colon ca and breast ca, the breast ca had fewer pos. nuclei than the colon ca but they were still there. Some days the positive nuclei are stronger in a sample that was just weakly positive before. Just want to understand what it is and what effects it. Thank you all for your ideas. Eva Permaul Georgetown University On Mon, Jul 23, 2012 at 7:16 PM, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: I should have added that this was from the workshop notes on a Hypotheticals Workshop I ran last year at our Australian National Meeting. 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 Tony Henwood (SCHN) Sent: Tuesday, 24 July 2012 9:00 AM To: 'Eva Permaul'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Secondary antibody causing nuclear staining? It is possible that this is due to Biotin nuclei where excess biotin is found in the nuclei of some cells, see below: Optically clear nuclei have been reported in endometrial epithelium associated with first and second trimester abortions (Sickel di Sant'Agnese 1994). Optically clear nuclei have also been found in different types of tissues of diverse organs such as ovary, thyroid and lung (Nakatani et al 1994, Mount Cooper 2001). The optically clear nuclei contain excess biotin. Endogenous biotin immunoreactivity is generally not visualized in formalin fixed, paraffin-embedded tissues unless a heat-induced antigen retrieval step has been introduced (Mount Cooper 2001). In this placental section, optically clear nuclei (containing biotin) bind to the streptavidin of the ABC technique giving a reaction similar to that seen with CMV containing cells. If a polymer method (or even the original Sternberger's PAP method) is used then this anomalous staining will disappear, thus allowing confident demonstration of CMV infected nuclei. The false-positive staining pattern caused by endogenous biotin can be cytoplasmic or nuclear. A report of positive immunoreactivity of hepatocellular carcinomas for inhibin was later determined to be a false-positive finding due to cytoplasmic endogenous biotin. Steroid cell tumours of the ovary were found to demonstrate endogenous biotin cytoplasmic staining in 36% of cases. Immunoreactivity for anti-Herpes virus immunohistochemical staining in a series of endometria was also later determined to be a false-positive result due to biotin. The prominent intranuclear inclusions, resembling herpes virus cytopathic effect, were caused by intranuclear biotin and not viral particles. Similar false positive staining for CMV in products of conception has also been reported (Mount Cooper 2001). False-positive staining can be cytoplasmic or nuclear. When cytoplasmic, the appearance of the false signal is that of a dull brown granular or fluffy staining pattern. If this quality of staining is observed with several different antibodies, endogenous staining by biotin should be considered. When nuclear, a false-positive reaction may be associated with optically clear nuclei identified on HE stained sections. False-positive staining due to endogenous biotin, however, does not occur in a cell membrane pattern (Mount Cooper 2001). Mount SL Cooper K (2001) Beware of biotin: a source of false-positive immunohistochemistry Current Diagnostic Pathology 7:161-167. Nakatani et al (1994) Am J Surg Pathol 18(6):637-642. Sickel di Sant'Agnese (1994) Arch Pathol Lab Med 118:831-833 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 Eva Permaul Sent: Monday, 23 July 2012 11:40 PM To:
Re: [Histonet] Secondary antibody causing nuclear staining?
We standard use a Citrate pH6. We do 20min at 98C followed by cooling in the citrate for 20min. Eva On Tue, Jul 24, 2012 at 8:29 AM, James Burchette Jr. james.burche...@duke.edu wrote: What is your heat retrieval process? Jim Burchette, HT(ASCP) QIHC Histologist and Fly Fishing Bum Orlando, Florida From: histonet-boun...@lists.utsouthwestern.edu [ histonet-boun...@lists.utsouthwestern.edu] on behalf of Eva Permaul [ e...@georgetown.edu] Sent: Tuesday, July 24, 2012 8:13 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Secondary antibody causing nuclear staining? I understand the point about the biotin and I should have said that when using the ABC method we have taken to always using an avidin/biotin blocking kit. We are using biotinylated secondary antibodies from Vector. I have seen the same problem occur in our anti-mouse, anti-rabbit and anti-goat. In my last run I had stomach fundus as well as skin melanoma, both had pos.nuclei in the negative (no primary). In another run I had colon ca and breast ca, the breast ca had fewer pos. nuclei than the colon ca but they were still there. Some days the positive nuclei are stronger in a sample that was just weakly positive before. Just want to understand what it is and what effects it. Thank you all for your ideas. Eva Permaul Georgetown University On Mon, Jul 23, 2012 at 7:16 PM, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: I should have added that this was from the workshop notes on a Hypotheticals Workshop I ran last year at our Australian National Meeting. 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 Tony Henwood (SCHN) Sent: Tuesday, 24 July 2012 9:00 AM To: 'Eva Permaul'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Secondary antibody causing nuclear staining? It is possible that this is due to Biotin nuclei where excess biotin is found in the nuclei of some cells, see below: Optically clear nuclei have been reported in endometrial epithelium associated with first and second trimester abortions (Sickel di Sant'Agnese 1994). Optically clear nuclei have also been found in different types of tissues of diverse organs such as ovary, thyroid and lung (Nakatani et al 1994, Mount Cooper 2001). The optically clear nuclei contain excess biotin. Endogenous biotin immunoreactivity is generally not visualized in formalin fixed, paraffin-embedded tissues unless a heat-induced antigen retrieval step has been introduced (Mount Cooper 2001). In this placental section, optically clear nuclei (containing biotin) bind to the streptavidin of the ABC technique giving a reaction similar to that seen with CMV containing cells. If a polymer method (or even the original Sternberger's PAP method) is used then this anomalous staining will disappear, thus allowing confident demonstration of CMV infected nuclei. The false-positive staining pattern caused by endogenous biotin can be cytoplasmic or nuclear. A report of positive immunoreactivity of hepatocellular carcinomas for inhibin was later determined to be a false-positive finding due to cytoplasmic endogenous biotin. Steroid cell tumours of the ovary were found to demonstrate endogenous biotin cytoplasmic staining in 36% of cases. Immunoreactivity for anti-Herpes virus immunohistochemical staining in a series of endometria was also later determined to be a false-positive result due to biotin. The prominent intranuclear inclusions, resembling herpes virus cytopathic effect, were caused by intranuclear biotin and not viral particles. Similar false positive staining for CMV in products of conception has also been reported (Mount Cooper 2001). False-positive staining can be cytoplasmic or nuclear. When cytoplasmic, the appearance of the false signal is that of a dull brown granular or fluffy staining pattern. If this quality of staining is observed with several different antibodies, endogenous staining by biotin should be considered. When nuclear, a false-positive reaction may be associated with optically clear nuclei identified on HE stained sections. False-positive staining due to endogenous biotin, however, does not occur in a cell membrane pattern (Mount Cooper 2001). Mount SL Cooper K (2001) Beware of biotin: a source of false-positive immunohistochemistry Current Diagnostic Pathology 7:161-167. Nakatani et al (1994) Am J Surg Pathol 18(6):637-642. Sickel di Sant'Agnese (1994) Arch Pathol
Re: [Histonet] Secondary antibody causing nuclear staining?
These samples were all human but I have seen it in Mouse mammary gland as well but those nuclei were lighter. Eva On Tue, Jul 24, 2012 at 8:51 AM, James Burchette Jr. james.burche...@duke.edu wrote: Thanks Eva. I don't know why you are having the nuclear staining problem. Your retrieval process isn't overly aggressive. I've used Vectors secondary and ABC reagents forever and have never had the issue you are describing. Human tissue? Jim Burchette, HT(ASCP) QIHC Histologist and Fly Fishing Bum Orlando, Florida From: histonet-boun...@lists.utsouthwestern.edu [ histonet-boun...@lists.utsouthwestern.edu] on behalf of Eva Permaul [ e...@georgetown.edu] Sent: Tuesday, July 24, 2012 8:41 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Secondary antibody causing nuclear staining? We standard use a Citrate pH6. We do 20min at 98C followed by cooling in the citrate for 20min. Eva On Tue, Jul 24, 2012 at 8:29 AM, James Burchette Jr. james.burche...@duke.edu wrote: What is your heat retrieval process? Jim Burchette, HT(ASCP) QIHC Histologist and Fly Fishing Bum Orlando, Florida From: histonet-boun...@lists.utsouthwestern.edu [ histonet-boun...@lists.utsouthwestern.edu] on behalf of Eva Permaul [ e...@georgetown.edu] Sent: Tuesday, July 24, 2012 8:13 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Secondary antibody causing nuclear staining? I understand the point about the biotin and I should have said that when using the ABC method we have taken to always using an avidin/biotin blocking kit. We are using biotinylated secondary antibodies from Vector. I have seen the same problem occur in our anti-mouse, anti-rabbit and anti-goat. In my last run I had stomach fundus as well as skin melanoma, both had pos.nuclei in the negative (no primary). In another run I had colon ca and breast ca, the breast ca had fewer pos. nuclei than the colon ca but they were still there. Some days the positive nuclei are stronger in a sample that was just weakly positive before. Just want to understand what it is and what effects it. Thank you all for your ideas. Eva Permaul Georgetown University On Mon, Jul 23, 2012 at 7:16 PM, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: I should have added that this was from the workshop notes on a Hypotheticals Workshop I ran last year at our Australian National Meeting. 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 Tony Henwood (SCHN) Sent: Tuesday, 24 July 2012 9:00 AM To: 'Eva Permaul'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Secondary antibody causing nuclear staining? It is possible that this is due to Biotin nuclei where excess biotin is found in the nuclei of some cells, see below: Optically clear nuclei have been reported in endometrial epithelium associated with first and second trimester abortions (Sickel di Sant'Agnese 1994). Optically clear nuclei have also been found in different types of tissues of diverse organs such as ovary, thyroid and lung (Nakatani et al 1994, Mount Cooper 2001). The optically clear nuclei contain excess biotin. Endogenous biotin immunoreactivity is generally not visualized in formalin fixed, paraffin-embedded tissues unless a heat-induced antigen retrieval step has been introduced (Mount Cooper 2001). In this placental section, optically clear nuclei (containing biotin) bind to the streptavidin of the ABC technique giving a reaction similar to that seen with CMV containing cells. If a polymer method (or even the original Sternberger's PAP method) is used then this anomalous staining will disappear, thus allowing confident demonstration of CMV infected nuclei. The false-positive staining pattern caused by endogenous biotin can be cytoplasmic or nuclear. A report of positive immunoreactivity of hepatocellular carcinomas for inhibin was later determined to be a false-positive finding due to cytoplasmic endogenous biotin. Steroid cell tumours of the ovary were found to demonstrate endogenous biotin cytoplasmic staining in 36% of cases. Immunoreactivity for anti-Herpes virus immunohistochemical staining in a series of endometria was also later determined to be a false-positive result due to biotin. The prominent intranuclear inclusions, resembling herpes virus cytopathic effect, were caused
[Histonet] Cyclin D1 IHC
Hello Histonetters, Has anyone done IHC using Cyclin D1 antibody? What vendor was used? What control tissue was used? Thank you. Donna Suresch - Merck Co. Donna L. Suresch Imaging Research Scientist Merck Research Laboratories Department of Imaging - West Point Campus Mail Stop: WP44KOffice: WP44-H129 770 Sumneytown Pike PO Box 4 West Point, PA 19486-0004 Phone: 215-652-7349 Fax: 215-993-6803 Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cyclin D1 IHC
We have used Neomarkers/Labvision cat.no. RM-9104 on mouse salivary glands and mammary glands. On Tue, Jul 24, 2012 at 11:16 AM, Suresch, Donna L. donna_sure...@merck.com wrote: Hello Histonetters, Has anyone done IHC using Cyclin D1 antibody? What vendor was used? What control tissue was used? Thank you. Donna Suresch - Merck Co. Donna L. Suresch Imaging Research Scientist Merck Research Laboratories Department of Imaging - West Point Campus Mail Stop: WP44KOffice: WP44-H129 770 Sumneytown Pike PO Box 4 West Point, PA 19486-0004 Phone: 215-652-7349 Fax: 215-993-6803 Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cyclin D1 IHC
Hi Donna, We use Dako #M3635 at 1:100. Mantle cell lymphoma is our control. Mark On Tue, Jul 24, 2012 at 8:16 AM, Suresch, Donna L. donna_sure...@merck.comwrote: Hello Histonetters, Has anyone done IHC using Cyclin D1 antibody? What vendor was used? What control tissue was used? Thank you. Donna Suresch - Merck Co. Donna L. Suresch Imaging Research Scientist Merck Research Laboratories Department of Imaging - West Point Campus Mail Stop: WP44KOffice: WP44-H129 770 Sumneytown Pike PO Box 4 West Point, PA 19486-0004 Phone: 215-652-7349 Fax: 215-993-6803 Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Unsubscribe
Please remove me from your list. Thanks, Stacey Crenshaw Human Resources Generalist Oregon Medical Group 1580 Valley River Drive, Suite 160 Eugene, OR 97401 Phone (541) 242-4339 Fax (541) 284-2038 Disclaimer: This electronic message may contain information that is Proprietary, Confidential, or legally privileged or protected. It is intended only for the use of the individual(s) and entity named in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from your computer. Do not deliver, distribute or copy this message and do not disclose its contents or take any action in reliance on the information it contains. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Question about waste storage temperature
Can I store (Xylene substitute, Isopropanol and Formalin) each, separated in 5 gallons (closed with a tight lid) plastic containers placed inside spill tray in a room that the temperature can rise up to 40°C in the summer? One container at the time, before it gets pick up by waste company. Thank you. Lin. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HistoTALK Show
Hi Everyone - This past HistoTALK http://www.histotalk.com/ show on Sunday, July 22nd had as its guest Wanda Jones from Emory University Hospital. Just wanted you all to know. Oh, and BTW, before that show #28, our guest was Billie Swisher! Two wonderful interviews. Yours, Dave ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Unsubscribe
OH NO! They're BACK! Saints preserve us! - Original Message - From: Stacey Crenshaw screns...@oregonmed.net To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu Sent: Tuesday, July 24, 2012 9:51:37 AM Subject: [Histonet] Unsubscribe Please remove me from your list. Thanks, Stacey Crenshaw Human Resources Generalist Oregon Medical Group 1580 Valley River Drive, Suite 160 Eugene, OR 97401 Phone (541) 242-4339 Fax (541) 284-2038 Disclaimer: This electronic message may contain information that is Proprietary, Confidential, or legally privileged or protected. It is intended only for the use of the individual(s) and entity named in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from your computer. Do not deliver, distribute or copy this message and do not disclose its contents or take any action in reliance on the information it contains. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CD200
I would be interested in this as well... Thank-you, Glen Dawson BS, HT(ASCP), QIHC Histology Technical Specialist Janesville, WI From: 41dm...@gmail.com Date: Fri, 20 Jul 2012 14:03:36 -0400 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD200 Is anyone out there running CD-200 with good results? If so, I would really appreciate it if you would email me privately so I can ask you a few questions. Thanks, Drew Sent from my iPhone ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Non-Histo. question
So, I go on maternity leave in 52 (I hope...) days...while out I will have one of those automatic messages that says I'm out...Since I get say 20ish emails a day from histonet I would assume that 20ish of these replys will go out to everyone, and I don't want to drive people bonkers. Maybe the moderator could answer...what do you want me to do? Cancel and rejoin when I get back, or is this just something that gets filtered out? Thanks Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] unsubscribe
Unsubscribe, please This message (including any attachments) may contain confidential, proprietary, privileged and/or private information. The information is intended to be for the use of the individual or entity designated above. If you are not the intended recipient of this message, please notify the sender immediately, and delete the message and any attachments. Any disclosure, reproduction, distribution or other use of this message or any attachments by an individual or entity other than the intended recipient is prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Non-Histo. question
Hi Sarah, I had a similar situation. If you go into your profile on the Histonet site you can click to not receive any of the Histonet messages while you are away. Then when you get back and want the messages again you just change it back. Eva On Tue, Jul 24, 2012 at 2:35 PM, Sarah Dysart sdys...@mirnarx.com wrote: So, I go on maternity leave in 52 (I hope...) days...while out I will have one of those automatic messages that says I'm out...Since I get say 20ish emails a day from histonet I would assume that 20ish of these replys will go out to everyone, and I don't want to drive people bonkers. Maybe the moderator could answer...what do you want me to do? Cancel and rejoin when I get back, or is this just something that gets filtered out? Thanks Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Secondary antibody causing nuclear staining?
Yes. The strength of the stained nuclei in the no primary slides are stronger on some days than others. On Tue, Jul 24, 2012 at 8:18 AM, Kim Donadio one_angel_sec...@yahoo.comwrote: Are you getting false positives and variations on the same control tissue for different days ? Sent from my iPhone On Jul 24, 2012, at 8:13 AM, Eva Permaul e...@georgetown.edu wrote: I understand the point about the biotin and I should have said that when using the ABC method we have taken to always using an avidin/biotin blocking kit. We are using biotinylated secondary antibodies from Vector. I have seen the same problem occur in our anti-mouse, anti-rabbit and anti-goat. In my last run I had stomach fundus as well as skin melanoma, both had pos.nuclei in the negative (no primary). In another run I had colon ca and breast ca, the breast ca had fewer pos. nuclei than the colon ca but they were still there. Some days the positive nuclei are stronger in a sample that was just weakly positive before. Just want to understand what it is and what effects it. Thank you all for your ideas. Eva Permaul Georgetown University On Mon, Jul 23, 2012 at 7:16 PM, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: I should have added that this was from the workshop notes on a Hypotheticals Workshop I ran last year at our Australian National Meeting. 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 Tony Henwood (SCHN) Sent: Tuesday, 24 July 2012 9:00 AM To: 'Eva Permaul'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Secondary antibody causing nuclear staining? It is possible that this is due to Biotin nuclei where excess biotin is found in the nuclei of some cells, see below: Optically clear nuclei have been reported in endometrial epithelium associated with first and second trimester abortions (Sickel di Sant'Agnese 1994). Optically clear nuclei have also been found in different types of tissues of diverse organs such as ovary, thyroid and lung (Nakatani et al 1994, Mount Cooper 2001). The optically clear nuclei contain excess biotin. Endogenous biotin immunoreactivity is generally not visualized in formalin fixed, paraffin-embedded tissues unless a heat-induced antigen retrieval step has been introduced (Mount Cooper 2001). In this placental section, optically clear nuclei (containing biotin) bind to the streptavidin of the ABC technique giving a reaction similar to that seen with CMV containing cells. If a polymer method (or even the original Sternberger's PAP method) is used then this anomalous staining will disappear, thus allowing confident demonstration of CMV infected nuclei. The false-positive staining pattern caused by endogenous biotin can be cytoplasmic or nuclear. A report of positive immunoreactivity of hepatocellular carcinomas for inhibin was later determined to be a false-positive finding due to cytoplasmic endogenous biotin. Steroid cell tumours of the ovary were found to demonstrate endogenous biotin cytoplasmic staining in 36% of cases. Immunoreactivity for anti-Herpes virus immunohistochemical staining in a series of endometria was also later determined to be a false-positive result due to biotin. The prominent intranuclear inclusions, resembling herpes virus cytopathic effect, were caused by intranuclear biotin and not viral particles. Similar false positive staining for CMV in products of conception has also been reported (Mount Cooper 2001). False-positive staining can be cytoplasmic or nuclear. When cytoplasmic, the appearance of the false signal is that of a dull brown granular or fluffy staining pattern. If this quality of staining is observed with several different antibodies, endogenous staining by biotin should be considered. When nuclear, a false-positive reaction may be associated with optically clear nuclei identified on HE stained sections. False-positive staining due to endogenous biotin, however, does not occur in a cell membrane pattern (Mount Cooper 2001). Mount SL Cooper K (2001) Beware of biotin: a source of false-positive immunohistochemistry Current Diagnostic Pathology 7:161-167. Nakatani et al (1994) Am J Surg Pathol 18(6):637-642. Sickel di Sant'Agnese (1994) Arch Pathol Lab Med 118:831-833 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
[Histonet] Bielschowsky questions, Luxol Fast Blue answers, and Protocol Drift.
Hi Everyone: I had a few questions regarding Bielschowsky silver stains. (1) What adhesive (if any) or type of slide do you use for the stain? (2) How do you clean the glassware? (3) When diluting the 40% formaldehyde when making up the developer, do you consider the 40% formaldehyde as 100% and then dilute it down by using 1 part formaldehyde and 9 parts distilled water? Or do you assume the 40% formaldehyde is 40% and then dilute it down using 1 part formaldehyde and 3 parts distilled water? (My protocol may have inadvertently changed from the first method to the second; I am not sure.) By the way, I want to thank everyone for helping me solve the problem of my Luxol Fast Blue staining the myelin too lightly. I discovered that somehow, I had started adding twice the amount of acetic acid to the Luxol staining solution as I should of. (This protocol drift, where a mistake can actually find its way into a written protocol, can be a real problem in a lab, especially when working for years by oneself, as I have) . But I also found that even reducing the acetic acid, while helping a lot, did not completely fix the problem. I needed to switch from staining the tissue for 2 hours at 60C to a full over-night (why I never needed to switch times before is a mystery). That did the trick beautifully. The myelin is staining perfectly again. Thanks again, Tim Wheelock Harvard Brain Bank Belmont, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Tennessee BB
Okay 30 years is too long in histology! I should have retired sooner than I did,. I saw this subject heading as a new stain for bacteria instead of an ad for a bed and breakfast in Tennessee. Rena Fail ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] 2012 Hard Tissue Forum Event in Bethesda, MD (August 18th!)
Hello! My name is Jack Ratliff and I am Chairman of the Hard Tissue Committee (HTC) for the National Society for Histotechnology (NSH). On behalf of the HTC and the NSH, we would like to invite you to attend and participate in the 2012 Hard Tissue Forum. This event will take place August 18th, 2012 and will be held at the Doubletree by Hilton in Bethesda, MD. We have put together an exciting program this year and we invite you to be a part of it! The program this year will focus on relevant histological and image analysis techniques associated with undemineralized bone and medical device implants. While the topics of microtomy (thin ground sectioning) and image analysis (microCT histomorphometry) will be highlighted, the sub-category of workshops will showcase a concentration of equipment, techniques, and technology that promises to be a unique single-day event demonstrating a hands-on application of specific techniques never before staged together in a single histology event! In fact, one of the highlights of this years program will be a first ever in the U.S and North America unveiling and demonstration of a non-contact laser microtome that has been developed to take thin sections of fresh and resin/plastic embedded tissues! To learn more about the Hard Tissue Forum, please visit http://s3.goeshow.com/nsh/2012HT/ereg572259.cfm?clear where you can find speaker related information, detailed workshop abstracts, and event registration information. If you have ANY questions, please feel free to contact Aubrey Wanner, Meeting Manager of the NSH (aub...@nsh.org) or myself (ratliffj...@hotmail.com) at your convenience. I look forward to seeing you in Bethesda! Yours Sincerely, Jack Jack L RatliffChairman, Hard Tissue Committee - National Society for Histotechnology ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Job Opportunity at Weill Cornell Medical College, NYC
Description The Tri-Institutional Laboratory of Comparative Pathology provides research pathology support to investigators at Weill Cornell Medical College, Memorial Sloan-Kettering Cancer Center, and The Rockefeller University, located in New York City. This position supports the mission of the Laboratory of Comparative Pathology by performing a range of histologic techniques on laboratory animal tissues for comparative pathologists, principal investigators and researchers; assists investigators in specimen requisition, handling and accessioning, following departmental protocols; Histology Services: Performs all steps of routine processing, paraffin-embedding (and OCT/plastic as needed) and microtomy (paraffin, frozen, plastic) on a variety of animal tissues (including eyes, bone, placenta and embryos); performs routine hematoxylin and eosin staining, as well as special histochemical staining (manual and automatic), cover slipping and immunohistochemistry as needed; performs special histologic techniques such as double-staining of embryos with Alcian blue and Alizarin red, and whole mount staining (mammary gland and lung); participates in quality control program for slides prepared, results entered and results released; orders reagent and supplies as needed; troubleshoots, cleans and performs routine maintenance of laboratory equipment related to histology and immunohistochemistry; performs preventive maintenance on histologic equipment; performs validation studies with introduction of new equipment/assays; performs other related duties as assigned. Qualifications At least three years experience working in a histology laboratory required; must be energetic, self-directed and self-motivated; computer proficiency necessary; Bachelors degree and HT, ASCP certified or eligible highly desired; excellent organization and communication skills required. For more information and to apply, see position #17920 at this link: https://cornellu.taleo.net/careersection/2000/jobsearch.ftl Sébastien Monette, DMV, MVSc, DACVP Head of Anatomic Pathology Tri-Institutional Laboratory of Comparative Pathology Memorial Sloan-Kettering Cancer Center Weill Cornell Medical College The Rockefeller University 1275 York Avenue, Box 270, New York NY 10065 Phone: 646-888-2420 Fax: 646-422-0139 Email MSKCC: monet...@mskcc.org Email WCMC: sem2...@med.cornell.edu Web site: http://www.mskcc.org/mskcc/html/92361.cfm = Please note that this e-mail and any files transmitted from Memorial Sloan-Kettering Cancer Center may be privileged, confidential, and protected from disclosure under applicable law. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any reading, dissemination, distribution, copying, or other use of this communication or any of its attachments is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting this message, any attachments, and all copies and backups from your computer. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] counter stain for PAS for fungus
Change all your reagents. Change periodic acid every week at least. Personally I see little advantage in fast green, and a common downside is overstaining in many labs, hiding fungus and basement membrane. Faint Hematoxylin is acceptable counterstain. For certain applications, e.g., dual staining for carbohydrate and mucin, adding AlcianBlue for 40 seconds is wickedly good. Dis/Advantage being dual staining methods impose two charges. Too bad. Instead of separate stains for mucin and carbohydrate, do them both every day and get good at it. We rarely do a straight PAS any more. Subtract the second charge where needed. The two analytes (carbohydrate and mucin) are different, yet the interpretation is synergistic. Where you see mucin glommed around a hypha-like structure it is. It also demonstrates internal structure in many fungi. AlcianBlue also stains bacteria, making it useful where polymicrobial (fungal and bacterial) infections are common. Call it your biofilm assay. So many fungi produce mucin that PAS-AB with hema is a superb method for detection. PAS-AB sine hema is considerably more sensitive, especially with pigmented fungal species but interpretation of PAS-AB without Hema is difficult for many pathologists. Steedman wrote the paper in 1951. Steve Steve A. McClain, MD McClain Labs, LLC 45 Manor Road, Smithtown, NY 11787 631 361 4000 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Unsubscribe
I vote for Raspberry! Claire From: histonet-boun...@lists.utsouthwestern.edu on behalf of nmhi...@comcast.net Sent: Tue 7/24/2012 11:36 AM To: Stacey Crenshaw Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Unsubscribe OH NO! They're BACK! Saints preserve us! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Secondary antibody causing nuclear staining?
Hello Eva I had a similar problem with an actin antibody many years ago. We eventually after many trials decided that it was an artifact caused by the heat retrieval possibly in relation to the length of fixation time. By playing with the dilution we eventually eliminated the problem by not retrieving that antibody at all and still got good antigenic staining. While advent of HIER has improved the staining of many antibodies it is not a necessity for all of them. regards Tony Tony Reilly B.App.Sc. , M.Sc. Chief Scientist, Anatomical Pathology Pathology Queensland-PA Laboratory Health Services Support Agency | Queensland Health Level 1, Building 15,Princess Alexandra Hospital Ipswich Road,WOOLLOONGABBA Qld4102 Ph: 07 3176 2412 Mob: 0402 139411 Fax: 07 3176 2930 Email: tony_rei...@health.qld.gov.au Web: www.health.qld.gov.au/qhcss/ Kim Donadio one_angel_sec...@yahoo.com 7/24/2012 10:18 pm Are you getting false positives and variations on the same control tissue for different days ? Sent from my iPhone On Jul 24, 2012, at 8:13 AM, Eva Permaul e...@georgetown.edu wrote: I understand the point about the biotin and I should have said that when using the ABC method we have taken to always using an avidin/biotin blocking kit. We are using biotinylated secondary antibodies from Vector. I have seen the same problem occur in our anti-mouse, anti-rabbit and anti-goat. In my last run I had stomach fundus as well as skin melanoma, both had pos.nuclei in the negative (no primary). In another run I had colon ca and breast ca, the breast ca had fewer pos. nuclei than the colon ca but they were still there. Some days the positive nuclei are stronger in a sample that was just weakly positive before. Just want to understand what it is and what effects it. Thank you all for your ideas. Eva Permaul Georgetown University On Mon, Jul 23, 2012 at 7:16 PM, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: I should have added that this was from the workshop notes on a Hypotheticals Workshop I ran last year at our Australian National Meeting. 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 Tony Henwood (SCHN) Sent: Tuesday, 24 July 2012 9:00 AM To: 'Eva Permaul'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Secondary antibody causing nuclear staining? It is possible that this is due to Biotin nuclei where excess biotin is found in the nuclei of some cells, see below: Optically clear nuclei have been reported in endometrial epithelium associated with first and second trimester abortions (Sickel di Sant'Agnese 1994). Optically clear nuclei have also been found in different types of tissues of diverse organs such as ovary, thyroid and lung (Nakatani et al 1994, Mount Cooper 2001). The optically clear nuclei contain excess biotin. Endogenous biotin immunoreactivity is generally not visualized in formalin fixed, paraffin-embedded tissues unless a heat-induced antigen retrieval step has been introduced (Mount Cooper 2001). In this placental section, optically clear nuclei (containing biotin) bind to the streptavidin of the ABC technique giving a reaction similar to that seen with CMV containing cells. If a polymer method (or even the original Sternberger's PAP method) is used then this anomalous staining will disappear, thus allowing confident demonstration of CMV infected nuclei. The false-positive staining pattern caused by endogenous biotin can be cytoplasmic or nuclear. A report of positive immunoreactivity of hepatocellular carcinomas for inhibin was later determined to be a false-positive finding due to cytoplasmic endogenous biotin. Steroid cell tumours of the ovary were found to demonstrate endogenous biotin cytoplasmic staining in 36% of cases. Immunoreactivity for anti-Herpes virus immunohistochemical staining in a series of endometria was also later determined to be a false-positive result due to biotin. The prominent intranuclear inclusions, resembling herpes virus cytopathic effect, were caused by intranuclear biotin and not viral particles. Similar false positive staining for CMV in products of conception has also been reported (Mount Cooper 2001). False-positive staining can be cytoplasmic or nuclear. When cytoplasmic, the appearance of the false signal is that of a dull brown granular or fluffy staining pattern. If this quality of staining is observed with several different antibodies, endogenous staining by biotin should be considered. When nuclear, a
[Histonet] How to UNSUBSCRIBE
From the computer that you receive your Histonet: Go to the bottom of any Histonet email. click on the link that end in listinfo/histonet Scroll to the bottom of the page Follow the directions to unsubscribe Keep this email, if you plan to re-subscribe after returning from a vacation or maternity leave. Peggy Wenk -Original Message- From: Lewin, Anne Sent: Tuesday, July 24, 2012 2:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] unsubscribe Unsubscribe, please This message (including any attachments) may contain confidential, proprietary, privileged and/or private information. The information is intended to be for the use of the individual or entity designated above. If you are not the intended recipient of this message, please notify the sender immediately, and delete the message and any attachments. Any disclosure, reproduction, distribution or other use of this message or any attachments by an individual or entity other than the intended recipient is prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
