From mike <@t> triagestaff.com Mon Jul 2 08:50:01 2012 From: mike <@t> triagestaff.com (Mike Comstock) Date: Mon Jul 2 08:50:18 2012 Subject: [Histonet] Histotechologist Needed for Florida Contract Position Message-ID: <4C0ECD9C1C22314ABB4FAA761F4EB13C04F75523DA@MBX.pldatacenter.local> Good Morning! Triage Staffing is in need of an experienced Histotechnologist with the HTL Florida license to assist one of our clients in primarily a grossing position. Contract length is 13 weeks and begins 7/23 or 7/30. Mon-Fri, Day Shift. *We offer a competitive hourly wage plus weekly non-taxed per diem, car allowance or car rental, and furnished private housing. *Triage also offers a comprehensive health insurance plan, dental, vision, 401K, and direct deposit every Friday. Interested candidates can email their resumes to mike@triagestaff.com or call 800.259.9897 x 204 for more information. We look forward to hearing from you! www.triagestaff.com From amber.mckenzie <@t> gastrodocs.net Mon Jul 2 12:17:20 2012 From: amber.mckenzie <@t> gastrodocs.net (Amber McKenzie) Date: Mon Jul 2 12:15:08 2012 Subject: [Histonet] Embedding beads In-Reply-To: <1340742010.84780.YahooMailNeo@web125802.mail.ne1.yahoo.com> References: <1340742010.84780.YahooMailNeo@web125802.mail.ne1.yahoo.com> Message-ID: <5A33C952BB67F4468AF1F36D739212BC308296E3@JERRY.Gia.com> How do you not cut thru the bead when sectioning? I'm intrigued by this process b/c I've never heard of this before. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kelly Boyd Sent: Tuesday, June 26, 2012 3:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Embedding beads I started using the embedding beads several months ago as a quick way to track who embedded what cassette. Each tech is assigned a number according to the number on their microtome. I get the beads from Cancer Diagnostics. You can probably find something similar at a craft store. These come in small round plastic containers that fit easily any where on the embedding center. They place the bead in a bottom corner of the cassette when topping it off with paraffin. They?have actually saved us time. Once the techs get used to it, it might add a few seconds to the embedding.?Before, the techs had to write down which blocks they embedded. (Very time consuming and often not complete).?If I needed to see who embedded a certain block, I had to go check that log book. Now I can see who embedded it just by looking at the block. Kelly?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From campbellj <@t> muhlbauerlab.com Mon Jul 2 13:29:33 2012 From: campbellj <@t> muhlbauerlab.com (Jennifer Campbell) Date: Mon Jul 2 13:29:41 2012 Subject: [Histonet] Embedding beads In-Reply-To: <5A33C952BB67F4468AF1F36D739212BC308296E3@JERRY.Gia.com> References: <1340742010.84780.YahooMailNeo@web125802.mail.ne1.yahoo.com> <5A33C952BB67F4468AF1F36D739212BC308296E3@JERRY.Gia.com> Message-ID: The identifier(whatever is using to distinguish who did the task) is not embedded with the specimen. It is put in the top portion of the cassette. At least that is what we have always done here. On Mon, Jul 2, 2012 at 1:17 PM, Amber McKenzie < amber.mckenzie@gastrodocs.net> wrote: > How do you not cut thru the bead when sectioning? I'm intrigued by this > process b/c I've never heard of this before. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto: > histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kelly Boyd > Sent: Tuesday, June 26, 2012 3:20 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Embedding beads > > I started using the embedding beads several months ago as a quick way to > track who embedded what cassette. Each tech is assigned a number according > to the number on their microtome. I get the beads from Cancer Diagnostics. > You can probably find something similar at a craft store. > These come in small round plastic containers that fit easily any where on > the embedding center. They place the bead in a bottom corner of the > cassette when topping it off with paraffin. > They have actually saved us time. Once the techs get used to it, it might > add a few seconds to the embedding. Before, the techs had to write down > which blocks they embedded. (Very time consuming and often not > complete). If I needed to see who embedded a certain block, I had to go > check that log book. Now I can see who embedded it just by looking at the > block. > > Kelly > _______________________________________________ > 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 > -- Jen Campbell, HT(ASCP) Supervisor of Technical Services Muhlbauer Dermatopathology Laboratory 61 Monroe Avenue, Ste B Pittsford NY 14534 P: 585.586.5166 F: 585.586.3137 IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. From aj.taylor <@t> blueyonder.co.uk Mon Jul 2 13:37:50 2012 From: aj.taylor <@t> blueyonder.co.uk (Alan Taylor) Date: Mon Jul 2 13:39:46 2012 Subject: [Histonet] Embedding beads References: <1340742010.84780.YahooMailNeo@web125802.mail.ne1.yahoo.com> <5A33C952BB67F4468AF1F36D739212BC308296E3@JERRY.Gia.com> Message-ID: <3A078ACBD81E4F39B2B2C8FAD4C6332F@merlin> Hi All The embedding bead goes into the cassette tray, into one corner, not into the stainless steel embedding mould with the tissue. The plastic cassette is placed on top of the mould, as usual and filled with molten wax, the coloured bead is quickly dropped in and the embedding mould is placed on the chiller to solidify in preparation for microtomy. The bead has no contact with the blade at any time, remaining at the back of the cassette tray. Hope this clarifies this easy process. Alan Taylor BSc(Hons), FRMS. Microtechnical Services 71 Sweetbrier Lane Heavitree Exeter. EX1 3AJ. Devon. UK. ----- Original Message ----- From: "Amber McKenzie" To: Sent: Monday, July 02, 2012 6:17 PM Subject: [Histonet] Embedding beads How do you not cut thru the bead when sectioning? I'm intrigued by this process b/c I've never heard of this before. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kelly Boyd Sent: Tuesday, June 26, 2012 3:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Embedding beads I started using the embedding beads several months ago as a quick way to track who embedded what cassette. Each tech is assigned a number according to the number on their microtome. I get the beads from Cancer Diagnostics. You can probably find something similar at a craft store. These come in small round plastic containers that fit easily any where on the embedding center. They place the bead in a bottom corner of the cassette when topping it off with paraffin. They have actually saved us time. Once the techs get used to it, it might add a few seconds to the embedding. Before, the techs had to write down which blocks they embedded. (Very time consuming and often not complete). If I needed to see who embedded a certain block, I had to go check that log book. Now I can see who embedded it just by looking at the block. Kelly _______________________________________________ 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 From hmarlatt26 <@t> gmail.com Mon Jul 2 14:17:01 2012 From: hmarlatt26 <@t> gmail.com (heather marlatt) Date: Mon Jul 2 14:17:04 2012 Subject: [Histonet] Slide Baskets???? Message-ID: I was wondering if anyone in histoland has any slide baskets for the old sakura DRS 601 stainer laying around??? I can't seem to find these for a decent price so I thought I'd ask. I only have one at the moment and as you might imagine that us really cramping my style. Thanks in advance Heather From Laurie <@t> blufrogpath.com Mon Jul 2 15:19:43 2012 From: Laurie <@t> blufrogpath.com (Laurie@blufrogpath.com) Date: Mon Jul 2 15:19:47 2012 Subject: [Histonet] Humidity Check Message-ID: <20120702131943.295dc6182df7e5cbb4f32bc101c30dcc.f604e52a69.wbe@email15.secureserver.net> Do others check the humidity of their Histo lab on a daily basis? Laurie Colbert From raj <@t> bluemarble.net Mon Jul 2 17:37:20 2012 From: raj <@t> bluemarble.net (Rebecca a. Johnson) Date: Mon Jul 2 17:37:36 2012 Subject: [Histonet] Mohs Message-ID: Need to know what Mohs techs are getting paid. Thanks Becky From adam.haberman <@t> oberlin.edu Tue Jul 3 07:45:53 2012 From: adam.haberman <@t> oberlin.edu (Adam Haberman) Date: Tue Jul 3 07:45:58 2012 Subject: [Histonet] What is this item called? Message-ID: Hello everyone, I am trying to identify a piece of glassware so that I can order a few more of it. It is a thick piece of glass with a large, wide depression in the middle. The item I am looking for is thicker and wider than a depression slide, and I use it as a dissecting dish. I have included a picture of the piece I am trying to identify, along with a depression slide for comparison. This is probably an item that is not widely used anymore, as it seems to predate all of the faculty here. However, it is perfect for the dissections I do with two pair of forceps under a dissecting microscope. It gives me much better working angles than the spot plates I resort to when I cannot find this item. If anyone can identify the item or, even better, tell me where I can order more, please let me know. Thank you very much. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 From jmacdonald <@t> mtsac.edu Tue Jul 3 08:13:37 2012 From: jmacdonald <@t> mtsac.edu (Jennifer MacDonald) Date: Tue Jul 3 08:13:47 2012 Subject: [Histonet] What is this item called? In-Reply-To: References: Message-ID: <2FB72470-1EA9-4BF5-8B04-4D613C5F06EE@mtsac.edu> A watch glass? On Jul 3, 2012, at 5:45 AM, Adam Haberman wrote: > Hello everyone, > > I am trying to identify a piece of glassware so that I can order a few more > of it. It is a thick piece of glass with a large, wide depression in the > middle. The item I am looking for is thicker and wider than a depression > slide, and I use it as a dissecting dish. I have included a picture of the > piece I am trying to identify, along with a depression slide for > comparison. This is probably an item that is not widely used anymore, as > it seems to predate all of the faculty here. However, it is perfect for > the dissections I do with two pair of forceps under a dissecting > microscope. It gives me much better working angles than the spot plates I > resort to when I cannot find this item. > > If anyone can identify the item or, even better, tell me where I can order > more, please let me know. Thank you very much. > > --Adam > > Adam Haberman > Visiting Assistant Professor of Biology > Oberlin College > adam.haberman@oberlin.edu > (440)775-6502 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Tue Jul 3 08:15:31 2012 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Tue Jul 3 08:18:40 2012 Subject: [Histonet] What is this item called? In-Reply-To: <2FB72470-1EA9-4BF5-8B04-4D613C5F06EE@mtsac.edu> References: , <2FB72470-1EA9-4BF5-8B04-4D613C5F06EE@mtsac.edu> Message-ID: <12A4DAFC2FEBB84B8DED5F5E9201B4E9179EC272E4@UTHCMS1.uthouston.edu> Is this a 3 by 1.25 inch glass slide approximately 3 mm thick with a 1 inch depression in the middle? Barry To: Adam Haberman Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] What is this item called? A watch glass? On Jul 3, 2012, at 5:45 AM, Adam Haberman wrote: > Hello everyone, > > I am trying to identify a piece of glassware so that I can order a few more > of it. It is a thick piece of glass with a large, wide depression in the > middle. The item I am looking for is thicker and wider than a depression > slide, and I use it as a dissecting dish. I have included a picture of the > piece I am trying to identify, along with a depression slide for > comparison. This is probably an item that is not widely used anymore, as > it seems to predate all of the faculty here. However, it is perfect for > the dissections I do with two pair of forceps under a dissecting > microscope. It gives me much better working angles than the spot plates I > resort to when I cannot find this item. > > If anyone can identify the item or, even better, tell me where I can order > more, please let me know. Thank you very much. > > --Adam > > Adam Haberman > Visiting Assistant Professor of Biology > Oberlin College > adam.haberman@oberlin.edu > (440)775-6502 > _______________________________________________ > 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 From Sandra.Harrison3 <@t> va.gov Tue Jul 3 08:22:50 2012 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Tue Jul 3 08:23:12 2012 Subject: [Histonet] Mohs In-Reply-To: References: Message-ID: Depends ;-) All kidding aside, there are factors that weigh in, such as geography, availability, training, experience, speed, etc. 10 years ago, when I worked as a Mohs' tech in the Denver, Colorado area, I received $25.00 per hour, plus benefits. I usually only worked 3 days per week, but on some rare occasions, worked till 9 p.m. trying to get to clear margins. I don't know what the current wages are. Sandy Harrison, HTL Histology Supervisor, Minneapolis VA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca a. Johnson Sent: Monday, July 02, 2012 5:37 PM To: histonet Subject: [Histonet] Mohs Need to know what Mohs techs are getting paid. Thanks Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandra.Harrison3 <@t> va.gov Tue Jul 3 08:23:52 2012 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Tue Jul 3 08:24:13 2012 Subject: [Histonet] Humidity Check In-Reply-To: <20120702131943.295dc6182df7e5cbb4f32bc101c30dcc.f604e52a69.wbe@email15.secureserver.net> References: <20120702131943.295dc6182df7e5cbb4f32bc101c30dcc.f604e52a69.wbe@email15.secureserver.net> Message-ID: Yes, we have a wireless temperature monitoring system, called Checkpoint, and each room has a sensor that monitors temp and humidity. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie@blufrogpath.com Sent: Monday, July 02, 2012 3:20 PM To: Histonet post Subject: [Histonet] Humidity Check Do others check the humidity of their Histo lab on a daily basis?= Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMaslanka <@t> stpetes.org Tue Jul 3 09:03:52 2012 From: JMaslanka <@t> stpetes.org (JMaslanka@stpetes.org) Date: Tue Jul 3 09:04:23 2012 Subject: [Histonet] Competency check list Message-ID: Would anyone be willing to share their IHC tech competency check list? Joe Maslanka BS, CT,HT (ASCP) Anatomical Pathology Technical Supervisor St Peter's Hospital,MT 59601 (P)(406) 447-2406 (F)(406)444-2126 Give thanks for ALL things..... From CIngles <@t> uwhealth.org Tue Jul 3 10:06:08 2012 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Tue Jul 3 10:06:50 2012 Subject: [Histonet] Humidity Check References: <20120702131943.295dc6182df7e5cbb4f32bc101c30dcc.f604e52a69.wbe@email15.secureserver.net> Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A866@UWHC-MAIL2.uwhis.hosp.wisc.edu> I think it is actually a requirement that temp and humidity are monitiored daily. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie@blufrogpath.com Sent: Mon 7/2/2012 3:19 PM To: Histonet post Subject: [Histonet] Humidity Check Do others check the humidity of their Histo lab on a daily basis?= Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CIngles <@t> uwhealth.org Tue Jul 3 10:07:13 2012 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Tue Jul 3 10:08:11 2012 Subject: [Histonet] Mohs References: Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A867@UWHC-MAIL2.uwhis.hosp.wisc.edu> I would love to know this too. I am getting about $23/hr but I have pretty good benefits. I'm at UWisc Madison Hospital. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rebecca a. Johnson Sent: Mon 7/2/2012 5:37 PM To: histonet Subject: [Histonet] Mohs Need to know what Mohs techs are getting paid. Thanks Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hardin <@t> oncology.wisc.edu Tue Jul 3 10:51:16 2012 From: hardin <@t> oncology.wisc.edu (Joe Hardin) Date: Tue Jul 3 10:51:25 2012 Subject: [Histonet] JB fixative Message-ID: <4FF314F4.5020707@oncology.wisc.edu> Hi All, I use a Zinc fixative called JB fixitive for tissues that I want to stain for CD antigens(see attachment). Does anyone know if this fixative kills all microbes and prions? From kim.tournear <@t> yahoo.com Tue Jul 3 10:59:53 2012 From: kim.tournear <@t> yahoo.com (Kim Tournear) Date: Tue Jul 3 11:10:03 2012 Subject: [Histonet] Mohs In-Reply-To: <064F1ACBAE8A78469AE2E41D533D87E505A867@UWHC-MAIL2.uwhis.hosp.wisc.edu> References: <064F1ACBAE8A78469AE2E41D533D87E505A867@UWHC-MAIL2.uwhis.hosp.wisc.edu> Message-ID: <5C862879-5A6D-4D83-A093-DACEC3AF0ACA@yahoo.com> You can get upwards of 30.00/hr in Arizona. Sent from the iPhone of Kim Tournear. On Jul 3, 2012, at 10:07 AM, "Ingles Claire " wrote: > I would love to know this too. I am getting about $23/hr but I have pretty good benefits. I'm at UWisc Madison Hospital. > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rebecca a. Johnson > Sent: Mon 7/2/2012 5:37 PM > To: histonet > Subject: [Histonet] Mohs > > > > > Need to know what Mohs techs are getting paid. > > > Thanks > Becky > _______________________________________________ > 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 From vtobias <@t> uw.edu Tue Jul 3 11:43:05 2012 From: vtobias <@t> uw.edu (Victor A. Tobias) Date: Tue Jul 3 11:43:52 2012 Subject: [Histonet] Billing 88342 Message-ID: Looking for other opinions from those who do consult/referral work. If a client sends in a request for a single antibody done on multiple blocks on a single specimen, do you bill the client for each tech component ? The client will do the interpretation. What happens in the above scenario if the request is to bill the patient? Knowing you get reimbursed for one, do you eat the other charges are make the client select the one block? We have run numbers on potential lost revenue and the number is significant. Victor Victor Tobias HT(ASCP) Clinical Applications Analyst Harborview Medical Center Dept of Pathology Room NJB244 Seattle, WA 98104 vtobias@u.washington.edu 206-744-2735 206-744-8240 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From jaylundgren <@t> gmail.com Tue Jul 3 12:05:27 2012 From: jaylundgren <@t> gmail.com (Jay Lundgren) Date: Tue Jul 3 12:05:35 2012 Subject: [Histonet] Billing 88342 In-Reply-To: References: Message-ID: I am neither a lawyer nor a health care administrator, but, in my experience, the Pathologist picks the (hopefully) most diagnostic blocks from the multiblock cases and submits them for IHC. If you do the requested IHC on, say, 4 blocks out of 30, you charge x4 for the technical fee. After all, you are using 4 times the supplies (buffer, antibody, etc.). Before you hit the cash paying patient with a bill, their primary care provider should warn them what it's going to cost. I have seen a good Pathologist only select one block for IHC when the clinician previously informed him that the patient had no insurance and was paying out of pocket. I think it's interesting that people control *their own* health care costs when no insurance company or the government is involved. Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) On Tue, Jul 3, 2012 at 11:43 AM, Victor A. Tobias wrote: > Looking for other opinions from those who do consult/referral work. > > If a client sends in a request for a single antibody done on multiple > blocks on a single specimen, do you bill the client for each tech component > ? The client will do the interpretation. > > What happens in the above scenario if the request is to bill the patient? > Knowing you get reimbursed for one, do you eat the other charges are make > the client select the one block? > > We have run numbers on potential lost revenue and the number is > significant. > > Victor > > > Victor Tobias HT(ASCP) > Clinical Applications Analyst > Harborview Medical Center > Dept of Pathology Room NJB244 > Seattle, WA 98104 > vtobias@u.washington.edu > 206-744-2735 > 206-744-8240 Fax > ================================================= > Privileged, confidential or patient identifiable information may be > contained in this message. This information is meant only for the use of > the intended recipients. If you are not the intended recipient, or if the > message has been addressed to you in error, do not read, disclose, > reproduce, distribute, disseminate or otherwise use this > transmission. Instead, please notify the sender by reply e-mail, and then > destroy all copies of the message and any attachments. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JMacDonald <@t> mtsac.edu Tue Jul 3 12:16:11 2012 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Tue Jul 3 12:16:58 2012 Subject: [Histonet] What is this item called? In-Reply-To: Message-ID: http://en.wikipedia.org/wiki/Watch_glass Adam Haberman Sent by: histonet-bounces@lists.utsouthwestern.edu 07/03/2012 05:47 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] What is this item called? Hello everyone, I am trying to identify a piece of glassware so that I can order a few more of it. It is a thick piece of glass with a large, wide depression in the middle. The item I am looking for is thicker and wider than a depression slide, and I use it as a dissecting dish. I have included a picture of the piece I am trying to identify, along with a depression slide for comparison. This is probably an item that is not widely used anymore, as it seems to predate all of the faculty here. However, it is perfect for the dissections I do with two pair of forceps under a dissecting microscope. It gives me much better working angles than the spot plates I resort to when I cannot find this item. If anyone can identify the item or, even better, tell me where I can order more, please let me know. Thank you very much. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Tue Jul 3 12:36:27 2012 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Tue Jul 3 12:36:35 2012 Subject: [Histonet] Billing 88342 In-Reply-To: References: Message-ID: <1341336987.69976.YahooMailNeo@web5702.biz.mail.ne1.yahoo.com> Where is the like button? Thumbs up! ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com ________________________________ From: Jay Lundgren To: Victor A. Tobias Cc: HISTONET Sent: Tuesday, July 3, 2012 12:05 PM Subject: Re: [Histonet] Billing 88342 ? ? I am neither a lawyer nor a health care administrator, but, in my experience, the Pathologist picks the (hopefully) most diagnostic blocks from the multiblock cases and submits them for IHC.? If you do the requested IHC on, say, 4 blocks out of 30, you charge x4 for the technical fee.? After all, you are using 4 times the supplies (buffer, antibody, etc.). ? ? Before you hit the cash paying patient with a bill, their primary care provider should warn them what it's going to cost. ? ? I have seen a good Pathologist only select one block for IHC when the clinician previously informed him that the patient had no insurance and was paying out of pocket. ? ? I think it's interesting that people control *their own* health care costs when no insurance company or the government is involved. ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? Sincerely, ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? Jay A. Lundgren, M.S., HTL (ASCP) On Tue, Jul 3, 2012 at 11:43 AM, Victor A. Tobias wrote: > Looking for other opinions from those who do consult/referral work. > > If a client sends in a request for a single antibody done on multiple > blocks on a single specimen, do you bill the client for each tech component > ? The client will do the interpretation. > > What happens in the above scenario if the request is to bill the patient? > Knowing you get reimbursed for one, do you eat the other charges are make > the client select the one block? > > We have run numbers on potential lost revenue and the number is > significant. > > Victor > > > Victor Tobias HT(ASCP) > Clinical Applications Analyst > Harborview Medical Center > Dept of Pathology Room NJB244 > Seattle, WA 98104 > vtobias@u.washington.edu > 206-744-2735 > 206-744-8240 Fax > ================================================= > Privileged, confidential or patient identifiable information may be > contained in this message. This information is meant only for the use of > the intended recipients. If you are not the intended recipient, or if the > message has been addressed to you in error, do not read, disclose, > reproduce, distribute, disseminate or otherwise use this > transmission. Instead, please notify the sender by reply e-mail, and then > destroy all copies of the message and any attachments. > > _______________________________________________ > 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 From hfedor <@t> jhmi.edu Tue Jul 3 12:40:14 2012 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Tue Jul 3 12:40:24 2012 Subject: [Histonet] Billing 88342 In-Reply-To: <1341336987.69976.YahooMailNeo@web5702.biz.mail.ne1.yahoo.com> References: <1341336987.69976.YahooMailNeo@web5702.biz.mail.ne1.yahoo.com> Message-ID: That is so true.! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, July 03, 2012 1:36 PM To: Jay Lundgren; Victor A. Tobias Cc: HISTONET Subject: Re: [Histonet] Billing 88342 Where is the like button? Thumbs up! ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com ________________________________ From: Jay Lundgren To: Victor A. Tobias Cc: HISTONET Sent: Tuesday, July 3, 2012 12:05 PM Subject: Re: [Histonet] Billing 88342 ? ? I am neither a lawyer nor a health care administrator, but, in my experience, the Pathologist picks the (hopefully) most diagnostic blocks from the multiblock cases and submits them for IHC.? If you do the requested IHC on, say, 4 blocks out of 30, you charge x4 for the technical fee.? After all, you are using 4 times the supplies (buffer, antibody, etc.). ? ? Before you hit the cash paying patient with a bill, their primary care provider should warn them what it's going to cost. ? ? I have seen a good Pathologist only select one block for IHC when the clinician previously informed him that the patient had no insurance and was paying out of pocket. ? ? I think it's interesting that people control *their own* health care costs when no insurance company or the government is involved. ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? Sincerely, ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? Jay A. Lundgren, M.S., HTL (ASCP) On Tue, Jul 3, 2012 at 11:43 AM, Victor A. Tobias wrote: > Looking for other opinions from those who do consult/referral work. > > If a client sends in a request for a single antibody done on multiple > blocks on a single specimen, do you bill the client for each tech > component ? The client will do the interpretation. > > What happens in the above scenario if the request is to bill the patient? > Knowing you get reimbursed for one, do you eat the other charges are > make the client select the one block? > > We have run numbers on potential lost revenue and the number is > significant. > > Victor > > > Victor Tobias HT(ASCP) > Clinical Applications Analyst > Harborview Medical Center > Dept of Pathology Room NJB244 > Seattle, WA 98104 > vtobias@u.washington.edu > 206-744-2735 > 206-744-8240 Fax > ================================================= > Privileged, confidential or patient identifiable information may be > contained in this message. This information is meant only for the use > of the intended recipients. If you are not the intended recipient, or > if the message has been addressed to you in error, do not read, > disclose, reproduce, distribute, disseminate or otherwise use this > transmission. Instead, please notify the sender by reply e-mail, and > then destroy all copies of the message and any attachments. > > _______________________________________________ > 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SHUNTER <@t> beaumont.edu Tue Jul 3 12:52:03 2012 From: SHUNTER <@t> beaumont.edu (Sue Hunter) Date: Tue Jul 3 12:52:07 2012 Subject: [Histonet] RE: Billing 88342 In-Reply-To: References: Message-ID: It is my understanding that you can only bill for one 88342 on a specimen - technical or professional, even if you perform the same stain on multiple blocks of that specimen. For A1, A2, A3 etc you can only charge once. For A1, B1, C1, etc you may charge a 88342 per specimen. Sue Sue Hunter, Supervisor Advanced Diagnostics Beaumont Health System Royal Oak MI 248-898-5146 shunter@beaumont.edu ----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Victor A. Tobias Sent: Tuesday, July 03, 2012 12:43 PM To: 'HISTONET' Subject: [Histonet] Billing 88342 Looking for other opinions from those who do consult/referral work. If a client sends in a request for a single antibody done on multiple blocks on a single specimen, do you bill the client for each tech component ? The client will do the interpretation. What happens in the above scenario if the request is to bill the patient? Knowing you get reimbursed for one, do you eat the other charges are make the client select the one block? We have run numbers on potential lost revenue and the number is significant. Victor Victor Tobias HT(ASCP) Clinical Applications Analyst Harborview Medical Center Dept of Pathology Room NJB244 Seattle, WA 98104 vtobias@u.washington.edu 206-744-2735 206-744-8240 Fax =================================================Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet To report this email as SPAM, please forward it to spam@websense.com. From TJohnson <@t> gnf.org Tue Jul 3 12:57:34 2012 From: TJohnson <@t> gnf.org (Teri Johnson) Date: Tue Jul 3 12:57:44 2012 Subject: [Histonet] RE: What is this item called? Message-ID: <9F3CFEE76E51B64991C7485270890B400CDC2518@EX4.lj.gnf.org> Dear Adam, Good luck finding it. There are some thick watch glasses that might work for your purposes but they aren't the size of glass slides. EMS has some well dishes that also might work, but they are also bigger. http://www.carolina.com/category/equipment+and+supplies/glass+and+plasticware/watch+glasses.do http://www.emsdiasum.com/microscopy/products/grids/accessories.aspx#71565 Hope this helps! Teri From rsrichmond <@t> gmail.com Tue Jul 3 13:10:12 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Tue Jul 3 13:10:16 2012 Subject: [Histonet] Re: JB fixative Message-ID: Joe Hardin (apparently at U of WI) asks: >>I use a zinc fixative called JB fixative for tissues that I want to stain for CD antigens (see attachment). Does anyone know if this fixative kills all microbes and prions?<< http://www.ihcworld.com/_protocols/histology/zinc_fixative.htm If this is the "fixative" (it doesn't contain any aldehyde) in question, I wouldn't depend on it to kill anything. You're depending on the alcohol in the processor to kill the beasties and probably also to fix the tissue. Attachments don't come through Histonet, by the way. Bob Richmond Samurai Pathologist Knoxville TN From arsenn <@t> hsh.org Tue Jul 3 13:24:36 2012 From: arsenn <@t> hsh.org (Senn, Amy R) Date: Tue Jul 3 13:24:44 2012 Subject: [Histonet] Humidity Message-ID: We do not check the temp/humidity in our room. We have 2 techs who feel it's always cold, and 2 techs who think it's way too hot L Amy, HT Holy Spirit Hospital Histology Laboratory 503 N. 21st Street, Camp Hill, PA 17011 Attention: This Message is intended only for the use of the individual or entity to which it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination or copying of this message or the taking of any action in reliance on the contents of this message is strictly prohibited. If you have received this message in error, please notify us immediately and destroy the original message. Thank you. From Longf1 <@t> LabCorp.com Tue Jul 3 13:27:29 2012 From: Longf1 <@t> LabCorp.com (Long, Florence) Date: Tue Jul 3 13:31:32 2012 Subject: [Histonet] What is this item called? In-Reply-To: References: Message-ID: Hi Adam, Would that be a modification of a "hanging-drop slide"? F. Long ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adam Haberman [adam.haberman@oberlin.edu] Sent: Tuesday, July 03, 2012 8:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] What is this item called? Hello everyone, I am trying to identify a piece of glassware so that I can order a few more of it. It is a thick piece of glass with a large, wide depression in the middle. The item I am looking for is thicker and wider than a depression slide, and I use it as a dissecting dish. I have included a picture of the piece I am trying to identify, along with a depression slide for comparison. This is probably an item that is not widely used anymore, as it seems to predate all of the faculty here. However, it is perfect for the dissections I do with two pair of forceps under a dissecting microscope. It gives me much better working angles than the spot plates I resort to when I cannot find this item. If anyone can identify the item or, even better, tell me where I can order more, please let me know. Thank you very much. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -This e-mail and any attachments may contain CONFIDENTIAL information, including PROTECTED HEALTH INFORMATION. If you are not the intended recipient, any use or disclosure of this information is STRICTLY PROHIBITED; you are requested to delete this e-mail and any attachments, notify the sender immediately, and notify the LabCorp Privacy Officer at privacyofficer@labcorp.com or call (877) 23-HIPAA / (877) 234-4722. From marktarango <@t> gmail.com Tue Jul 3 13:34:07 2012 From: marktarango <@t> gmail.com (Mark Tarango) Date: Tue Jul 3 13:34:10 2012 Subject: [Histonet] Billing 88342 In-Reply-To: References: Message-ID: I would think that if you're billing the client and not the insurance that you could charge per block for the technical. After all you're just providing the stain to them. In my opinion, the client should eat this cost. I would let the client know that you'd be billing this way before staining the slides. They could order 25 stains on a single part and you would be in the red big time. If you're billing the patient directly, you would have to follow the rule of charging only once per specimen. Mark On Tue, Jul 3, 2012 at 9:43 AM, Victor A. Tobias wrote: > Looking for other opinions from those who do consult/referral work. > > If a client sends in a request for a single antibody done on multiple > blocks on a single specimen, do you bill the client for each tech component > ? The client will do the interpretation. > > What happens in the above scenario if the request is to bill the patient? > Knowing you get reimbursed for one, do you eat the other charges are make > the client select the one block? > > We have run numbers on potential lost revenue and the number is > significant. > > Victor > > > Victor Tobias HT(ASCP) > Clinical Applications Analyst > Harborview Medical Center > Dept of Pathology Room NJB244 > Seattle, WA 98104 > vtobias@u.washington.edu > 206-744-2735 > 206-744-8240 Fax > ================================================= > Privileged, confidential or patient identifiable information may be > contained in this message. This information is meant only for the use of > the intended recipients. If you are not the intended recipient, or if the > message has been addressed to you in error, do not read, disclose, > reproduce, distribute, disseminate or otherwise use this > transmission. Instead, please notify the sender by reply e-mail, and then > destroy all copies of the message and any attachments. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gayle.callis <@t> bresnan.net Tue Jul 3 13:36:13 2012 From: gayle.callis <@t> bresnan.net (gayle callis) Date: Tue Jul 3 13:36:23 2012 Subject: [Histonet] JB fixative Message-ID: <000001cd594a$bb578720$32069560$@bresnan.net> Hi All, You wrote: I use a Zinc fixative called JB fixative for tissues that I want to stain for CD antigens(see attachment). Does anyone know if this fixative kills all microbes and prions? You did not say what species you are working with? The answer to your question is no for both microbes and prions. JB (acronym for Jay Beckstead) fixative is formalin/aldehyde free, and is for CD markers. Beckstead developed this fixative when studying CD markers in human lymphomas. Beckstead JH (1994) A simple technique for preservation of fixation-sensitive antigens in paraffin-embedded tissues. J Histochem Cytochem. 42(8):1127-34. Free online Beckstead JH (1995) A simple technique for preservation of fixation-sensitive antigens in paraffin-embedded tissues: addendum. J Histochem Cytochem. 43(3):345. Free online Did your attachment contain the information below (via IHCworld)? Zinc Fixative (JB Fixative) Formalin Free 0.1M Tris Buffer, pH 7.4 Tris Base -------------------------------- 12.1 g (TRIZMA) 1N HCL ----------------------------------- 81.5 ml Distilled water -------------------------- 900 ml Mix to dissolve. Adjust pH to 7.4 Zinc Fixative Calcium Acetate ---------------------- 0.5 g Zinc Acetate -------------------------- 5.0 g Zinc Chloride -------------------------- 5.0 g 0.1M Tris Buffer made above ------ 1000 ml Mix to dissolve. The final pH will be approximately 6.5-7.0. Do not readjust the pH, as this will cause the zinc to come out of solution. Store Zinc Fixative at room temperature. Fix tissues for 24 to 48 hours. Fixation longer than 48 hours may make the tissue brittle and difficult to cut. Description: Tissues fixed in this solution followed by paraffin embedding and sectioning results in antigen preservation comparable to that in frozen sections with antibodies to these cell surface markers (CD1, CD3, CD4, CD7, CD8, CD19, CD31). Morphological preservation was also comparable to formalin-fixed sections. As killing microbes, and I presume bacteria, virus or fungi, you should use neutral buffered formalin. Prions are a whole other story since the only way to totally eradicate these particles is incineration. CDC has guidelines on handling prions in a laboratory. Gayle M. Callis HTL/HT/MT(ASCP) 1 Home | About | Disclaimer | Privacy | Contact | Advertise | Site Mapratory. From jaylundgren <@t> gmail.com Tue Jul 3 14:21:06 2012 From: jaylundgren <@t> gmail.com (Jay Lundgren) Date: Tue Jul 3 14:21:11 2012 Subject: [Histonet] RE: Billing 88342 In-Reply-To: References: Message-ID: Sue, As I said, I don't know the specific regulation, and I don't remember specific block numbers of cases that I have charged. The Laboratory Director (in most cases a Pathologist) has final say about what gets charged anyway (in most places). What's interesting, is that if what you say about A1, B1, C1 vs. A1, A2, and A3 is correct, *one could gross in a case with an eye towards recompensation*. If it were about the money. Which it's not. Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) From adam.haberman <@t> oberlin.edu Tue Jul 3 14:23:43 2012 From: adam.haberman <@t> oberlin.edu (Adam Haberman) Date: Tue Jul 3 14:23:51 2012 Subject: [Histonet] What is this item called? In-Reply-To: References: Message-ID: Barry, It is 3" x 1 3/4", and the depression has a 1 1/2" diameter. Thanks for all the ideas so far. It isn't a hanging drop slide or a watch glass. You can see a picture of it next to a depression slide here: http://imgur.com/yBwvw I don't see a better way to send a picture on this list. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 On Tue, Jul 3, 2012 at 2:27 PM, Long, Florence wrote: > Hi Adam, > Would that be a modification of a "hanging-drop slide"? > F. Long > > ________________________________________ > From: histonet-bounces@lists.utsouthwestern.edu [ > histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adam Haberman [ > adam.haberman@oberlin.edu] > Sent: Tuesday, July 03, 2012 8:45 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] What is this item called? > > Hello everyone, > > I am trying to identify a piece of glassware so that I can order a few more > of it. It is a thick piece of glass with a large, wide depression in the > middle. The item I am looking for is thicker and wider than a depression > slide, and I use it as a dissecting dish. I have included a picture of the > piece I am trying to identify, along with a depression slide for > comparison. This is probably an item that is not widely used anymore, as > it seems to predate all of the faculty here. However, it is perfect for > the dissections I do with two pair of forceps under a dissecting > microscope. It gives me much better working angles than the spot plates I > resort to when I cannot find this item. > > If anyone can identify the item or, even better, tell me where I can order > more, please let me know. Thank you very much. > > --Adam > > Adam Haberman > Visiting Assistant Professor of Biology > Oberlin College > adam.haberman@oberlin.edu > (440)775-6502 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -This e-mail and any attachments may contain CONFIDENTIAL information, > including PROTECTED HEALTH INFORMATION. If you are not the intended > recipient, any use or disclosure of this information is STRICTLY > PROHIBITED; you are requested to delete this e-mail and any attachments, > notify the sender immediately, and notify the LabCorp Privacy Officer at > privacyofficer@labcorp.com or call (877) 23-HIPAA / (877) 234-4722. > > From Laurie <@t> blufrogpath.com Tue Jul 3 14:24:47 2012 From: Laurie <@t> blufrogpath.com (Laurie@blufrogpath.com) Date: Tue Jul 3 14:24:52 2012 Subject: [Histonet] Eyewash Requirements Message-ID: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> What is the time requirement for flushing the eyewash? We u flush for 30 seconds and then I was told by our safety officer that it should be 3 minutes. How often do you test your shower? Laurie Colbert From sdysart <@t> mirnarx.com Tue Jul 3 13:59:02 2012 From: sdysart <@t> mirnarx.com (Sarah Dysart) Date: Tue Jul 3 14:31:11 2012 Subject: [Histonet] Eosin Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D500BF339B7@BL2PRD0710MB363.namprd07.prod.outlook.com> So I have always used Eosin-Y stain...can someone tell me which is better...alcohol or phloxine?? Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 From jaylundgren <@t> gmail.com Tue Jul 3 14:32:06 2012 From: jaylundgren <@t> gmail.com (Jay Lundgren) Date: Tue Jul 3 14:32:11 2012 Subject: [Histonet] What is this item called? In-Reply-To: References: Message-ID: Brilliant way to send a picture. I <3 interwebs. Whatever it is, I want one. So it's made of optical quality glass, like a slide or a coverslip? Does it have a company's name on it? I'll bet you it's expensive. Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) From jaylundgren <@t> gmail.com Tue Jul 3 14:34:26 2012 From: jaylundgren <@t> gmail.com (Jay Lundgren) Date: Tue Jul 3 14:34:30 2012 Subject: [Histonet] Eyewash Requirements In-Reply-To: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> References: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> Message-ID: I test my shower every morning. 8) Sorry, couldn't resist. From sowmyakedarnath <@t> gmail.com Tue Jul 3 14:37:44 2012 From: sowmyakedarnath <@t> gmail.com (Sowmya Kedarnath) Date: Tue Jul 3 14:37:47 2012 Subject: [Histonet] Used Decloaker Message-ID: Hello Histonetters! Anybody interested in selling a used Decloaker kindly email me with your contact details.Would greatly appreciate the help. Best From rjr6 <@t> psu.edu Tue Jul 3 14:46:07 2012 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Tue Jul 3 14:46:33 2012 Subject: [Histonet] Eyewash Requirements In-Reply-To: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> References: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> Message-ID: We have to test the eyewash weekly and let it run for at least 30 seconds. The shower is tested by the university's safety people I think just once a year. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie@blufrogpath.com Sent: Tuesday, July 03, 2012 3:25 PM To: Histonet post Subject: [Histonet] Eyewash Requirements What is the time requirement for flushing the eyewash? We u=d to flush for 30 seconds and then I was told by our safety officer that it should be 3 minutes. How often do you test your shower? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Tue Jul 3 14:58:57 2012 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Tue Jul 3 14:59:00 2012 Subject: [Histonet] Eyewash Requirements In-Reply-To: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> Message-ID: <14E2C6176416974295479C64A11CB9AE0113A9690BF2@SBS2K8.premierlab.local> We run/test weekly both the safety shower and eyewash. (I have seen where you need to do a weekly assessment and then a monthy test, we just decided to do it weekly that way we know that the water has not been the pipes for a long time). We leave the eyewash on for about 5 minutes, its easy to do that since it drains, but the safety shower we collect in a 5 gallon bucket and that fills up in really quickly, so about 30 seconds, I'm not sure there is a testing system that could hold 3 minutes of water from the shower unless you are running into a large garbage can. You also need to test the temp of the water it should not exceed 100 degrees F. We also fill out a safety shower/eyewash program assessment yearly. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie@blufrogpath.com Sent: Tuesday, July 03, 2012 1:25 PM To: Histonet post Subject: [Histonet] Eyewash Requirements What is the time requirement for flushing the eyewash? We u=d to flush for 30 seconds and then I was told by our safety officer that it should be 3 minutes. How often do you test your shower? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pkromund <@t> gundluth.org Tue Jul 3 14:59:36 2012 From: pkromund <@t> gundluth.org (Romundstad, Pamela K) Date: Tue Jul 3 15:00:50 2012 Subject: [Histonet] Automated Special Stainers Message-ID: Hello, I'm interested in opinions on what Histologist prefer in their automated special stain instruments. Is it primarily between DAKO and Ventana? Any imput would be appreciated. Pamela Romundstad HT, QIHC Gundersen Lutheran ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of histonet-request@lists.utsouthwestern.edu [histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, July 03, 2012 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 104, 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. Embedding beads (Amber McKenzie) 2. Re: Embedding beads (Jennifer Campbell) 3. Re: Embedding beads (Alan Taylor) 4. Slide Baskets???? (heather marlatt) 5. Humidity Check (Laurie@blufrogpath.com) 6. Mohs (Rebecca a. Johnson) 7. What is this item called? (Adam Haberman) 8. Re: What is this item called? (Jennifer MacDonald) 9. RE: What is this item called? (Rittman, Barry R) 10. RE: Mohs (Harrison, Sandra C.) 11. RE: Humidity Check (Harrison, Sandra C.) 12. Competency check list (JMaslanka@stpetes.org) 13. RE: Humidity Check (Ingles Claire ) 14. RE: Mohs (Ingles Claire ) 15. JB fixative (Joe Hardin) 16. Re: Mohs (Kim Tournear) 17. Billing 88342 (Victor A. Tobias) ---------------------------------------------------------------------- Message: 1 Date: Mon, 2 Jul 2012 17:17:20 +0000 From: Amber McKenzie Subject: [Histonet] Embedding beads To: "histonet@lists.utsouthwestern.edu" Message-ID: <5A33C952BB67F4468AF1F36D739212BC308296E3@JERRY.Gia.com> Content-Type: text/plain; charset="iso-8859-1" How do you not cut thru the bead when sectioning? I'm intrigued by this process b/c I've never heard of this before. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kelly Boyd Sent: Tuesday, June 26, 2012 3:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Embedding beads I started using the embedding beads several months ago as a quick way to track who embedded what cassette. Each tech is assigned a number according to the number on their microtome. I get the beads from Cancer Diagnostics. You can probably find something similar at a craft store. These come in small round plastic containers that fit easily any where on the embedding center. They place the bead in a bottom corner of the cassette when topping it off with paraffin. They?have actually saved us time. Once the techs get used to it, it might add a few seconds to the embedding.?Before, the techs had to write down which blocks they embedded. (Very time consuming and often not complete).?If I needed to see who embedded a certain block, I had to go check that log book. Now I can see who embedded it just by looking at the block. Kelly?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Mon, 2 Jul 2012 14:29:33 -0400 From: Jennifer Campbell Subject: Re: [Histonet] Embedding beads To: Amber McKenzie Cc: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset=ISO-8859-1 The identifier(whatever is using to distinguish who did the task) is not embedded with the specimen. It is put in the top portion of the cassette. At least that is what we have always done here. On Mon, Jul 2, 2012 at 1:17 PM, Amber McKenzie < amber.mckenzie@gastrodocs.net> wrote: > How do you not cut thru the bead when sectioning? I'm intrigued by this > process b/c I've never heard of this before. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto: > histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kelly Boyd > Sent: Tuesday, June 26, 2012 3:20 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Embedding beads > > I started using the embedding beads several months ago as a quick way to > track who embedded what cassette. Each tech is assigned a number according > to the number on their microtome. I get the beads from Cancer Diagnostics. > You can probably find something similar at a craft store. > These come in small round plastic containers that fit easily any where on > the embedding center. They place the bead in a bottom corner of the > cassette when topping it off with paraffin. > They have actually saved us time. Once the techs get used to it, it might > add a few seconds to the embedding. Before, the techs had to write down > which blocks they embedded. (Very time consuming and often not > complete). If I needed to see who embedded a certain block, I had to go > check that log book. Now I can see who embedded it just by looking at the > block. > > Kelly > _______________________________________________ > 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 > -- Jen Campbell, HT(ASCP) Supervisor of Technical Services Muhlbauer Dermatopathology Laboratory 61 Monroe Avenue, Ste B Pittsford NY 14534 P: 585.586.5166 F: 585.586.3137 IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. ------------------------------ Message: 3 Date: Mon, 2 Jul 2012 19:37:50 +0100 From: "Alan Taylor" Subject: Re: [Histonet] Embedding beads To: "Amber McKenzie" , Message-ID: <3A078ACBD81E4F39B2B2C8FAD4C6332F@merlin> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Hi All The embedding bead goes into the cassette tray, into one corner, not into the stainless steel embedding mould with the tissue. The plastic cassette is placed on top of the mould, as usual and filled with molten wax, the coloured bead is quickly dropped in and the embedding mould is placed on the chiller to solidify in preparation for microtomy. The bead has no contact with the blade at any time, remaining at the back of the cassette tray. Hope this clarifies this easy process. Alan Taylor BSc(Hons), FRMS. Microtechnical Services 71 Sweetbrier Lane Heavitree Exeter. EX1 3AJ. Devon. UK. ----- Original Message ----- From: "Amber McKenzie" To: Sent: Monday, July 02, 2012 6:17 PM Subject: [Histonet] Embedding beads How do you not cut thru the bead when sectioning? I'm intrigued by this process b/c I've never heard of this before. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kelly Boyd Sent: Tuesday, June 26, 2012 3:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Embedding beads I started using the embedding beads several months ago as a quick way to track who embedded what cassette. Each tech is assigned a number according to the number on their microtome. I get the beads from Cancer Diagnostics. You can probably find something similar at a craft store. These come in small round plastic containers that fit easily any where on the embedding center. They place the bead in a bottom corner of the cassette when topping it off with paraffin. They have actually saved us time. Once the techs get used to it, it might add a few seconds to the embedding. Before, the techs had to write down which blocks they embedded. (Very time consuming and often not complete). If I needed to see who embedded a certain block, I had to go check that log book. Now I can see who embedded it just by looking at the block. Kelly _______________________________________________ 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 ------------------------------ Message: 4 Date: Mon, 2 Jul 2012 12:17:01 -0700 From: heather marlatt Subject: [Histonet] Slide Baskets???? To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset=ISO-8859-1 I was wondering if anyone in histoland has any slide baskets for the old sakura DRS 601 stainer laying around??? I can't seem to find these for a decent price so I thought I'd ask. I only have one at the moment and as you might imagine that us really cramping my style. Thanks in advance Heather ------------------------------ Message: 5 Date: Mon, 02 Jul 2012 13:19:43 -0700 From: Subject: [Histonet] Humidity Check To: "Histonet post" Message-ID: <20120702131943.295dc6182df7e5cbb4f32bc101c30dcc.f604e52a69.wbe@email15.secureserver.net> Content-Type: text/plain; charset="utf-8" Do others check the humidity of their Histo lab on a daily basis? Laurie Colbert ------------------------------ Message: 6 Date: Mon, 2 Jul 2012 18:37:20 -0400 From: "Rebecca a. Johnson" Subject: [Histonet] Mohs To: "histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Need to know what Mohs techs are getting paid. Thanks Becky ------------------------------ Message: 7 Date: Tue, 3 Jul 2012 08:45:53 -0400 From: Adam Haberman Subject: [Histonet] What is this item called? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hello everyone, I am trying to identify a piece of glassware so that I can order a few more of it. It is a thick piece of glass with a large, wide depression in the middle. The item I am looking for is thicker and wider than a depression slide, and I use it as a dissecting dish. I have included a picture of the piece I am trying to identify, along with a depression slide for comparison. This is probably an item that is not widely used anymore, as it seems to predate all of the faculty here. However, it is perfect for the dissections I do with two pair of forceps under a dissecting microscope. It gives me much better working angles than the spot plates I resort to when I cannot find this item. If anyone can identify the item or, even better, tell me where I can order more, please let me know. Thank you very much. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 ------------------------------ Message: 8 Date: Tue, 3 Jul 2012 06:13:37 -0700 From: Jennifer MacDonald Subject: Re: [Histonet] What is this item called? To: Adam Haberman Cc: "histonet@lists.utsouthwestern.edu" Message-ID: <2FB72470-1EA9-4BF5-8B04-4D613C5F06EE@mtsac.edu> Content-Type: text/plain; charset=us-ascii A watch glass? On Jul 3, 2012, at 5:45 AM, Adam Haberman wrote: > Hello everyone, > > I am trying to identify a piece of glassware so that I can order a few more > of it. It is a thick piece of glass with a large, wide depression in the > middle. The item I am looking for is thicker and wider than a depression > slide, and I use it as a dissecting dish. I have included a picture of the > piece I am trying to identify, along with a depression slide for > comparison. This is probably an item that is not widely used anymore, as > it seems to predate all of the faculty here. However, it is perfect for > the dissections I do with two pair of forceps under a dissecting > microscope. It gives me much better working angles than the spot plates I > resort to when I cannot find this item. > > If anyone can identify the item or, even better, tell me where I can order > more, please let me know. Thank you very much. > > --Adam > > Adam Haberman > Visiting Assistant Professor of Biology > Oberlin College > adam.haberman@oberlin.edu > (440)775-6502 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 3 Jul 2012 08:15:31 -0500 From: "Rittman, Barry R" Subject: RE: [Histonet] What is this item called? To: "histonet@lists.utsouthwestern.edu" Message-ID: <12A4DAFC2FEBB84B8DED5F5E9201B4E9179EC272E4@UTHCMS1.uthouston.edu> Content-Type: text/plain; charset="us-ascii" Is this a 3 by 1.25 inch glass slide approximately 3 mm thick with a 1 inch depression in the middle? Barry To: Adam Haberman Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] What is this item called? A watch glass? On Jul 3, 2012, at 5:45 AM, Adam Haberman wrote: > Hello everyone, > > I am trying to identify a piece of glassware so that I can order a few more > of it. It is a thick piece of glass with a large, wide depression in the > middle. The item I am looking for is thicker and wider than a depression > slide, and I use it as a dissecting dish. I have included a picture of the > piece I am trying to identify, along with a depression slide for > comparison. This is probably an item that is not widely used anymore, as > it seems to predate all of the faculty here. However, it is perfect for > the dissections I do with two pair of forceps under a dissecting > microscope. It gives me much better working angles than the spot plates I > resort to when I cannot find this item. > > If anyone can identify the item or, even better, tell me where I can order > more, please let me know. Thank you very much. > > --Adam > > Adam Haberman > Visiting Assistant Professor of Biology > Oberlin College > adam.haberman@oberlin.edu > (440)775-6502 > _______________________________________________ > 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 ------------------------------ Message: 10 Date: Tue, 3 Jul 2012 08:22:50 -0500 From: "Harrison, Sandra C." Subject: RE: [Histonet] Mohs To: "Rebecca a. Johnson" , "histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Depends ;-) All kidding aside, there are factors that weigh in, such as geography, availability, training, experience, speed, etc. 10 years ago, when I worked as a Mohs' tech in the Denver, Colorado area, I received $25.00 per hour, plus benefits. I usually only worked 3 days per week, but on some rare occasions, worked till 9 p.m. trying to get to clear margins. I don't know what the current wages are. Sandy Harrison, HTL Histology Supervisor, Minneapolis VA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca a. Johnson Sent: Monday, July 02, 2012 5:37 PM To: histonet Subject: [Histonet] Mohs Need to know what Mohs techs are getting paid. Thanks Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 3 Jul 2012 08:23:52 -0500 From: "Harrison, Sandra C." Subject: RE: [Histonet] Humidity Check To: , "Histonet post" Message-ID: Content-Type: text/plain; charset="utf-8" Yes, we have a wireless temperature monitoring system, called Checkpoint, and each room has a sensor that monitors temp and humidity. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie@blufrogpath.com Sent: Monday, July 02, 2012 3:20 PM To: Histonet post Subject: [Histonet] Humidity Check Do others check the humidity of their Histo lab on a daily basis?= Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Tue, 3 Jul 2012 08:03:52 -0600 From: JMaslanka@stpetes.org Subject: [Histonet] Competency check list To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Would anyone be willing to share their IHC tech competency check list? Joe Maslanka BS, CT,HT (ASCP) Anatomical Pathology Technical Supervisor St Peter's Hospital,MT 59601 (P)(406) 447-2406 (F)(406)444-2126 Give thanks for ALL things..... ------------------------------ Message: 13 Date: Tue, 3 Jul 2012 10:06:08 -0500 From: "Ingles Claire " Subject: RE: [Histonet] Humidity Check To: "Histonet post" Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A866@UWHC-MAIL2.uwhis.hosp.wisc.edu> Content-Type: text/plain; charset="iso-8859-1" I think it is actually a requirement that temp and humidity are monitiored daily. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie@blufrogpath.com Sent: Mon 7/2/2012 3:19 PM To: Histonet post Subject: [Histonet] Humidity Check Do others check the humidity of their Histo lab on a daily basis?= Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Tue, 3 Jul 2012 10:07:13 -0500 From: "Ingles Claire " Subject: RE: [Histonet] Mohs To: "histonet" Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A867@UWHC-MAIL2.uwhis.hosp.wisc.edu> Content-Type: text/plain; charset="iso-8859-1" I would love to know this too. I am getting about $23/hr but I have pretty good benefits. I'm at UWisc Madison Hospital. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rebecca a. Johnson Sent: Mon 7/2/2012 5:37 PM To: histonet Subject: [Histonet] Mohs Need to know what Mohs techs are getting paid. Thanks Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Tue, 03 Jul 2012 10:51:16 -0500 From: Joe Hardin Subject: [Histonet] JB fixative To: histonet@lists.utsouthwestern.edu Message-ID: <4FF314F4.5020707@oncology.wisc.edu> Content-Type: text/plain; charset="us-ascii" Hi All, I use a Zinc fixative called JB fixitive for tissues that I want to stain for CD antigens(see attachment). Does anyone know if this fixative kills all microbes and prions? ------------------------------ Message: 16 Date: Tue, 3 Jul 2012 10:59:53 -0500 From: Kim Tournear Subject: Re: [Histonet] Mohs To: Ingles Claire Cc: histonet Message-ID: <5C862879-5A6D-4D83-A093-DACEC3AF0ACA@yahoo.com> Content-Type: text/plain; charset=us-ascii You can get upwards of 30.00/hr in Arizona. Sent from the iPhone of Kim Tournear. On Jul 3, 2012, at 10:07 AM, "Ingles Claire " wrote: > I would love to know this too. I am getting about $23/hr but I have pretty good benefits. I'm at UWisc Madison Hospital. > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rebecca a. Johnson > Sent: Mon 7/2/2012 5:37 PM > To: histonet > Subject: [Histonet] Mohs > > > > > Need to know what Mohs techs are getting paid. > > > Thanks > Becky > _______________________________________________ > 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 ------------------------------ Message: 17 Date: Tue, 3 Jul 2012 16:43:05 +0000 From: "Victor A. Tobias" Subject: [Histonet] Billing 88342 To: "'HISTONET'" Message-ID: Content-Type: text/plain; charset="us-ascii" Looking for other opinions from those who do consult/referral work. If a client sends in a request for a single antibody done on multiple blocks on a single specimen, do you bill the client for each tech component ? The client will do the interpretation. What happens in the above scenario if the request is to bill the patient? Knowing you get reimbursed for one, do you eat the other charges are make the client select the one block? We have run numbers on potential lost revenue and the number is significant. Victor Victor Tobias HT(ASCP) Clinical Applications Analyst Harborview Medical Center Dept of Pathology Room NJB244 Seattle, WA 98104 vtobias@u.washington.edu 206-744-2735 206-744-8240 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 104, Issue 3 **************************************** From adam.haberman <@t> oberlin.edu Tue Jul 3 15:10:43 2012 From: adam.haberman <@t> oberlin.edu (Adam Haberman) Date: Tue Jul 3 15:10:48 2012 Subject: [Histonet] What is this item called? In-Reply-To: References: Message-ID: Jay, there are no markings on it at all. They are making it hard for me to give them money. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 On Tue, Jul 3, 2012 at 3:32 PM, Jay Lundgren wrote: > > Brilliant way to send a picture. I <3 interwebs. Whatever it is, I want > one. So it's made of optical quality glass, like a slide or a coverslip? > Does it have a company's name on it? I'll bet you it's expensive. > > Sincerely, > > Jay A. Lundgren, > M.S., HTL (ASCP) > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > From mgriffo <@t> u.washington.edu Tue Jul 3 16:36:34 2012 From: mgriffo <@t> u.washington.edu (Maureen Griffo) Date: Tue Jul 3 16:37:05 2012 Subject: [Histonet] Test Message-ID: <4FF365E2.1030602@u.washington.edu> Hello, Testing this email to see if my message is being viewed. Please shoot me a quick "yes" if you see this. Thank you, Maureen From Tony_Reilly <@t> health.qld.gov.au Tue Jul 3 18:33:33 2012 From: Tony_Reilly <@t> health.qld.gov.au (Tony Reilly) Date: Tue Jul 3 18:34:14 2012 Subject: [Histonet] What is this item called? In-Reply-To: References: Message-ID: <4FF40DEC.411C.0039.0@health.qld.gov.au> Hello I have not seen one of these for many years. I may be wrong but in the back of my mind I seem to remember using these at university for growing cell cultures. That might be an area to focus your search though they have most likely been superceded by now. regards Tony Tony Reilly B.App.Sc. , M.Sc. Chief Scientist, Anatomical Pathology Pathology Queensland-PA Laboratory _________________________________________________ Clinical and Statewide Services Division| QueenslandHealth Level 1, Building 15,Princess Alexandra Hospital Ipswich Road,WOOLLOONGABBA Qld4102 Ph: 07 3176 2412 Mob: 0402 139411 Fax: 07 3176 2930 Email: tony_reilly@health.qld.gov.au Web: www.health.qld.gov.au/qhcss/ >>> Adam Haberman 7/4/2012 5:23 am >>> Barry, It is 3" x 1 3/4", and the depression has a 1 1/2" diameter. Thanks for all the ideas so far. It isn't a hanging drop slide or a watch glass. You can see a picture of it next to a depression slide here: http://imgur.com/yBwvw I don't see a better way to send a picture on this list. --Adam Adam Haberman Visiting Assistant Professor of Biology Oberlin College adam.haberman@oberlin.edu (440)775-6502 On Tue, Jul 3, 2012 at 2:27 PM, Long, Florence wrote: > Hi Adam, > Would that be a modification of a "hanging-drop slide"? > F. Long > > ________________________________________ > From: histonet-bounces@lists.utsouthwestern.edu [ > histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adam Haberman [ > adam.haberman@oberlin.edu] > Sent: Tuesday, July 03, 2012 8:45 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] What is this item called? > > Hello everyone, > > I am trying to identify a piece of glassware so that I can order a few more > of it. It is a thick piece of glass with a large, wide depression in the > middle. The item I am looking for is thicker and wider than a depression > slide, and I use it as a dissecting dish. I have included a picture of the > piece I am trying to identify, along with a depression slide for > comparison. This is probably an item that is not widely used anymore, as > it seems to predate all of the faculty here. However, it is perfect for > the dissections I do with two pair of forceps under a dissecting > microscope. It gives me much better working angles than the spot plates I > resort to when I cannot find this item. > > If anyone can identify the item or, even better, tell me where I can order > more, please let me know. Thank you very much. > > --Adam > > Adam Haberman > Visiting Assistant Professor of Biology > Oberlin College > adam.haberman@oberlin.edu > (440)775-6502 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -This e-mail and any attachments may contain CONFIDENTIAL information, > including PROTECTED HEALTH INFORMATION. If you are not the intended > recipient, any use or disclosure of this information is STRICTLY > PROHIBITED; you are requested to delete this e-mail and any attachments, > notify the sender immediately, and notify the LabCorp Privacy Officer at > privacyofficer@labcorp.com or call (877) 23-HIPAA / (877) 234-4722. > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ******************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. ********************************************************************************** From c_m_hernandezjr <@t> yahoo.com Tue Jul 3 23:34:22 2012 From: c_m_hernandezjr <@t> yahoo.com (Carlos Hernandez) Date: Tue Jul 3 23:34:26 2012 Subject: [Histonet] Immuno Controls Message-ID: <1341376462.63284.YahooMailNeo@web31804.mail.mud.yahoo.com> Hi All! I was wondering if there is anybody out there that has an over abundance of controls for mart/melan a, s100, hmb45, pan keratin, ki-67, and mitf that they are will to share or sell? ?Please let me know if you can help me out. Thanks, Carlos From daniela.bodemer <@t> mcri.edu.au Wed Jul 4 06:02:56 2012 From: daniela.bodemer <@t> mcri.edu.au (Daniela Bodemer) Date: Wed Jul 4 06:03:08 2012 Subject: [Histonet] Frozens and antigen retrieval Message-ID: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad ______________________________________________________________________ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com ______________________________________________________________________ From histotalk <@t> yahoo.com Wed Jul 4 06:18:44 2012 From: histotalk <@t> yahoo.com (David Kemler) Date: Wed Jul 4 06:18:52 2012 Subject: [Histonet] Mohs In-Reply-To: References: Message-ID: <1341400724.34324.YahooMailNeo@web121501.mail.ne1.yahoo.com> I assume you are talking about HT(ASCP) Registered techs. The majority of Mohs Techs are "off-the-street" people. Here in FL for Registered Licensed techs, between $25 - $30 / hour. The "off-the-street" folks" $14 - $18, sometimes as low as $12.00 / hr. ? Yours, Dave ________________________________ From: Rebecca a. Johnson To: histonet Sent: Monday, July 2, 2012 6:37 PM Subject: [Histonet] Mohs ? ? ? ? Need to know what Mohs techs are getting paid.? ? ? Thanks ? ? Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Wed Jul 4 06:28:48 2012 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Wed Jul 4 06:28:57 2012 Subject: [Histonet] Frozens and antigen retrieval In-Reply-To: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> References: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> Message-ID: Usually not. Fixation cross-links the proteins, which can mask the epitope (=antibody binding site of the antigen). So antigen retrieval breaks the fixative cross-links, exposing the epitope. If there's no fixation, there's no cross-links, so the epitope is usually exposed and available to easily bind to the antibody. Plus, there's no destruction of tissue morphology if you're not using antigen retrieval, so the quality of the section looks much nicer. That being said, there may be some antibody out there that still needs antigen retrieval on frozen section, but then the company's protocol probably wouldn't say "optional". So try it the first time without antigen retrieval. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 The opinions expressed are mine, and do not reflect Beaumont. -----Original Message----- From: Daniela Bodemer Sent: Wednesday, July 04, 2012 7:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozens and antigen retrieval Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad ______________________________________________________________________ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com ______________________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ibernard <@t> uab.edu Wed Jul 4 08:32:47 2012 From: ibernard <@t> uab.edu (Ian R Bernard) Date: Wed Jul 4 08:32:57 2012 Subject: [Histonet] Immuno Controls In-Reply-To: <1341376462.63284.YahooMailNeo@web31804.mail.mud.yahoo.com> References: <1341376462.63284.YahooMailNeo@web31804.mail.mud.yahoo.com> Message-ID: Looking for the same Ian -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carlos Hernandez Sent: Tuesday, July 03, 2012 11:34 PM To: Histonet Subject: [Histonet] Immuno Controls Hi All! I was wondering if there is anybody out there that has an over abundance of controls for mart/melan a, s100, hmb45, pan keratin, ki-67, and mitf that they are will to share or sell? ?Please let me know if you can help me out. Thanks, Carlos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From W.E.J.Hoekert <@t> olvg.nl Wed Jul 4 10:09:03 2012 From: W.E.J.Hoekert <@t> olvg.nl (Hoekert, W.E.J.) Date: Wed Jul 4 10:09:14 2012 Subject: [Histonet] How to make HP control tissue? References: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> Message-ID: <1190CB05C44B13409483514729C2FC3601F84261@PAIT42.olvg.nl> Hi Histonetters, Does anybody has experience in making your own HP control tissue? I am tired of using positive biopsies of patients since they are always almost finished. I have heard of a procedure on making HP controls but I am not exactly sure how it is done. I have tried the following: I went to the microbiology department and asked for some freshly grown HP bacteria. We scraped them of the petridishes and put them in formalin. I had them fixed for 24 hours and then I injected them into some already fixed lung tissue and processed it as normal. But if I do a HP stain on that tissue, I don't see any bacteria. Probably they rinse off during the process. Can anyone tell me how to do it? Thanks in advance, Willem Hoekert OLVG The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. From chapcl <@t> yahoo.com Wed Jul 4 11:10:51 2012 From: chapcl <@t> yahoo.com (William) Date: Wed Jul 4 11:11:02 2012 Subject: [Histonet] How to make HP control tissue? In-Reply-To: <1190CB05C44B13409483514729C2FC3601F84261@PAIT42.olvg.nl> References: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> <1190CB05C44B13409483514729C2FC3601F84261@PAIT42.olvg.nl> Message-ID: It may be hard to find someone to help you in the States. Doing that is against regulations. We must use human tissue with a natural infection I we are going to be using it as a control for human medical testing. That being said, you are right, your hp is washing out of the tissue. My suggestion is to not fix the hp culture, inject it fresh into unfixed tissue. Preferable fresh autopsy stomach or intestine. Allow it to culture (and attach) for about an hour or 2 and then fix. Be careful too long out of fixative and autolysis will occur. I hope that works. I have no experience with lung. It should work, but I do not know if the environment would cause the bacteria to attach and grow. Will Sent from my iPhone On Jul 4, 2012, at 11:09 AM, "Hoekert, W.E.J." wrote: > Hi Histonetters, > > Does anybody has experience in making your own HP control tissue? I am tired of using positive biopsies of patients since they are always almost finished. I have heard of a procedure on making HP controls but I am not exactly sure how it is done. > > I have tried the following: I went to the microbiology department and asked for some freshly grown HP bacteria. We scraped them of the petridishes and put them in formalin. I had them fixed for 24 hours and then I injected them into some already fixed lung tissue and processed it as normal. But if I do a HP stain on that tissue, I don't see any bacteria. Probably they rinse off during the process. > > Can anyone tell me how to do it? > > Thanks in advance, > > Willem Hoekert > OLVG > The Netherlands > > > Disclaimer: > > Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. > In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marktarango <@t> gmail.com Wed Jul 4 11:40:27 2012 From: marktarango <@t> gmail.com (Mark Tarango) Date: Wed Jul 4 11:40:31 2012 Subject: [Histonet] How to make HP control tissue? In-Reply-To: References: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> <1190CB05C44B13409483514729C2FC3601F84261@PAIT42.olvg.nl> Message-ID: Hi Willem, H. pylori needs the low pH of the stomach to survive and grow. It won't naturally be in lung tissue. When looking for HP, it's found on the mucosa and gastric pits of the stomach. Since you're looking for it in a specific place, I think putting it in lung is a bad idea. Your best bet for control tissue is a positive gastrectomy specimen. You could chop that up into a thousand pieces and never run out. We've had several cases like this. Send me your address and a FedEx account number and I'll see about send you a block or two. Mark On Wednesday, July 4, 2012, Hoekert, W.E.J. wrote: > Hi Histonetters, > > Does anybody has experience in making your own HP control tissue? I am > tired of using positive biopsies of patients since they are always almost > finished. I have heard of a procedure on making HP controls but I am not > exactly sure how it is done. > > I have tried the following: I went to the microbiology department and > asked for some freshly grown HP bacteria. We scraped them of the > petridishes and put them in formalin. I had them fixed for 24 hours and > then I injected them into some already fixed lung tissue and processed it > as normal. But if I do a HP stain on that tissue, I don't see any bacteria. > Probably they rinse off during the process. > > Can anyone tell me how to do it? > > Thanks in advance, > > Willem Hoekert > OLVG > The Netherlands > > > Disclaimer: > > Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). > Verstrekking aan en gebruik door anderen dan geadresseerden is niet > toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de > verzender hiervan op de hoogte te stellen en het bericht te verwijderen. > In verband met electronische verzending kunnen aan dit e-mail bericht geen > rechten worden ontleend. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ewolfe118 <@t> gmail.com Wed Jul 4 14:21:50 2012 From: ewolfe118 <@t> gmail.com (E Wolfe) Date: Wed Jul 4 14:21:55 2012 Subject: [Histonet] PCNA sc-7907? Message-ID: Hello Histonet - Has anyone had any success staining with PCNA sc-7907 on frozen, paraformaldehyde-fixed tissue? I am experiencing a lot of background, and so far diluting down to 1:5000 hasn't helped at all. Any advice would be very much appreciated! From fbozkurt <@t> gmail.com Wed Jul 4 15:00:01 2012 From: fbozkurt <@t> gmail.com (Mehmet Fatih BOZKURT) Date: Wed Jul 4 15:00:05 2012 Subject: [Histonet] PCNA sc-7907? In-Reply-To: References: Message-ID: dilute antibody with %1 BSA.. and use %1 BSA for serum blocking... On Wed, Jul 4, 2012 at 10:59 PM, Mehmet Fatih BOZKURT wrote: > dilute antibody with %1 BSA.. and use %1 BSA for serum blocking... > > On Wed, Jul 4, 2012 at 10:21 PM, E Wolfe wrote: > >> Hello Histonet - >> >> Has anyone had any success staining with PCNA sc-7907 on frozen, >> paraformaldehyde-fixed tissue? I am experiencing a lot of background, and >> so far diluting down to 1:5000 hasn't helped at all. Any advice would be >> very much appreciated! >> _______________________________________________ >> 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: +902722281312-173/237 > > -- Mehmet Fatih BOZKURT, DVM, PhD Afyon Kocatepe University Faculty of Veterinary Medicine Department of Pathology 03030, ANS Campus Afyonkarahisar-TURKEY Tel: +902722281312-173/237 From pvalente <@t> sbcglobal.net Wed Jul 4 21:15:03 2012 From: pvalente <@t> sbcglobal.net (Patricia Valente) Date: Wed Jul 4 21:15:14 2012 Subject: [Histonet] (no subject) Message-ID: <1341454503.40538.YahooMailNeo@web181315.mail.ne1.yahoo.com> http://newmodeluk.com/time.php?eight207.jpg From MWhite <@t> mhs.net Thu Jul 5 05:36:08 2012 From: MWhite <@t> mhs.net (White, Marcia) Date: Thu Jul 5 05:36:16 2012 Subject: [Histonet] RE: Billing 88342 In-Reply-To: References: Message-ID: <8B4B5BF3FCB6D24CBEAA63ACA5FDAD3B045D04A5@MHSEXMB04.mhs.net> Sue is correct you can only bill once even if you perform the same stain on multiple blocks. Marcia M White Pathology Department Memorial Hospital West 954-844-4197 954-967-7627(fax) -----Original Message----- From: Sue Hunter [mailto:SHUNTER@beaumont.edu] Sent: Tuesday, July 03, 2012 1:52 PM To: Victor A. Tobias; 'HISTONET' Subject: [Histonet] RE: Billing 88342 It is my understanding that you can only bill for one 88342 on a specimen - technical or professional, even if you perform the same stain on multiple blocks of that specimen. For A1, A2, A3 etc you can only charge once. For A1, B1, C1, etc you may charge a 88342 per specimen. Sue Sue Hunter, Supervisor Advanced Diagnostics Beaumont Health System Royal Oak MI 248-898-5146 shunter@beaumont.edu ----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Victor A. Tobias Sent: Tuesday, July 03, 2012 12:43 PM To: 'HISTONET' Subject: [Histonet] Billing 88342 Looking for other opinions from those who do consult/referral work. If a client sends in a request for a single antibody done on multiple blocks on a single specimen, do you bill the client for each tech component ? The client will do the interpretation. What happens in the above scenario if the request is to bill the patient? Knowing you get reimbursed for one, do you eat the other charges are make the client select the one block? We have run numbers on potential lost revenue and the number is significant. Victor Victor Tobias HT(ASCP) Clinical Applications Analyst Harborview Medical Center Dept of Pathology Room NJB244 Seattle, WA 98104 vtobias@u.washington.edu 206-744-2735 206-744-8240 Fax =================================================Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet To report this email as SPAM, please forward it to spam@websense.com. From Farnana <@t> nehealth.com Thu Jul 5 06:42:56 2012 From: Farnana <@t> nehealth.com (Amy Farnan) Date: Thu Jul 5 06:43:07 2012 Subject: [Histonet] 88342 billing Message-ID: <4FF5457B.26ED.00D9.1@nehealth.com> Hi Victor, I am coming in late on this conversation regarding billing of 88342. You can only bill a single specimen once for an antibody. If you were performing multiple antibodies then you can charge 88342 x the number of antibodies. If you charge more it can be considered fraud so be careful. A great publication for these types of questions is called Coding Alert. It talks about all these types of issues and new cpt's etc. One other suggestion that I noted in a response is if its a self bill or I should say a client bill is to bill them the number of times you stained it as long as its not insurance. This is also fraud. You can not change your billing depending on if a patient's insurance is being billed or not. Amy Disclaimer: The information in this message is confidential. If you are not the intended recipient, do not disclose, copy, or distribute this message, and please immediately contact the sender. From gmartin <@t> marshallmedical.org Thu Jul 5 10:40:35 2012 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Thu Jul 5 10:40:40 2012 Subject: [Histonet] Grandfather TC Change Message-ID: <6ED9D4252F278841A0593D3D788AF24C14931B1E@mailsvr.MARSHMED.local> I'm interested if anyone is dealing with the expired Grandfather TC rule. I'm mainly interested in how you may be negotiating with your hospital concerning the reimbursement difference. Any thoughts would be helpful. Gary Martin Director of Operations El Dorado Pathology Med. Grp. Marshall Med. Ctr. Pathology Dept. 530-626-2608 gmaritn@marshallhospital.org This e-mail is only intended for the person 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 then the intended recipient is strictly prohibited. If you receive this communication in error, please contact the sender and destroy all forms of this communication. From Timothy.Morken <@t> ucsfmedctr.org Thu Jul 5 10:58:56 2012 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Timothy) Date: Thu Jul 5 10:58:30 2012 Subject: [Histonet] Nerve tease specimen holding Message-ID: <8D7C2D242DBD45498006B21122072BF8B5A84283@MCINFRWEM003.ucsfmedicalcenter.org> For all you doing nerve teasing, do you hold the specimen in buffer (after fixation) until teasing is requested or go ahead and process through osmium and glycerol and then hold? Thanks! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.morken@ucsfmedctr.org From Joyce.Weems <@t> emoryhealthcare.org Thu Jul 5 11:09:39 2012 From: Joyce.Weems <@t> emoryhealthcare.org (Weems, Joyce K.) Date: Thu Jul 5 11:09:54 2012 Subject: [Histonet] RE: Grandfather TC Change In-Reply-To: <6ED9D4252F278841A0593D3D788AF24C14931B1E@mailsvr.MARSHMED.local> References: <6ED9D4252F278841A0593D3D788AF24C14931B1E@mailsvr.MARSHMED.local> Message-ID: There's no negotiation that I know of.. we will probably just lose money because the DRG will not include those costs. We have good client pricing with our reference labs, but we won't know how this will affect us all until the dust settles. Maybe the DRG fairies will adjust the reimbursement to include those tests!!??? Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martin, Gary Sent: Thursday, July 05, 2012 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grandfather TC Change I'm interested if anyone is dealing with the expired Grandfather TC rule. I'm mainly interested in how you may be negotiating with your hospital concerning the reimbursement difference. Any thoughts would be helpful. Gary Martin Director of Operations El Dorado Pathology Med. Grp. Marshall Med. Ctr. Pathology Dept. 530-626-2608 gmaritn@marshallhospital.org This e-mail is only intended for the person 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 then the intended recipient is strictly prohibited. If you receive this communication in error, please contact the sender and destroy all forms of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). From brian <@t> prometheushealthcare.com Thu Jul 5 11:19:34 2012 From: brian <@t> prometheushealthcare.com (Brian-Prometheus) Date: Thu Jul 5 11:19:44 2012 Subject: [Histonet] Histology Openings in NY Message-ID: <020001cd5ac9$f9b3f820$ed1be860$@com> Good Morning! My name is Brian Feldman with Prometheus Healthcare and I specialize in the recruitment of lab professionals nationwide. I just wanted to reach out to you about a few histology openings I have in the NY area. -Histotechs needed at a reference lab in Port Chester NY, night shift histotechs and mid shift grosser* -Grosser and histotechs needed at a private lab in Westchester county NY, overnight position* -Histotech needed at a hospital in NYC, must have ASCP* -Histotechs needed at a private lab in Suffern, NY - all shifts* *need state license All positions are full time and permanent and offer competitive salaries and full benefits. If you know anyone that would be interested in these positions, please feel free to pass along this information. I would appreciate any help and we do offer a referral bonus. Or, for immediate consideration, they can reply back with their most up to date resume. Thank you so much for your time! Brian Feldman Principal Prometheus Healthcare Office 301-693-9057 Fax 301-368-2478 brian@prometheushealthcare.com www.prometheushealthcare.com *** Stay up to date on the newest positions and healthcare trends nationwide on Twitter!*** http://twitter.com/PrometheusBlog From estellamireles <@t> gmail.com Thu Jul 5 11:41:42 2012 From: estellamireles <@t> gmail.com (Stella Mireles) Date: Thu Jul 5 11:41:46 2012 Subject: [Histonet] Nerve tease specimen holding In-Reply-To: <8D7C2D242DBD45498006B21122072BF8B5A84283@MCINFRWEM003.ucsfmedicalcenter.org> References: <8D7C2D242DBD45498006B21122072BF8B5A84283@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: When I was doing nerve teasing, we would hold in glycerol in the refrig. On Thu, Jul 5, 2012 at 10:58 AM, Morken, Timothy < Timothy.Morken@ucsfmedctr.org> wrote: > For all you doing nerve teasing, do you hold the specimen in buffer (after > fixation) until teasing is requested or go ahead and process through osmium > and glycerol and then hold? > > Thanks! > > Tim Morken > Supervisor, Electron Microscopy/Neuromuscular Special Studies > Department of Pathology > UC San Francisco Medical Center > 505 Parnassus Ave, Box 1656 > Room S570 > San Francisco, CA 94143 > > (415) 353-1266 (ph) > (415) 514-3403 (fax) > tim.morken@ucsfmedctr.org > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Nancy_Schmitt <@t> pa-ucl.com Thu Jul 5 12:31:20 2012 From: Nancy_Schmitt <@t> pa-ucl.com (Nancy Schmitt) Date: Thu Jul 5 12:31:24 2012 Subject: [Histonet] Eyewash Requirements In-Reply-To: <20120703200431.DEC0A1AA036@mail.pa-ucl.com> References: <20120703200431.DEC0A1AA036@mail.pa-ucl.com> Message-ID: <906B4DA90ED1DB4DB6C7E94D7CEE6C36E1F990@PEITHA.wad.pa-ucl.com> Laurie- Our safety office also states 3 minutes for eyewash - we test and document weekly. We test our shower spring and fall for time and amount (50 liters in 1 minute) Nancy ------------------------------------------ Message: 14 Date: Tue, 03 Jul 2012 12:24:47 -0700 From: Subject: [Histonet] Eyewash Requirements To: "Histonet post" Message-ID: <20120703122447.295dc6182df7e5cbb4f32bc101c30dcc.e345099a2e.wbe@email15.secureserver.net> Content-Type: text/plain; charset="utf-8" What is the time requirement for flushing the eyewash? We u flush for 30 seconds and then I was told by our safety officer that it should be 3 minutes. How often do you test your shower? Laurie Colbert NOTICE: This email may contain legally privileged information. The information is for the use of only the intended recipient(s) even if addressed incorrectly. If you are not the intended recipient, please notify the sender that you have received it in error and then delete it along with any attachments. Thank you. From one_angel_secret <@t> yahoo.com Thu Jul 5 12:52:32 2012 From: one_angel_secret <@t> yahoo.com (Kim Donadio) Date: Thu Jul 5 12:52:37 2012 Subject: [Histonet] Frozens and antigen retrieval In-Reply-To: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> References: <6CABD9E9-578E-4027-89DE-3057ADA41990@mcri.edu.au> Message-ID: <1341510752.86543.YahooMailNeo@web112303.mail.gq1.yahoo.com> You should not have to use retreival. Retrieval either by heat/solution or enzyme digestion is usually needed when the antigen/antibody site has been blocked/masked by the fixation process. ? Kim D? ________________________________ From: Daniela Bodemer To: histonet@lists.utsouthwestern.edu Sent: Wednesday, July 4, 2012 7:02 AM Subject: [Histonet] Frozens and antigen retrieval Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad ______________________________________________________________________ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com ______________________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Margaret.Perry <@t> sdstate.edu Thu Jul 5 14:59:42 2012 From: Margaret.Perry <@t> sdstate.edu (Perry, Margaret) Date: Thu Jul 5 14:59:49 2012 Subject: [Histonet] what is this item called Message-ID: <25F4FBA34BE9D142964ECC4525B82AEE039F48@SDSU-EX03.jacks.local> I found something at tedpella.com. It's a glass bottom dish, catalog number 14026. There are several sizes. It looks similar to the one you posted. Margaret Perry HT(ASCP) Dept of Veterinary and Biomedical services Box 2175 South Dakota State University Brookings SD 57007 605-688-5638 From JadeRyan83 <@t> yahoo.com Thu Jul 5 23:39:40 2012 From: JadeRyan83 <@t> yahoo.com (Jade Ryan) Date: Thu Jul 5 23:39:44 2012 Subject: [Histonet] Opening lab, need consultation. Message-ID: <34D25E4E-4B3F-4E9D-993C-7330ACAC9FE4@yahoo.com> Hello Everyone, Our dermatologists wants to pursue in opening a derm lab in Roseville, Ca area. We need someone to be the point person for this position as a consult. Since the position is a consult, you dont need to be in our facility all throughtout the process. However, we do need you for the initial, middle, and final phase of project specially the training of the employee for the equipments. The duties include applying for clia license, ca lab license, equipment consult, lab layout & design, writing the procedural manual, etc. About our company, we do about 8k cases a year and all derm tissues. So right now, we only need the lab to be routine H/E with a couple of special stains for fungus and bacteria. Email me with your bids & what I can expect from you. Jade Ryan, MHA, BSN Sent from my iPad From kcruise <@t> path.wustl.edu Fri Jul 6 11:28:24 2012 From: kcruise <@t> path.wustl.edu (Cruise, Karen) Date: Fri Jul 6 11:28:30 2012 Subject: [Histonet] (no subject) Message-ID: Please remove me from your mailing list. Karen E. Cruise Histologist (Research Technician II) Washington University School of Medicine Laboratory for Translational Pathology 216 S. Kingshighway Room 2332 St. Louis, MO 63110 (314) 454-8636 Phone (314) 454-5525 Fax kcruise@path.wustl.edu The materials in this email are private and may contain Protected Health Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return email. From dea.leslie <@t> gmail.com Fri Jul 6 12:05:33 2012 From: dea.leslie <@t> gmail.com (Deanna Leslie) Date: Fri Jul 6 12:05:41 2012 Subject: [Histonet] Florida HT Licensure Message-ID: Hi all! I am desperate and hope someone here can help me out. I am ASCP HT certified with over 20 years in histology. I am planning to relocate to Florida this fall. I have been to their DOH homepage to try to figure out how to get my Florida licensure. I cannot make heads or tails of that page and actually cannot find HT or HTL license application. If you can shed some light on this issue to help me out it would be most greatly appreciated!! Deanna Leslie HT ASCP 937-369-9913 From Laurie <@t> blufrogpath.com Fri Jul 6 12:07:06 2012 From: Laurie <@t> blufrogpath.com (Laurie@blufrogpath.com) Date: Fri Jul 6 12:07:11 2012 Subject: [Histonet] Light Green Counterstain Message-ID: <20120706100706.295dc6182df7e5cbb4f32bc101c30dcc.8bc86b1aa6.wbe@email15.secureserver.net> Can I use fast green counterstain in place of light green for GMS and fungal PAS? What is the difference between the two counterstains? Laurie Colbert From anna.c.hughes <@t> gsk.com Fri Jul 6 12:10:23 2012 From: anna.c.hughes <@t> gsk.com (Anna Hughes) Date: Fri Jul 6 12:10:34 2012 Subject: [Histonet] Re: Frozens and antigen retrieval Message-ID: <56B8D7633148A9419B559EBE8E9CF5904221B9@019-SN1MPN1-033.019D.MGD.MSFT.NET> I typically fix my frozen slides in 4% PFA or 10% NBF for 30min at 4C before staining. Antigen retrieval isn't usually necessary for frozens, even fixing them before staining - but it still depends on the antigen....for some targets in frozen brain sections of mice - I have used a 5% Formic Acid retrieval. Also, for other targets on frozen sections - if needed - I like the Uni-Trieve low temperature retrieval solution. It works nicely and doesn't affect the tissue. Anna C. Hughes, BS, HTL (ASCP) anna.c.hughes@gsk.com From Valerie.Hannen <@t> parrishmed.com Fri Jul 6 12:10:53 2012 From: Valerie.Hannen <@t> parrishmed.com (Hannen, Valerie) Date: Fri Jul 6 12:10:58 2012 Subject: [Histonet] Light Green Counterstain In-Reply-To: <20120706100706.295dc6182df7e5cbb4f32bc101c30dcc.8bc86b1aa6.wbe@email15.secureserver.net> Message-ID: <450B7A81EDA0C54E97C53D60F00776C322E7D40E24@isexstore03> I do use a Fast Green Substitute for our Light Green counterstain. Valerie Hannen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie@blufrogpath.com Sent: Friday, July 06, 2012 1:07 PM To: Histonet post Subject: [Histonet] Light Green Counterstain Can I use fast green counterstain in place of light green for GMS and fungal PAS? What is the difference between the two counterstains? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ************************************************************** "This email is intended solely for the use of the individual to whom it is addressed and may contain information that is privileged, confidential or otherwise exempt from disclosure under applicable law. If the reader of this email is not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please immediately delete this message. Thank you" ************************************************************** From thiggins <@t> cddmedical.com Fri Jul 6 12:15:29 2012 From: thiggins <@t> cddmedical.com (Tim Higgins) Date: Fri Jul 6 12:15:32 2012 Subject: [Histonet] Re: Frozens and antigen retrieval In-Reply-To: <20120706170312.D552F1B0C663@barracuda.crvinc.net> References: <20120706170312.D552F1B0C663@barracuda.crvinc.net> Message-ID: Agree, only if the antigen has been bound by a fixative. -----Original ________________________________ From: Daniela Bodemer To: histonet@lists.utsouthwestern.edu Sent: Wednesday, July 4, 2012 7:02 AM Subject: [Histonet] Frozens and antigen retrieval Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad From Valerie.Hannen <@t> parrishmed.com Fri Jul 6 13:07:28 2012 From: Valerie.Hannen <@t> parrishmed.com (Hannen, Valerie) Date: Fri Jul 6 13:07:31 2012 Subject: [Histonet] Florida HT Licensure In-Reply-To: Message-ID: <450B7A81EDA0C54E97C53D60F00776C322E7D40E25@isexstore03> Deanna, Follow the following steps to get the application: go to: 1) Florida Department of Health then click on 2) Health Professionals then 3) Licensing and Regulations then 4) Health Care Professions then 5) Clincial Laboratory Personnel then 6) Apply for License then 7) Applications and Forms then you click on what ever level that you qualify for...scroll down and print the application info as needed. I hope this helps!! Valerie A. Hannen, MLT(ASCP),HTL,SU(FL) Histology Section Chief Parrish Medical Center 951 N. Washington Ave. Titusville, Florida 32976 Phone:(321) 268-6333 ext. 7506 Fax: (321) 268-6149 valerie.hannen@parrishmed.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Deanna Leslie Sent: Friday, July 06, 2012 1:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Florida HT Licensure Hi all! I am desperate and hope someone here can help me out. I am ASCP HT certified with over 20 years in histology. I am planning to relocate to Florida this fall. I have been to their DOH homepage to try to figure out how to get my Florida licensure. I cannot make heads or tails of that page and actually cannot find HT or HTL license application. If you can shed some light on this issue to help me out it would be most greatly appreciated!! Deanna Leslie HT ASCP 937-369-9913 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ************************************************************** "This email is intended solely for the use of the individual to whom it is addressed and may contain information that is privileged, confidential or otherwise exempt from disclosure under applicable law. If the reader of this email is not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please immediately delete this message. Thank you" ************************************************************** From LouroP <@t> Princeton.Huntingdon.com Fri Jul 6 13:34:02 2012 From: LouroP <@t> Princeton.Huntingdon.com (Louro, Pedro) Date: Fri Jul 6 13:34:11 2012 Subject: [Histonet] Human paraffin block Message-ID: Hi everyone, Where can I purchase a human tonsil paraffin block to use in a GLP laboratory? My QA department needs paperwork that accompanies where the tissue came from. Thanks for the help Pedro ******************************************************************************************** Our Values: Excellence, Drive, Ownership, Challenge, Teamwork, Respect ******************************************************************************************** LEGAL NOTICE This message is confidential and contains information which may be legally privileged. It is intended for the stated addressee(s) only. Access to this email by anyone else is unauthorised. 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No. GB425507072 Registered Office: Woolley Road, Alconbury, Huntingdon, Cambridgeshire PE28 4HS ********************************************************************** From one_angel_secret <@t> yahoo.com Fri Jul 6 15:31:43 2012 From: one_angel_secret <@t> yahoo.com (Kim Donadio) Date: Fri Jul 6 15:31:47 2012 Subject: [Histonet] Florida HT Licensure In-Reply-To: References: Message-ID: <1341606703.67463.YahooMailNeo@web112305.mail.gq1.yahoo.com> http://doh.state.fl.us/mqa/ClinLab/clp_lic_req.html ? ? ________________________________ From: Deanna Leslie To: histonet@lists.utsouthwestern.edu Sent: Friday, July 6, 2012 1:05 PM Subject: [Histonet] Florida HT Licensure Hi all! I am desperate and hope someone here can help me out. I am ASCP HT certified with over 20 years in histology.? I am planning to relocate to Florida this fall. I have been to their DOH homepage to try to figure out how to get my Florida licensure. I cannot make heads or tails of that page and actually cannot find HT or HTL license application. If you can shed some light on this issue to help me out it would be most greatly appreciated!! Deanna Leslie HT ASCP 937-369-9913 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sabishop123 <@t> hotmail.com Fri Jul 6 16:33:12 2012 From: sabishop123 <@t> hotmail.com (Sarah Bishop) Date: Fri Jul 6 16:33:17 2012 Subject: [Histonet] FFPE Control Blocks Message-ID: Hello Histonet,I'm looking for Pneumocystis and Helicobacter FFPE Blocks. I am willing to pay for any processing or procurement fees associated with these blocks. If you are able to help me please send me an email at sabishop123@hotmail.com. Thank you so much in advance for the help. Sarah From rsrichmond <@t> gmail.com Sat Jul 7 07:51:19 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Sat Jul 7 07:51:24 2012 Subject: [Histonet] refreshing perspective on formaldehyde Message-ID: This "Stone Soup" comic strip gives you a different view of formaldehyde: http://www.gocomics.com/stonesoup/2012/07/07 though tykes this size should pickle their frogs in 70% alcohol (reagent alcohol or isopropanol). Bob Richmond Samurai Pathologist Knoxville TN From argonautro <@t> yahoo.com Sat Jul 7 09:50:10 2012 From: argonautro <@t> yahoo.com (Pirici Daniel) Date: Sat Jul 7 09:51:15 2012 Subject: [Histonet] A neuronal membrane marker? Message-ID: <1341672610.93930.YahooMailNeo@web113214.mail.gq1.yahoo.com> Hi to all Histoneters, I am searching for some while now for a good antibody against a specific neuronal membrane target. Also, it should??see? the neurites? membranes, not only the perykarions'.. I know tubulin III and NF are very good, but for cytoskeleton. Anti-acetylcholine receptors and anti-Na+/K+ ATPase antibodies might work but are too selective (acetylcholine R will not show all neuronal populations) or not specific enough (I have found some papers stating that Na+/K+ ATPase stains glial membranes too). ? So, please may I have a word of advice? Many many thanks in advance! Sincerely, Daniel ________________________________________________________________________________________________ Pirici Nicolae Daniel, MD, PhD Department of Histology University of Medicine and Pharmacy Craiova Petru Rares Street 2, 200349 Craiova, Dolj Romania From LPaveli1 <@t> hurleymc.com Sat Jul 7 12:34:03 2012 From: LPaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Sat Jul 7 12:34:12 2012 Subject: [Histonet] FFPE Control Blocks In-Reply-To: References: Message-ID: <89F4666A496DC949A819ECC40E11C8670C6517@EXCHANGEMB1.hmc.hurleymc.com> By joining the membership of the Michigan society, you have access to a large control bank, free of charge. Anyone can join! www.mihisto.org Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Sarah Bishop [sabishop123@hotmail.com] Sent: Friday, July 06, 2012 5:33 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FFPE Control Blocks Hello Histonet,I'm looking for Pneumocystis and Helicobacter FFPE Blocks. I am willing to pay for any processing or procurement fees associated with these blocks. If you are able to help me please send me an email at sabishop123@hotmail.com. Thank you so much in advance for the help. Sarah _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rsrichmond <@t> gmail.com Sat Jul 7 12:41:55 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Sat Jul 7 12:41:58 2012 Subject: [Histonet] Re: Light Green Counterstain Message-ID: Light green SF and Fast green FCF are extremely similar triarylmethyl dyes that are almost the same color, and fast green can be substituted as a counterstain. Light green may have gone out of manufacture. Triarylmethyl dyes are potent carcinogens, and they could become hard to get or work with in the future. Bob Richmond Samurai Pathologist Knoxville TN From rsrichmond <@t> gmail.com Sun Jul 8 13:26:01 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Sun Jul 8 13:26:08 2012 Subject: [Histonet] Re: [PATHO-L] refreshing perspective on formaldehyde In-Reply-To: <11A898B32EBD0649AB3E81FF75A146D50593F747@ccrexbe02.ccr.cchcs.org> References: <11A898B32EBD0649AB3E81FF75A146D50593F747@ccrexbe02.ccr.cchcs.org> Message-ID: A couple of provocative replies about formaldehyde outside the pathology laboratory. In one hospital the specimens have to be submitted in saline and transferred to formalin in the lab, while in the other glyoxal (a different aldehyde fixative) was successfully substituted. First of all, since where does a nursing supervisor or a hospital administrator get the authority to practice pathology? And what pathologist (a lot, alas) is so ignorant of the basics of managing tissue as to think that biopsy specimens can be put in saline? (And I've seen worse - in one hospital the nurses insisted that bronchoscopic specimens be sent out dry, and left in a receiving area with no one present there.) If I were not allowed an aldehyde fixative at all, I'd resort to the transport media often misrepresented as fixatives, and move the specimen to formaldehyde in the lab. Though I'd prefer glyoxal if I could slip it by the MBAs, though as Don Ross notes you have to readjust all your immunohistochemistry for the different fixative. If you're going to do histology, you've got to know your materials. Bob Richmond Samurai Pathologist Knoxville TN **************************************** On Sun, Jul 8, 2012 at 1:13 PM, Don Ross MD wrote: > An unnecessary nuisance, I agree. A similar concern prompted us to switch to Prefer (TM), which is alcoholic glyoxal. A byproduct has been faster, better fixation, and easier impox (much less antigen retrieval). The validations for ER, PR and HER2 were a bother, but we got through it alright. > > -----Original Message----- > From: patho-l-bounces@mailman.srv.ualberta.ca on behalf of roupen dekmezian > Sent: Sun 7/8/2012 11:31 AM > To: patho-l@mailman.srv.ualberta.ca; histonet@lists.utsouthwestern.edu > Subject: Re: [PATHO-L] refreshing perspective on formaldehyde > > Funny but not so funny when OSHA hits you with a fine and bans formaldehyde from virtually everywhere in the hospital, except the pathology lab. It all started with a leak on one of the hospital floors due to a careless person who didn't close the tap tight after using the formalin container. This led to an evacuation of the entire floor leading to all the hooplas of the media, fire and police departments. Of course this was followed by the Gov. inspectors who assessed the fines and determined that to be able to keep formalin (even the small bottles for G.I. biopsies) anywhere but the lab, we have to train the entire hospital staff for spill response. Now we use saline instead, until the specimens arrive in pathology where formalin is added. Till now we haven't had a disaster of fixation but I know it's around the corner. Soon many of you will have to contend with the "enforcers". Reminds me of the prescient statement: > 'A listing...does not by itself mean that a substance will cause cancer,'" said Dr. John Bucher, associate director of the National > Toxicology Program. Moreover, Dr. Bucher said the updated listings "do not trigger any immediate new restrictions on the substances, but other > government agencies may use the information in the future as part of their regulatory decisions." (Thank you Adel Assaad) > > Dekmezian, Houston > >> Date: Sat, 7 Jul 2012 08:51:19 -0400 >> From: rsrichmond@gmail.com >> To: patho-l@mailman.srv.ualberta.ca; histonet@lists.utsouthwestern.edu >> Subject: [PATHO-L] refreshing perspective on formaldehyde >> >> This "Stone Soup" comic strip gives you a different view of formaldehyde: >> >> http://www.gocomics.com/stonesoup/2012/07/07 >> >> though tykes this size should pickle their frogs in 70% alcohol >> (reagent alcohol or isopropanol). >> >> Bob Richmond >> Samurai Pathologist >> Knoxville TN From tgenade <@t> gmail.com Mon Jul 9 02:16:19 2012 From: tgenade <@t> gmail.com (Tyrone Genade) Date: Mon Jul 9 02:16:31 2012 Subject: [Histonet] PFA: how long per mass/type tissue? Message-ID: Hello, Is there a guideline as to how long to fix in PFA based on the mass of the tissue or type of tissue? I am working with fish. I have had been experimenting with Davidson's Fixative but it gave too much background under the fluorescence microscope. I have tried fixing in PFA and then decalcifying in Kristenson's and this worked very well but I have lost some antigens. I know, from past experience, that a fixing the dissected out brain in PFA for 10 minutes on ice did not destroy the antigens so I am now wonder if the problem is over-fixation? What I did this time round (because it is very difficult to dissect out the entire brain without losing the hypothalamus and damaging the tectal surface) was to cut away the excess tissue and fix the cranium with eyes overnight at 4 oC in PFA and the decalcify in Kristenson's for 2 days at 4 oC. I got good cryo-sections and the fine structure was well preserved. The cranium is small, about 12 x 6 mm in total and about 4 mm thick, so I guess a mass of about 200 mg. I'm going to try a 2 hour fix* at 4 oC and see how that goes. Any suggestions? I don't want to kill fish needlessly. *I have read of perfusion fixed rat brain being post-fixed for two hours at 4 oC and the published sections look quite good. I can't perfusion fix. I can't find a vessel large enough for a canula. Thanks -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@gmail.com tel: +27-84-632-1925 (c) ******************************************************************************** Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord. To find out how to receive this FREE gift visit http://www.alpha.org. From BDeBrosse-Serra <@t> isisph.com Mon Jul 9 09:55:16 2012 From: BDeBrosse-Serra <@t> isisph.com (Bea DeBrosse-Serra) Date: Mon Jul 9 09:55:40 2012 Subject: [Histonet] Need help with CD40 on frozen sections Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FBFB@EXCHMB01.isis.local> Histonetters, I need some help with CD40 on frozen sections (spleen, non-treated). I cut my frozens, let them briefly air dry and keep them at -20?C. When I use them for staining, I air dry them, fix them in cold acetone for 5 minutes and start the staining process. I use the primary AB from BD Pharmingen, # 550285 at various dilution, incubate them overnight at 4?C, use the secondary HRP conjugated antibody, DAB, counterstain and don't get any staining. Does anyone have any pointers, suggestions? Thank you in advance, Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 From jcox90 <@t> yahoo.com Mon Jul 9 10:32:37 2012 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Mon Jul 9 10:32:43 2012 Subject: [Histonet] Formalin Fixation issues Message-ID: <1341847957.52395.YahooMailNeo@web161604.mail.bf1.yahoo.com> Hi Histonetters, Is anyone having problems with breast fixation?prior to?processing in formalin? For a couple of months now our breast specimens aren't fixing very well before gross.?Our Pathologist thinks they have changed something in the formalin itself.?We?utilize 15/1 ratio and in some cases let fix over the weekend. This has only been happening over the last couple of months and can't seem to figure this out. Any advise or similar problems, would love to hear from you.. Thanks, Jill From relia1 <@t> earthlink.net Mon Jul 9 11:23:12 2012 From: relia1 <@t> earthlink.net (Pam Barker) Date: Mon Jul 9 11:23:20 2012 Subject: [Histonet] RELIA Histology Opportunity Weekly UPdate: 7/9/2012. Exciting new Opportunities. Not just your average histology job !!! Message-ID: <014801cd5def$2358c700$6a0a5500$@earthlink.net> Hi Histonetters, I hope everyone's week is kicking off to a great start. I know mine is!! Over the past few days yes this weekend even!! I have been given some exciting new positions to work on. The clients are ready to hire ASAP and offering excellent compensation packages. These are all permanent full time positions. Each of them has a unique characteristic that makes it NOT YOUR AVERAGE HISTOLOGY JOB. Here is a list of my current openings: Mohs Tech - Sarasota, FL Lead Tech - South Bend, IN Night Shift Tech - Charlotte, NC HT/HTL - Collegeville, PA Histology Tech - Austin, TX Histology Tech - Portland, ME Histotech - Bristol, TN Electron Microscopy Tech - Long Island, NY (NY Lic req.) **** More positions coming in all of the time so if you are looking let me know where and I will keep you posted. ****** REMEMBER. It never hurts to look! If you would like to know what is so special about any or all of these positions then Let's talk. You can reach me toll free at 866-607-3542 until 6 EST and anytime on my cell phone at 407-353-5070, or shoot me an email at relia1@earthlink.net to schedule a time to talk. Thank You! Pam Barker President/Senior Recruiting Specialist-Histology RELIA Solutions Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.facebook.com /PamBarkerRELIA www.linkedin.com/in/reliasolutions www.twitter.com/pamatrelia From rsrichmond <@t> gmail.com Mon Jul 9 12:25:05 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Mon Jul 9 12:25:14 2012 Subject: [Histonet] Re: Formalin Fixation issues Message-ID: Jill Cox asks: >>Is anyone having problems with breast fixation prior to processing in formalin? For a couple of months now our breast specimens aren't fixing very well before gross. Our pathologist thinks they have changed something in the formalin itself. We utilize 15/1 ratio and in some cases let fix over the weekend. This has only been happening over the last couple of months and can't seem to figure this out. Any advice or similar problems, would love to hear from you.<< Breast specimens shouldn't be expected to fix before they're grossed, or at least before they've been cut into thin slices. Delaying fixation compromises immunostaining, to say nothing of H & E. They should be grossed as promptly as possible after they're received, and should never sit over the weekend without dissection. I seriously doubt that anything in the formalin has changed. It's your technique that needs to change. Unfortunately, this is a difficult task to delegate. Bob Richmond Samurai Pathologist Knoxville TN From TJohnson <@t> gnf.org Mon Jul 9 12:46:25 2012 From: TJohnson <@t> gnf.org (Teri Johnson) Date: Mon Jul 9 12:46:31 2012 Subject: [Histonet] RE: Need help with CD40 on frozen sections Message-ID: <9F3CFEE76E51B64991C7485270890B400CDC4A3B@EX4.lj.gnf.org> Hi Bea! I just left you a voicemail message. I looked up this antibody data sheet and to say the least the staining on the image is underwhelming! I would try fixing the cryosections with Beckstead's zinc prior to staining, and you can even consider fixing the spleens with Zinc and then cryoprotecting with sucrose prior to freezing. It appears they recommend using a biotinylated secondary and then streptavidin-HRP for staining. You might see if that helps as well. Teri Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 From rsrichmond <@t> gmail.com Mon Jul 9 12:51:10 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Mon Jul 9 12:51:18 2012 Subject: [Histonet] Re: Formalin Fixation issues Message-ID: At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be fixed > before grossing, it's also not getting fixed after processing. But thanks > for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN From melissa <@t> alliedsearchpartners.com Mon Jul 9 12:54:26 2012 From: melissa <@t> alliedsearchpartners.com (Melissa Phelan) Date: Mon Jul 9 12:54:35 2012 Subject: [Histonet] Day Shift Histotech Job Near Ithaca, NY Message-ID: Hi Everyone, I have a Day Shift Monday-Friday Histotech job opening about 45 miles Southeast of Ithaca, NY (About an hour North of Scranton, PA). This position requires a NY license and ASCP certification. Please message me for details if you are interested! To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php Melissa Phelan LinkedIn: http://www.linkedin.com/in/melissaphelan President, Laboratory Staffing Allied Search Partners P: 888.388.7571 F: 888.388.7572 M: 407.697.1175 www.alliedsearchpartners.com From jcox90 <@t> yahoo.com Mon Jul 9 13:00:41 2012 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Mon Jul 9 13:00:45 2012 Subject: [Histonet] Re: Formalin Fixation issues In-Reply-To: References: Message-ID: <1341856841.88220.YahooMailNeo@web161606.mail.bf1.yahoo.com> Yes Bob, the sections are good and thin. We use Scientific Products, 10% NBF. Just seems like the Formalin is watered down. Going to try a different brand. I have worked for Pathologist's that cram cassettes but this is not the case here. From: Bob Richmond To: histonet@lists.utsouthwestern.edu Sent: Monday, July 9, 2012 10:51 AM Subject: [Histonet] Re: Formalin Fixation issues At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be fixed > before grossing, it's also not getting fixed after processing. But thanks > for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Joyce.Weems <@t> emoryhealthcare.org Mon Jul 9 13:33:56 2012 From: Joyce.Weems <@t> emoryhealthcare.org (Weems, Joyce K.) Date: Mon Jul 9 13:34:03 2012 Subject: [Histonet] Re: Formalin Fixation issues In-Reply-To: <1341856841.88220.YahooMailNeo@web161606.mail.bf1.yahoo.com> References: <1341856841.88220.YahooMailNeo@web161606.mail.bf1.yahoo.com> Message-ID: I would also call the company to see if they have had others with problems. Has it all been the same lot number? Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, July 09, 2012 2:01 PM To: Bob Richmond; Histonet@Lists. Edu Subject: Re: [Histonet] Re: Formalin Fixation issues Yes Bob, the sections are good and thin. We use Scientific Products, 10% NBF. Just seems like the Formalin is watered down. Going to try a different brand. I have worked for Pathologist's that cram cassettes but this is not the case here. From: Bob Richmond To: histonet@lists.utsouthwestern.edu Sent: Monday, July 9, 2012 10:51 AM Subject: [Histonet] Re: Formalin Fixation issues At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be > fixed before grossing, it's also not getting fixed after processing. > But thanks for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ 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 ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). From liz <@t> premierlab.com Mon Jul 9 13:54:10 2012 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Mon Jul 9 13:54:17 2012 Subject: [Histonet] Re: Formalin Fixation issues In-Reply-To: Message-ID: <14E2C6176416974295479C64A11CB9AE0113A9690C43@SBS2K8.premierlab.local> There is a real nice article on fixation and handling of breast lumpectomy samples in the Journal of Histotechnology by Dr. Stephen Ruby. It goes over a nice technique we used to use on those samples, worked great. It did use alcoholic formalin which I suspect can't be used currently if you have not validated for that fixative for ER/PR etc. But the overall technique can be used with 10% NBF. The thickness of samples is key as Bob pointed out. We had a new pathologist on gross and the next day we had about 20 or 30 recuts on their breast samples. That number of reacts was uncommon. I needed to figure out what was the issue and we found out it was the size of tissue that was placed into the cassettes. I explained that they needed to place smaller and thinner pieces of tissue in the cassette. Once I did that we did not have anymore problems with their samples. The article information is below. Paper Towel "Sandwich" - Alcoholic Formalin Fixation for Breast Biopsies Author: Ruby, Stephen G. Source: Journal of Histotechnology, Number 1, March 1999 , pp. 49-51(3) Publisher: Maney Publishing Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bob Richmond Sent: Monday, July 09, 2012 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Formalin Fixation issues At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be fixed > before grossing, it's also not getting fixed after processing. But thanks > for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Jul 9 15:32:58 2012 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Mon Jul 9 15:33:05 2012 Subject: [Histonet] GBS IHC Message-ID: <4FFB07BA.7400.0077.1@harthosp.org> Does anyone know of a good antibody to Group B Streptococci (GBS) that works on formalin-fixed, paraffin embedded tissue? I had a wonderful mAb from a company in Canada, but they went out of business. Thank you. Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax From Ramona_Nelson <@t> bd.com Tue Jul 10 06:50:59 2012 From: Ramona_Nelson <@t> bd.com (Ramona_Nelson@bd.com) Date: Tue Jul 10 06:51:09 2012 Subject: [Histonet] AUTO: Ramona Nelson is out of the office. (returning 07/16/2012) Message-ID: I am out of the office until 07/16/20 I Note: This is an automated response to your message &quo t;Histonet Digest, Vol 104, Issue 10" sent on 7/9/2012 1:00:01 PM. This is the only notification you will receive whi person is away. _________________________________________________________________ ************************************************************* ****** IMPORTANT MESSAGE FOR RECIPIENTS IN THE U.S.A.: This messa services or message and you wou advertisements or solicitations from this e-mail to optoutbygroup@bd.com. * ****************************************************************** T his m the designated proprietary information an attorney-client privilege or other confidentialit If you are not a designated recipient, you may not review or distribute this message. If you received this in error, pl notify the sender by reply e-mail and delete this message. Thank you . **************************************************************** *** and Compan ******************* ************************************************ From Lisa.White3 <@t> va.gov Tue Jul 10 08:45:31 2012 From: Lisa.White3 <@t> va.gov (White, Lisa M.) Date: Tue Jul 10 08:46:07 2012 Subject: [Histonet] Subject: Re: [PATHO-L] refreshing perspective on formaldehyde Message-ID: <2B2ECF33934F5D4996D8BE03EFDF397609E9E660@VHAV09MSGA3.v09.med.va.gov> For the paranoid out there: imagine what a lawyer will do to a hospital in court when Formalin (or other proper fixative) was not allowed to be utilized and the specimen is ruined. The lawsuit settlement will make a fine from OSHA look like buying a candy bar. More than likely it will be a member of or government or high society type who is "wronged" before changes are made. It is hard to believe that a hospital would allow the fixative to be removed from all areas. Yes everyone who is utilizing the fixative needs to be educated in spill clean-up and have spill kits maintained in their area. It is a small price to pay to protect the integrity of the specimen. What if it was your loved ones specimen? I have heard of others leaving the taps open. The remedy was to buy bottles with screw caps with the bonus of the fixative not being as heavy. Lisa White, HT(ASCP) Supervisory HT James H. Quillen VAMC PO Box 4000 Corner of Veterans Way and Lamont PLMS 113 Mountain Home, TN 37684 423-979-3567 423-979-3401 fax From sayeeduddin <@t> sbcglobal.net Tue Jul 10 09:02:17 2012 From: sayeeduddin <@t> sbcglobal.net (Mohammad Sayeeduddin) Date: Tue Jul 10 09:02:24 2012 Subject: [Histonet] Beecher tissue array 1 mm. punch set. Message-ID: <1341928937.86834.YahooMailClassic@web81506.mail.mud.yahoo.com> Hi Histonet members, I am in trouble. I need 1 mm. tissue array punch set ( Beecher manual instrument ). Either you can sell us one or exchange for a box of ten sets of 0.6mm size punch sets. Please contact me at sayeeduddin@sbcglobal.net Thanks in advance for your response. Sayeed. From dreynold <@t> mdanderson.org Tue Jul 10 09:32:51 2012 From: dreynold <@t> mdanderson.org (Reynolds,Donna M) Date: Tue Jul 10 09:30:07 2012 Subject: [Histonet] CD40 Message-ID: <785BBF0C5F49CE41BA74460A43A08F0232223A8783@DCPWVMBXC0VS3.mdanderson.edu> What are you using for Endogenous blocking? If you are using methanol with H2O2 that may be your problem. Many CD antibodies are very sensitive to the methanol. Try PBS or whatever buffer you are using to dilute the H202. I would also try going straight form the -20 into cold acetone without air drying and fix for10 min. Donna Reynolds HT (ASCP) U. T. M.D. Anderson Cancer Center Chief histology Tech, Department Cancer Biology 713-792-8106 ------------------------------ Message: 3 Date: Mon, 9 Jul 2012 07:55:16 -0700 From: Bea DeBrosse-Serra Subject: [Histonet] Need help with CD40 on frozen sections To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FBFB@EXCHMB01.isis.local> Content-Type: text/plain; charset="iso-8859-1" Histonetters, I need some help with CD40 on frozen sections (spleen, non-treated). I cut my frozens, let them briefly air dry and keep them at -20?C. When I use them for staining, I air dry them, fix them in cold acetone for 5 minutes and start the staining process. I use the primary AB from BD Pharmingen, # 550285 at various dilution, incubate them overnight at 4?C, use the secondary HRP conjugated antibody, DAB, counterstain and don't get any staining. Does anyone have any pointers, suggestions? Thank you in advance, Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 *********************************** From cathy.crumpton <@t> tuality.org Tue Jul 10 10:51:00 2012 From: cathy.crumpton <@t> tuality.org (Cathy Crumpton) Date: Tue Jul 10 10:47:48 2012 Subject: [Histonet] IVD P63 Message-ID: I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 From kmerriam2003 <@t> yahoo.com Tue Jul 10 10:48:54 2012 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Tue Jul 10 10:49:01 2012 Subject: [Histonet] human vimentin in mouse tissue Message-ID: <1341935334.82981.YahooMailNeo@web130104.mail.mud.yahoo.com> Greetings! ? I want to stain for human vimentin in mouse tissues (human fibroblasts).? I many years ago, I used?a biotinylated V9 clone for just this purpose and it worked great.??I tried the?biotiylated V9 clone from abcam, but I am not getting any staining at all.? Any suggestions? ? Kim ? Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA? From one_angel_secret <@t> yahoo.com Tue Jul 10 11:02:38 2012 From: one_angel_secret <@t> yahoo.com (Kim Donadio) Date: Tue Jul 10 11:02:43 2012 Subject: [Histonet] IVD P63 In-Reply-To: References: Message-ID: <1341936158.34785.YahooMailNeo@web112318.mail.gq1.yahoo.com> Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. ________________________________ From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody.? I would like to purchase IVD P63 from another vendor, who else sells it?? I have having a few problems finding good info just using Google.? What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CThornton <@t> dahlchase.com Tue Jul 10 11:48:20 2012 From: CThornton <@t> dahlchase.com (Clare Thornton) Date: Tue Jul 10 11:48:29 2012 Subject: [Histonet] IVD P63 In-Reply-To: <1341936158.34785.YahooMailNeo@web112318.mail.gq1.yahoo.com> References: <1341936158.34785.YahooMailNeo@web112318.mail.gq1.yahoo.com> Message-ID: I just spoke with my Biocare rep. He said it is untrue at this time, and they are still selling p63. Clare J. Thornton, HTL(ASCP),QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthornton@dahlchase.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Tuesday, July 10, 2012 12:03 PM To: Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IVD P63 Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. ________________________________ From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody.? I would like to purchase IVD P63 from another vendor, who else sells it?? I have having a few problems finding good info just using Google.? What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 _______________________________________________ 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 From patrick.lewis <@t> seattlechildrens.org Tue Jul 10 12:09:23 2012 From: patrick.lewis <@t> seattlechildrens.org (Lewis, Patrick) Date: Tue Jul 10 12:09:32 2012 Subject: [Histonet] Methanol in H2O2 explanation Message-ID: <3903BE18914F4440834F0E620415FFCA0BCC7461@PPWEXD01B.childrens.sea.kids> Hi everyone, Can someone refresh/enlighten me as to why Methanol is used in H2O2 blocking. Let's assume that I take my slides through the xylene/ethanol steps and that after the 70% etoh wash step, they are now in TBST. I could see adding methanol if you were doing the H2O2 blocking as part of the xylene/etoh process and maybe you had the methanol as a substitute for the 70% etoh. My xylene/ETOH looks like this. 3 x 5 min xylene. 2 x 5 min 100% etoh 2 x 5 min 95% etoh 1 x 5 min 70% Then my slides are washed and then epitope retrieved. After epitope retrieval they are again washed and then I do my H2O2 block without methanol. Thanks Patrick. CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From aamador <@t> pcnm.com Tue Jul 10 12:11:44 2012 From: aamador <@t> pcnm.com (Amanda Amador) Date: Tue Jul 10 12:12:07 2012 Subject: [Histonet] Re: Formalin Fixation issues (Bob Richmond) Message-ID: <11AB4FFC26B3144D8346B10630E534D51B07E8C8@S10MAILD001N3.SH10.lan> If you have large specimens, it is best to "open" them as soon as possible. If it is a breast, at least do your prelim measurements and slice it open, leaving it still attached, so that the formain can penetrate the tissue. If you go ahead and slice it up, at least wrap it, in order with paper towels, to keep orientation while it fixes. You can always block it and then let it fix too. You just need to be sure to slice it thin so that it fixes; and make sure you have adequate spacing between cassettes. You just need to figure out what way is best for your facility. Amanda Amador, AAS, ASCP(CM) Sr. Histotechnician Pathology Consultants of New Mexico 600 N. Richardson Roswell, NM 88202 575-622-5600, ext 218 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of histonet-request@lists.utsouthwestern.edu [histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, July 10, 2012 11:02 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 104, Issue 11 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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: Formalin Fixation issues (Bob Richmond) 2. RE: Need help with CD40 on frozen sections (Teri Johnson) 3. Re: Formalin Fixation issues (Bob Richmond) 4. Day Shift Histotech Job Near Ithaca, NY (Melissa Phelan) 5. Re: Re: Formalin Fixation issues (Jill Cox) 6. RE: Re: Formalin Fixation issues (Weems, Joyce K.) 7. RE: Re: Formalin Fixation issues (Elizabeth Chlipala) 8. GBS IHC (Richard Cartun) 9. AUTO: Ramona Nelson is out of the office. (returning 07/16/2012) (Ramona_Nelson@bd.com) 10. Subject: Re: [PATHO-L] refreshing perspective on formaldehyde (White, Lisa M.) 11. Beecher tissue array 1 mm. punch set. (Mohammad Sayeeduddin) 12. CD40 (Reynolds,Donna M) 13. IVD P63 (Cathy Crumpton) 14. human vimentin in mouse tissue (Kim Merriam) 15. Re: IVD P63 (Kim Donadio) 16. RE: IVD P63 (Clare Thornton) ---------------------------------------------------------------------- Message: 1 Date: Mon, 9 Jul 2012 13:25:05 -0400 From: Bob Richmond Subject: [Histonet] Re: Formalin Fixation issues To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Jill Cox asks: >>Is anyone having problems with breast fixation prior to processing in formalin? For a couple of months now our breast specimens aren't fixing very well before gross. Our pathologist thinks they have changed something in the formalin itself. We utilize 15/1 ratio and in some cases let fix over the weekend. This has only been happening over the last couple of months and can't seem to figure this out. Any advice or similar problems, would love to hear from you.<< Breast specimens shouldn't be expected to fix before they're grossed, or at least before they've been cut into thin slices. Delaying fixation compromises immunostaining, to say nothing of H & E. They should be grossed as promptly as possible after they're received, and should never sit over the weekend without dissection. I seriously doubt that anything in the formalin has changed. It's your technique that needs to change. Unfortunately, this is a difficult task to delegate. Bob Richmond Samurai Pathologist Knoxville TN ------------------------------ Message: 2 Date: Mon, 9 Jul 2012 17:46:25 +0000 From: Teri Johnson Subject: [Histonet] RE: Need help with CD40 on frozen sections To: "histonet@lists.utsouthwestern.edu" Cc: "Bea DeBrosse-Serra \(BDeBrosse-Serra@isisph.com\)" Message-ID: <9F3CFEE76E51B64991C7485270890B400CDC4A3B@EX4.lj.gnf.org> Content-Type: text/plain; charset="us-ascii" Hi Bea! I just left you a voicemail message. I looked up this antibody data sheet and to say the least the staining on the image is underwhelming! I would try fixing the cryosections with Beckstead's zinc prior to staining, and you can even consider fixing the spleens with Zinc and then cryoprotecting with sucrose prior to freezing. It appears they recommend using a biotinylated secondary and then streptavidin-HRP for staining. You might see if that helps as well. Teri Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 ------------------------------ Message: 3 Date: Mon, 9 Jul 2012 13:51:10 -0400 From: Bob Richmond Subject: [Histonet] Re: Formalin Fixation issues To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be fixed > before grossing, it's also not getting fixed after processing. But thanks > for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN ------------------------------ Message: 4 Date: Mon, 09 Jul 2012 13:54:26 -0400 From: Melissa Phelan Subject: [Histonet] Day Shift Histotech Job Near Ithaca, NY To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi Everyone, I have a Day Shift Monday-Friday Histotech job opening about 45 miles Southeast of Ithaca, NY (About an hour North of Scranton, PA). This position requires a NY license and ASCP certification. Please message me for details if you are interested! To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php Melissa Phelan LinkedIn: http://www.linkedin.com/in/melissaphelan President, Laboratory Staffing Allied Search Partners P: 888.388.7571 F: 888.388.7572 M: 407.697.1175 www.alliedsearchpartners.com ------------------------------ Message: 5 Date: Mon, 9 Jul 2012 11:00:41 -0700 (PDT) From: Jill Cox Subject: Re: [Histonet] Re: Formalin Fixation issues To: Bob Richmond , "Histonet@Lists. Edu" Message-ID: <1341856841.88220.YahooMailNeo@web161606.mail.bf1.yahoo.com> Content-Type: text/plain; charset=us-ascii Yes Bob, the sections are good and thin. We use Scientific Products, 10% NBF. Just seems like the Formalin is watered down. Going to try a different brand. I have worked for Pathologist's that cram cassettes but this is not the case here. From: Bob Richmond To: histonet@lists.utsouthwestern.edu Sent: Monday, July 9, 2012 10:51 AM Subject: [Histonet] Re: Formalin Fixation issues At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be fixed > before grossing, it's also not getting fixed after processing. But thanks > for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 9 Jul 2012 18:33:56 +0000 From: "Weems, Joyce K." Subject: RE: [Histonet] Re: Formalin Fixation issues To: "'Jill Cox'" , Bob Richmond , "Histonet@Lists. Edu" Message-ID: Content-Type: text/plain; charset="us-ascii" I would also call the company to see if they have had others with problems. Has it all been the same lot number? Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, July 09, 2012 2:01 PM To: Bob Richmond; Histonet@Lists. Edu Subject: Re: [Histonet] Re: Formalin Fixation issues Yes Bob, the sections are good and thin. We use Scientific Products, 10% NBF. Just seems like the Formalin is watered down. Going to try a different brand. I have worked for Pathologist's that cram cassettes but this is not the case here. From: Bob Richmond To: histonet@lists.utsouthwestern.edu Sent: Monday, July 9, 2012 10:51 AM Subject: [Histonet] Re: Formalin Fixation issues At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be > fixed before grossing, it's also not getting fixed after processing. > But thanks for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ 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 ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). ------------------------------ Message: 7 Date: Mon, 9 Jul 2012 12:54:10 -0600 From: Elizabeth Chlipala Subject: RE: [Histonet] Re: Formalin Fixation issues To: 'Bob Richmond' , "histonet@lists.utsouthwestern.edu" Message-ID: <14E2C6176416974295479C64A11CB9AE0113A9690C43@SBS2K8.premierlab.local> Content-Type: text/plain; charset="us-ascii" There is a real nice article on fixation and handling of breast lumpectomy samples in the Journal of Histotechnology by Dr. Stephen Ruby. It goes over a nice technique we used to use on those samples, worked great. It did use alcoholic formalin which I suspect can't be used currently if you have not validated for that fixative for ER/PR etc. But the overall technique can be used with 10% NBF. The thickness of samples is key as Bob pointed out. We had a new pathologist on gross and the next day we had about 20 or 30 recuts on their breast samples. That number of reacts was uncommon. I needed to figure out what was the issue and we found out it was the size of tissue that was placed into the cassettes. I explained that they needed to place smaller and thinner pieces of tissue in the cassette. Once I did that we did not have anymore problems with their samples. The article information is below. Paper Towel "Sandwich" - Alcoholic Formalin Fixation for Breast Biopsies Author: Ruby, Stephen G. Source: Journal of Histotechnology, Number 1, March 1999 , pp. 49-51(3) Publisher: Maney Publishing Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bob Richmond Sent: Monday, July 09, 2012 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Formalin Fixation issues At this point I'm not sure I understand what Jill Cox's problem is. Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk. Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies. Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently? Bob Richmond Samurai Pathologist Knoxville TN *************************************************** On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox wrote: > I think you misunderstood what I meant. I know it's not supposed to be fixed > before grossing, it's also not getting fixed after processing. But thanks > for your two cents.. > > From: Bob Richmond > To: histonet@lists.utsouthwestern.edu > Sent: Monday, July 9, 2012 10:25 AM > Subject: [Histonet] Re: Formalin Fixation issues > > Jill Cox asks: >>Is anyone having problems with breast fixation prior > to processing in formalin? For a couple of months now our breast > specimens aren't fixing very well before gross. Our pathologist thinks > they have changed something in the formalin itself. We utilize 15/1 > ratio and in some cases let fix over the weekend. This has only been > happening over the last couple of months and can't seem to figure this > out. Any advice or similar problems, would love to hear from you.<< > > Breast specimens shouldn't be expected to fix before they're grossed, > or at least before they've been cut into thin slices. Delaying > fixation compromises immunostaining, to say nothing of H & E. They > should be grossed as promptly as possible after they're received, and > should never sit over the weekend without dissection. > > I seriously doubt that anything in the formalin has changed. It's your > technique that needs to change. Unfortunately, this is a difficult > task to delegate. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 09 Jul 2012 16:32:58 -0400 From: "Richard Cartun" Subject: [Histonet] GBS IHC To: "Histonet" Message-ID: <4FFB07BA.7400.0077.1@harthosp.org> Content-Type: text/plain; charset=US-ASCII Does anyone know of a good antibody to Group B Streptococci (GBS) that works on formalin-fixed, paraffin embedded tissue? I had a wonderful mAb from a company in Canada, but they went out of business. Thank you. Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax ------------------------------ Message: 9 Date: Tue, 10 Jul 2012 07:50:59 -0400 From: Ramona_Nelson@bd.com Subject: [Histonet] AUTO: Ramona Nelson is out of the office. (returning 07/16/2012) To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I am out of the office until 07/16/20 I Note: This is an automated response to your message &quo t;Histonet Digest, Vol 104, Issue 10" sent on 7/9/2012 1:00:01 PM. This is the only notification you will receive whi person is away. _________________________________________________________________ ************************************************************* ****** IMPORTANT MESSAGE FOR RECIPIENTS IN THE U.S.A.: This messa services or message and you wou advertisements or solicitations from this e-mail to optoutbygroup@bd.com. * ****************************************************************** T his m the designated proprietary information an attorney-client privilege or other confidentialit If you are not a designated recipient, you may not review or distribute this message. If you received this in error, pl notify the sender by reply e-mail and delete this message. Thank you . **************************************************************** *** and Compan ******************* ************************************************ ------------------------------ Message: 10 Date: Tue, 10 Jul 2012 09:45:31 -0400 From: "White, Lisa M." Subject: [Histonet] Subject: Re: [PATHO-L] refreshing perspective on formaldehyde To: Message-ID: <2B2ECF33934F5D4996D8BE03EFDF397609E9E660@VHAV09MSGA3.v09.med.va.gov> Content-Type: text/plain; charset="us-ascii" For the paranoid out there: imagine what a lawyer will do to a hospital in court when Formalin (or other proper fixative) was not allowed to be utilized and the specimen is ruined. The lawsuit settlement will make a fine from OSHA look like buying a candy bar. More than likely it will be a member of or government or high society type who is "wronged" before changes are made. It is hard to believe that a hospital would allow the fixative to be removed from all areas. Yes everyone who is utilizing the fixative needs to be educated in spill clean-up and have spill kits maintained in their area. It is a small price to pay to protect the integrity of the specimen. What if it was your loved ones specimen? I have heard of others leaving the taps open. The remedy was to buy bottles with screw caps with the bonus of the fixative not being as heavy. Lisa White, HT(ASCP) Supervisory HT James H. Quillen VAMC PO Box 4000 Corner of Veterans Way and Lamont PLMS 113 Mountain Home, TN 37684 423-979-3567 423-979-3401 fax ------------------------------ Message: 11 Date: Tue, 10 Jul 2012 07:02:17 -0700 (PDT) From: Mohammad Sayeeduddin Subject: [Histonet] Beecher tissue array 1 mm. punch set. To: histonet@lists.utsouthwestern.edu Message-ID: <1341928937.86834.YahooMailClassic@web81506.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi Histonet members, I am in trouble. I need 1 mm. tissue array punch set ( Beecher manual instrument ). Either you can sell us one or exchange for a box of ten sets of 0.6mm size punch sets. Please contact me at sayeeduddin@sbcglobal.net Thanks in advance for your response. Sayeed. ------------------------------ Message: 12 Date: Tue, 10 Jul 2012 09:32:51 -0500 From: "Reynolds,Donna M" Subject: [Histonet] CD40 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <785BBF0C5F49CE41BA74460A43A08F0232223A8783@DCPWVMBXC0VS3.mdanderson.edu> Content-Type: text/plain; charset="us-ascii" What are you using for Endogenous blocking? If you are using methanol with H2O2 that may be your problem. Many CD antibodies are very sensitive to the methanol. Try PBS or whatever buffer you are using to dilute the H202. I would also try going straight form the -20 into cold acetone without air drying and fix for10 min. Donna Reynolds HT (ASCP) U. T. M.D. Anderson Cancer Center Chief histology Tech, Department Cancer Biology 713-792-8106 ------------------------------ Message: 3 Date: Mon, 9 Jul 2012 07:55:16 -0700 From: Bea DeBrosse-Serra Subject: [Histonet] Need help with CD40 on frozen sections To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FBFB@EXCHMB01.isis.local> Content-Type: text/plain; charset="iso-8859-1" Histonetters, I need some help with CD40 on frozen sections (spleen, non-treated). I cut my frozens, let them briefly air dry and keep them at -20?C. When I use them for staining, I air dry them, fix them in cold acetone for 5 minutes and start the staining process. I use the primary AB from BD Pharmingen, # 550285 at various dilution, incubate them overnight at 4?C, use the secondary HRP conjugated antibody, DAB, counterstain and don't get any staining. Does anyone have any pointers, suggestions? Thank you in advance, Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 *********************************** ------------------------------ Message: 13 Date: Tue, 10 Jul 2012 15:51:00 +0000 From: Cathy Crumpton Subject: [Histonet] IVD P63 To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="iso-8859-1" I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 ------------------------------ Message: 14 Date: Tue, 10 Jul 2012 08:48:54 -0700 (PDT) From: Kim Merriam Subject: [Histonet] human vimentin in mouse tissue To: Histonet Message-ID: <1341935334.82981.YahooMailNeo@web130104.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Greetings! ? I want to stain for human vimentin in mouse tissues (human fibroblasts).? I many years ago, I used?a biotinylated V9 clone for just this purpose and it worked great.??I tried the?biotiylated V9 clone from abcam, but I am not getting any staining at all.? Any suggestions? ? Kim ? Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA? ------------------------------ Message: 15 Date: Tue, 10 Jul 2012 09:02:38 -0700 (PDT) From: Kim Donadio Subject: Re: [Histonet] IVD P63 To: Cathy Crumpton , "histonet@lists.utsouthwestern.edu" Message-ID: <1341936158.34785.YahooMailNeo@web112318.mail.gq1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. ________________________________ From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody.? I would like to purchase IVD P63 from another vendor, who else sells it?? I have having a few problems finding good info just using Google.? What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Tue, 10 Jul 2012 12:48:20 -0400 From: Clare Thornton Subject: RE: [Histonet] IVD P63 To: 'Kim Donadio' , Cathy Crumpton , "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="iso-8859-1" I just spoke with my Biocare rep. He said it is untrue at this time, and they are still selling p63. Clare J. Thornton, HTL(ASCP),QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthornton@dahlchase.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Tuesday, July 10, 2012 12:03 PM To: Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IVD P63 Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. ________________________________ From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody.? I would like to purchase IVD P63 from another vendor, who else sells it?? I have having a few problems finding good info just using Google.? What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 104, Issue 11 ***************************************** From Timothy.Morken <@t> ucsfmedctr.org Tue Jul 10 12:39:07 2012 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Timothy) Date: Tue Jul 10 12:38:43 2012 Subject: [Histonet] microscope ocular questions Message-ID: <8D7C2D242DBD45498006B21122072BF8B5B1455A@MCINFRWEM003.ucsfmedicalcenter.org> Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.morken@ucsfmedctr.org From LPaveli1 <@t> hurleymc.com Tue Jul 10 13:13:08 2012 From: LPaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Tue Jul 10 13:13:19 2012 Subject: [Histonet] IVD P63 In-Reply-To: References: <1341936158.34785.YahooMailNeo@web112318.mail.gq1.yahoo.com>, Message-ID: <89F4666A496DC949A819ECC40E11C8670C6B16@EXCHANGEMB1.hmc.hurleymc.com> Clare, Just got off the phone with my Biocare rep, Lissa Wall. I'm afraid that it IS true. Long story short, legal issues going on, and Biocare will stop selling P63 TODAY, 7/10/12. Please talk to your rep ASAP to get those last minute orders in today. I well remember when the last (no name) company received sole rights to an antibody and the pricing quadrupled overnight! HURRY!! Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Clare Thornton [CThornton@dahlchase.com] Sent: Tuesday, July 10, 2012 12:48 PM To: 'Kim Donadio'; Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IVD P63 I just spoke with my Biocare rep. He said it is untrue at this time, and they are still selling p63. Clare J. Thornton, HTL(ASCP),QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthornton@dahlchase.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Tuesday, July 10, 2012 12:03 PM To: Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IVD P63 Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. ________________________________ From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cathy.crumpton <@t> tuality.org Tue Jul 10 13:18:18 2012 From: cathy.crumpton <@t> tuality.org (Cathy Crumpton) Date: Tue Jul 10 13:18:21 2012 Subject: [Histonet] IVD P63 Message-ID: Yes, it is true. I just talked with Biogenex Laboratories and they will be carrying both the 4A4 and 4B1E12 clones in the foreseeable future. Other places I have talked to are depleting their stock and not carrying P63 anymore. Ventana wants $800+ dollars for 50 tests worth and do not carry concentrates. What a racket! Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 From lpwenk <@t> sbcglobal.net Tue Jul 10 13:26:30 2012 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Jul 10 13:26:38 2012 Subject: [Histonet] microscope ocular questions Message-ID: <37465B9E0CE145209FB848F59FB9FFE2@HP2010> Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -----Original Message----- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.morken@ucsfmedctr.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Timothy.Morken <@t> ucsfmedctr.org Tue Jul 10 13:49:02 2012 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Timothy) Date: Tue Jul 10 13:48:33 2012 Subject: [Histonet] microscope ocular questions In-Reply-To: <37465B9E0CE145209FB848F59FB9FFE2@HP2010> References: <37465B9E0CE145209FB848F59FB9FFE2@HP2010> Message-ID: <8D7C2D242DBD45498006B21122072BF8B5B14656@MCINFRWEM003.ucsfmedicalcenter.org> Thanks Peggy, That is clear. I used microscopes for years with one fixed ocular and one focusable ocular. I was wondering about why now both oculars are "focusable" yet one has more usability than the other. Maybe to accomodate greater variation? Or maybe is due to the advent of parfocal microscopes I found some instructions on parfocal adjustment that refers to setting both oculars to zero when doing the initial focus at high magnification, then setting the ocular adjustment for each eye at low magnification. So that makes sense for individualistic adjustment. However, I was asked why one ocular has easier use and more graduations that the other and I didn't have a good answer to that...The person thought the oculars were not the same so there was some problem with the microscope. Tim -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 10, 2012 11:27 AM To: Histonet Subject: RE: [Histonet] microscope ocular questions Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -----Original Message----- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.morken@ucsfmedctr.org _______________________________________________ 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 From ltougas <@t> dawsoncollege.qc.ca Tue Jul 10 13:57:41 2012 From: ltougas <@t> dawsoncollege.qc.ca (Liette Tougas) Date: Tue Jul 10 13:57:51 2012 Subject: [Histonet] direct IF on "normal" animal tissue Message-ID: <7618BC6D53F39149B7B57615C218AADF2CA286538C@EXCHANGE.ad.dawsoncollege.qc.ca> I teach immunology and histology to biomedical laboratory technology students and I would like to perform a simple direct immunofluorescence technique on "normal" animal tissue obtained from the butcher. I believe I could easily obtain either kidney, heart or liver either from pig or beef. Therefore, I was wondering which fluorescent antibody to purchase in order to demonstrate an antigen normally present in such animal tissues and that would not produce too much background, considering an expectable certain degree of autolysis. I appreciate any suggestion and detailed information if possible. Thank you in advance, Liette Tougas, RT, B.Sc., M.Sc. Biomedical Laboratory Technology Department Dawson College, Montreal, Qc From lpwenk <@t> sbcglobal.net Tue Jul 10 14:55:06 2012 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Jul 10 14:55:10 2012 Subject: Fw: [Histonet] microscope ocular questions Message-ID: <4AB0F0ACB50D48B8919713FC130A84EC@HP2010> Tim, You are welcome, but I'm not Peggy. I'm Lee, her husband. I'm very familiar with binoculars, so I only assumed that they are the same as microscopes. Lee Wenk -----Original Message----- From: Morken, Timothy Sent: Tuesday, July 10, 2012 2:49 PM To: Lee & Peggy Wenk ; Histonet Subject: RE: [Histonet] microscope ocular questions Thanks Peggy, That is clear. I used microscopes for years with one fixed ocular and one focusable ocular. I was wondering about why now both oculars are "focusable" yet one has more usability than the other. Maybe to accomodate greater variation? Or maybe is due to the advent of parfocal microscopes I found some instructions on parfocal adjustment that refers to setting both oculars to zero when doing the initial focus at high magnification, then setting the ocular adjustment for each eye at low magnification. So that makes sense for individualistic adjustment. However, I was asked why one ocular has easier use and more graduations that the other and I didn't have a good answer to that...The person thought the oculars were not the same so there was some problem with the microscope. Tim -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 10, 2012 11:27 AM To: Histonet Subject: RE: [Histonet] microscope ocular questions Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -----Original Message----- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.morken@ucsfmedctr.org _______________________________________________ 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 From billodonnell <@t> catholichealth.net Tue Jul 10 15:14:39 2012 From: billodonnell <@t> catholichealth.net (O'Donnell, Bill) Date: Tue Jul 10 15:15:25 2012 Subject: [Histonet] microscope ocular questions In-Reply-To: <8D7C2D242DBD45498006B21122072BF8B5B14656@MCINFRWEM003.ucsfmedicalcenter.org> References: <37465B9E0CE145209FB848F59FB9FFE2@HP2010> <8D7C2D242DBD45498006B21122072BF8B5B14656@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <4940DF6D1C5FDF48931B6966AAEF93956854F4@chimsx08.CHI.catholichealth.net> Tim, The duel-adjustable eyepiece vs the single adjustable one, I believe has to do with the type of prism that is used. Mis-matched eyepieces can be a problem, especially if from different manufacturers or even magnification. Magnification differences would be somewhat obvious as you will never get them to focus for two eyes. In order to use a binocular telescope (really cool instrument, once had the chance to use one of those) you need, not only matched eyepieces, but ideally from the same lot #. However, focusing two ten-inch telescopes to the same focal plane is a bit touchier than a microscope. Its one of the reasons you don't see too many of them. Have you tried simply swapping them (left to right/right to left)? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy Sent: Tuesday, July 10, 2012 1:49 PM To: Lee & Peggy Wenk; Histonet Subject: RE: [Histonet] microscope ocular questions Thanks Peggy, That is clear. I used microscopes for years with one fixed ocular and one focusable ocular. I was wondering about why now both oculars are "focusable" yet one has more usability than the other. Maybe to accomodate greater variation? Or maybe is due to the advent of parfocal microscopes I found some instructions on parfocal adjustment that refers to setting both oculars to zero when doing the initial focus at high magnification, then setting the ocular adjustment for each eye at low magnification. So that makes sense for individualistic adjustment. However, I was asked why one ocular has easier use and more graduations that the other and I didn't have a good answer to that...The person thought the oculars were not the same so there was some problem with the microscope. Tim -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 10, 2012 11:27 AM To: Histonet Subject: RE: [Histonet] microscope ocular questions Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -----Original Message----- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.morken@ucsfmedctr.org _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. From laurie.reilly <@t> jcu.edu.au Tue Jul 10 18:21:56 2012 From: laurie.reilly <@t> jcu.edu.au (Reilly, Laurie) Date: Tue Jul 10 18:22:21 2012 Subject: [Histonet] Another Microscope Question Message-ID: Dear All, My microscope training has been mainly on brightfield microscopes using Koehler illumination. Recently, in discussions with Veterinary Pathologists, they have been advocating winding the condenser way down to the field diaphragm when they are examining urine samples. This goes against the microscopy principles that I have been taught and now teach to our students. Can anyone enlighten me on the value of this practice? Could the same effect be obtained by closing the aperture diaphragm? Thanks in advance for your wisdom. Regards, Laurie. Mr. Laurie REILLY Histopathology School of Veterinary and Biomedical Sciences James Cook University Townsville Qld. 4811 Australia. Phone 07 4781 4468 From one_angel_secret <@t> yahoo.com Tue Jul 10 19:28:29 2012 From: one_angel_secret <@t> yahoo.com (Kim Donadio) Date: Tue Jul 10 19:28:35 2012 Subject: [Histonet] direct IF on "normal" animal tissue In-Reply-To: <7618BC6D53F39149B7B57615C218AADF2CA286538C@EXCHANGE.ad.dawsoncollege.qc.ca> References: <7618BC6D53F39149B7B57615C218AADF2CA286538C@EXCHANGE.ad.dawsoncollege.qc.ca> Message-ID: <1341966509.97867.YahooMailNeo@web112304.mail.gq1.yahoo.com> Hi Liette, ?????????????? I didnt see if you got help yet. Ill give it a try. First I would go with the pig kidney,. and this antibody looks like it should work . ? http://www.abcam.com/Pig-IgG-secondary-antibody-H-L-ab6911.html ? This is my best guess off the top of my head. If you try it and it works, let us know, ? oh and you can look on that site for procedures too. and i bet you could call and get some good info. ? hope this helps, good luck :) ? Kim D ? ________________________________ From: Liette Tougas To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 2:57 PM Subject: [Histonet] direct IF on "normal" animal tissue I teach immunology and histology to biomedical laboratory technology students and I would like to perform a simple direct immunofluorescence technique on "normal" animal tissue obtained from the butcher.? I believe I could easily obtain either kidney, heart or liver either from pig or beef.? Therefore, I was wondering which fluorescent antibody to purchase in order to demonstrate an antigen normally present in such animal tissues and that would not produce too much background, considering an expectable certain degree of autolysis. I appreciate any suggestion and detailed information if possible. Thank you in advance, Liette Tougas, RT, B.Sc., M.Sc. Biomedical Laboratory Technology Department Dawson College, Montreal, Qc _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tony.henwood <@t> health.nsw.gov.au Tue Jul 10 20:25:42 2012 From: tony.henwood <@t> health.nsw.gov.au (Tony Henwood (SCHN)) Date: Tue Jul 10 20:25:58 2012 Subject: [Histonet] RE: Another Microscope Question In-Reply-To: References: Message-ID: <6D6BD1DE8A5571489398B392A38A71579D1C5BF3@xmdb02.nch.kids> Hi Laurie, How are you? The idea of "pushing out of Koehler" as you described gives you a "poor-man's", pseudo-phase effect. I use it for unstained frozen sections of renal and skin biopsies that are destined for immunofluorescence. And then I get mean and request the students who I have demonstrated this technique to, to return the microscope to "Koehler perfection". I suppose it teaches the student to be familiar with the microscopy results and to recognise when a better image is achievable. I am such a mean person. A useful reference?: Head ES. Tschen EH. (1982) Unstained negative image patterns. A checkpoint for quality slides. American Journal of Dermatopathology. 4(2):143-8. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Reilly, Laurie Sent: Wednesday, 11 July 2012 9:22 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Another Microscope Question Dear All, My microscope training has been mainly on brightfield microscopes using Koehler illumination. Recently, in discussions with Veterinary Pathologists, they have been advocating winding the condenser way down to the field diaphragm when they are examining urine samples. This goes against the microscopy principles that I have been taught and now teach to our students. Can anyone enlighten me on the value of this practice? Could the same effect be obtained by closing the aperture diaphragm? Thanks in advance for your wisdom. Regards, Laurie. Mr. Laurie REILLY Histopathology School of Veterinary and Biomedical Sciences James Cook University Townsville Qld. 4811 Australia. Phone 07 4781 4468 ********************************************************************************* 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. ********************************************************************************* From carl.hobbs <@t> kcl.ac.uk Tue Jul 10 22:31:56 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Tue Jul 10 22:32:08 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation Message-ID: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 From tony.henwood <@t> health.nsw.gov.au Tue Jul 10 23:20:52 2012 From: tony.henwood <@t> health.nsw.gov.au (Tony Henwood (SCHN)) Date: Tue Jul 10 23:21:03 2012 Subject: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> References: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> Message-ID: <6D6BD1DE8A5571489398B392A38A71579D1C5E14@xmdb02.nch.kids> Hi Carl, What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, 11 July 2012 1:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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. ********************************************************************************* From Mandy.Bell <@t> chomp.org Wed Jul 11 06:31:44 2012 From: Mandy.Bell <@t> chomp.org (Bell, Mandy) Date: Wed Jul 11 06:32:20 2012 Subject: [Histonet] congenital syphilis Message-ID: <87583127C45A5B4994B162C67437826518A3A8F5@EXMAIL1P.chomp.org> Hi, I was hoping someone would have a suggestion for a stain for congenital syphilis on a placenta. Our pathologist wants this done- I know a warthinstarry or steiner would work, but we do not do either of these stains. Would a diff-quik stain work? Thanks, Mandy Bell HTL(ASCP) Community Hospital of the Monterey Peninsula 2 Harris Court Suite B3/B4 Monterey, CA 93940 (831) 647-4791 Confidentiality Notice: This is a transmission from Community Hospital of the Monterey Peninsula. This message and any attached documents may be confidential and contain information protected by state and federal medical privacy statutes. They are intended only for the use of the addressee. If you are not the intended recipient, any disclosure, copying, or distribution of this information is strictly prohibited. If you received this transmission in error, please accept our apologies and notify the sender. Thank you. From Farnana <@t> nehealth.com Wed Jul 11 06:53:58 2012 From: Farnana <@t> nehealth.com (Amy Farnan) Date: Wed Jul 11 06:54:07 2012 Subject: [Histonet] Consulting Message-ID: <4FFD3111.26ED.00D9.1@nehealth.com> Ryan Jade, I am inquiring if you may have found a consultant to help you with your dermatology laboratory? Laboratory Concepts, LLC can help you with your needs. I would be happy to speak with you in detail offline. Regards, Amy Farnan A.farnan@yahoo.com Disclaimer: The information in this message is confidential. If you are not the intended recipient, do not disclose, copy, or distribute this message, and please immediately contact the sender. From settembr <@t> umdnj.edu Wed Jul 11 08:31:25 2012 From: settembr <@t> umdnj.edu (Settembre, Dana) Date: Wed Jul 11 08:31:40 2012 Subject: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> References: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> Message-ID: Carl, H202 mixed with Buffer here.(instead of aq.) That's OK too, right? Dana -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Tuesday, July 10, 2012 11:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed Jul 11 09:13:49 2012 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Jul 11 09:14:01 2012 Subject: [Histonet] congenital syphilis In-Reply-To: <87583127C45A5B4994B162C67437826518A3A8F5@EXMAIL1P.chomp.org> References: <87583127C45A5B4994B162C67437826518A3A8F5@EXMAIL1P.chomp.org> Message-ID: <4FFD51DD.7400.0077.1@harthosp.org> Immunohistochemical testing for Treponema pallidum is the preferred method of identification. There are labs (including my own) that offer this. Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax >>> "Bell, Mandy" 7/11/2012 7:31 AM >>> Hi, I was hoping someone would have a suggestion for a stain for congenital syphilis on a placenta. Our pathologist wants this done- I know a warthinstarry or steiner would work, but we do not do either of these stains. Would a diff-quik stain work? Thanks, Mandy Bell HTL(ASCP) Community Hospital of the Monterey Peninsula 2 Harris Court Suite B3/B4 Monterey, CA 93940 (831) 647-4791 Confidentiality Notice: This is a transmission from Community Hospital of the Monterey Peninsula. This message and any attached documents may be confidential and contain information protected by state and federal medical privacy statutes. They are intended only for the use of the addressee. If you are not the intended recipient, any disclosure, copying, or distribution of this information is strictly prohibited. If you received this transmission in error, please accept our apologies and notify the sender. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Wed Jul 11 09:17:03 2012 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Wed Jul 11 09:17:09 2012 Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <6D6BD1DE8A5571489398B392A38A71579D1C5E14@xmdb02.nch.kids> References: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> <6D6BD1DE8A5571489398B392A38A71579D1C5E14@xmdb02.nch.kids> Message-ID: <002b01cd5f6f$d85f5f70$891e1e50$@gmx.at> For the enzymatic activity of peroxidase it needs an electron-donator (or receptor - I can't find the literature...) in the vicinity; therefore H2O2 and DAB are added ad once, and DAB is oxidized and transformed into the insoluble, amorphe substance through polymerization. Without the donator H2O2 in excess works as inhibitor and blocks the activation-side of the enzyme. I think H2O2 in methanol was primarly preferred, because the frozen slides were fixed at the same time. Rehydration after dewaxing depends on the following reagens. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood (SCHN) Gesendet: Mittwoch, 11. Juli 2012 06:21 An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation Hi Carl, What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, 11 July 2012 1:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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. **************************************************************************** ***** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Jul 11 09:33:37 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jul 11 09:33:42 2012 Subject: [Histonet] Another Microscope Question In-Reply-To: References: Message-ID: <1342017217.82429.YahooMailNeo@web121406.mail.ne1.yahoo.com> Laurie: K?hler illumination is not just observing objects in bright field, it implies the use 2 diaphragms; one near the light source and focused in a way that the diaphragm closest to the light source covers (illuminates)?only the area to be observed. With the condenser diaphragm you then increase/decrease the intensity of the light. It is most useful for photomicrography and to obtain the maximum resolution of the numerical aperture (NA) of your objectives. If you look at the slide side wise, you will see that the illuminated area corresponds only to the working filed of each objective. That is why when using K?hler illumination you have to adjust the light source diaphragm every time you change the objective under use. What your pathologists are trying to do is just to increase/reduce the amount of light reaching the object by increasing/reducing the distance of the condenser from the object to be observed. Many years ago and starting in the mid 1920s, Carl Zeiss (and also Ernst Leitz) designed the condensers of their research microscopes with the condenser diaphragm mounted on a slide with a knob. By doing so these condensers could move the condenser diaphragm side wise in a way that the light entered into the condenser not straight from the light source obliquously, and the objects were illuminated side wise. This illumination permitted to observe unstained slides and produced the illusion of "three dimension" and permitted to observe unstained structures. This type of illumination was quite popular in hematology labs to observe blood smears In 1935 the same Carl Zeiss started to manufacture objectives/condenser sytems using Zernike's principle, and phase microscopy was born. The method of lowering the condenser used by your pathologists has to produce a very poor quality image. You would be better off if you cut a piece of black paper half the diameter of the filter ring of your condenser and place it there. With this opaque filter the light will get to your object side wise, the same as if you have moved the condenser side wise. Ren? J. ________________________________ From: "Reilly, Laurie" To: "Histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 7:21 PM Subject: [Histonet] Another Microscope Question Dear All, My microscope training has been mainly on brightfield microscopes using Koehler illumination. Recently, in discussions with Veterinary Pathologists, they have been advocating winding the condenser way down to the field diaphragm when they are examining urine samples. This goes against the microscopy principles that I have been taught and now teach to our students. Can anyone enlighten me on the value of this practice? Could the same effect be obtained by closing the aperture diaphragm? Thanks in advance for your wisdom. Regards,? Laurie. Mr. Laurie REILLY Histopathology School of? Veterinary and Biomedical Sciences James Cook University Townsville? Qld.? 4811 Australia. Phone 07 4781 4468 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Wed Jul 11 09:50:05 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Wed Jul 11 09:50:24 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation Message-ID: <11D9615B89C10747B1C985966A63D7CA386254787E@KCL-MAIL04.kclad.ds.kcl.ac.uk> I made a mistake....I DO rehydrate before staining, as Tony pointed out. I just don't use graded alcohols between the xylene and water ( x4 changes 74OP IMS). Carl Hobbs Histology Manager Wolfson Centre for Age-Related Diseases School of Biomedical Sciences King's College London Guys Campus London SE1 1UL Tel.020 7848 6810 fax 020 7848 6816 carl.hobbs@kcl.ac.uk From BDeBrosse-Serra <@t> isisph.com Wed Jul 11 10:13:49 2012 From: BDeBrosse-Serra <@t> isisph.com (Bea DeBrosse-Serra) Date: Wed Jul 11 10:13:59 2012 Subject: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <11D9615B89C10747B1C985966A63D7CA386254787E@KCL-MAIL04.kclad.ds.kcl.ac.uk> References: <11D9615B89C10747B1C985966A63D7CA386254787E@KCL-MAIL04.kclad.ds.kcl.ac.uk> Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FC19@EXCHMB01.isis.local> And the reason for that is.......??? Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, July 11, 2012 7:50 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Re: Methanol in H2O2 explanation I made a mistake....I DO rehydrate before staining, as Tony pointed out. I just don't use graded alcohols between the xylene and water ( x4 changes 74OP IMS). Carl Hobbs Histology Manager Wolfson Centre for Age-Related Diseases School of Biomedical Sciences King's College London Guys Campus London SE1 1UL Tel.020 7848 6810 fax 020 7848 6816 carl.hobbs@kcl.ac.uk From Timothy.Morken <@t> ucsfmedctr.org Wed Jul 11 11:12:04 2012 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Timothy) Date: Wed Jul 11 11:11:37 2012 Subject: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <002b01cd5f6f$d85f5f70$891e1e50$@gmx.at> References: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> <6D6BD1DE8A5571489398B392A38A71579D1C5E14@xmdb02.nch.kids> <002b01cd5f6f$d85f5f70$891e1e50$@gmx.at> Message-ID: <8D7C2D242DBD45498006B21122072BF8B5BBA179@MCINFRWEM003.ucsfmedicalcenter.org> It's funny how histology lore gets passed down, passed around and misunderstood over the decades. When I ask questions in labs about why they do a certain procedure they can rarely name a source (besides a particular long-retired particular tech who wrote the original procedure) or give reason as to why it is done that way. Even if it works well, at least people could take the time to understand why! When I was researching buffer compositions long ago I found a dozen variations of PBS alone (and only a dozen because I just stopped looking). When you delve into these things it becomes clear that people made their own solutions for a particular purpose. Maybe they wrote a paper, taught a course or wrote a book. Then that formula was copied by many others and their particular formulation became the "state of the art" even if the "art" practiced by someone else had little to do with the original pursuit (the Sheehan book is full of variations of stains that were originally for specific research purposes. There is precious little guidance in any books about what is good for a particular use!). The reason for using methanol / H2O2 that I was told long ago was prevent frozen sections from being damaged by H2O2 bubbling. In fact, we would fix frozens in cold acetone and THEN put the slides in cold methanol/H2O2 (double fixation?) In that lab we never used methanol for deparaffinized sections. Since I had no exposure to the clin lab I never knew about using methanol for blood smear fixation. From literature searches it seems methanol, rather than ethanol, is used primarily because it is cheaper. In other words, it does not appear to be a magical substance! The questions asked here are good ones. However, hopefully those reading and responsible for producing procedure manuals will include the reasoning for each step of the method and not just assume it is necessary, or "common knowledge," just because it has been handed down through generations of long suffering techs! Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, July 11, 2012 7:17 AM To: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation For the enzymatic activity of peroxidase it needs an electron-donator (or receptor - I can't find the literature...) in the vicinity; therefore H2O2 and DAB are added ad once, and DAB is oxidized and transformed into the insoluble, amorphe substance through polymerization. Without the donator H2O2 in excess works as inhibitor and blocks the activation-side of the enzyme. I think H2O2 in methanol was primarly preferred, because the frozen slides were fixed at the same time. Rehydration after dewaxing depends on the following reagens. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood (SCHN) Gesendet: Mittwoch, 11. Juli 2012 06:21 An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation Hi Carl, What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, 11 July 2012 1:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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. **************************************************************************** ***** _______________________________________________ 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 From b-frederick <@t> northwestern.edu Wed Jul 11 11:13:39 2012 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Wed Jul 11 11:13:47 2012 Subject: [Histonet] Beecher Message-ID: <62C639732D3F274DACED033EBDF6ADAF1E26B8AE@evcspmbx3.ads.northwestern.edu> All ,I know this came up before,but who took over Beecher? We need to order some more punchers. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-frederick@northwestern.edu From LLoss <@t> dermwisconsin.com Wed Jul 11 11:22:50 2012 From: LLoss <@t> dermwisconsin.com (Lee Loss) Date: Wed Jul 11 11:23:02 2012 Subject: [Histonet] job opening Message-ID: <2B99CB40EC5CC7409888A2961CBF9688CBB5F8@EX2010.DERM.LOCAL> We are currently looking for a histotech to join our rapidly growing practice and explore the exciting world of dermatopathology in a new state-of-the-art laboratory overlooking beautiful Lake Michigan. At Dermatology Associates of Wisconsin you will enjoy performing a wide variety of tests including grossing, DIFs, special stains, and IHC. You will experience working 4 days per week for a dermatology group averaging over 30% growth annually for the past 10 years. Benefits include: * 401k match of 100% of the first 4% of employee contribution * Company profit sharing contribution of 7% of employee earnings * Company funded cash balance plan that is a yearly contribution of 2% of employee earnings * Immediate PTO accrual * Leadership that enjoys teaching * A great team atmosphere * Employee discounts * Opportunities for professional growth For more details contact: Human Resources 801 York Street Manitowoc, WI 54220 HR@dermwisconsin.com Office: (920)683-5278 Fax:(920)663-9004 ________________________________ The materials and information in this e-mail are confidential and may contain Protected Health Information covered under the HIPAA Privacy Rule. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution, or action taken in reliance on the contents of this information is strictly forbidden by law. If you have received this e-mail in error, please notify me by reply e-mail and then delete this message. Do not pass any of this information to anyone else. Thank you for your cooperation. From hfedor <@t> jhmi.edu Wed Jul 11 11:42:53 2012 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Wed Jul 11 11:44:31 2012 Subject: [Histonet] RE: Beecher In-Reply-To: <62C639732D3F274DACED033EBDF6ADAF1E26B8AE@evcspmbx3.ads.northwestern.edu> References: <62C639732D3F274DACED033EBDF6ADAF1E26B8AE@evcspmbx3.ads.northwestern.edu> Message-ID: http://www.pathologydevices.com/TMArrayer.htm You can contact Ron Gebing at Pathology Devices. Helen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Wednesday, July 11, 2012 12:14 PM To: Fellow HistoNetters Subject: [Histonet] Beecher All ,I know this came up before,but who took over Beecher? We need to order some more punchers. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-frederick@northwestern.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Jul 11 11:50:00 2012 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Jul 11 11:48:30 2012 Subject: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <8D7C2D242DBD45498006B21122072BF8B5BBA179@MCINFRWEM003.ucsfmedicalcenter.org> References: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> <6D6BD1DE8A5571489398B392A38A71579D1C5E14@xmdb02.nch.kids> <002b01cd5f6f$d85f5f70$891e1e50$@gmx.at> <8D7C2D242DBD45498006B21122072BF8B5BBA179@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <4FFDAEB8.2050302@umdnj.edu> I agree, lots of "lore" gets passed down without question or understanding. Many years ago a new PhD who had trained in a good lab came to my lab looking for some "NaH." I tried to explain that there was no such chemical when she produced a xeroxed lab protocol calling for NaH to make a malete buffer for EM (that shows you how long ago this was!). So I found the exact recipe but with NaOH. The techs all knew it was a typo but she did not. Geoff On 7/11/2012 12:12 PM, Morken, Timothy wrote: > It's funny how histology lore gets passed down, passed around and misunderstood over the decades. When I ask questions in labs about why they do a certain procedure they can rarely name a source (besides a particular long-retired particular tech who wrote the original procedure) or give reason as to why it is done that way. Even if it works well, at least people could take the time to understand why! > > When I was researching buffer compositions long ago I found a dozen variations of PBS alone (and only a dozen because I just stopped looking). When you delve into these things it becomes clear that people made their own solutions for a particular purpose. Maybe they wrote a paper, taught a course or wrote a book. Then that formula was copied by many others and their particular formulation became the "state of the art" even if the "art" practiced by someone else had little to do with the original pursuit (the Sheehan book is full of variations of stains that were originally for specific research purposes. There is precious little guidance in any books about what is good for a particular use!). > > The reason for using methanol / H2O2 that I was told long ago was prevent frozen sections from being damaged by H2O2 bubbling. In fact, we would fix frozens in cold acetone and THEN put the slides in cold methanol/H2O2 (double fixation?) In that lab we never used methanol for deparaffinized sections. Since I had no exposure to the clin lab I never knew about using methanol for blood smear fixation. From literature searches it seems methanol, rather than ethanol, is used primarily because it is cheaper. In other words, it does not appear to be a magical substance! > > The questions asked here are good ones. However, hopefully those reading and responsible for producing procedure manuals will include the reasoning for each step of the method and not just assume it is necessary, or "common knowledge," just because it has been handed down through generations of long suffering techs! > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang > Sent: Wednesday, July 11, 2012 7:17 AM > To: histonet@lists.utsouthwestern.edu > Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation > > For the enzymatic activity of peroxidase it needs an electron-donator (or receptor - I can't find the literature...) in the vicinity; therefore H2O2 and DAB are added ad once, and DAB is oxidized and transformed into the insoluble, amorphe substance through polymerization. > Without the donator H2O2 in excess works as inhibitor and blocks the activation-side of the enzyme. > I think H2O2 in methanol was primarly preferred, because the frozen slides were fixed at the same time. > > Rehydration after dewaxing depends on the following reagens. > > Gudrun > > -----Urspr?ngliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood (SCHN) > Gesendet: Mittwoch, 11. Juli 2012 06:21 > An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu > Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation > > Hi Carl, > > What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. > > Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. > > I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. > > > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager& Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl > Sent: Wednesday, 11 July 2012 1:32 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Methanol in H2O2 explanation > > Why do some people use methanolic H2O2? > Why do people rehydrate after dewaxing? > Both are unnecessary, under usual conditions. > > Methanol was used in the early days as a peroxidase blocker by itself. > The combination was devised as a "belt and bracer" method. > As you stated, you use aq H2O2 effectively. > So do I and many others. > > However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... > > After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... > > Carl Hobbs > Histology and Imaging Manager > Wolfson CARD > School of Biomedical Sciences > Kings College London > Guys Campus > SE1 1UL > Tel: 020 78486813 > Fax: 020 78486816 > 020 78486813 > > > _______________________________________________ > 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. > **************************************************************************** > ***** > > _______________________________________________ > 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcauliff@umdnj.edu ********************************************** From brett_connolly <@t> merck.com Wed Jul 11 12:18:16 2012 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Wed Jul 11 12:18:20 2012 Subject: [Histonet] RE: Re: Methanol in H2O2 explanation In-Reply-To: <4FFDAEB8.2050302@umdnj.edu> References: <11D9615B89C10747B1C985966A63D7CA386254754E@KCL-MAIL04.kclad.ds.kcl.ac.uk> <6D6BD1DE8A5571489398B392A38A71579D1C5E14@xmdb02.nch.kids> <002b01cd5f6f$d85f5f70$891e1e50$@gmx.at> <8D7C2D242DBD45498006B21122072BF8B5BBA179@MCINFRWEM003.ucsfmedicalcenter.org> <4FFDAEB8.2050302@umdnj.edu> Message-ID: Now that's funny! Even if it makes we PhDs look bad. I was once given some tissue in "formaldehyde" from one of my colleagues in another department to process and stain. The resulting slides looked horrendous and I suspected the fix was the problem. I went back to their lab and asked to see the bottle of "formaldehyde" which turned out to be formamide. Then I got the shoulder shrug and "We thought it was close enough" response. Arrrrrgghh. Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Geoff McAuliffe Sent: Wednesday, July 11, 2012 12:50 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Re: Methanol in H2O2 explanation I agree, lots of "lore" gets passed down without question or understanding. Many years ago a new PhD who had trained in a good lab came to my lab looking for some "NaH." I tried to explain that there was no such chemical when she produced a xeroxed lab protocol calling for NaH to make a malete buffer for EM (that shows you how long ago this was!). So I found the exact recipe but with NaOH. The techs all knew it was a typo but she did not. Geoff On 7/11/2012 12:12 PM, Morken, Timothy wrote: > It's funny how histology lore gets passed down, passed around and misunderstood over the decades. When I ask questions in labs about why they do a certain procedure they can rarely name a source (besides a particular long-retired particular tech who wrote the original procedure) or give reason as to why it is done that way. Even if it works well, at least people could take the time to understand why! > > When I was researching buffer compositions long ago I found a dozen variations of PBS alone (and only a dozen because I just stopped looking). When you delve into these things it becomes clear that people made their own solutions for a particular purpose. Maybe they wrote a paper, taught a course or wrote a book. Then that formula was copied by many others and their particular formulation became the "state of the art" even if the "art" practiced by someone else had little to do with the original pursuit (the Sheehan book is full of variations of stains that were originally for specific research purposes. There is precious little guidance in any books about what is good for a particular use!). > > The reason for using methanol / H2O2 that I was told long ago was prevent frozen sections from being damaged by H2O2 bubbling. In fact, we would fix frozens in cold acetone and THEN put the slides in cold methanol/H2O2 (double fixation?) In that lab we never used methanol for deparaffinized sections. Since I had no exposure to the clin lab I never knew about using methanol for blood smear fixation. From literature searches it seems methanol, rather than ethanol, is used primarily because it is cheaper. In other words, it does not appear to be a magical substance! > > The questions asked here are good ones. However, hopefully those reading and responsible for producing procedure manuals will include the reasoning for each step of the method and not just assume it is necessary, or "common knowledge," just because it has been handed down through generations of long suffering techs! > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang > Sent: Wednesday, July 11, 2012 7:17 AM > To: histonet@lists.utsouthwestern.edu > Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation > > For the enzymatic activity of peroxidase it needs an electron-donator (or receptor - I can't find the literature...) in the vicinity; therefore H2O2 and DAB are added ad once, and DAB is oxidized and transformed into the insoluble, amorphe substance through polymerization. > Without the donator H2O2 in excess works as inhibitor and blocks the activation-side of the enzyme. > I think H2O2 in methanol was primarly preferred, because the frozen slides were fixed at the same time. > > Rehydration after dewaxing depends on the following reagens. > > Gudrun > > -----Urspr?ngliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood (SCHN) > Gesendet: Mittwoch, 11. Juli 2012 06:21 > An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu > Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation > > Hi Carl, > > What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. > > Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. > > I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. > > > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager& Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl > Sent: Wednesday, 11 July 2012 1:32 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Methanol in H2O2 explanation > > Why do some people use methanolic H2O2? > Why do people rehydrate after dewaxing? > Both are unnecessary, under usual conditions. > > Methanol was used in the early days as a peroxidase blocker by itself. > The combination was devised as a "belt and bracer" method. > As you stated, you use aq H2O2 effectively. > So do I and many others. > > However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... > > After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... > > Carl Hobbs > Histology and Imaging Manager > Wolfson CARD > School of Biomedical Sciences > Kings College London > Guys Campus > SE1 1UL > Tel: 020 78486813 > Fax: 020 78486816 > 020 78486813 > > > _______________________________________________ > 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. > **************************************************************************** > ***** > > _______________________________________________ > 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. From mary.helie <@t> yale.edu Wed Jul 11 12:36:45 2012 From: mary.helie <@t> yale.edu (Helie, Mary) Date: Wed Jul 11 12:36:54 2012 Subject: [Histonet] Antibody Inventory Message-ID: Good Afternoon Does anyone have any suggestions for primary antibody inventory management? Thank you Mary Helie From Karen.Heckford <@t> DignityHealth.org Wed Jul 11 12:37:56 2012 From: Karen.Heckford <@t> DignityHealth.org (Heckford, Karen - SMMC-SF) Date: Wed Jul 11 12:38:02 2012 Subject: [Histonet] P63 Message-ID: <3328693C43A557458850CC37CE16CD1877954A52@chw-msg-829.chw.edu> I guess since Biocare cannot sell their P63 antibody who is carrying it then? Dako, Ventana etc? Karen Heckford HT ASCP CE Lead Histology Technician St. Mary's Medical Center 450 Stanyan St. San Francisco, Ca. 94117 415-668-1000 ext. 6167 karen.heckford@dignityhealth.org Caution: This email message, including all content and attachments, is CONFIDENTIAL and may be of a nature that is LEGALLY PRIVILEGED. The information contained in this email message is intended only for the use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you have received this document in error. Any further review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately by reply email. Thank you." From Loralee_Mcmahon <@t> URMC.Rochester.edu Wed Jul 11 12:41:43 2012 From: Loralee_Mcmahon <@t> URMC.Rochester.edu (McMahon, Loralee A) Date: Wed Jul 11 12:42:20 2012 Subject: [Histonet] RE: Beecher In-Reply-To: <62C639732D3F274DACED033EBDF6ADAF1E26B8AE@evcspmbx3.ads.northwestern.edu> References: <62C639732D3F274DACED033EBDF6ADAF1E26B8AE@evcspmbx3.ads.northwestern.edu> Message-ID: This is where I get my stuff for my TMA machine from http://www.estigen.com/ Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick [b-frederick@northwestern.edu] Sent: Wednesday, July 11, 2012 12:13 PM To: Fellow HistoNetters Subject: [Histonet] Beecher All ,I know this came up before,but who took over Beecher? We need to order some more punchers. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-frederick@northwestern.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Wed Jul 11 13:22:38 2012 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Wed Jul 11 13:22:45 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at> Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter enzymehistochemistry - the thing, that an electron-donator is used for demonstration. And he states, that methanol alone also inhibits peroxidase - perhaps by the fixation = denaturation effect on the protein? In the following publication one can see the equations for the peroxidase-reaction and this phrase: "In this study, we have used a steady state kinetics approach to demonstrate that MPO is irreversibly inactivated by excess H2O2 in the absence of reducing substrate." The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of Inactivation Rate by Organic Donor Substrates It's rather exhausting to prove the own statements ;-) puh. Gudrun -----Urspr?ngliche Nachricht----- Von: Morken, Timothy [mailto:Timothy.Morken@ucsfmedctr.org] Gesendet: Mittwoch, 11. Juli 2012 18:12 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] RE: Re: Methanol in H2O2 explanation It's funny how histology lore gets passed down, passed around and misunderstood over the decades. When I ask questions in labs about why they do a certain procedure they can rarely name a source (besides a particular long-retired particular tech who wrote the original procedure) or give reason as to why it is done that way. Even if it works well, at least people could take the time to understand why! When I was researching buffer compositions long ago I found a dozen variations of PBS alone (and only a dozen because I just stopped looking). When you delve into these things it becomes clear that people made their own solutions for a particular purpose. Maybe they wrote a paper, taught a course or wrote a book. Then that formula was copied by many others and their particular formulation became the "state of the art" even if the "art" practiced by someone else had little to do with the original pursuit (the Sheehan book is full of variations of stains that were originally for specific research purposes. There is precious little guidance in any books about what is good for a particular use!). The reason for using methanol / H2O2 that I was told long ago was prevent frozen sections from being damaged by H2O2 bubbling. In fact, we would fix frozens in cold acetone and THEN put the slides in cold methanol/H2O2 (double fixation?) In that lab we never used methanol for deparaffinized sections. Since I had no exposure to the clin lab I never knew about using methanol for blood smear fixation. From literature searches it seems methanol, rather than ethanol, is used primarily because it is cheaper. In other words, it does not appear to be a magical substance! The questions asked here are good ones. However, hopefully those reading and responsible for producing procedure manuals will include the reasoning for each step of the method and not just assume it is necessary, or "common knowledge," just because it has been handed down through generations of long suffering techs! Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, July 11, 2012 7:17 AM To: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation For the enzymatic activity of peroxidase it needs an electron-donator (or receptor - I can't find the literature...) in the vicinity; therefore H2O2 and DAB are added ad once, and DAB is oxidized and transformed into the insoluble, amorphe substance through polymerization. Without the donator H2O2 in excess works as inhibitor and blocks the activation-side of the enzyme. I think H2O2 in methanol was primarly preferred, because the frozen slides were fixed at the same time. Rehydration after dewaxing depends on the following reagens. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood (SCHN) Gesendet: Mittwoch, 11. Juli 2012 06:21 An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation Hi Carl, What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, 11 July 2012 1:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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. **************************************************************************** ***** _______________________________________________ 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 From LPaveli1 <@t> hurleymc.com Wed Jul 11 13:26:14 2012 From: LPaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Wed Jul 11 13:26:26 2012 Subject: [Histonet] RE: Antibody Inventory In-Reply-To: References: Message-ID: <89F4666A496DC949A819ECC40E11C8670C6D39@EXCHANGEMB1.hmc.hurleymc.com> We made up an excel spread sheet that contains: Antibody/Ancillary solutions(buffer, detection kits) Company Source Catalog # Lot # Expiration Date When we open an new antibody, we clear out the Lot # & Expiration date and add it to the "re-order list, ensuring that we never run out. Then when the new antibody comes in, we can quickly add it to the supply list. Additionally, the techs in the department found that it helps them to assign a color to each expiration month. So the expiration date "cell" will be; blue for January, red for February........and so on to give a visual help when watching for upcoming expiring antibodies. They also made a "cheat sheet" of colors that correspond to each month. I have also found that this list helps me immensly when inventory comes around!! Always did hate fishing around in the 'frig searching and counting antibodies!! I'm lucky, my department is very good about keeping up with the incoming/outgoing stock. hope this helps! :) Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Helie, Mary [mary.helie@yale.edu] Sent: Wednesday, July 11, 2012 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Antibody Inventory Good Afternoon Does anyone have any suggestions for primary antibody inventory management? Thank you Mary Helie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Loralee_Mcmahon <@t> URMC.Rochester.edu Wed Jul 11 13:35:48 2012 From: Loralee_Mcmahon <@t> URMC.Rochester.edu (McMahon, Loralee A) Date: Wed Jul 11 13:37:20 2012 Subject: [Histonet] RE: P63 In-Reply-To: <3328693C43A557458850CC37CE16CD1877954A52@chw-msg-829.chw.edu> References: <3328693C43A557458850CC37CE16CD1877954A52@chw-msg-829.chw.edu> Message-ID: They are still selling it. But they do not know for how long. And because it is a legal issue they can't tell me whom is going to be the sole provider either. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heckford, Karen - SMMC-SF [Karen.Heckford@DignityHealth.org] Sent: Wednesday, July 11, 2012 1:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] P63 I guess since Biocare cannot sell their P63 antibody who is carrying it then? Dako, Ventana etc? Karen Heckford HT ASCP CE Lead Histology Technician St. Mary's Medical Center 450 Stanyan St. San Francisco, Ca. 94117 415-668-1000 ext. 6167 karen.heckford@dignityhealth.org Caution: This email message, including all content and attachments, is CONFIDENTIAL and may be of a nature that is LEGALLY PRIVILEGED. The information contained in this email message is intended only for the use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you have received this document in error. Any further review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately by reply email. Thank you." _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rosenfeldtek <@t> hotmail.com Wed Jul 11 14:14:21 2012 From: rosenfeldtek <@t> hotmail.com (Jerry Ricks) Date: Wed Jul 11 14:14:29 2012 Subject: [Histonet] Methanol in H2O2 explanation--Summary In-Reply-To: <000f01cd5f92$271cb870$75562950$@gmx.at> References: <000f01cd5f92$271cb870$75562950$@gmx.at> Message-ID: Hi Histonetters, This has been a fun conversation. I called up a technical support scientist at Vector Labs with the question and was told basically "let me get back to you on that." :) But with some digging and the reference from Gudrun (thanks!) I've pieced together the following narrative. 1) Excess H2O2 leads to accumulation of Peroxidase compound III. The Third Intermediate Compound of Horseradish Peroxidase and Hydrogen Peroxide Philip George J. Biol. Chem. 1953 201: 427-434. www.jbc.org/content/201/1/427.full.pdf 2) Compound III is inactive as a peroxidase and can transfer of electron to hydrogen peroxide forming hydroxyl radical which attacks the heme porphoryin ring causing irreversible inactivation of the enzyme. Femji et-al The Open Enzyme Inhibition Journal, 2009, 2, 28-35 3) Methanol itself effectively (ethanol not so much) cleaves Heme groups from Horseradish peroxidase. Werner Straus J Histochem Cytochem, 1974 22:908 4) As the Vector Labs Quenching Protocols cheat sheet explains: "Methanol accelerates the destruction of the heme groups so a lower concentration of H2O2 can be used for a longer period of time." www.vectorlabs.com/Protocols/Supprotocols/quenchHRP.pdf So Methanol actually is a "magic chemical" in this case! Jerry Ricks Research Scientist University of Washington Department of Pathology > From: gu.lang@gmx.at > To: histonet@lists.utsouthwestern.edu > Date: Wed, 11 Jul 2012 20:22:38 +0200 > Subject: [Histonet] Re: Methanol in H2O2 explanation > > Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter > enzymehistochemistry - the thing, that an electron-donator is used for > demonstration. > And he states, that methanol alone also inhibits peroxidase - perhaps by > the fixation = denaturation effect on the protein? > > In the following publication one can see the equations for the > peroxidase-reaction and this phrase: > "In this study, we have used a steady state kinetics approach to demonstrate > that MPO is irreversibly inactivated by excess H2O2 in the absence of > reducing substrate." > The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based > Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of > Inactivation Rate by Organic Donor Substrates > > It's rather exhausting to prove the own statements ;-) puh. > > Gudrun > > > -----Urspr?ngliche Nachricht----- > Von: Morken, Timothy [mailto:Timothy.Morken@ucsfmedctr.org] > Gesendet: Mittwoch, 11. Juli 2012 18:12 > An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu > Betreff: RE: [Histonet] RE: Re: Methanol in H2O2 explanation > > It's funny how histology lore gets passed down, passed around and > misunderstood over the decades. When I ask questions in labs about why they > do a certain procedure they can rarely name a source (besides a particular > long-retired particular tech who wrote the original procedure) or give > reason as to why it is done that way. Even if it works well, at least people > could take the time to understand why! > > When I was researching buffer compositions long ago I found a dozen > variations of PBS alone (and only a dozen because I just stopped looking). > When you delve into these things it becomes clear that people made their own > solutions for a particular purpose. Maybe they wrote a paper, taught a > course or wrote a book. Then that formula was copied by many others and > their particular formulation became the "state of the art" even if the "art" > practiced by someone else had little to do with the original pursuit (the > Sheehan book is full of variations of stains that were originally for > specific research purposes. There is precious little guidance in any books > about what is good for a particular use!). > > The reason for using methanol / H2O2 that I was told long ago was prevent > frozen sections from being damaged by H2O2 bubbling. In fact, we would fix > frozens in cold acetone and THEN put the slides in cold methanol/H2O2 > (double fixation?) In that lab we never used methanol for deparaffinized > sections. Since I had no exposure to the clin lab I never knew about using > methanol for blood smear fixation. From literature searches it seems > methanol, rather than ethanol, is used primarily because it is cheaper. In > other words, it does not appear to be a magical substance! > > The questions asked here are good ones. However, hopefully those reading and > responsible for producing procedure manuals will include the reasoning for > each step of the method and not just assume it is necessary, or "common > knowledge," just because it has been handed down through generations of long > suffering techs! > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang > Sent: Wednesday, July 11, 2012 7:17 AM > To: histonet@lists.utsouthwestern.edu > Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation > > For the enzymatic activity of peroxidase it needs an electron-donator (or > receptor - I can't find the literature...) in the vicinity; therefore H2O2 > and DAB are added ad once, and DAB is oxidized and transformed into the > insoluble, amorphe substance through polymerization. > Without the donator H2O2 in excess works as inhibitor and blocks the > activation-side of the enzyme. > I think H2O2 in methanol was primarly preferred, because the frozen slides > were fixed at the same time. > > Rehydration after dewaxing depends on the following reagens. > > Gudrun > > -----Urspr?ngliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony > Henwood (SCHN) > Gesendet: Mittwoch, 11. Juli 2012 06:21 > An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu > Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation > > Hi Carl, > > What do you mean by "Why do people rehydrate after dewaxing?" Do you really > mean that slides do not require rehydration or do you mean that slides can > be left to dry after de-waxing prior to staining. > > Re-hydration is necessary, otherwise xylene will prevent aqueous stains from > doing their thing efficiently. > > I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, > the reactive oxygen reacted with the methanol to then degrade the enzyme, > but I need to look closer at this chemistry. > > > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, > Westmead NSW 2145, AUSTRALIA > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl > Sent: Wednesday, 11 July 2012 1:32 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Methanol in H2O2 explanation > > Why do some people use methanolic H2O2? > Why do people rehydrate after dewaxing? > Both are unnecessary, under usual conditions. > > Methanol was used in the early days as a peroxidase blocker by itself. > The combination was devised as a "belt and bracer" method. > As you stated, you use aq H2O2 effectively. > So do I and many others. > > However, for unfixed frozen sections, I would never use aq H2O2, if I wanted > sections remaining on my slides.... > > After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px > block while you make up you AR solutions.... > > Carl Hobbs > Histology and Imaging Manager > Wolfson CARD > School of Biomedical Sciences > Kings College London > Guys Campus > SE1 1UL > Tel: 020 78486813 > Fax: 020 78486816 > 020 78486813 > > > _______________________________________________ > 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. > **************************************************************************** > ***** > > _______________________________________________ > 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brannon <@t> alliedsearchpartners.com Wed Jul 11 14:47:25 2012 From: brannon <@t> alliedsearchpartners.com (Brannon Owens) Date: Wed Jul 11 14:47:39 2012 Subject: [Histonet] Mohs Histotech Opening- 4 Day Work Week- Sarasota, FL Message-ID: Allied Search Partners is working with a laboratory in the area of Sarasota, FL looking for a qualified and licensed Histotechnologist or Histotechnician with at least one year of experience with Mohs. Also available in the state of FL- a Histology Manager position. Visit our Careers page for details! http://www.alliedsearchpartners.com/careers.php To Apply Please email or fax resume to Brannon@alliedsearchpartners.com or fax to 888 388 7572 . No other information is given about location or the organization at this time. Please send resume for review by our recruiters and all qualified candidates will be submitted to HR for further review. Once we have had time to further consider your application, then more information will be made available. Thank you! Position: Histotechnician or Histotechnologist Schedule: Full time/4 day work week Location: Sarasota, FL area Excellent pay and benefits! Requirements: At least 1 year working experience with Mohs and processing of permanent sections HT/HTL certification by ASCP FL Clinical Lab License -- Brannon Owens Recruitment Manager Allied Search Partners T: 888.388.7571 ext. 106 F: 888.388.7572 To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php LinkedIn: http://www.linkedin.com/pub/brannon-owens/28/528/823 *If you wish to no longer receive emails from Allied Search Partners please respond to this email message with "remove." This email including its attachments is intended only for the confidential use of the individual to whom it is addressed. If you are not the intended recipient, any use, dissemination, distribution or copying of this message or its attachments is prohibited. If you have received this message in error, please notify us immediately, and delete this message and its attachments permanently from your system. From Diane.Tokugawa <@t> kp.org Wed Jul 11 16:56:14 2012 From: Diane.Tokugawa <@t> kp.org (Diane.Tokugawa@kp.org) Date: Wed Jul 11 16:56:27 2012 Subject: [Histonet] Diane Tokugawa/CA/KAIPERM is out of the office. Message-ID: I will be out of the office starting 07/11/2012 and will not return until 07/18/2012. Note: For Cytology issues, please call Molly at 8-421-5487 or Wanda 8-421-5426 For Histology issues, please call Barbara at 8-421-4959, IHC/Histo issues Kiran at 8-421-5404, or general histology client service at 8-421-5408. From BDeBrosse-Serra <@t> isisph.com Wed Jul 11 16:58:16 2012 From: BDeBrosse-Serra <@t> isisph.com (Bea DeBrosse-Serra) Date: Wed Jul 11 16:58:25 2012 Subject: [Histonet] Thank you for CD40 suggestions! Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FC21@EXCHMB01.isis.local> Thank you everybody who gave me excellent suggestions and feedback on CD40 on frozen sections, especially Gayle! I will keep the individuals posted on how everything goes. Thanks, Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 From davmpec <@t> hotmail.com Wed Jul 11 17:10:52 2012 From: davmpec <@t> hotmail.com (David M. Peck) Date: Wed Jul 11 17:10:55 2012 Subject: [Histonet] (no subject) Message-ID: http://linkgolfscore.com/wp-admin/likeit.php?unit72.html From tony.henwood <@t> health.nsw.gov.au Wed Jul 11 18:06:15 2012 From: tony.henwood <@t> health.nsw.gov.au (Tony Henwood (SCHN)) Date: Wed Jul 11 18:06:32 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation In-Reply-To: <000f01cd5f92$271cb870$75562950$@gmx.at> References: <000f01cd5f92$271cb870$75562950$@gmx.at> Message-ID: <6D6BD1DE8A5571489398B392A38A71579D1C726F@xmdb02.nch.kids> So, the thinking at the time was to kill the endogenous peroxidase twice! No one can say that us oldies weren't thorough! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, 12 July 2012 4:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter enzymehistochemistry - the thing, that an electron-donator is used for demonstration. And he states, that methanol alone also inhibits peroxidase - perhaps by the fixation = denaturation effect on the protein? In the following publication one can see the equations for the peroxidase-reaction and this phrase: "In this study, we have used a steady state kinetics approach to demonstrate that MPO is irreversibly inactivated by excess H2O2 in the absence of reducing substrate." The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of Inactivation Rate by Organic Donor Substrates It's rather exhausting to prove the own statements ;-) puh. Gudrun -----Urspr?ngliche Nachricht----- Von: Morken, Timothy [mailto:Timothy.Morken@ucsfmedctr.org] Gesendet: Mittwoch, 11. Juli 2012 18:12 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] RE: Re: Methanol in H2O2 explanation It's funny how histology lore gets passed down, passed around and misunderstood over the decades. When I ask questions in labs about why they do a certain procedure they can rarely name a source (besides a particular long-retired particular tech who wrote the original procedure) or give reason as to why it is done that way. Even if it works well, at least people could take the time to understand why! When I was researching buffer compositions long ago I found a dozen variations of PBS alone (and only a dozen because I just stopped looking). When you delve into these things it becomes clear that people made their own solutions for a particular purpose. Maybe they wrote a paper, taught a course or wrote a book. Then that formula was copied by many others and their particular formulation became the "state of the art" even if the "art" practiced by someone else had little to do with the original pursuit (the Sheehan book is full of variations of stains that were originally for specific research purposes. There is precious little guidance in any books about what is good for a particular use!). The reason for using methanol / H2O2 that I was told long ago was prevent frozen sections from being damaged by H2O2 bubbling. In fact, we would fix frozens in cold acetone and THEN put the slides in cold methanol/H2O2 (double fixation?) In that lab we never used methanol for deparaffinized sections. Since I had no exposure to the clin lab I never knew about using methanol for blood smear fixation. From literature searches it seems methanol, rather than ethanol, is used primarily because it is cheaper. In other words, it does not appear to be a magical substance! The questions asked here are good ones. However, hopefully those reading and responsible for producing procedure manuals will include the reasoning for each step of the method and not just assume it is necessary, or "common knowledge," just because it has been handed down through generations of long suffering techs! Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, July 11, 2012 7:17 AM To: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] RE: Re: Methanol in H2O2 explanation For the enzymatic activity of peroxidase it needs an electron-donator (or receptor - I can't find the literature...) in the vicinity; therefore H2O2 and DAB are added ad once, and DAB is oxidized and transformed into the insoluble, amorphe substance through polymerization. Without the donator H2O2 in excess works as inhibitor and blocks the activation-side of the enzyme. I think H2O2 in methanol was primarly preferred, because the frozen slides were fixed at the same time. Rehydration after dewaxing depends on the following reagens. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood (SCHN) Gesendet: Mittwoch, 11. Juli 2012 06:21 An: 'Hobbs, Carl'; histonet@lists.utsouthwestern.edu Betreff: [Histonet] RE: Re: Methanol in H2O2 explanation Hi Carl, What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, 11 July 2012 1:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides.... After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions.... Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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. **************************************************************************** ***** _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jonathan.Cremer <@t> med.kuleuven.be Thu Jul 12 04:23:44 2012 From: Jonathan.Cremer <@t> med.kuleuven.be (Jonathan Cremer) Date: Thu Jul 12 04:23:58 2012 Subject: [Histonet] human vimentin in mouse tissue In-Reply-To: <1341935334.82981.YahooMailNeo@web130104.mail.mud.yahoo.com> References: <1341935334.82981.YahooMailNeo@web130104.mail.mud.yahoo.com> Message-ID: Hi Kim, I use Sigma V4630 in mouse tissue, but it is sold as an anti-human antibody (but reactive to many species). I use it at a dilution of 1:2500 with a PerkinElmer Cy3 TSA kit, with a Jackson HRP-conjugated secondary. Oh, before I forget, standard AR with citrate, pH 6. Hope this helps. Jonathan --- Jonathan Cremer Laboratory Technician TARGID - KU Leuven ________________________________________ Van: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] namens Kim Merriam [kmerriam2003@yahoo.com] Verzonden: dinsdag 10 juli 2012 17:48 To: Histonet Onderwerp: [Histonet] human vimentin in mouse tissue Greetings! I want to stain for human vimentin in mouse tissues (human fibroblasts). I many years ago, I used a biotinylated V9 clone for just this purpose and it worked great. I tried the biotiylated V9 clone from abcam, but I am not getting any staining at all. Any suggestions? Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From patpxs <@t> gmail.com Thu Jul 12 07:54:14 2012 From: patpxs <@t> gmail.com (Paula Sicurello) Date: Thu Jul 12 07:54:18 2012 Subject: [Histonet] Epic Beaker module Message-ID: Good Morning One and All, Does anyone out there have any experience with the Beaker module of the LIS system EPIC? We have tests that require tables and charts to be inserted into the final report, pathologists need to have a case queue and other items. If you are using Epic/Beaker- please let me know if your experiences and what if anything have you had Epic build into it for you. Thanks, Paula -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 From njohnson <@t> kcskincenter.com Thu Jul 12 11:36:33 2012 From: njohnson <@t> kcskincenter.com (Work KCSCC) Date: Thu Jul 12 11:35:21 2012 Subject: [Histonet] Ventana symphony Message-ID: Can anyone give me information on their experience with the Ventana Symphony stainer? Nacaela Sent from my iPhone From mward <@t> wakehealth.edu Thu Jul 12 12:04:09 2012 From: mward <@t> wakehealth.edu (Martha Ward-Pathology) Date: Thu Jul 12 12:06:07 2012 Subject: [Histonet] Epic Beaker module In-Reply-To: References: Message-ID: We would interested in any replies as well. ? Martha Ward, MT (ASCP) QIHC Manager Molecular Diagnostics Lab Medical Center Boulevard ?\? Winston-Salem, NC 27157 p 336.716.2109 ?\? f 336.716.5890 ? mward@wakehealth.edu ? ? ? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Thursday, July 12, 2012 8:54 AM To: HistoNet Subject: [Histonet] Epic Beaker module Good Morning One and All, Does anyone out there have any experience with the Beaker module of the LIS system EPIC? We have tests that require tables and charts to be inserted into the final report, pathologists need to have a case queue and other items. If you are using Epic/Beaker- please let me know if your experiences and what if anything have you had Epic build into it for you. Thanks, Paula -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Thu Jul 12 12:23:39 2012 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Jul 12 12:23:53 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation Message-ID: Hi, There is a paper from 1972 (!) by Streefkerk entitled: "Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxidase following methanol. JHC 30:504. So perhaps the origin of methanol is not that mysterious. I cannot imagine that methanol is still considered for cryo's when performing IHC: it certainly kill many epitopes! Cheers, Chris Chris van der Loos Academic Medical Center Dept. of Pathology Amsterdam The Netherlands Date: Wed, 11 Jul 2012 20:22:38 +0200 From: "Gudrun Lang" Subject: [Histonet] Re: Methanol in H2O2 explanation To: Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at> Content-Type: text/plain; charset="iso-8859-1" Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter enzymehistochemistry - the thing, that an electron-donator is used for demonstration. And he states, that methanol alone also inhibits peroxidase - perhaps by the fixation = denaturation effect on the protein? In the following publication one can see the equations for the peroxidase-reaction and this phrase: "In this study, we have used a steady state kinetics approach to demonstrate that MPO is irreversibly inactivated by excess H2O2 in the absence of reducing substrate." The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of Inactivation Rate by Organic Donor Substrates It's rather exhausting to prove the own statements ;-) puh. Gudrun De informatie opgenomen in dit bericht kan vertrouwelijk zijn en is uitsluitend bestemd voor de geadresseerde. Indien u dit bericht onterecht ontvangt, wordt u verzocht de inhoud niet te gebruiken en de afzender direct te informeren door het bericht te retourneren. Het Academisch Medisch Centrum is een publiekrechtelijke rechtspersoon in de zin van de W.H.W. (Wet Hoger Onderwijs en Wetenschappelijk Onderzoek) en staat geregistreerd bij de Kamer van Koophandel voor Amsterdam onder nr. 34362777. De Algemene Inkoop Voorwaarden van het AMC zijn van toepassing op en maken integraal onderdeel uit van alle rechtsbetrekkingen, daaronder mede verstaan alle inkoop opdrachten en overeenkomsten, tussen AMC en derden. Deze voorwaarden zijn te raadplegen op www.amc.nl en worden op verzoek toegezonden. ________________________________ This message may contain confidential information and is intended exclusively for the addressee. If you receive this message unintentionally, please do not use the contents but notify the sender immediately by return e-mail. Academic Medical Center is a legal person by public law and is registered at the Chamber of Commerce for Amsterdam under no. 34362777. The AMC General Purchase Terms constitute an integral part of all legal relations, including but not limited to all purchase orders and contracts, between the AMC and third parties. These terms can be downloaded at the website www.amc.nl and will be sent to you at your request. From brannon <@t> alliedsearchpartners.com Thu Jul 12 13:48:21 2012 From: brannon <@t> alliedsearchpartners.com (Brannon Owens) Date: Thu Jul 12 13:48:39 2012 Subject: [Histonet] HT/HTL Full Time Opportunities in New York state! Message-ID: Allied Search Partners is working with different laboratories in the state of New York, looking for qualified and licensed Histotechnologists or Histotechnicians. To Apply Please email or fax resume to Brannon@alliedsearchpartners.com or fax to 888 388 7572. No other information is given about location or the organization at this time. Please send resume for review by our recruiters and all qualified candidates will be submitted to HR for further review. Once we have had time to further consider your application, then more information will be made available. Thank you! Position 1: Immunohistochemistry Technician/Technologist (IHC) Schedule: Tuesday-Saturday 2pm-10:30pm Location: Port Chester, NY Requirements: experience in IHC, a NY license, ASCP HT/HTL certification Position 2: Histotechnician or Histotechnologist Schedule: Tuesday-Saturday 7am-3:30pm Location: Port Chester, NY Requirements: NY Licensed, ASCP HT/HTL certification Position 3: Histotechnician or Histotechnologist Schedule: Full time/Permanent Day Shift Location: Binghamton, NY Requirements: NY Licensed, ASCP HT/HTL certification -- Brannon Owens Recruitment Manager Allied Search Partners T: 888.388.7571 ext. 106 F: 888.388.7572 To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php LinkedIn: http://www.linkedin.com/pub/brannon-owens/28/528/823 From lyork <@t> kwbpathology.com Thu Jul 12 14:06:27 2012 From: lyork <@t> kwbpathology.com (Leigh York) Date: Thu Jul 12 14:06:31 2012 Subject: [Histonet] ER, Clone SP1 Message-ID: Dako is going to discontinue the ER , SP1 clone. We really want to use the SP1 clone, but do not want to use an ASR. We can get it from Ventana only we are told. Can anyone elaborate on how they are handling this in their lab or if they have switched to an ASR? Thanks, Leigh York From carl.hobbs <@t> kcl.ac.uk Thu Jul 12 14:27:37 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Thu Jul 12 14:28:07 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation Message-ID: <11D9615B89C10747B1C985966A63D7CA38626A5736@KCL-MAIL04.kclad.ds.kcl.ac.uk> "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?" Right. An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer. Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply? Re use of methanol/ethanol....I use IMS. Always have done. NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs. We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction. "For the enzymatic activity of peroxidase it needs an electron-donator" The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas. However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 From patpxs <@t> gmail.com Thu Jul 12 15:04:51 2012 From: patpxs <@t> gmail.com (Paula Sicurello) Date: Thu Jul 12 15:04:54 2012 Subject: [Histonet] Beaker module in EPIC Message-ID: Hello all Listers, I've already asked the Histonet but I'm wondering about a different thing this time. Does anyone have experience with Beaker in conjunction with Anatomic Pathology, Surgical Pathology or the multiple other clinical labs that are in hospitals today? I work in Anatomic/Surgical Pathology which encompasses all the Clinical Laboratory services here at Duke. We have been told to determine what we need for the Beaker module to be useful to us. I've seen a brief demo. as to how it will work for the ordering clinicians (they can order every test STAT!). Please, send me any of the pluses or minuses that you've experienced using Beaker. Thanks, Paula :-) -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 From jmyers1 <@t> aol.com Thu Jul 12 16:19:35 2012 From: jmyers1 <@t> aol.com (Jmyers1) Date: Thu Jul 12 16:19:58 2012 Subject: [Histonet] p63 and Biocare Medical Message-ID: <8CF2E8B2C0332D7-7DC-410D@webmail-d128.sysops.aol.com> Fellow Histonetters: Over the past few days there has been some discussion on this forum regarding Biocare Medical's ability to continue offering the p63 antibody, some of which has been correct and some of it incorrect. Please be assured that Biocare still has the right to sell products that contain p63, but future U.S. sales rights are being negotiated and are subject to change. At present, only Biocare and Ventana Medical Systems are licensed to distribute IVD-labeled products that contain p63. If you have any concerns about this situation, please direct them to the Biocare sales representative that is responsible for the area in which your laboratory is located. Sincerely, Joe Myers, M.S., CT(ASCP)QIHC Senior Technical Sales Specialist ? Biocare Medical, LLC (and Chair, IHC Resource Group ? National Society for Histotechnology) From pruegg <@t> ihctech.net Thu Jul 12 16:45:20 2012 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Jul 12 16:45:29 2012 Subject: [Histonet] ER, Clone SP1 In-Reply-To: References: Message-ID: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> >From what I understand the SP1 Er clone has also been discontinued in USA by Leica/Novacastra, I am told it is still being sold by Leica in Europe? Must be another licensing issue because some are soooo greedy. I knew this was going to happen and stock piled some of the ER SP1, but it won't last forever. Patsy Ruegg, HT(ASCP)QIHC Ruegg Consulting 40864 Arkansas Ave Bennett, CO 80102 Phone: 303-644-4538 Fax: 720-859-4110 pruegg@ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh York Sent: Thursday, July 12, 2012 1:06 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] ER, Clone SP1 Dako is going to discontinue the ER , SP1 clone. We really want to use the SP1 clone, but do not want to use an ASR. We can get it from Ventana only we are told. Can anyone elaborate on how they are handling this in their lab or if they have switched to an ASR? Thanks, Leigh York _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From one_angel_secret <@t> yahoo.com Thu Jul 12 16:54:31 2012 From: one_angel_secret <@t> yahoo.com (Kim Donadio) Date: Thu Jul 12 16:54:44 2012 Subject: [Histonet] ER, Clone SP1 In-Reply-To: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> Message-ID: <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com> I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? Sent from my iPhone On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: >> From what I understand the SP1 Er clone has also been discontinued in USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? Must > be another licensing issue because some are soooo greedy. I knew this was > going to happen and stock piled some of the ER SP1, but it won't last > forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana only > we are told. Can anyone elaborate on how they are handling this in their > lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > 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 From Timothy.Morken <@t> ucsfmedctr.org Thu Jul 12 17:05:17 2012 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Timothy) Date: Thu Jul 12 17:04:40 2012 Subject: [Histonet] ER, Clone SP1 In-Reply-To: <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com> References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com> Message-ID: <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Thursday, July 12, 2012 2:55 PM To: Patsy Ruegg Cc: Subject: Re: [Histonet] ER, Clone SP1 I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? Sent from my iPhone On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: >> From what I understand the SP1 Er clone has also been discontinued in >> USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? > Must be another licensing issue because some are soooo greedy. I knew > this was going to happen and stock piled some of the ER SP1, but it > won't last forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh > York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to > use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana > only we are told. Can anyone elaborate on how they are handling this > in their lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kim.osullivan <@t> monash.edu Thu Jul 12 18:20:25 2012 From: kim.osullivan <@t> monash.edu (Kim O'Sullivan) Date: Thu Jul 12 18:20:30 2012 Subject: [Histonet] Gamma delta T cell and CD T cell detection on FFPE tissue Message-ID: Hi everyone, Can anyone tell me of a commercially available antibody that works on FFPE human tissue for gamma delta T cells ( TCR)? Secondly does anyone have a protocol for CD4 T cells on human tissue, I have tried both DAKO mouse anti human and both clones available from Novacastra (4B12 and 1F6) using microwave antigen retrieval (citrate buffer pH 6, EDTA pH 8 and TRIS HCL pH 9) and pressure cooker with the same buffers for different time periods ( 1, 2,3,4,and 5 minutes). Any suggestions, protocols or alternative antibodies would be appreciated! Regards From pruegg <@t> ihctech.net Thu Jul 12 21:44:33 2012 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Jul 12 21:44:36 2012 Subject: [Histonet] ER, Clone SP1 In-Reply-To: <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com> <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <0227518F256543848B5009F06BDF4EEB@Patsyoffice> Yea you are right Tim, I just wonder what ever happened to trying to get the best ab for the patient, SP1 is in my opinion the best ER ab but now the patient suffers because we can't use what is best for them, there is something wrong with that picture. Patsy Ruegg, HT(ASCP)QIHC Ruegg Consulting 40864 Arkansas Ave Bennett, CO 80102 Phone: 303-644-4538 Fax: 720-859-4110 pruegg@ihctech.net -----Original Message----- From: Morken, Timothy [mailto:Timothy.Morken@ucsfmedctr.org] Sent: Thursday, July 12, 2012 4:05 PM To: Kim Donadio; Patsy Ruegg Cc: Subject: RE: [Histonet] ER, Clone SP1 It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Thursday, July 12, 2012 2:55 PM To: Patsy Ruegg Cc: Subject: Re: [Histonet] ER, Clone SP1 I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? Sent from my iPhone On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: >> From what I understand the SP1 Er clone has also been discontinued in >> USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? > Must be another licensing issue because some are soooo greedy. I knew > this was going to happen and stock piled some of the ER SP1, but it > won't last forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh > York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to > use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana > only we are told. Can anyone elaborate on how they are handling this > in their lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jonathan.Cremer <@t> med.kuleuven.be Fri Jul 13 01:45:20 2012 From: Jonathan.Cremer <@t> med.kuleuven.be (Jonathan Cremer) Date: Fri Jul 13 01:45:34 2012 Subject: [Histonet] ER, Clone SP1 In-Reply-To: <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com>, <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree. It's ridiculous! --- ________________________________________ Van: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] namens Morken, Timothy [Timothy.Morken@ucsfmedctr.org] Verzonden: vrijdag 13 juli 2012 0:05 To: Kim Donadio; Patsy Ruegg Cc: Onderwerp: RE: [Histonet] ER, Clone SP1 It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Thursday, July 12, 2012 2:55 PM To: Patsy Ruegg Cc: Subject: Re: [Histonet] ER, Clone SP1 I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? Sent from my iPhone On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: >> From what I understand the SP1 Er clone has also been discontinued in >> USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? > Must be another licensing issue because some are soooo greedy. I knew > this was going to happen and stock piled some of the ER SP1, but it > won't last forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh > York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to > use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana > only we are told. Can anyone elaborate on how they are handling this > in their lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > 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 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 From mward <@t> wakehealth.edu Fri Jul 13 06:42:01 2012 From: mward <@t> wakehealth.edu (Martha Ward-Pathology) Date: Fri Jul 13 06:42:17 2012 Subject: [Histonet] Beaker module in EPIC In-Reply-To: References: Message-ID: Again, we would be interested in any replies as well. Our institution is going live with Epic this fall but the labs will not be using Beaker until later on. We are still trying to figure out how all this will work and welcome all information. Paula, how soon will Duke be up and using Beaker? Martha Ward Wake Forest Baptist Health 336-716-2109 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Thursday, July 12, 2012 4:05 PM To: HistoNet; Microscopy@microscopy.com Subject: [Histonet] Beaker module in EPIC Hello all Listers, I've already asked the Histonet but I'm wondering about a different thing this time. Does anyone have experience with Beaker in conjunction with Anatomic Pathology, Surgical Pathology or the multiple other clinical labs that are in hospitals today? I work in Anatomic/Surgical Pathology which encompasses all the Clinical Laboratory services here at Duke. We have been told to determine what we need for the Beaker module to be useful to us. I've seen a brief demo. as to how it will work for the ordering clinicians (they can order every test STAT!). Please, send me any of the pluses or minuses that you've experienced using Beaker. Thanks, Paula :-) -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NMP <@t> stowers.org Fri Jul 13 07:06:34 2012 From: NMP <@t> stowers.org (Marsh, Nannette) Date: Fri Jul 13 07:06:48 2012 Subject: [Histonet] ER, Clone SP1 In-Reply-To: References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com>, <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <2C40E43D1F7A56408C4463FD245DDDF9AE46A699@EXCHMB-02.stowers-institute.org> I love the comparison :-) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jonathan Cremer Sent: Friday, July 13, 2012 1:45 AM To: Morken, Timothy; Kim Donadio; Patsy Ruegg Cc: Subject: RE: [Histonet] ER, Clone SP1 That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree. It's ridiculous! --- ________________________________________ Van: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] namens Morken, Timothy [Timothy.Morken@ucsfmedctr.org] Verzonden: vrijdag 13 juli 2012 0:05 To: Kim Donadio; Patsy Ruegg Cc: Onderwerp: RE: [Histonet] ER, Clone SP1 It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Thursday, July 12, 2012 2:55 PM To: Patsy Ruegg Cc: Subject: Re: [Histonet] ER, Clone SP1 I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? Sent from my iPhone On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: >> From what I understand the SP1 Er clone has also been discontinued in >> USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? > Must be another licensing issue because some are soooo greedy. I knew > this was going to happen and stock piled some of the ER SP1, but it > won't last forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh > York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to > use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana > only we are told. Can anyone elaborate on how they are handling this > in their lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > 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 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nelsonrnch <@t> verizon.net Fri Jul 13 07:19:18 2012 From: nelsonrnch <@t> verizon.net (PATTI NELSON HT(ASCP)) Date: Fri Jul 13 07:19:24 2012 Subject: [Histonet] Different Avenues To Become Certified Message-ID: <1342181958.20737.YahooMailNeo@web84511.mail.ne1.yahoo.com> ?Good morning Histo-Land, A question was asked to me by my Medical Director the other day, so I ask for everyones expert answers to this question.? Is there any other entities other than ASCP to be certified by for HT/HTL ? ? I thank everyone in advance. Have a great weekend. BEST REGARDS, ? PATTI RUBEN-NELSON? H.T.(ASCP) PNP LABORATORY CONSULTANTS SUPERVISOR/DGC P.O. BOX 412 CABAZON, CA. 92230 cell (909) 841-9761 nelsonrnch@verizon.net ? CONFIDENTIALITY NOTICE: This message and any included attachments are from Patti Nelson, PNP Laboratory Consultants? ?and are intended only for the addressee.? The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or exemption from disclosure under applicable law.? Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful.? If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call? 909-841-9761. From mary-sturm <@t> uiowa.edu Fri Jul 13 08:18:06 2012 From: mary-sturm <@t> uiowa.edu (Sturm, Mary Therese M) Date: Fri Jul 13 08:18:11 2012 Subject: [Histonet] TRAP staining Message-ID: <17B53F9D0552174BAB6F33E0B69BBD9910847B70@hc-mailboxc1-n3.healthcare.uiowa.edu> Currently we are trying to use Sigma Kit 386A for Trap staining of FFPE rabbit spine.....and getting no staining , only tons of gold background. Anyone have experience with this procedure? Thanks Mary Sturm ________________________________ Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you. ________________________________ From Loralee_Mcmahon <@t> URMC.Rochester.edu Fri Jul 13 08:33:23 2012 From: Loralee_Mcmahon <@t> URMC.Rochester.edu (McMahon, Loralee A) Date: Fri Jul 13 08:33:39 2012 Subject: [Histonet] RE: TRAP staining In-Reply-To: <17B53F9D0552174BAB6F33E0B69BBD9910847B70@hc-mailboxc1-n3.healthcare.uiowa.edu> References: <17B53F9D0552174BAB6F33E0B69BBD9910847B70@hc-mailboxc1-n3.healthcare.uiowa.edu> Message-ID: What was your tissue decaled in? Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sturm, Mary Therese M [mary-sturm@uiowa.edu] Sent: Friday, July 13, 2012 9:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TRAP staining Currently we are trying to use Sigma Kit 386A for Trap staining of FFPE rabbit spine.....and getting no staining , only tons of gold background. Anyone have experience with this procedure? Thanks Mary Sturm ________________________________ Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you. ________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Jul 13 08:55:15 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jul 13 08:55:23 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation In-Reply-To: References: Message-ID: <1342187715.62239.YahooMailNeo@web121405.mail.ne1.yahoo.com> This is the real explanation. Mind that the title refers to erythrocyte pseudoperoxidase inhibition and in the title of the article is the procedural sequence: 1- fix the blood smear with methanol, which is?the standard procedure, and 2- apply H2O2 to quench the peroxidase activity. Ren? J. ________________________________ From: C.M. van der Loos To: "histonet netserver (histonet@lists.utsouthwestern.edu)" Sent: Thursday, July 12, 2012 1:23 PM Subject: [Histonet] Re: Methanol in H2O2 explanation Hi, There is a paper from 1972 (!) by Streefkerk entitled: "Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxidase following methanol. JHC 30:504. So perhaps the origin of methanol is not that mysterious. I cannot imagine that methanol is still considered for cryo's when performing IHC: it certainly kill many epitopes! Cheers, Chris Chris van der Loos Academic Medical Center Dept. of Pathology Amsterdam The Netherlands Date: Wed, 11 Jul 2012 20:22:38 +0200 From: "Gudrun Lang" Subject: [Histonet] Re: Methanol in H2O2 explanation To: Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at> Content-Type: text/plain;? ? ? charset="iso-8859-1" Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter enzymehistochemistry - the thing, that an electron-donator is used for demonstration. And he states, that methanol alone also inhibits peroxidase - perhaps by the fixation = denaturation effect on the protein? In the following publication one can see the equations for the peroxidase-reaction and this phrase: "In this study, we have used a steady state kinetics approach to demonstrate that MPO is irreversibly inactivated by excess H2O2 in the absence of reducing substrate." The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of Inactivation Rate by Organic Donor Substrates It's rather exhausting to prove the own statements? ;-) puh. Gudrun De informatie opgenomen in dit bericht kan vertrouwelijk zijn en is uitsluitend bestemd voor de geadresseerde. Indien u dit bericht onterecht ontvangt, wordt u verzocht de inhoud niet te gebruiken en de afzender direct te informeren door het bericht te retourneren. Het Academisch Medisch Centrum is een publiekrechtelijke rechtspersoon in de zin van de W.H.W. (Wet Hoger Onderwijs en Wetenschappelijk Onderzoek) en staat geregistreerd bij de Kamer van Koophandel voor Amsterdam onder nr. 34362777. De Algemene Inkoop Voorwaarden van het AMC zijn van toepassing op en maken integraal onderdeel uit van alle rechtsbetrekkingen, daaronder mede verstaan alle inkoop opdrachten en overeenkomsten, tussen AMC en derden. Deze voorwaarden zijn te raadplegen op http://www.amc.nl/ en worden op verzoek toegezonden. ________________________________ This message may contain confidential information and is intended exclusively for the addressee. If you receive this message unintentionally, please do not use the contents but notify the sender immediately by return e-mail. Academic Medical Center is a legal person by public law and is registered at the Chamber of Commerce for Amsterdam under no. 34362777. The AMC General Purchase Terms constitute an integral part of all legal relations, including but not limited to all purchase orders and contracts, between the AMC and third parties. These terms can be downloaded at the website www.amc.nl and will be sent to you at your request. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Jul 13 09:07:42 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jul 13 09:07:49 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation In-Reply-To: <11D9615B89C10747B1C985966A63D7CA38626A5736@KCL-MAIL04.kclad.ds.kcl.ac.uk> References: <11D9615B89C10747B1C985966A63D7CA38626A5736@KCL-MAIL04.kclad.ds.kcl.ac.uk> Message-ID: <1342188462.30207.YahooMailNeo@web121406.mail.ne1.yahoo.com> I have been fascinated by this long discussion about H2O2 and its use?in IHC procedures. We have to start from the fact that ALL cells contain peroxidase because ALL cells respire and ALL cells need to use oxygen they receive from blood (or hemolymph in crustaceans and other arthropods). The initial IHC procedure were peroxidase-anti-peroxidase (PAP) methods (and many remain as such). A fixed cell = a dead cell BUT in spite of being dead, the peroxidase they contain is still?able of reacting with the PAP complex. The cell, being dead, cannot make more peroxidase, but there is still peroxidase in it after any type of fixation, even with NBF, as Carl points out. IF that peroxidase is not "quenched" = INHIBITED the use of a PAP IHC procedure on that uninhibited peroxidase = excessive background?in the section after the IHC method is?completed. So, how to avoid that background? By "quenching" or inhibiting the peroxidase contained in the cell. How? With a substance rich in oxygen that will inactivate the peroxidase by "spending" it?and that substance is H2O2 used?at the start of the IHC protocol. Ren? J.?? ________________________________ From: "Hobbs, Carl" To: "histonet@lists.utsouthwestern.edu" Sent: Thursday, July 12, 2012 3:27 PM Subject: [Histonet] Re: Methanol in H2O2 explanation "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?" Right. An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer. Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply? Re use of methanol/ethanol....I use IMS. Always have done. NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs. We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize? sites of AB:Ag interaction. "For the enzymatic activity of peroxidase it needs an electron-donator" The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas. However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Loralee_Mcmahon <@t> URMC.Rochester.edu Fri Jul 13 09:18:13 2012 From: Loralee_Mcmahon <@t> URMC.Rochester.edu (McMahon, Loralee A) Date: Fri Jul 13 09:20:51 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation In-Reply-To: <1342188462.30207.YahooMailNeo@web121406.mail.ne1.yahoo.com> References: <11D9615B89C10747B1C985966A63D7CA38626A5736@KCL-MAIL04.kclad.ds.kcl.ac.uk>, <1342188462.30207.YahooMailNeo@web121406.mail.ne1.yahoo.com> Message-ID: I think that the original question was why is it recommended in some procedures that H202 be diluted in methanol. And others say distilled water or buffer. What makes one better than the other? Some people I have worked with insist on getting 30% h202 and diluting it in methanol to a 3% solution. Others use 3% that you buy from the drug store. No one seems to know why except that is how it has always been done. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa [rjbuesa@yahoo.com] Sent: Friday, July 13, 2012 10:07 AM To: Hobbs, Carl; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Methanol in H2O2 explanation I have been fascinated by this long discussion about H2O2 and its use in IHC procedures. We have to start from the fact that ALL cells contain peroxidase because ALL cells respire and ALL cells need to use oxygen they receive from blood (or hemolymph in crustaceans and other arthropods). The initial IHC procedure were peroxidase-anti-peroxidase (PAP) methods (and many remain as such). A fixed cell = a dead cell BUT in spite of being dead, the peroxidase they contain is still able of reacting with the PAP complex. The cell, being dead, cannot make more peroxidase, but there is still peroxidase in it after any type of fixation, even with NBF, as Carl points out. IF that peroxidase is not "quenched" = INHIBITED the use of a PAP IHC procedure on that uninhibited peroxidase = excessive background in the section after the IHC method is completed. So, how to avoid that background? By "quenching" or inhibiting the peroxidase contained in the cell. How? With a substance rich in oxygen that will inactivate the peroxidase by "spending" it and that substance is H2O2 used at the start of the IHC protocol. Ren? J. ________________________________ From: "Hobbs, Carl" To: "histonet@lists.utsouthwestern.edu" Sent: Thursday, July 12, 2012 3:27 PM Subject: [Histonet] Re: Methanol in H2O2 explanation "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?" Right. An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer. Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply? Re use of methanol/ethanol....I use IMS. Always have done. NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs. We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction. "For the enzymatic activity of peroxidase it needs an electron-donator" The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas. However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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 From ree3 <@t> leicester.ac.uk Fri Jul 13 09:45:02 2012 From: ree3 <@t> leicester.ac.uk (Edwards, Richard E.) Date: Fri Jul 13 09:45:14 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation In-Reply-To: References: <11D9615B89C10747B1C985966A63D7CA38626A5736@KCL-MAIL04.kclad.ds.kcl.ac.uk>, <1342188462.30207.YahooMailNeo@web121406.mail.ne1.yahoo.com> Message-ID: <7722595275A4DD4FA225B92CDBF174A101A648733121@EXC-MBX3.cfs.le.ac.uk> We use hydrogen peroxide in a sodium azide solution,(on acetone fixed GMA processed tissue) works fine. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of McMahon, Loralee A Sent: 13 July 2012 15:18 To: Rene J Buesa; Hobbs, Carl; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Methanol in H2O2 explanation I think that the original question was why is it recommended in some procedures that H202 be diluted in methanol. And others say distilled water or buffer. What makes one better than the other? Some people I have worked with insist on getting 30% h202 and diluting it in methanol to a 3% solution. Others use 3% that you buy from the drug store. No one seems to know why except that is how it has always been done. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa [rjbuesa@yahoo.com] Sent: Friday, July 13, 2012 10:07 AM To: Hobbs, Carl; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Methanol in H2O2 explanation I have been fascinated by this long discussion about H2O2 and its use in IHC procedures. We have to start from the fact that ALL cells contain peroxidase because ALL cells respire and ALL cells need to use oxygen they receive from blood (or hemolymph in crustaceans and other arthropods). The initial IHC procedure were peroxidase-anti-peroxidase (PAP) methods (and many remain as such). A fixed cell = a dead cell BUT in spite of being dead, the peroxidase they contain is still able of reacting with the PAP complex. The cell, being dead, cannot make more peroxidase, but there is still peroxidase in it after any type of fixation, even with NBF, as Carl points out. IF that peroxidase is not "quenched" = INHIBITED the use of a PAP IHC procedure on that uninhibited peroxidase = excessive background in the section after the IHC method is completed. So, how to avoid that background? By "quenching" or inhibiting the peroxidase contained in the cell. How? With a substance rich in oxygen that will inactivate the peroxidase by "spending" it and that substance is H2O2 used at the start of the IHC protocol. Ren? J. ________________________________ From: "Hobbs, Carl" To: "histonet@lists.utsouthwestern.edu" Sent: Thursday, July 12, 2012 3:27 PM Subject: [Histonet] Re: Methanol in H2O2 explanation "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?" Right. An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer. Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply? Re use of methanol/ethanol....I use IMS. Always have done. NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs. We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction. "For the enzymatic activity of peroxidase it needs an electron-donator" The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas. However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From patrick.lewis <@t> seattlechildrens.org Fri Jul 13 11:16:25 2012 From: patrick.lewis <@t> seattlechildrens.org (Lewis, Patrick) Date: Fri Jul 13 11:16:36 2012 Subject: [Histonet] H2O2 Recep Message-ID: <3903BE18914F4440834F0E620415FFCA0BCC774C@PPWEXD01B.childrens.sea.kids> So what I'm gathering from all this. Thanks by the way for the discussion. Methanol is unnecessary for the H2O2, reaction, but it can make it slightly faster since both methods will be working in concert. If you have tissue that has more peroxidase than usual, then extra time is required. Also, If you use straight aq H2O2 on frozen sections the bubbling may damage the tissue. CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From TJohnson <@t> gnf.org Fri Jul 13 12:33:36 2012 From: TJohnson <@t> gnf.org (Teri Johnson) Date: Fri Jul 13 12:33:41 2012 Subject: [Histonet] RE: Methanol in H2O2 explanation Message-ID: <9F3CFEE76E51B64991C7485270890B400CDC58FF@EX4.lj.gnf.org> Patrick Lewis wrote: So what I'm gathering from all this. Thanks by the way for the discussion. Methanol is unnecessary for the H2O2, reaction, but it can make it slightly faster since both methods will be working in concert. If you have tissue that has more peroxidase than usual, then extra time is required. Also, If you use straight aq H2O2 on frozen sections the bubbling may damage the tissue. I too am loving the discussion. It truly highlights what we continue to do due to tradition without thinking and how to back up and research where things started and why. It also highlights what we thought we knew and what we know now. Good stuff! As for H2O2 on frozen sections, regardless of the solvent (MeOH vs buffer vs aq) we always brought the H2O2 concentration down by 10% (0.3% final) and increased the time to 30 minutes to avoid the tissue damage issue. Because of its known effect on some antigens, I discontinued using methanol many years ago for both paraffin and frozen section. To my understanding, the idea of diluting 30% to working concentration allowed for more consistency vs buying 3% off the pharmacy shelf. Who knew how long the bottle had been sitting on the shelf? After practicing both options, I found no difference in efficacy in quenching endogenous peroxidase. Some add sodium azide to the quenching solution because it is known to inhibit peroxidase and is believed to be a more "complete" peroxidase blocker. I have never adopted this practice but would love to see data that shows it is better in some circumstances. I think the practice does not hurt anything but falls under the heading "just in case." Gayle Callis, among others, may be able to shed some additional light on this. Happy Friday! Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 From mike <@t> pathview.com Fri Jul 13 13:03:27 2012 From: mike <@t> pathview.com (Michael Mihalik) Date: Fri Jul 13 13:04:09 2012 Subject: [Histonet] Beaker module in EPIC In-Reply-To: References: Message-ID: <00b101cd6121$e08c5280$a1a4f780$@pathview.com> I'm curious. Are ANY labs using Epic for their APLIS? The last I heard was a sales presentation a couple of years ago where the sales person declined to show the APLIS "because it wasn't ready". I do know of hospitals using Epic for order entry, though. Michael Mihalik PathView Systems |?cell: 214.733.7688?| 800.798.3540 | fax: 952.241.7369 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martha Ward-Pathology Sent: Friday, July 13, 2012 7:42 AM To: Paula Sicurello; HistoNet; Microscopy@microscopy.com Subject: RE: [Histonet] Beaker module in EPIC Again, we would be interested in any replies as well. Our institution is going live with Epic this fall but the labs will not be using Beaker until later on. We are still trying to figure out how all this will work and welcome all information. Paula, how soon will Duke be up and using Beaker? Martha Ward Wake Forest Baptist Health 336-716-2109 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Thursday, July 12, 2012 4:05 PM To: HistoNet; Microscopy@microscopy.com Subject: [Histonet] Beaker module in EPIC Hello all Listers, I've already asked the Histonet but I'm wondering about a different thing this time. Does anyone have experience with Beaker in conjunction with Anatomic Pathology, Surgical Pathology or the multiple other clinical labs that are in hospitals today? I work in Anatomic/Surgical Pathology which encompasses all the Clinical Laboratory services here at Duke. We have been told to determine what we need for the Beaker module to be useful to us. I've seen a brief demo. as to how it will work for the ordering clinicians (they can order every test STAT!). Please, send me any of the pluses or minuses that you've experienced using Beaker. Thanks, Paula :-) -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 _______________________________________________ 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 From rjbuesa <@t> yahoo.com Fri Jul 13 13:16:49 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jul 13 13:16:54 2012 Subject: [Histonet] RE: Methanol in H2O2 explanation In-Reply-To: <9F3CFEE76E51B64991C7485270890B400CDC58FF@EX4.lj.gnf.org> References: <9F3CFEE76E51B64991C7485270890B400CDC58FF@EX4.lj.gnf.org> Message-ID: <1342203409.21483.YahooMailNeo@web121405.mail.ne1.yahoo.com> The only problem with using "pure" 30% H2O2 is that it has to be refrigerated and used with great caution because on contact with the skin can produce very nasty chemical burns. That is why I always used 3% "commercial" H2O2. You have to consider r also that at a 3% dilution the amount of H2O2 is in excess of what is needed to quench the tissues peroxidase. Ren?? J. ________________________________ From: Teri Johnson To: "histonet@lists.utsouthwestern.edu" Sent: Friday, July 13, 2012 1:33 PM Subject: [Histonet] RE: Methanol in H2O2 explanation Patrick Lewis wrote: So what I'm gathering from all this. Thanks by the way for the discussion. Methanol is unnecessary for the H2O2, reaction, but it can make it slightly faster since both methods will be working in concert.? If you have tissue that has more peroxidase than usual, then extra time is required. Also, If you use straight aq H2O2 on frozen sections the bubbling may damage the tissue. I too am loving the discussion. It truly highlights what we continue to do due to tradition without thinking and how to back up and research where things started and why. It also highlights what we thought we knew and what we know now. Good stuff! As for H2O2 on frozen sections, regardless of the solvent (MeOH vs buffer vs aq) we always brought the H2O2 concentration down by 10% (0.3% final) and increased the time to 30 minutes to avoid the tissue damage issue. Because of its known effect on some antigens, I discontinued using methanol many years ago for both paraffin and frozen section. To my understanding, the idea of diluting 30% to working concentration allowed for more consistency vs buying 3% off the pharmacy shelf. Who knew how long the bottle had been sitting on the shelf? After practicing both options, I found no difference in efficacy in quenching endogenous peroxidase. Some add sodium azide to the quenching solution because it is known to inhibit peroxidase and is believed to be a more "complete" peroxidase blocker. I have never adopted this practice but would love to see data that shows it is better in some circumstances. I think the practice does not hurt anything but falls under the heading "just in case." Gayle Callis, among others, may be able to shed some additional light on this. Happy Friday! Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Jul 13 13:21:23 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jul 13 13:21:27 2012 Subject: [Histonet] H2O2 Recep In-Reply-To: <3903BE18914F4440834F0E620415FFCA0BCC774C@PPWEXD01B.childrens.sea.kids> References: <3903BE18914F4440834F0E620415FFCA0BCC774C@PPWEXD01B.childrens.sea.kids> Message-ID: <1342203683.64573.YahooMailNeo@web121406.mail.ne1.yahoo.com> The bubbling of H2O2 frozen tissue is caused by the chemical destruction of the tissues. Do you remember when you apply 3% H2O2 to a skin wound? Don't you remember the bubbling it produces? That is the oxygen destruction of the white blood cells that constitute the pus in any wound, as well as the chemical destruction (oxidation) of the live skin cells surrounding the wound. The addition of methanol does not affect the oxidation of the peroxidase. It is just "a way of doing things" those things that we use to do without any specific (scientific) reason. Ren? J. ________________________________ From: "Lewis, Patrick" To: "'Histonet@lists.utsouthwestern.edu'" Sent: Friday, July 13, 2012 12:16 PM Subject: [Histonet] H2O2 Recep So what I'm gathering from all this. Thanks by the way for the discussion. Methanol is unnecessary for the H2O2, reaction, but it can make it slightly faster since both methods will be working in concert.? If you have tissue that has more peroxidase than usual, then extra time is required. Also, If you use straight aq H2O2 on frozen sections the bubbling may damage the tissue. CONFIDENTIALITY NOTICE:? This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law.? Any unauthorized review, use, disclosure or distribution is prohibited.? If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mward <@t> wakehealth.edu Fri Jul 13 14:13:07 2012 From: mward <@t> wakehealth.edu (Martha Ward-Pathology) Date: Fri Jul 13 14:13:25 2012 Subject: [Histonet] Beaker module in EPIC In-Reply-To: <00b101cd6121$e08c5280$a1a4f780$@pathview.com> References: <00b101cd6121$e08c5280$a1a4f780$@pathview.com> Message-ID: So far we have not had any responses so I assume the answer is no. ? Martha Ward, MT (ASCP) QIHC Manager Molecular Diagnostics Lab Medical Center Boulevard ?\? Winston-Salem, NC 27157 p 336.716.2109 ?\? f 336.716.5890 ? mward@wakehealth.edu ? ? ? -----Original Message----- From: Michael Mihalik [mailto:mike@pathview.com] Sent: Friday, July 13, 2012 2:03 PM To: Martha Ward-Pathology; 'Paula Sicurello'; 'HistoNet'; Microscopy@microscopy.com Subject: RE: [Histonet] Beaker module in EPIC I'm curious. Are ANY labs using Epic for their APLIS? The last I heard was a sales presentation a couple of years ago where the sales person declined to show the APLIS "because it wasn't ready". I do know of hospitals using Epic for order entry, though. Michael Mihalik PathView Systems |?cell: 214.733.7688?| 800.798.3540 | fax: 952.241.7369 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martha Ward-Pathology Sent: Friday, July 13, 2012 7:42 AM To: Paula Sicurello; HistoNet; Microscopy@microscopy.com Subject: RE: [Histonet] Beaker module in EPIC Again, we would be interested in any replies as well. Our institution is going live with Epic this fall but the labs will not be using Beaker until later on. We are still trying to figure out how all this will work and welcome all information. Paula, how soon will Duke be up and using Beaker? Martha Ward Wake Forest Baptist Health 336-716-2109 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Thursday, July 12, 2012 4:05 PM To: HistoNet; Microscopy@microscopy.com Subject: [Histonet] Beaker module in EPIC Hello all Listers, I've already asked the Histonet but I'm wondering about a different thing this time. Does anyone have experience with Beaker in conjunction with Anatomic Pathology, Surgical Pathology or the multiple other clinical labs that are in hospitals today? I work in Anatomic/Surgical Pathology which encompasses all the Clinical Laboratory services here at Duke. We have been told to determine what we need for the Beaker module to be useful to us. I've seen a brief demo. as to how it will work for the ordering clinicians (they can order every test STAT!). Please, send me any of the pluses or minuses that you've experienced using Beaker. Thanks, Paula :-) -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 _______________________________________________ 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 From madeleinehuey <@t> gmail.com Fri Jul 13 15:31:53 2012 From: madeleinehuey <@t> gmail.com (Madeleine Huey) Date: Fri Jul 13 15:32:02 2012 Subject: [Histonet] Re: Histonet Digest, Vol 104, Issue 15 In-Reply-To: <50002bdb.6710b60a.2b1a.ffffa39bSMTPIN_ADDED@mx.google.com> References: <50002bdb.6710b60a.2b1a.ffffa39bSMTPIN_ADDED@mx.google.com> Message-ID: All, ER, clone SP1 is availuable in www.epitomics.com (cat. #4200-1) for $205. Hope this help! Madeleine Huey Supervisor - Pathology (IPOX & Histology) madeleine_h@elcaminohospital.org On Fri, Jul 13, 2012 at 7:08 AM, wrote: > 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-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@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: Epic Beaker module (Martha Ward-Pathology) > 2. Re: Methanol in H2O2 explanation (C.M. van der Loos) > 3. HT/HTL Full Time Opportunities in New York state! (Brannon Owens) > 4. ER, Clone SP1 (Leigh York) > 5. Re: Methanol in H2O2 explanation (Hobbs, Carl) > 6. Beaker module in EPIC (Paula Sicurello) > 7. p63 and Biocare Medical (Jmyers1) > 8. RE: ER, Clone SP1 (Patsy Ruegg) > 9. Re: ER, Clone SP1 (Kim Donadio) > 10. RE: ER, Clone SP1 (Morken, Timothy) > 11. Gamma delta T cell and CD T cell detection on FFPE tissue > (Kim O'Sullivan) > 12. RE: ER, Clone SP1 (Patsy Ruegg) > 13. RE: ER, Clone SP1 (Jonathan Cremer) > 14. RE: Beaker module in EPIC (Martha Ward-Pathology) > 15. RE: ER, Clone SP1 (Marsh, Nannette) > 16. Different Avenues To Become Certified (PATTI NELSON HT(ASCP)) > 17. TRAP staining (Sturm, Mary Therese M) > 18. RE: TRAP staining (McMahon, Loralee A) > 19. Re: Re: Methanol in H2O2 explanation (Rene J Buesa) > 20. Re: Re: Methanol in H2O2 explanation (Rene J Buesa) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 12 Jul 2012 17:04:09 +0000 > From: Martha Ward-Pathology > Subject: RE: [Histonet] Epic Beaker module > To: Paula Sicurello , HistoNet > > Message-ID: > > > Content-Type: text/plain; charset="iso-8859-1" > > We would interested in any replies as well. > > > > Martha Ward, MT (ASCP) QIHC > Manager > > Molecular Diagnostics Lab > Medical Center Boulevard \ Winston-Salem, NC 27157 > p 336.716.2109 \ f 336.716.5890 > mward@wakehealth.edu > > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello > Sent: Thursday, July 12, 2012 8:54 AM > To: HistoNet > Subject: [Histonet] Epic Beaker module > > Good Morning One and All, > > Does anyone out there have any experience with the Beaker module of the LIS system EPIC? We have tests that require tables and charts to be inserted into the final report, pathologists need to have a case queue and other items. > > If you are using Epic/Beaker- please let me know if your experiences and what if anything have you had Epic build into it for you. > > Thanks, > > Paula > > -- > Paula Sicurello, HTL (ASCP) > Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 > P: 919.684.2091 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 2 > Date: Thu, 12 Jul 2012 17:23:39 +0000 > From: "C.M. van der Loos" > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: "histonet netserver (histonet@lists.utsouthwestern.edu)" > > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Hi, > > There is a paper from 1972 (!) by Streefkerk entitled: "Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxidase following methanol. JHC 30:504. So perhaps the origin of methanol is not that mysterious. > > I cannot imagine that methanol is still considered for cryo's when performing IHC: it certainly kill many epitopes! > > > > Cheers, Chris > > > > Chris van der Loos > > Academic Medical Center > > Dept. of Pathology > > Amsterdam > > The Netherlands > > > > Date: Wed, 11 Jul 2012 20:22:38 +0200 > From: "Gudrun Lang" > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: > Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at> > Content-Type: text/plain; charset="iso-8859-1" > > Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter > enzymehistochemistry - the thing, that an electron-donator is used for > demonstration. > And he states, that methanol alone also inhibits peroxidase - perhaps by > the fixation = denaturation effect on the protein? > > In the following publication one can see the equations for the > peroxidase-reaction and this phrase: > "In this study, we have used a steady state kinetics approach to demonstrate > that MPO is irreversibly inactivated by excess H2O2 in the absence of > reducing substrate." > The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based > Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of > Inactivation Rate by Organic Donor Substrates > > It's rather exhausting to prove the own statements ;-) puh. > > Gudrun > > > De informatie opgenomen in dit bericht kan vertrouwelijk zijn en is uitsluitend bestemd voor de geadresseerde. Indien u dit bericht onterecht ontvangt, wordt u verzocht de inhoud niet te gebruiken en de afzender direct te informeren door het bericht te retourneren. Het Academisch Medisch Centrum is een publiekrechtelijke rechtspersoon in de zin van de W.H.W. (Wet Hoger Onderwijs en Wetenschappelijk Onderzoek) en staat geregistreerd bij de Kamer van Koophandel voor Amsterdam onder nr. 34362777. De Algemene Inkoop Voorwaarden van het AMC zijn van toepassing op en maken integraal onderdeel uit van alle rechtsbetrekkingen, daaronder mede verstaan alle inkoop opdrachten en overeenkomsten, tussen AMC en derden. Deze voorwaarden zijn te raadplegen op www.amc.nl en worden op verzoek toegezonden. > > ________________________________ > > This message may contain confidential information and is intended exclusively for the addressee. If you receive this message unintentionally, please do not use the contents but notify the sender immediately by return e-mail. Academic Medical Center is a legal person by public law and is registered at the Chamber of Commerce for Amsterdam under no. 34362777. The AMC General Purchase Terms constitute an integral part of all legal relations, including but not limited to all purchase orders and contracts, between the AMC and third parties. These terms can be downloaded at the website www.amc.nl and will be sent to you at your request. > > > ------------------------------ > > Message: 3 > Date: Thu, 12 Jul 2012 14:48:21 -0400 > From: Brannon Owens > Subject: [Histonet] HT/HTL Full Time Opportunities in New York state! > To: "histonet@lists.utsouthwestern.edu" > > Message-ID: > Content-Type: text/plain; charset="US-ASCII" > > Allied Search Partners is working with different laboratories in the state > of New York, looking for qualified and licensed Histotechnologists or > Histotechnicians. > > To Apply Please email or fax resume to Brannon@alliedsearchpartners.com > or fax to 888 388 7572. No other > information is given about location or the organization at this time. Please > send resume for review by our recruiters and all qualified candidates will > be submitted to HR for further review. Once we have had time to further > consider your application, then more information will be made available. > Thank you! > > Position 1: > Immunohistochemistry Technician/Technologist (IHC) > Schedule: Tuesday-Saturday 2pm-10:30pm > Location: Port Chester, NY > Requirements: experience in IHC, a NY license, ASCP HT/HTL certification > > Position 2: > Histotechnician or Histotechnologist > Schedule: Tuesday-Saturday 7am-3:30pm > Location: Port Chester, NY > Requirements: NY Licensed, ASCP HT/HTL certification > > Position 3: > Histotechnician or Histotechnologist > Schedule: Full time/Permanent Day Shift > Location: Binghamton, NY > Requirements: NY Licensed, ASCP HT/HTL certification > > -- > Brannon Owens > Recruitment Manager > Allied Search Partners > > > T: 888.388.7571 ext. 106 > > F: 888.388.7572 > > > > To view a complete list of Allied Search Partners current openings go to: > http://www.alliedsearchpartners.com/careers.php > > LinkedIn: http://www.linkedin.com/pub/brannon-owens/28/528/823 > > > > > > > > ------------------------------ > > Message: 4 > Date: Thu, 12 Jul 2012 15:06:27 -0400 > From: "Leigh York" > Subject: [Histonet] ER, Clone SP1 > To: > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Dako is going to discontinue the ER , SP1 clone. We really want to use the SP1 clone, but do not want to use an ASR. We can get it from Ventana only we are told. Can anyone elaborate on how they are handling this in their lab or if they have switched to an ASR? > > Thanks, > > Leigh York > > ------------------------------ > > Message: 5 > Date: Thu, 12 Jul 2012 20:27:37 +0100 > From: "Hobbs, Carl" > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: "histonet@lists.utsouthwestern.edu" > > Message-ID: > <11D9615B89C10747B1C985966A63D7CA38626A5736@KCL-MAIL04.kclad.ds.kcl.ac.uk> > > Content-Type: text/plain; charset="us-ascii" > > "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?" > > Right. > > An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer. > Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply? > > Re use of methanol/ethanol....I use IMS. Always have done. > NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs. > We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction. > > "For the enzymatic activity of peroxidase it needs an electron-donator" > The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas. > However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that > > > Carl Hobbs > Histology Manager > Wolfson CARD > School of Biomedical Sciences > Kings College London > Guys Campus > SE1 1UL > Tel: 020 78486813 > Fax: 020 78486816 > 020 78486813 > > > > > ------------------------------ > > Message: 6 > Date: Thu, 12 Jul 2012 16:04:51 -0400 > From: Paula Sicurello > Subject: [Histonet] Beaker module in EPIC > To: HistoNet , > Microscopy@microscopy.com > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Hello all Listers, > > I've already asked the Histonet but I'm wondering about a different thing > this time. > > Does anyone have experience with Beaker in conjunction with Anatomic > Pathology, Surgical Pathology or the multiple other clinical labs that are > in hospitals today? > > I work in Anatomic/Surgical Pathology which encompasses all the Clinical > Laboratory services here at Duke. We have been told to determine what we > need for the Beaker module to be useful to us. I've seen a brief demo. as > to how it will work for the ordering clinicians (they can order every test > STAT!). > > Please, send me any of the pluses or minuses that you've experienced using > Beaker. > > Thanks, > > Paula :-) > > -- > Paula Sicurello, HTL (ASCP) > Supervisor, Clinical Electron Microscopy Laboratory > Duke University Health System > Rm.#251M, Duke South, Green Zone > Durham, North Carolina 27710 > P: 919.684.2091 > > > ------------------------------ > > Message: 7 > Date: Thu, 12 Jul 2012 17:19:35 -0400 (EDT) > From: Jmyers1 > Subject: [Histonet] p63 and Biocare Medical > To: histonet@lists.utsouthwestern.edu, jmyers@biocare.net > Message-ID: <8CF2E8B2C0332D7-7DC-410D@webmail-d128.sysops.aol.com> > Content-Type: text/plain; charset="utf-8" > > > Fellow Histonetters: > Over the past few days there has been some discussion on this forum regarding Biocare Medical's ability to continue offering the p63 antibody, some of which has been correct and some of it incorrect. Please be assured that Biocare still has the right to sell products that contain p63, but future U.S. sales rights are being negotiated and are subject to change. At present, only Biocare and Ventana Medical Systems are licensed to distribute IVD-labeled products that contain p63. If you have any concerns about this situation, please direct them to the Biocare sales representative that is responsible for the area in which your laboratory is located. > Sincerely, > Joe Myers, M.S., CT(ASCP)QIHC > Senior Technical Sales Specialist ??? Biocare Medical, LLC > (and Chair, IHC Resource Group ??? National Society for Histotechnology) > > > ------------------------------ > > Message: 8 > Date: Thu, 12 Jul 2012 15:45:20 -0600 > From: "Patsy Ruegg" > Subject: RE: [Histonet] ER, Clone SP1 > To: "'Leigh York'" , > > Message-ID: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> > Content-Type: text/plain; charset="us-ascii" > > >From what I understand the SP1 Er clone has also been discontinued in USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? Must > be another licensing issue because some are soooo greedy. I knew this was > going to happen and stock piled some of the ER SP1, but it won't last > forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana only > we are told. Can anyone elaborate on how they are handling this in their > lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 9 > Date: Thu, 12 Jul 2012 17:54:31 -0400 > From: Kim Donadio > Subject: Re: [Histonet] ER, Clone SP1 > To: Patsy Ruegg > Cc: "" > > Message-ID: <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com> > Content-Type: text/plain; charset=us-ascii > > I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? > > Sent from my iPhone > > On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: > >>> From what I understand the SP1 Er clone has also been discontinued in USA by >> Leica/Novacastra, I am told it is still being sold by Leica in Europe? Must >> be another licensing issue because some are soooo greedy. I knew this was >> going to happen and stock piled some of the ER SP1, but it won't last >> forever. >> >> Patsy Ruegg, HT(ASCP)QIHC >> Ruegg Consulting >> 40864 Arkansas Ave >> Bennett, CO 80102 >> Phone: 303-644-4538 >> Fax: 720-859-4110 >> pruegg@ihctech.net >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] ER, Clone SP1 >> >> Dako is going to discontinue the ER , SP1 clone. We really want to use the >> SP1 clone, but do not want to use an ASR. We can get it from Ventana only >> we are told. Can anyone elaborate on how they are handling this in their >> lab or if they have switched to an ASR? >> >> Thanks, >> >> Leigh York >> _______________________________________________ >> 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 > > > > ------------------------------ > > Message: 10 > Date: Thu, 12 Jul 2012 15:05:17 -0700 > From: "Morken, Timothy" > Subject: RE: [Histonet] ER, Clone SP1 > To: "Kim Donadio" , "Patsy Ruegg" > > Cc: "" > > Message-ID: > <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> > > Content-Type: text/plain; charset=us-ascii > > It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. > > For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: > Subject: Re: [Histonet] ER, Clone SP1 > > I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? > > Sent from my iPhone > > On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: > >>> From what I understand the SP1 Er clone has also been discontinued in >>> USA by >> Leica/Novacastra, I am told it is still being sold by Leica in Europe? >> Must be another licensing issue because some are soooo greedy. I knew >> this was going to happen and stock piled some of the ER SP1, but it >> won't last forever. >> >> Patsy Ruegg, HT(ASCP)QIHC >> Ruegg Consulting >> 40864 Arkansas Ave >> Bennett, CO 80102 >> Phone: 303-644-4538 >> Fax: 720-859-4110 >> pruegg@ihctech.net >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] ER, Clone SP1 >> >> Dako is going to discontinue the ER , SP1 clone. We really want to >> use the >> SP1 clone, but do not want to use an ASR. We can get it from Ventana >> only we are told. Can anyone elaborate on how they are handling this >> in their lab or if they have switched to an ASR? >> >> Thanks, >> >> Leigh York >> _______________________________________________ >> 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 11 > Date: Fri, 13 Jul 2012 09:20:25 +1000 > From: "Kim O'Sullivan" > Subject: [Histonet] Gamma delta T cell and CD T cell detection on FFPE > tissue > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Hi everyone, > > Can anyone tell me of a commercially available antibody that works on FFPE > human tissue for gamma delta T cells ( TCR)? Secondly does anyone have a > protocol for CD4 T cells on human tissue, I have tried both DAKO mouse anti > human and both clones available from Novacastra (4B12 and 1F6) using > microwave antigen retrieval (citrate buffer pH 6, EDTA pH 8 and TRIS HCL pH > 9) and pressure cooker with the same buffers for different time periods ( > 1, 2,3,4,and 5 minutes). Any suggestions, protocols or alternative > antibodies would be appreciated! > > Regards > > > ------------------------------ > > Message: 12 > Date: Thu, 12 Jul 2012 20:44:33 -0600 > From: "Patsy Ruegg" > Subject: RE: [Histonet] ER, Clone SP1 > To: "'Morken, Timothy'" , "'Kim > Donadio'" > Cc: Histonet@lists.utsouthwestern.edu > Message-ID: <0227518F256543848B5009F06BDF4EEB@Patsyoffice> > Content-Type: text/plain; charset="us-ascii" > > Yea you are right Tim, I just wonder what ever happened to trying to get the > best ab for the patient, SP1 is in my opinion the best ER ab but now the > patient suffers because we can't use what is best for them, there is > something wrong with that picture. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: Morken, Timothy [mailto:Timothy.Morken@ucsfmedctr.org] > Sent: Thursday, July 12, 2012 4:05 PM > To: Kim Donadio; Patsy Ruegg > Cc: > Subject: RE: [Histonet] ER, Clone SP1 > > It is most likely licensing issues between companies. A company may carry it > under license from another company but the terms change and the licensee > either loses the license (the other company won't license it), can't afford > it or other issues. > > For example, when I worked in the antibody industry we had several > antibodies for a certain target but another company got a patent on the > target and all antibodies to the target (still not sure why that is > possible!). They finally decided to enforce their patent and wanted a > licensing fee that was higher than our annual sales of all the antibodies > put together. Obviously not a viable deal for our company. > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: > Subject: Re: [Histonet] ER, Clone SP1 > > I'm curious as to why all if a sudden some of these can be sold else where > and not in the USA. Is it a FDA issue? > > Sent from my iPhone > > On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: > >>> From what I understand the SP1 Er clone has also been discontinued in >>> USA by >> Leica/Novacastra, I am told it is still being sold by Leica in Europe? >> Must be another licensing issue because some are soooo greedy. I knew >> this was going to happen and stock piled some of the ER SP1, but it >> won't last forever. >> >> Patsy Ruegg, HT(ASCP)QIHC >> Ruegg Consulting >> 40864 Arkansas Ave >> Bennett, CO 80102 >> Phone: 303-644-4538 >> Fax: 720-859-4110 >> pruegg@ihctech.net >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] ER, Clone SP1 >> >> Dako is going to discontinue the ER , SP1 clone. We really want to >> use the >> SP1 clone, but do not want to use an ASR. We can get it from Ventana >> only we are told. Can anyone elaborate on how they are handling this >> in their lab or if they have switched to an ASR? >> >> Thanks, >> >> Leigh York >> _______________________________________________ >> 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ------------------------------ > > Message: 13 > Date: Fri, 13 Jul 2012 06:45:20 +0000 > From: Jonathan Cremer > Subject: RE: [Histonet] ER, Clone SP1 > To: "Morken, Timothy" , Kim Donadio > , Patsy Ruegg > Cc: "" > > Message-ID: > > > Content-Type: text/plain; charset="us-ascii" > > That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree. > It's ridiculous! > --- > > > > ________________________________________ > Van: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] namens Morken, Timothy [Timothy.Morken@ucsfmedctr.org] > Verzonden: vrijdag 13 juli 2012 0:05 > To: Kim Donadio; Patsy Ruegg > Cc: > Onderwerp: RE: [Histonet] ER, Clone SP1 > > It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. > > For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: > Subject: Re: [Histonet] ER, Clone SP1 > > I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? > > Sent from my iPhone > > On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: > >>> From what I understand the SP1 Er clone has also been discontinued in >>> USA by >> Leica/Novacastra, I am told it is still being sold by Leica in Europe? >> Must be another licensing issue because some are soooo greedy. I knew >> this was going to happen and stock piled some of the ER SP1, but it >> won't last forever. >> >> Patsy Ruegg, HT(ASCP)QIHC >> Ruegg Consulting >> 40864 Arkansas Ave >> Bennett, CO 80102 >> Phone: 303-644-4538 >> Fax: 720-859-4110 >> pruegg@ihctech.net >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] ER, Clone SP1 >> >> Dako is going to discontinue the ER , SP1 clone. We really want to >> use the >> SP1 clone, but do not want to use an ASR. We can get it from Ventana >> only we are told. Can anyone elaborate on how they are handling this >> in their lab or if they have switched to an ASR? >> >> Thanks, >> >> Leigh York >> _______________________________________________ >> 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 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 > > > > ------------------------------ > > Message: 14 > Date: Fri, 13 Jul 2012 11:42:01 +0000 > From: Martha Ward-Pathology > Subject: RE: [Histonet] Beaker module in EPIC > To: Paula Sicurello , HistoNet > , "Microscopy@microscopy.com" > > Message-ID: > > > Content-Type: text/plain; charset="us-ascii" > > Again, we would be interested in any replies as well. Our institution is going live with Epic this fall but the labs will not be using Beaker until later on. We are still trying to figure out how all this will work and welcome all information. > Paula, how soon will Duke be up and using Beaker? > > Martha Ward > Wake Forest Baptist Health > 336-716-2109 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello > Sent: Thursday, July 12, 2012 4:05 PM > To: HistoNet; Microscopy@microscopy.com > Subject: [Histonet] Beaker module in EPIC > > Hello all Listers, > > I've already asked the Histonet but I'm wondering about a different thing this time. > > Does anyone have experience with Beaker in conjunction with Anatomic Pathology, Surgical Pathology or the multiple other clinical labs that are in hospitals today? > > I work in Anatomic/Surgical Pathology which encompasses all the Clinical Laboratory services here at Duke. We have been told to determine what we need for the Beaker module to be useful to us. I've seen a brief demo. as to how it will work for the ordering clinicians (they can order every test STAT!). > > Please, send me any of the pluses or minuses that you've experienced using Beaker. > > Thanks, > > Paula :-) > > -- > Paula Sicurello, HTL (ASCP) > Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 > P: 919.684.2091 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 15 > Date: Fri, 13 Jul 2012 07:06:34 -0500 > From: "Marsh, Nannette" > Subject: RE: [Histonet] ER, Clone SP1 > To: "'Jonathan Cremer'" , "Morken, > Timothy" , Kim Donadio > , Patsy Ruegg > Cc: "" > > Message-ID: > <2C40E43D1F7A56408C4463FD245DDDF9AE46A699@EXCHMB-02.stowers-institute.org> > > Content-Type: text/plain; charset="us-ascii" > > I love the comparison :-) > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jonathan Cremer > Sent: Friday, July 13, 2012 1:45 AM > To: Morken, Timothy; Kim Donadio; Patsy Ruegg > Cc: > Subject: RE: [Histonet] ER, Clone SP1 > > That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree. > It's ridiculous! > --- > > > > ________________________________________ > Van: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] namens Morken, Timothy [Timothy.Morken@ucsfmedctr.org] > Verzonden: vrijdag 13 juli 2012 0:05 > To: Kim Donadio; Patsy Ruegg > Cc: > Onderwerp: RE: [Histonet] ER, Clone SP1 > > It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. > > For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio > Sent: Thursday, July 12, 2012 2:55 PM > To: Patsy Ruegg > Cc: > Subject: Re: [Histonet] ER, Clone SP1 > > I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? > > Sent from my iPhone > > On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: > >>> From what I understand the SP1 Er clone has also been discontinued in >>> USA by >> Leica/Novacastra, I am told it is still being sold by Leica in Europe? >> Must be another licensing issue because some are soooo greedy. I knew >> this was going to happen and stock piled some of the ER SP1, but it >> won't last forever. >> >> Patsy Ruegg, HT(ASCP)QIHC >> Ruegg Consulting >> 40864 Arkansas Ave >> Bennett, CO 80102 >> Phone: 303-644-4538 >> Fax: 720-859-4110 >> pruegg@ihctech.net >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh >> York >> Sent: Thursday, July 12, 2012 1:06 PM >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] ER, Clone SP1 >> >> Dako is going to discontinue the ER , SP1 clone. We really want to >> use the >> SP1 clone, but do not want to use an ASR. We can get it from Ventana >> only we are told. Can anyone elaborate on how they are handling this >> in their lab or if they have switched to an ASR? >> >> Thanks, >> >> Leigh York >> _______________________________________________ >> 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 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 16 > Date: Fri, 13 Jul 2012 05:19:18 -0700 (PDT) > From: "PATTI NELSON HT\(ASCP\)" > Subject: [Histonet] Different Avenues To Become Certified > To: Histonet > Message-ID: > <1342181958.20737.YahooMailNeo@web84511.mail.ne1.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Good morning Histo-Land, > > A question was asked to me by my Medical Director the other day, so I ask for everyones expert answers to this question. > Is there any other entities other than ASCP to be certified by for HT/HTL ? > > I thank everyone in advance. Have a great weekend. > > BEST REGARDS, > > PATTI RUBEN-NELSON H.T.(ASCP) > PNP LABORATORY CONSULTANTS > SUPERVISOR/DGC > P.O. BOX 412 > CABAZON, CA. 92230 > cell (909) 841-9761 > nelsonrnch@verizon.net > > > CONFIDENTIALITY NOTICE: > This message and any included attachments are from Patti Nelson, PNP Laboratory Consultants > and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or exemption from disclosure under applicable > law. Unauthorized forwarding, printing, copying, distribution, or use of > such information is strictly prohibited and may be unlawful. If you are not > the addressee, please promptly delete this message and notify the sender of > the delivery error by e-mail or you may call 909-841-9761. > > ------------------------------ > > Message: 17 > Date: Fri, 13 Jul 2012 13:18:06 +0000 > From: "Sturm, Mary Therese M" > Subject: [Histonet] TRAP staining > To: "histonet@lists.utsouthwestern.edu" > > Message-ID: > <17B53F9D0552174BAB6F33E0B69BBD9910847B70@hc-mailboxc1-n3.healthcare.uiowa.edu> > > Content-Type: text/plain; charset="us-ascii" > > Currently we are trying to use Sigma Kit 386A for Trap staining of FFPE rabbit spine.....and getting no staining , only tons of gold background. Anyone have experience with this procedure? > > Thanks > Mary Sturm > > > > > ________________________________ > Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you. > ________________________________ > > > ------------------------------ > > Message: 18 > Date: Fri, 13 Jul 2012 09:33:23 -0400 > From: "McMahon, Loralee A" > Subject: [Histonet] RE: TRAP staining > To: "Sturm, Mary Therese M" , > "histonet@lists.utsouthwestern.edu" > > Message-ID: > > > Content-Type: text/plain; charset="us-ascii" > > What was your tissue decaled in? > > Loralee McMahon, HTL (ASCP) > Immunohistochemistry Supervisor > Strong Memorial Hospital > Department of Surgical Pathology > (585) 275-7210 > ________________________________________ > From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sturm, Mary Therese M [mary-sturm@uiowa.edu] > Sent: Friday, July 13, 2012 9:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] TRAP staining > > Currently we are trying to use Sigma Kit 386A for Trap staining of FFPE rabbit spine.....and getting no staining , only tons of gold background. Anyone have experience with this procedure? > > Thanks > Mary Sturm > > > > > ________________________________ > Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you. > ________________________________ > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ > > Message: 19 > Date: Fri, 13 Jul 2012 06:55:15 -0700 (PDT) > From: Rene J Buesa > Subject: Re: [Histonet] Re: Methanol in H2O2 explanation > To: "C.M. van der Loos" , "histonet > netserver \(histonet@lists.utsouthwestern.edu\)" > > Message-ID: > <1342187715.62239.YahooMailNeo@web121405.mail.ne1.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > This is the real explanation. > Mind that the title refers to erythrocyte pseudoperoxidase inhibition and in the title of the article is the procedural sequence: > 1- fix the blood smear with methanol, which is the standard procedure, and > 2- apply H2O2 to quench the peroxidase activity. > Ren? J. > > > ________________________________ > From: C.M. van der Loos > To: "histonet netserver (histonet@lists.utsouthwestern.edu)" > Sent: Thursday, July 12, 2012 1:23 PM > Subject: [Histonet] Re: Methanol in H2O2 explanation > > Hi, > > There is a paper from 1972 (!) by Streefkerk entitled: "Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxidase following methanol. JHC 30:504. So perhaps the origin of methanol is not that mysterious. > > I cannot imagine that methanol is still considered for cryo's when performing IHC: it certainly kill many epitopes! > > > > Cheers, Chris > > > > Chris van der Loos > > Academic Medical Center > > Dept. of Pathology > > Amsterdam > > The Netherlands > > > > Date: Wed, 11 Jul 2012 20:22:38 +0200 > From: "Gudrun Lang" > Subject: [Histonet] Re: Methanol in H2O2 explanation > To: > Message-ID: <000f01cd5f92$271cb870$75562950$@gmx.at> > Content-Type: text/plain; charset="iso-8859-1" > > Ha! I've found it. It can be found in Dr. Kiernan's book in the chapter > enzymehistochemistry - the thing, that an electron-donator is used for > demonstration. > And he states, that methanol alone also inhibits peroxidase - perhaps by > the fixation = denaturation effect on the protein? > > In the following publication one can see the equations for the > peroxidase-reaction and this phrase: > "In this study, we have used a steady state kinetics approach to demonstrate > that MPO is irreversibly inactivated by excess H2O2 in the absence of > reducing substrate." > The Open Enzyme Inhibition Journal, 2009, 2, 28-35 Mechanism-Based > Inhibition of Myeloperoxidase by Hydrogen Peroxide: Enhancement of > Inactivation Rate by Organic Donor Substrates > > It's rather exhausting to prove the own statements ;-) puh. > > Gudrun > > > De informatie opgenomen in dit bericht kan vertrouwelijk zijn en is uitsluitend bestemd voor de geadresseerde. Indien u dit bericht onterecht ontvangt, wordt u verzocht de inhoud niet te gebruiken en de afzender direct te informeren door het bericht te retourneren. Het Academisch Medisch Centrum is een publiekrechtelijke rechtspersoon in de zin van de W.H.W. (Wet Hoger Onderwijs en Wetenschappelijk Onderzoek) en staat geregistreerd bij de Kamer van Koophandel voor Amsterdam onder nr. 34362777. De Algemene Inkoop Voorwaarden van het AMC zijn van toepassing op en maken integraal onderdeel uit van alle rechtsbetrekkingen, daaronder mede verstaan alle inkoop opdrachten en overeenkomsten, tussen AMC en derden. Deze voorwaarden zijn te raadplegen op http://www.amc.nl/ en worden op verzoek toegezonden. > > ________________________________ > > This message may contain confidential information and is intended exclusively for the addressee. If you receive this message unintentionally, please do not use the contents but notify the sender immediately by return e-mail. Academic Medical Center is a legal person by public law and is registered at the Chamber of Commerce for Amsterdam under no. 34362777. The AMC General Purchase Terms constitute an integral part of all legal relations, including but not limited to all purchase orders and contracts, between the AMC and third parties. These terms can be downloaded at the website www.amc.nl and will be sent to you at your request. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > Message: 20 > Date: Fri, 13 Jul 2012 07:07:42 -0700 (PDT) > From: Rene J Buesa > Subject: Re: [Histonet] Re: Methanol in H2O2 explanation > To: "Hobbs, Carl" , > "histonet@lists.utsouthwestern.edu" > > Message-ID: > <1342188462.30207.YahooMailNeo@web121406.mail.ne1.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > I have been fascinated by this long discussion about H2O2 and its use in IHC procedures. > We have to start from the fact that ALL cells contain peroxidase because ALL cells respire and ALL cells need to use oxygen they receive from blood (or hemolymph in crustaceans and other arthropods). > The initial IHC procedure were peroxidase-anti-peroxidase (PAP) methods (and many remain as such). > A fixed cell = a dead cell BUT in spite of being dead, the peroxidase they contain is still able of reacting with the PAP complex. > The cell, being dead, cannot make more peroxidase, but there is still peroxidase in it after any type of fixation, even with NBF, as Carl points out. > IF that peroxidase is not "quenched" = INHIBITED the use of a PAP IHC procedure on that uninhibited peroxidase = excessive background in the section after the IHC method is completed. > So, how to avoid that background? By "quenching" or inhibiting the peroxidase contained in the cell. > How? With a substance rich in oxygen that will inactivate the peroxidase by "spending" it and that substance is H2O2 used at the start of the IHC protocol. > Ren? J. > > > ________________________________ > From: "Hobbs, Carl" > To: "histonet@lists.utsouthwestern.edu" > Sent: Thursday, July 12, 2012 3:27 PM > Subject: [Histonet] Re: Methanol in H2O2 explanation > > "H202 mixed with Buffer here.(instead of aq.) That's OK too, right?" > > Right. > > An addition to the misconceptions that Tim interestingly elucidates: some use H2O2 in buffer because they think that , as the enzyme is performing a physiologic catalytic reaction, it should be at a physiological pH, in a physiological buffer. > Well....if it's a frozen section, it's been fixed; if it's a P.wax section, it's been fixed in Formalin, fixed in alcohol, subjected to 60C heat.....so, the fact that endog. Peroxidases survive those assaults, indicates that physiologic conditions no longer apply? > > Re use of methanol/ethanol....I use IMS. Always have done. > NB: great excess of substrate can " kill " enzymes ( irreversible substrate inhibition): that is why we add a great excess of H2O2 ( substrate) to "kill" endog. Pxs. > We then use a stoichiometric quantity of the same reagent ( H2O2) in the presence of DAB/AEC, to visualize sites of AB:Ag interaction. > > "For the enzymatic activity of peroxidase it needs an electron-donator" > The enzymatic activity of peroxidase needs no electron donor, as regards Immuno.....it will "digest" H2O2 into water and oxygen gas. > However, if you wish to use HRP as a reporter molecule, you must also add a suitable chromogen ( DAB, AEC) that > > > Carl Hobbs > Histology Manager > Wolfson CARD > School of Biomedical Sciences > Kings College London > Guys Campus > SE1 1UL > Tel: 020 78486813 > Fax: 020 78486816 > 020 78486813 > > > _______________________________________________ > 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 > > End of Histonet Digest, Vol 104, Issue 15 > ***************************************** From Rebecca.Riesen <@t> hma.com Fri Jul 13 15:41:22 2012 From: Rebecca.Riesen <@t> hma.com (Riesen, Rebecca) Date: Fri Jul 13 15:41:26 2012 Subject: [Histonet] slide/block combo mailer Message-ID: What company is making/distributing the new One Slide/One Block combo mailer? From pruegg <@t> ihctech.net Fri Jul 13 16:33:56 2012 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Jul 13 16:34:00 2012 Subject: [Histonet] ER, Clone SP1 References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice> <876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com> <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <32D34B81446E4031B84AF579CC454049@Patsyoffice> I need to correct my statement about ER Sp1 being sold in Europe by Leica after taking it off the market in the USA, I was confusing this ab with the HER2 CB11 that Leica was still selling in Europe and not the USA, they had to take it off the market in US to get FDA approval which they apparently just got. I apologize for this misinformation, I am getting old and can't keep things straight anymore, and it was the HER2 CB11 I stock piled not the ER SP1. Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg Consulting 40864 Arkansas Ave Bennett, CO 80102 Phone: 303-644-4538 Fax: 720-859-4110 pruegg@ihctech.net -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Thursday, July 12, 2012 8:45 PM To: 'Morken, Timothy'; 'Kim Donadio' Cc: '' Subject: RE: [Histonet] ER, Clone SP1 Yea you are right Tim, I just wonder what ever happened to trying to get the best ab for the patient, SP1 is in my opinion the best ER ab but now the patient suffers because we can't use what is best for them, there is something wrong with that picture. Patsy Ruegg, HT(ASCP)QIHC Ruegg Consulting 40864 Arkansas Ave Bennett, CO 80102 Phone: 303-644-4538 Fax: 720-859-4110 pruegg@ihctech.net -----Original Message----- From: Morken, Timothy [mailto:Timothy.Morken@ucsfmedctr.org] Sent: Thursday, July 12, 2012 4:05 PM To: Kim Donadio; Patsy Ruegg Cc: Subject: RE: [Histonet] ER, Clone SP1 It is most likely licensing issues between companies. A company may carry it under license from another company but the terms change and the licensee either loses the license (the other company won't license it), can't afford it or other issues. For example, when I worked in the antibody industry we had several antibodies for a certain target but another company got a patent on the target and all antibodies to the target (still not sure why that is possible!). They finally decided to enforce their patent and wanted a licensing fee that was higher than our annual sales of all the antibodies put together. Obviously not a viable deal for our company. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Thursday, July 12, 2012 2:55 PM To: Patsy Ruegg Cc: Subject: Re: [Histonet] ER, Clone SP1 I'm curious as to why all if a sudden some of these can be sold else where and not in the USA. Is it a FDA issue? Sent from my iPhone On Jul 12, 2012, at 5:45 PM, "Patsy Ruegg" wrote: >> From what I understand the SP1 Er clone has also been discontinued in >> USA by > Leica/Novacastra, I am told it is still being sold by Leica in Europe? > Must be another licensing issue because some are soooo greedy. I knew > this was going to happen and stock piled some of the ER SP1, but it > won't last forever. > > Patsy Ruegg, HT(ASCP)QIHC > Ruegg Consulting > 40864 Arkansas Ave > Bennett, CO 80102 > Phone: 303-644-4538 > Fax: 720-859-4110 > pruegg@ihctech.net > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leigh > York > Sent: Thursday, July 12, 2012 1:06 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] ER, Clone SP1 > > Dako is going to discontinue the ER , SP1 clone. We really want to > use the > SP1 clone, but do not want to use an ASR. We can get it from Ventana > only we are told. Can anyone elaborate on how they are handling this > in their lab or if they have switched to an ASR? > > Thanks, > > Leigh York > _______________________________________________ > 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jaderyan83 <@t> yahoo.com Fri Jul 13 16:53:28 2012 From: jaderyan83 <@t> yahoo.com (JadeRyan) Date: Fri Jul 13 16:53:33 2012 Subject: [Histonet] Histology Project Message-ID: <248965B1-DD7F-431F-9C68-35591893FACC@yahoo.com> Hi everyone, I need someone to do these things for our future Derm lab. Please email me the cost and how long it would take. Chemical Hygiene Manual, the Exposure Control Manual, the Procedure Manual and the Q.C. Manual to CLIA standards. Regards, Jade Ryan, MHA, BSN Sent from my iPhone From chrispjaz <@t> yahoo.com Fri Jul 13 20:30:51 2012 From: chrispjaz <@t> yahoo.com (Christopher Jacobs) Date: Fri Jul 13 20:30:56 2012 Subject: [Histonet] Negative controls and CAP Message-ID: <1342229451.62515.YahooMailNeo@web120104.mail.ne1.yahoo.com> Histonetters, I have been made the IHC lead in a fairly large laboratory that is seeking to become CAP accredited. I am definitely a newbie when it comes to CAP. A question was brought up today about how we do our negative controls. Specifically, should we run another negative control with any IHCs that need to be repeated or if we should run another negative control if at a later date a pathologist orders additional IHCs on a case. With our current protocol, we run one negative control per detection kit used, per case. Consequently, most cases end up with just one negative control slide. We do NOT run another negative control if we have to repeat one of a group of immunos or if a pathologist orders additional stains at a later point. I am wondering if this is good practice, and acceptable with CAP. I would love to find some literature that could help me make a case either way on this. Actually, any literature or pointers regarding IHCs and becoming CAP accredited will probably help save what little hair I have left. Thank you, Chris Jacobs, HT(ASCP)QIHC From amario3 <@t> uic.edu Sat Jul 14 11:53:22 2012 From: amario3 <@t> uic.edu (Andrea Marion) Date: Sat Jul 14 11:53:28 2012 Subject: [Histonet] What is this item called? Message-ID: <920cd1aed7d6e5f472d2105872523ca9.squirrel@webmail.uic.edu> Hi Adam, The photo was helpful. I've used them before in an embryology lab, for dissecting out Drosophila imaginal discs. We had single-well and 3-well versions. The 3 well version was slightly longer than the one you showed, and the wells were smaller, but the general proportions were the same (gently sloping sides, not steep slides like a spot plate). I did some searching because I'd like to have a few for our lab too. The best I could come up with is: 1 - Corning and Fisher Scientific used to carry 3-well versions but both are now discontinued. Corning #7223. Fisher #21-379. 2 - Fisher also carried single well versions (referred to in a number of Drosophila dissection protocols), but no product # mentioned. 3 - these are similar, but the depression is not sloped: http://www.leaveonlybubbles.com/catalog/product_info.php?products_id=63 4 - Not sure if you are using glass or ceramic spot plates, but there are glass spot plates with more gradually sloping sides that might fit your needs: http://lab-crunch.ecrater.com/p/11132699/corning-pyrex-9-well-spot-plate http://www.mccronemicroscopes.com/store/catalog.asp?item=345&category_id=0 You might try sending your photo to Fisher/Corning etc customer service and see what they come up with. Let me know if you find anything. Good luck! Andrea Andrea Marion Graduate Student University of Illinois at Chicago From carl.hobbs <@t> kcl.ac.uk Sat Jul 14 13:23:12 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Sat Jul 14 13:24:07 2012 Subject: [Histonet] Re: Methanol in H2O2 explanation Message-ID: <11D9615B89C10747B1C985966A63D7CA38626A5A9A@KCL-MAIL04.kclad.ds.kcl.ac.uk> I thought that I answered the original Q? Imho, 3% H2O2 in distilled water is as good as any other "recipe", given time and concentration. Sure, use a lower conc.... IF you achieve negative rbcs. I use water because it is cheaper. A lower concentration will take longer....why bother?? Kill that enzyme. If you are concerned re frozen sections....well, use Glucose oxidase system. Far more expensive. Good in some very messy bloody situations... Non-reversible ( irreversible) denaturation/inactivation of endogenous Pxs is achieved by "feeding" them a massive overdose of H2O2. This is not suicide...it's murder! Well, depends on your definition of Life ;-) NB: inactivation can be reversible/non-reversible so, imho, not a good term to use. Use "irreversible inactivation"? Them peroxidases have evolved over millions of years to survive and...we come along with hair bleach and kill them! Re methanol/H2O2 adversely affecting Pwax sections: I say no way. The tissue has been processed via alcohols for several Hrs. Bottom line: Water/Methanol/azide/H2O2.....are your rbcs/neutrophils colourless ?? Have you got a good section? If yes, then your H2O2 blocking method is OK!! So...if you use an expensive commercial H2O2 blocking reagent...why? To be sure.....I completely subscribe to the Skool that says that if you do get some non-specific staining but, get very good specific staining....ignore the former. Eg: if you get rbc staining with a GFAP Ab....so what? It is only aesthetics..... Understand what you are doing and...why. Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 From carl.hobbs <@t> kcl.ac.uk Sat Jul 14 13:27:10 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Sat Jul 14 13:28:07 2012 Subject: [Histonet] RE: Methanol in H2O2 explanation Message-ID: <11D9615B89C10747B1C985966A63D7CA38626A5A9B@KCL-MAIL04.kclad.ds.kcl.ac.uk> "If you have tissue that has more peroxidase than usual, then extra time is required." Depends on the concentration of H2O2 used ;-) A massive excess of substrate ( 3%) is good. Carl From wilson6848 <@t> yahoo.com Sat Jul 14 18:11:32 2012 From: wilson6848 <@t> yahoo.com (Wilson A) Date: Sat Jul 14 18:11:35 2012 Subject: [Histonet] Histotech Training Message-ID: <1342307492.23532.YahooMailNeo@web120905.mail.ne1.yahoo.com> Hi, ??? I have a question about the training requirement for the HT Certification Test. My question is, will the ASCP allow a guy who works as a lab assistant in the histology lab andintends to train as an histotech for one and half hour a day for twelve months to sit for the test? ? ? Thanks, Wilson From LBUSTAMANTE <@t> cvm.tamu.edu Sat Jul 14 18:14:17 2012 From: LBUSTAMANTE <@t> cvm.tamu.edu (Bustamante, Lin) Date: Sat Jul 14 18:14:20 2012 Subject: [Histonet] fo a small GI lab.... Question Message-ID: <94B6DC15AAF2F046BF847D4C1CA9AAC97307C00C@CVMMB02.cvm.tamu.edu> How do you store your waste before been picked up by any company. I need to know what is acceptable by CAP Inspectors; temperature for the room. Our containers are outside lab, etc. Any information will be very much appreciated. Thank you. Lin Bustamante From jon2038433 <@t> maricopa.edu Sat Jul 14 19:04:23 2012 From: jon2038433 <@t> maricopa.edu (Jon Hannasch) Date: Sat Jul 14 18:54:10 2012 Subject: [Histonet] Looking for histotech job Message-ID: Hello everyone, I am a newly certified histotechnician. I live in Arizona. I recently completed a NAACLS certified program through Phoenix college and just got my ASCP certification. I am looking for a job as a histotech. If you have any openings or know of any that I could apply for please contact me at Jon2038433@maricopa.edu that way I can send you my resume. Thank you and I hope you talk to you soon! From cls71877 <@t> gmail.com Sat Jul 14 22:32:10 2012 From: cls71877 <@t> gmail.com (Cristi Rigazio) Date: Sat Jul 14 22:32:17 2012 Subject: [Histonet] fo a small GI lab.... Question In-Reply-To: <94B6DC15AAF2F046BF847D4C1CA9AAC97307C00C@CVMMB02.cvm.tamu.edu> References: <94B6DC15AAF2F046BF847D4C1CA9AAC97307C00C@CVMMB02.cvm.tamu.edu> Message-ID: We have a metal drum, 55 gallon that is stored at rm temp. We have a weekly check that we document. Sent from my iPhone On Jul 14, 2012, at 4:14 PM, "Bustamante, Lin" wrote: > How do you store your waste before been picked up by any company. I need to know what is acceptable by CAP Inspectors; temperature for the room. Our containers are outside lab, etc. Any information will be very much appreciated. > Thank you. > Lin Bustamante > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joelleweaver <@t> hotmail.com Sun Jul 15 08:04:57 2012 From: joelleweaver <@t> hotmail.com (joelle weaver) Date: Sun Jul 15 08:05:03 2012 Subject: [Histonet] Histotech Training In-Reply-To: <1342307492.23532.YahooMailNeo@web120905.mail.ne1.yahoo.com> References: <1342307492.23532.YahooMailNeo@web120905.mail.ne1.yahoo.com> Message-ID: Take a look at the ASCP BOC eligibility criteria at http://www.ascp.org/Board-of-Certification/GetCertifiedin a nutshell it is pretty much this at this time: Candidates must have completed a NAACLS Histotechnician program in the last 5 years. Or, Candidates must have acquired a minimum of 90 quarter hours or 60 semester hours in an accredited university or college, to include 12 semester hours of chemistry and biology, or possess an Associate degree from an accredited university or college with a combination of 18 quarter chemistry and of biology, plus a 1 year experience in histopathology Candidates must also have acquired experience within the past 10 years in any of the following areas; Fixation, Processing, Microtomy and Staining.They define "FT" employment and sliding scale for PT work in the materials.What is on the exam is outlined completely on the BOC page and in this guideline/outline- http://www.ascp.org/PDF/ExaminationContentGuidelinesHT.aspx Joelle Weaver MAOM, HTL (ASCP) QIHC > Date: Sat, 14 Jul 2012 16:11:32 -0700 > From: wilson6848@yahoo.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histotech Training > > Hi, > I have a question about the training requirement for the HT Certification Test. My question is, will the ASCP allow a guy who works as a lab assistant in the histology lab andintends to train as an histotech for one and half hour a day for twelve months to sit for the test? > > Thanks, > Wilson > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joelleweaver <@t> hotmail.com Sun Jul 15 08:16:08 2012 From: joelleweaver <@t> hotmail.com (joelle weaver) Date: Sun Jul 15 08:16:12 2012 Subject: [Histonet] Different Avenues To Become Certified In-Reply-To: <1342181958.20737.YahooMailNeo@web84511.mail.ne1.yahoo.com> References: <1342181958.20737.YahooMailNeo@web84511.mail.ne1.yahoo.com> Message-ID: Recently the other relatively recognized certifying organization, NCA, had merged with the BOR ( now BOC- board of certification). I do not believe that there is any other nationally recognized certifying body other than the ASCP for histotechnicians and histotechnologists. They are the only body who grants the HT/HTL credentials or their equivalent to my knowledge. If there are other certifications out there, to my knowledge, they are not well known or recognized. Here is the link to the ASCP/BOC eligibility-routes.http://www.ascp.org/Board-of-Certification/GetCertified Joelle Weaver MAOM, HTL (ASCP) QIHC > Date: Fri, 13 Jul 2012 05:19:18 -0700 > From: nelsonrnch@verizon.net > To: Histonet@lists.utsouthwestern.edu > CC: > Subject: [Histonet] Different Avenues To Become Certified > > Good morning Histo-Land, > > A question was asked to me by my Medical Director the other day, so I ask for everyones expert answers to this question. > Is there any other entities other than ASCP to be certified by for HT/HTL ? > > I thank everyone in advance. Have a great weekend. > > BEST REGARDS, > > PATTI RUBEN-NELSON H.T.(ASCP) > PNP LABORATORY CONSULTANTS > SUPERVISOR/DGC > P.O. BOX 412 > CABAZON, CA. 92230 > cell (909) 841-9761 > nelsonrnch@verizon.net > > > CONFIDENTIALITY NOTICE: > This message and any included attachments are from Patti Nelson, PNP Laboratory Consultants > and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or exemption from disclosure under applicable > law. Unauthorized forwarding, printing, copying, distribution, or use of > such information is strictly prohibited and may be unlawful. If you are not > the addressee, please promptly delete this message and notify the sender of > the delivery error by e-mail or you may call 909-841-9761. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From w_alkadhumi <@t> yahoo.com Sun Jul 15 11:25:21 2012 From: w_alkadhumi <@t> yahoo.com (wassan alkadhumi) Date: Sun Jul 15 11:25:25 2012 Subject: [Histonet] Buckling Artifact Message-ID: <1342369521.70552.YahooMailNeo@web45204.mail.sp1.yahoo.com> Dear members, I have a recent problem in our IHC sectioning " the buckling artifact ", and it shows in charged slides only not in normal slides. Our pathologists are finding a problem in reading the slides because of it, dose any one please have an idea in how to get red of it?? I had this buckling artifact for the last 4 weeks so please help. thanks Wassan Histo-technician Sulaymanyah From carl.hobbs <@t> kcl.ac.uk Sun Jul 15 12:30:00 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Sun Jul 15 12:31:05 2012 Subject: [Histonet] RE: Buckling Artifact Message-ID: <11D9615B89C10747B1C985966A63D7CA38626A5AB0@KCL-MAIL04.kclad.ds.kcl.ac.uk> Can you submit an image? I can imagine what this is but, no point until I see an image? What are you doing that is different, if this is a new problem? Curiously, Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 From lpwenk <@t> sbcglobal.net Sun Jul 15 12:52:22 2012 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sun Jul 15 12:52:28 2012 Subject: [Histonet] Histotech Training In-Reply-To: References: <1342307492.23532.YahooMailNeo@web120905.mail.ne1.yahoo.com> Message-ID: <9DB648563BD34B2EB6FFAF550FB2E9C6@HP2010> Note: The exact wording under HT eligibility is: ". . . AND one year full time acceptable experience in a histopathology (clinical, veterinary, industry or research) laboratory in the U.S., Canada or an accredited laboratory* within the last ten years." Very important to note the "One year full time experience". There is another page that talks about full-time vs. part-time experience. http://ascp.org/Board-of-Certification/GetCertified/Step-2/Verify-your-experience.html "Full-time experience is defined as a minimum of thirty-five (35) hours per week. Individuals who have part-time experience may be permitted to utilize prorated part-time experience to meet the work experience requirements. For example, if you are employed 20 hours per week for one year, your experience would be computed as 20 divided by 35 multiplied by 52 weeks, or the equivalent of 29.7 weeks of full time employment." Therefore, to qualify to take the exam, your person would need to work a minimum of 35 hours/week for 52 weeks, to be equal to 1 year full time experience. Your person, who is working 1.5 hours/day = 7.5 hours/week (1.5 x 5) Therefore, his full time equivalent would be: (7.5 divided by 35) multiplied by 52 weeks = 11.1 weeks of full time work Therefore, your person will have to work 4.7 years, at 1.5 hours/day, to be equivalent to 1 year full time experience. Now, that being said, some of his hours as a lab assistant MIGHT, just MIGHT, be allowed to be counted as histotechnology experience, for example, if he changes the solutions on the tissue processor, or runs the automated H&E or coverslipper. Some of these MIGHT be considered histotech responsibilities. The problem is, ASCP won't say over the phone whether some of the experience will or will not count, and how much of it will (or will not) count. You can ask, but they usually say to apply and then a decision will be made. And if ASCP decides it doesn't count, and the person doesn't have enough hours for 1 year full time, they are denied being allowed to take the exam, and they do NOT get their money back ($200 right now for HT). So, I suggest having MORE THAN half of his hours being histotech job responsibilities only - embedding, sectioning, special stains, IHC stains, troubleshooting, making solutions, etc. And then LESS THAN half being the blurred areas where histotechs or lab assistants might do it, depending upon staffing (processors, coverslipper, etc.). So not all his lab assistant job responsibilities can be counted. That's still over 2.4 years of histotech responsibility (at 1.5 hours/day), PLUS the number of hours/weeks he has as lab assistant that MIGHT, just MIGHT qualify as histotech responsibilities. But don't quote me. This is really gray, shaky ground. It's a lot easier when the person is working 20 hours/week as a histotech, and it takes 2 years to qualify. Or 35-40 hours/week as a histotech and it takes 1 year to quality. Also, frankly, in my opinion, working 1.5 hours/day, or 7.5 hours/week for 1 year is NOT enough time for him to get the technical knowledge to be able to pass the exam. It's $200 to apply for the HT exam. Please give him enough time to learn all the material, so he doesn't have to take the exam again. And again. That's a lot of money to waste when not adequately prepared. Peggy A. Wenk, HTL(ASCP)SLS -----Original Message----- From: joelle weaver Sent: Sunday, July 15, 2012 9:04 AM To: wilson6848@yahoo.com ; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotech Training Take a look at the ASCP BOC eligibility criteria at http://www.ascp.org/Board-of-Certification/GetCertifiedin a nutshell it is pretty much this at this time: Candidates must have completed a NAACLS Histotechnician program in the last 5 years. Or, Candidates must have acquired a minimum of 90 quarter hours or 60 semester hours in an accredited university or college, to include 12 semester hours of chemistry and biology, or possess an Associate degree from an accredited university or college with a combination of 18 quarter chemistry and of biology, plus a 1 year experience in histopathology Candidates must also have acquired experience within the past 10 years in any of the following areas; Fixation, Processing, Microtomy and Staining.They define "FT" employment and sliding scale for PT work in the materials.What is on the exam is outlined completely on the BOC page and in this guideline/outline- http://www.ascp.org/PDF/ExaminationContentGuidelinesHT.aspx Joelle Weaver MAOM, HTL (ASCP) QIHC > Date: Sat, 14 Jul 2012 16:11:32 -0700 > From: wilson6848@yahoo.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histotech Training > > Hi, > I have a question about the training requirement for the HT > Certification Test. My question is, will the ASCP allow a guy who works as > a lab assistant in the histology lab andintends to train as an histotech > for one and half hour a day for twelve months to sit for the test? > > Thanks, > Wilson > _______________________________________________ > 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 From settembr <@t> umdnj.edu Mon Jul 16 07:22:23 2012 From: settembr <@t> umdnj.edu (Settembre, Dana) Date: Mon Jul 16 07:22:35 2012 Subject: [Histonet] Negative controls and CAP In-Reply-To: <1342229451.62515.YahooMailNeo@web120104.mail.ne1.yahoo.com> References: <1342229451.62515.YahooMailNeo@web120104.mail.ne1.yahoo.com> Message-ID: Hi Chris, CAP actually has a specific checklist question on Negative Controls. The question is ANP.22570 and if I remember correctly it is quite extensive. Most of the CAP questions are about 7 to 10 lines. The Negative control question is more than a page long. Get your hands a copy of the most current checklist for anatomic pathology - That will include Immunohistochemistry. By the way, the rule is that a negative control must be included with each run and there is much, much more. ANP.22570 Good Luck, Dana Settembre Immunohistochemistry Lab University Hospital - UMDNJ Newark, NJ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christopher Jacobs Sent: Friday, July 13, 2012 9:31 PM To: histonet@lists.utsouthwestern.edu Cc: cjacobs@clinpath.com Subject: [Histonet] Negative controls and CAP Histonetters, I have been made the IHC lead in a fairly large laboratory that is seeking to become CAP accredited. I am definitely a newbie when it comes to CAP. A question was brought up today about how we do our negative controls. Specifically, should we run another negative control with any IHCs that need to be repeated or if we should run another negative control if at a later date a pathologist orders additional IHCs on a case. With our current protocol, we run one negative control per detection kit used, per case. Consequently, most cases end up with just one negative control slide. We do NOT run another negative control if we have to repeat one of a group of immunos or if a pathologist orders additional stains at a later point. I am wondering if this is good practice, and acceptable with CAP. I would love to find some literature that could help me make a case either way on this. Actually, any literature or pointers regarding IHCs and becoming CAP accredited will probably help save what little hair I have left. Thank you, Chris Jacobs, HT(ASCP)QIHC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From trevor.mornan <@t> avantik-us.com Mon Jul 16 09:39:18 2012 From: trevor.mornan <@t> avantik-us.com (Trevor Mornan) Date: Mon Jul 16 09:39:01 2012 Subject: [Histonet] Summer Savings from Avantik Biogroup Message-ID: Sent By: Avantik Biogroup 32 Commerce Street P.O. Box 1299 Springfield NJ 07081 United States To view as a web page press on or copy this link into your browsers address bar https://u01.spsend.com/speasapage.aspx?X=5P143NSOI4O1O9G400VDWO If you prefer not to receive future e-mails of this type, please copy to your browser or press on this link "http://u01.spsend.com/SpeSupIt.aspx?X=5P143NSOI4O1O9G400VDWO&Addr=histonet~~2lists.utsouthwestern.edu" to unsubscribe. From sdysart <@t> mirnarx.com Mon Jul 16 10:13:19 2012 From: sdysart <@t> mirnarx.com (Sarah Dysart) Date: Mon Jul 16 10:13:28 2012 Subject: [Histonet] Conferences? Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D501077DE79@BL2PRD0710MB363.namprd07.prod.outlook.com> Does anyone know of any conferences or conventions going on in the histology world in the next 8 weeks?? I am 8 months pregnant and was just told that I need to go to a conference this year...unfortunately I cannot fly. The conference would have to be somewhere drive-able from Austin, Texas (preferably no more than 10 hours or so...we Texas people are used to long drives to get places...). That would open up LA, OK, TX, MO, etc. 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 From l.kammili <@t> yahoo.com Mon Jul 16 10:13:33 2012 From: l.kammili <@t> yahoo.com (Lakshmi Kammili) Date: Mon Jul 16 10:13:41 2012 Subject: [Histonet] Split of cost per slide Message-ID: <1342451613.13988.YahooMailNeo@web113719.mail.gq1.yahoo.com> Hi, I am working for a Pathology core lab in George Washington University. We are looking for the split of cost per slide. How much it costs in materials (gloves, reagents, plastic wear) and time (technical time and hourly use of machines) to process a tissue sample from formalin to a stained slide for light microscopy or EM.? If any one of you have any idea about this, Since we were asked to submit this way for NIH funding to the lab, that would be very kind of you if you can share this information with us. Thank you so much, Lakshmi Kammili, Sr. Research Associate, Pathology Core lab, GWU school of medicine, Washington?DC. From lpwenk <@t> sbcglobal.net Mon Jul 16 10:26:50 2012 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Mon Jul 16 10:27:00 2012 Subject: [Histonet] Conferences? In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D501077DE79@BL2PRD0710MB363.namprd07.prod.outlook.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D501077DE79@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: <90C4FFF325284FCF8B1D38DDC6F3A56F@HP2010> Most of the state symposiums are in the spring, since the NSH symposium is in the fall. I just checked the NSH meetings calendar, and I don't see any state meetings for the next several months. Any chance you can go to the NSH Symposium in Vancouver, Canada (which is Region IX). Sept. 29-Oct. 3? Don't know where in the 8th month PG you are. http://www.histoconvention.org/ How about a series of NSH Teleconferences? If you can't listen to it the day it's given, that's OK. Every lab that pays for a teleconference gets a CD about 4-6 weeks later, with the PowerPoint and speaker's voice, and can listen to it up to 2 years later and still get CE. Plus, everyone else in your lab can listen and get CE. Not as much fun as going away to a meeting, but if you have money - and being pregnant limits your ability to attend a meeting - this might be the way to go. They are $125 each. http://www.nsh.org/content/nsh-teleconference-series Peggy A. Wenk, HTL(ASCP)SLS (Disclosure - I'm the NSH Teleconference Coordinator, but I don't get paid for this volunteer position.) -----Original Message----- From: Sarah Dysart Sent: Monday, July 16, 2012 11:13 AM To: histonet Subject: [Histonet] Conferences? Does anyone know of any conferences or conventions going on in the histology world in the next 8 weeks?? I am 8 months pregnant and was just told that I need to go to a conference this year...unfortunately I cannot fly. The conference would have to be somewhere drive-able from Austin, Texas (preferably no more than 10 hours or so...we Texas people are used to long drives to get places...). That would open up LA, OK, TX, MO, etc. 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 From rjbuesa <@t> yahoo.com Mon Jul 16 11:01:35 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jul 16 11:01:42 2012 Subject: [Histonet] Split of cost per slide In-Reply-To: <1342451613.13988.YahooMailNeo@web113719.mail.gq1.yahoo.com> References: <1342451613.13988.YahooMailNeo@web113719.mail.gq1.yahoo.com> Message-ID: <1342454495.56188.YahooMailNeo@web121401.mail.ne1.yahoo.com> Under separate cover I am sending you something on this subject. Ren? J. ________________________________ From: Lakshmi Kammili To: "histonet@lists.utsouthwestern.edu" Sent: Monday, July 16, 2012 11:13 AM Subject: [Histonet] Split of cost per slide Hi, I am working for a Pathology core lab in George Washington University. We are looking for the split of cost per slide. How much it costs in materials (gloves, reagents, plastic wear) and time (technical time and hourly use of machines) to process a tissue sample from formalin to a stained slide for light microscopy or EM.? If any one of you have any idea about this, Since we were asked to submit this way for NIH funding to the lab, that would be very kind of you if you can share this information with us. Thank you so much, Lakshmi Kammili, Sr. Research Associate, Pathology Core lab, GWU school of medicine, Washington?DC. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DSiena <@t> statlab.com Mon Jul 16 11:47:32 2012 From: DSiena <@t> statlab.com (Debra Siena) Date: Mon Jul 16 11:47:36 2012 Subject: [Histonet] PMMA bone question Message-ID: Hi All, I need to ask if a gram stain can be performed on a PMMA embedded bone sections? If so are there modifications to a routine gram that must be done for the stain to work? Thank you all for any information that can be provided and if the exact procedure is needed, I can provide that as well. Best wishes Debbie Siena, HT(ASCP)QIHC StatLab Medical Products Technical Support Manager 407 Interchange Street | McKinney, TX 75071 t:?800.442.3573 ext. 229 | f: 972.436.1369 dsiena@statlab.com | www.statlab.com From Ken_Marissael <@t> vwr.com Mon Jul 16 12:04:17 2012 From: Ken_Marissael <@t> vwr.com (Ken_Marissael@vwr.com) Date: Mon Jul 16 12:06:09 2012 Subject: [Histonet] Ken Marissael is out of the office Message-ID: I will be out of the office starting 07/16/2012 and will not return until 08/01/2012. I will be on vacation, but will not be travelling, I will have access to e-mails sporadically in the evening. Please contact my manager Scott Nuetzel for emergencies at 410-302-3045, or for routine items... healthcareservice.com or customer service at 877-881-1192. From Brenda <@t> nsh.org Mon Jul 16 13:04:34 2012 From: Brenda <@t> nsh.org (Brenda Royce) Date: Mon Jul 16 13:04:42 2012 Subject: [Histonet] Re: Conferences Message-ID: <507923F3D62F404D919F15ECD68D12FE173939@NSH-SRVR01.nsh.local> Hi Sarah - NSH has teleconferences every month where you can earn contact hours. We also have recorded sessions from our Symposium/Convention online for purchase so you can listen to material, receive handout and earn contact hours. That may be your best option so you don't have to travel right now. Visit NSH website: www.nsh.org under events and professional development tabs or Click here for Teleconferences http://s3.goeshow.com/nsh/NSHTC2012/ereg949997.cfm?pg=home Click here for Online Learning Center http://www.softconference.com/NSH/generic.asp?ID=7372 Please feel free to contact me if you have any other questions. Kind Regards, Brenda Royce Membership Manager National Society for Histotechnology 8850 Stanford Blvd, Suite 2900 Columbia, Maryland 21045 Ph# 443-535-4064?? Fax #443-535-4055 Visit us at www.nsh.org Come see us in Vancouver this year for our Symposium/Convention. Visit www.histoconvention.org for details Looking for continuing education? Visit the NSH website. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, July 16, 2012 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 104, Issue 20 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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: Buckling Artifact (Hobbs, Carl) 2. Re: Histotech Training (Lee & Peggy Wenk) 3. RE: Negative controls and CAP (Settembre, Dana) 4. Summer Savings from Avantik Biogroup (Trevor Mornan) 5. Conferences? (Sarah Dysart) 6. Split of cost per slide (Lakshmi Kammili) 7. Re: Conferences? (Lee & Peggy Wenk) 8. Re: Split of cost per slide (Rene J Buesa) 9. PMMA bone question (Debra Siena) ---------------------------------------------------------------------- Message: 1 Date: Sun, 15 Jul 2012 18:30:00 +0100 From: "Hobbs, Carl" Subject: [Histonet] RE: Buckling Artifact To: "histonet@lists.utsouthwestern.edu" Message-ID: <11D9615B89C10747B1C985966A63D7CA38626A5AB0@KCL-MAIL04.kclad.ds.kcl.ac.uk> Content-Type: text/plain; charset="us-ascii" Can you submit an image? I can imagine what this is but, no point until I see an image? What are you doing that is different, if this is a new problem? Curiously, Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ------------------------------ Message: 2 Date: Sun, 15 Jul 2012 13:52:22 -0400 From: "Lee & Peggy Wenk" Subject: Re: [Histonet] Histotech Training To: "joelle weaver" , , Message-ID: <9DB648563BD34B2EB6FFAF550FB2E9C6@HP2010> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Note: The exact wording under HT eligibility is: ". . . AND one year full time acceptable experience in a histopathology (clinical, veterinary, industry or research) laboratory in the U.S., Canada or an accredited laboratory* within the last ten years." Very important to note the "One year full time experience". There is another page that talks about full-time vs. part-time experience. http://ascp.org/Board-of-Certification/GetCertified/Step-2/Verify-your-experience.html "Full-time experience is defined as a minimum of thirty-five (35) hours per week. Individuals who have part-time experience may be permitted to utilize prorated part-time experience to meet the work experience requirements. For example, if you are employed 20 hours per week for one year, your experience would be computed as 20 divided by 35 multiplied by 52 weeks, or the equivalent of 29.7 weeks of full time employment." Therefore, to qualify to take the exam, your person would need to work a minimum of 35 hours/week for 52 weeks, to be equal to 1 year full time experience. Your person, who is working 1.5 hours/day = 7.5 hours/week (1.5 x 5) Therefore, his full time equivalent would be: (7.5 divided by 35) multiplied by 52 weeks = 11.1 weeks of full time work Therefore, your person will have to work 4.7 years, at 1.5 hours/day, to be equivalent to 1 year full time experience. Now, that being said, some of his hours as a lab assistant MIGHT, just MIGHT, be allowed to be counted as histotechnology experience, for example, if he changes the solutions on the tissue processor, or runs the automated H&E or coverslipper. Some of these MIGHT be considered histotech responsibilities. The problem is, ASCP won't say over the phone whether some of the experience will or will not count, and how much of it will (or will not) count. You can ask, but they usually say to apply and then a decision will be made. And if ASCP decides it doesn't count, and the person doesn't have enough hours for 1 year full time, they are denied being allowed to take the exam, and they do NOT get their money back ($200 right now for HT). So, I suggest having MORE THAN half of his hours being histotech job responsibilities only - embedding, sectioning, special stains, IHC stains, troubleshooting, making solutions, etc. And then LESS THAN half being the blurred areas where histotechs or lab assistants might do it, depending upon staffing (processors, coverslipper, etc.). So not all his lab assistant job responsibilities can be counted. That's still over 2.4 years of histotech responsibility (at 1.5 hours/day), PLUS the number of hours/weeks he has as lab assistant that MIGHT, just MIGHT qualify as histotech responsibilities. But don't quote me. This is really gray, shaky ground. It's a lot easier when the person is working 20 hours/week as a histotech, and it takes 2 years to qualify. Or 35-40 hours/week as a histotech and it takes 1 year to quality. Also, frankly, in my opinion, working 1.5 hours/day, or 7.5 hours/week for 1 year is NOT enough time for him to get the technical knowledge to be able to pass the exam. It's $200 to apply for the HT exam. Please give him enough time to learn all the material, so he doesn't have to take the exam again. And again. That's a lot of money to waste when not adequately prepared. Peggy A. Wenk, HTL(ASCP)SLS -----Original Message----- From: joelle weaver Sent: Sunday, July 15, 2012 9:04 AM To: wilson6848@yahoo.com ; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotech Training Take a look at the ASCP BOC eligibility criteria at http://www.ascp.org/Board-of-Certification/GetCertifiedin a nutshell it is pretty much this at this time: Candidates must have completed a NAACLS Histotechnician program in the last 5 years. Or, Candidates must have acquired a minimum of 90 quarter hours or 60 semester hours in an accredited university or college, to include 12 semester hours of chemistry and biology, or possess an Associate degree from an accredited university or college with a combination of 18 quarter chemistry and of biology, plus a 1 year experience in histopathology Candidates must also have acquired experience within the past 10 years in any of the following areas; Fixation, Processing, Microtomy and Staining.They define "FT" employment and sliding scale for PT work in the materials.What is on the exam is outlined completely on the BOC page and in this guideline/outline- http://www.ascp.org/PDF/ExaminationContentGuidelinesHT.aspx Joelle Weaver MAOM, HTL (ASCP) QIHC > Date: Sat, 14 Jul 2012 16:11:32 -0700 > From: wilson6848@yahoo.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histotech Training > > Hi, > I have a question about the training requirement for the HT > Certification Test. My question is, will the ASCP allow a guy who works as > a lab assistant in the histology lab andintends to train as an histotech > for one and half hour a day for twelve months to sit for the test? > > Thanks, > Wilson > _______________________________________________ > 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 ------------------------------ Message: 3 Date: Mon, 16 Jul 2012 08:22:23 -0400 From: "Settembre, Dana" Subject: RE: [Histonet] Negative controls and CAP To: Christopher Jacobs , "histonet@lists.utsouthwestern.edu" Cc: "cjacobs@clinpath.com" Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Chris, CAP actually has a specific checklist question on Negative Controls. The question is ANP.22570 and if I remember correctly it is quite extensive. Most of the CAP questions are about 7 to 10 lines. The Negative control question is more than a page long. Get your hands a copy of the most current checklist for anatomic pathology - That will include Immunohistochemistry. By the way, the rule is that a negative control must be included with each run and there is much, much more. ANP.22570 Good Luck, Dana Settembre Immunohistochemistry Lab University Hospital - UMDNJ Newark, NJ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christopher Jacobs Sent: Friday, July 13, 2012 9:31 PM To: histonet@lists.utsouthwestern.edu Cc: cjacobs@clinpath.com Subject: [Histonet] Negative controls and CAP Histonetters, I have been made the IHC lead in a fairly large laboratory that is seeking to become CAP accredited. I am definitely a newbie when it comes to CAP. A question was brought up today about how we do our negative controls. Specifically, should we run another negative control with any IHCs that need to be repeated or if we should run another negative control if at a later date a pathologist orders additional IHCs on a case. With our current protocol, we run one negative control per detection kit used, per case. Consequently, most cases end up with just one negative control slide. We do NOT run another negative control if we have to repeat one of a group of immunos or if a pathologist orders additional stains at a later point. I am wondering if this is good practice, and acceptable with CAP. I would love to find some literature that could help me make a case either way on this. Actually, any literature or pointers regarding IHCs and becoming CAP accredited will probably help save what little hair I have left. Thank you, Chris Jacobs, HT(ASCP)QIHC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: 16 Jul 2012 08:39:18 -0600 From: "Trevor Mornan" Subject: [Histonet] Summer Savings from Avantik Biogroup To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=utf-8 Sent By: Avantik Biogroup 32 Commerce Street P.O. Box 1299 Springfield NJ 07081 United States To view as a web page press on or copy this link into your browsers address bar https://u01.spsend.com/speasapage.aspx?X=5P143NSOI4O1O9G400VDWO If you prefer not to receive future e-mails of this type, please copy to your browser or press on this link "http://u01.spsend.com/SpeSupIt.aspx?X=5P143NSOI4O1O9G400VDWO&Addr=histonet~~2lists.utsouthwestern.edu" to unsubscribe. ------------------------------ Message: 5 Date: Mon, 16 Jul 2012 15:13:19 +0000 From: Sarah Dysart Subject: [Histonet] Conferences? To: histonet Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D501077DE79@BL2PRD0710MB363.namprd07.prod.outlook.com> Content-Type: text/plain; charset="us-ascii" Does anyone know of any conferences or conventions going on in the histology world in the next 8 weeks?? I am 8 months pregnant and was just told that I need to go to a conference this year...unfortunately I cannot fly. The conference would have to be somewhere drive-able from Austin, Texas (preferably no more than 10 hours or so...we Texas people are used to long drives to get places...). That would open up LA, OK, TX, MO, etc. 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 ------------------------------ Message: 6 Date: Mon, 16 Jul 2012 08:13:33 -0700 (PDT) From: Lakshmi Kammili Subject: [Histonet] Split of cost per slide To: "histonet@lists.utsouthwestern.edu" Message-ID: <1342451613.13988.YahooMailNeo@web113719.mail.gq1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi, I am working for a Pathology core lab in George Washington University. We are looking for the split of cost per slide. How much it costs in materials (gloves, reagents, plastic wear) and time (technical time and hourly use of machines) to process a tissue sample from formalin to a stained slide for light microscopy or EM.? If any one of you have any idea about this, Since we were asked to submit this way for NIH funding to the lab, that would be very kind of you if you can share this information with us. Thank you so much, Lakshmi Kammili, Sr. Research Associate, Pathology Core lab, GWU school of medicine, Washington?DC. ------------------------------ Message: 7 Date: Mon, 16 Jul 2012 11:26:50 -0400 From: "Lee & Peggy Wenk" Subject: Re: [Histonet] Conferences? To: "Sarah Dysart" , "histonet" Message-ID: <90C4FFF325284FCF8B1D38DDC6F3A56F@HP2010> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Most of the state symposiums are in the spring, since the NSH symposium is in the fall. I just checked the NSH meetings calendar, and I don't see any state meetings for the next several months. Any chance you can go to the NSH Symposium in Vancouver, Canada (which is Region IX). Sept. 29-Oct. 3? Don't know where in the 8th month PG you are. http://www.histoconvention.org/ How about a series of NSH Teleconferences? If you can't listen to it the day it's given, that's OK. Every lab that pays for a teleconference gets a CD about 4-6 weeks later, with the PowerPoint and speaker's voice, and can listen to it up to 2 years later and still get CE. Plus, everyone else in your lab can listen and get CE. Not as much fun as going away to a meeting, but if you have money - and being pregnant limits your ability to attend a meeting - this might be the way to go. They are $125 each. http://www.nsh.org/content/nsh-teleconference-series Peggy A. Wenk, HTL(ASCP)SLS (Disclosure - I'm the NSH Teleconference Coordinator, but I don't get paid for this volunteer position.) -----Original Message----- From: Sarah Dysart Sent: Monday, July 16, 2012 11:13 AM To: histonet Subject: [Histonet] Conferences? Does anyone know of any conferences or conventions going on in the histology world in the next 8 weeks?? I am 8 months pregnant and was just told that I need to go to a conference this year...unfortunately I cannot fly. The conference would have to be somewhere drive-able from Austin, Texas (preferably no more than 10 hours or so...we Texas people are used to long drives to get places...). That would open up LA, OK, TX, MO, etc. 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 ------------------------------ Message: 8 Date: Mon, 16 Jul 2012 09:01:35 -0700 (PDT) From: Rene J Buesa Subject: Re: [Histonet] Split of cost per slide To: Lakshmi Kammili , "histonet@lists.utsouthwestern.edu" Message-ID: <1342454495.56188.YahooMailNeo@web121401.mail.ne1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Under separate cover I am sending you something on this subject. Ren? J. ________________________________ From: Lakshmi Kammili To: "histonet@lists.utsouthwestern.edu" Sent: Monday, July 16, 2012 11:13 AM Subject: [Histonet] Split of cost per slide Hi, I am working for a Pathology core lab in George Washington University. We are looking for the split of cost per slide. How much it costs in materials (gloves, reagents, plastic wear) and time (technical time and hourly use of machines) to process a tissue sample from formalin to a stained slide for light microscopy or EM.? If any one of you have any idea about this, Since we were asked to submit this way for NIH funding to the lab, that would be very kind of you if you can share this information with us. Thank you so much, Lakshmi Kammili, Sr. Research Associate, Pathology Core lab, GWU school of medicine, Washington?DC. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Mon, 16 Jul 2012 11:47:32 -0500 From: Debra Siena Subject: [Histonet] PMMA bone question To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi All, I need to ask if a gram stain can be performed on a PMMA embedded bone sections? If so are there modifications to a routine gram that must be done for the stain to work? Thank you all for any information that can be provided and if the exact procedure is needed, I can provide that as well. Best wishes Debbie Siena, HT(ASCP)QIHC StatLab Medical Products Technical Support Manager 407 Interchange Street | McKinney, TX 75071 t:?800.442.3573 ext. 229 | f: 972.436.1369 dsiena@statlab.com | www.statlab.com ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 104, Issue 20 ***************************************** From CIngles <@t> uwhealth.org Mon Jul 16 14:57:55 2012 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Mon Jul 16 14:58:32 2012 Subject: [Histonet] ER, Clone SP1 References: <6959269F7DA142FCB3EEEC3CFA29B6E2@Patsyoffice><876C9001-87DB-4723-8F71-D99F14D303EA@yahoo.com>, <8D7C2D242DBD45498006B21122072BF8B5BBAC03@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A86F@UWHC-MAIL2.uwhis.hosp.wisc.edu> Don't give anyone any ideas!! Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jonathan Cremer Sent: Fri 7/13/2012 1:45 AM To: Morken, Timothy; Kim Donadio; Patsy Ruegg Cc: Subject: RE: [Histonet] ER, Clone SP1 That drives me up the wall as well. How can you possibly patent something that has been made by nature, and in theory is present in every single human being (I'm talking about any random gene here)? That would be like Newton patenting gravity and then going around and charging everyone a licensing fee for staying on earth instead of floating off in space, or claiming every apple that has fallen off a tree. It's ridiculous! --- ________________________________________ From pvalente <@t> sbcglobal.net Mon Jul 16 20:21:31 2012 From: pvalente <@t> sbcglobal.net (Patricia Valente) Date: Mon Jul 16 20:21:35 2012 Subject: [Histonet] (no subject) Message-ID: <1342488091.52177.YahooMailNeo@web181106.mail.ne1.yahoo.com> http://autogoblog.com/wp-admin/searchn.php?library227.jpg From JMacDonald <@t> mtsac.edu Mon Jul 16 22:55:20 2012 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Mon Jul 16 22:54:50 2012 Subject: [Histonet] Conferences? In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D501077DE79@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: Will an on-line 'workshop' work? ASCP and NSH have some. Sarah Dysart Sent by: histonet-bounces@lists.utsouthwestern.edu 07/16/2012 08:15 AM To histonet cc Subject [Histonet] Conferences? Does anyone know of any conferences or conventions going on in the histology world in the next 8 weeks?? I am 8 months pregnant and was just told that I need to go to a conference this year...unfortunately I cannot fly. The conference would have to be somewhere drive-able from Austin, Texas (preferably no more than 10 hours or so...we Texas people are used to long drives to get places...). That would open up LA, OK, TX, MO, etc. 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 From mw <@t> personifysearch.com Tue Jul 17 07:00:19 2012 From: mw <@t> personifysearch.com (Matt Ward) Date: Tue Jul 17 07:00:27 2012 Subject: [Histonet] New Histology Opportunities Message-ID: Good Morning Histonet, We are currently recruiting for some great new histology opportunities. We are currently searching for Histotechs who are looking for a rewarding career working for an industry leader. Our client offers a strong compensation package, benefits, and upward mobility as they are growing at rapid rate. Currently looking for Histotechs in the following areas: DC/MD, VA, NC, SC, GA, FL, NJ/NY, IL If you are a histotech and looking to join a world leader, please send me a copy of your resume and let me know when you would be available to speak in more detail. Have a great day! Regards, Matt Ward *Account Executive* *Personify* 5020 Weston Parkway Suite 315 Cary NC 27513 (Tel) 800.875.6188 direct ext 103 (Fax) 919.460.0642 www.personifysearch.com From Tobias.Nolden <@t> fli.bund.de Tue Jul 17 08:09:08 2012 From: Tobias.Nolden <@t> fli.bund.de (Nolden, Tobias) Date: Tue Jul 17 08:09:17 2012 Subject: [Histonet] Microcut H1200 Biorad request Message-ID: <8AC171303505BD4285780C39FB34FFE266D9EB775C@FLI-RIE-MBC035.intern.fli.bund.local> Dear Histonetters, I'm send this exceptional request hoping that someone can help me. In future experiments, I want to use organotypic cell culture techniques of mouse/rat brain slices. For this purposes I found a rather old vibrotome (Biorad Microcut H1200) in our institute. We got the vibrotome years ago from the University of Amsterdam but unfortunately nobody worked with this device here and there was no manual available. Since a lot of people still work with the device - there are recent publications where the vibrotome is mentioned in the MM section - I ask you, if anyone has got the manual and can send me a copy. This would help me a lot because Biorad stopped to sell this item years ago and they do not have any information for me after a dozen of phone calls. I am looking forward to hearing from you hoping that somebody can help me with a copy of the manual. Best wishes Tobias **************************************************** Dr. Tobias Nolden, Postdoc Friedrich-Loeffler-Institut Bundesforschungsinstitut f?r Tiergesundheit Federal Research Institute for Animal Health S?dufer 10 17493 Greifswald - Insel Riems, Germany * +49-38351-7-1265 Fax: +49-38351-7-1275 http://www.fli.bund.de **************************************************** From kdboydhisto <@t> yahoo.com Tue Jul 17 12:16:33 2012 From: kdboydhisto <@t> yahoo.com (Kelly Boyd) Date: Tue Jul 17 12:16:36 2012 Subject: [Histonet] (no subject) Message-ID: <1342545393.60586.YahooMailNeo@web125805.mail.ne1.yahoo.com> Since there is?discussion of formalin and methanol, I would like to ask everyone.....Why would a vendor add methanol to their reagent alcohol and their 10% buffered formalin?? What are the advantages/disadvantages to the tissue especially for processing? Kelly? From Gina.Rodriguez <@t> leica-microsystems.com Tue Jul 17 13:49:13 2012 From: Gina.Rodriguez <@t> leica-microsystems.com (Gina.Rodriguez@leica-microsystems.com) Date: Tue Jul 17 13:49:21 2012 Subject: [Histonet] AUTO: Gina Rodriguez is out of the office. (returning 07/19/2012) Message-ID: I am out of the office until 07/19/2012. I will respond to your message when I return. If you need immediate assistance please contact 800-248-0123 or Tech.support@leica-microsystems.com Note: This is an automated response to your message "Histonet Digest, Vol 104, Issue 21" sent on 7/17/2012 12:02:42 PM. This is the only notification you will receive while this person is away. _____________________________________________________________________ This e-mail has been scanned for viruses by Verizon Business Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.verizonbusiness.com/uk From gloria.cole <@t> usa.net Tue Jul 17 14:03:08 2012 From: gloria.cole <@t> usa.net (Gloria Cole) Date: Tue Jul 17 14:03:14 2012 Subject: [Histonet] Histology/Grossing position Message-ID: <672qgqTci6784S04.1342551788@web04.cms.usa.net> Hello Everybody, I have a histology/grossing day shift position opening soon in Irving, Texas. Must possess AA degree or enough hours to satisfy CLIA high complexity testing requirements. Please send resumes to gcole@nueterrapathology.com Thanks, Gloria Cole Supervisor/Manager Nueterra Pathology 214-681-2240 From rjbuesa <@t> yahoo.com Tue Jul 17 15:28:26 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jul 17 15:28:30 2012 Subject: [Histonet] (no subject) In-Reply-To: <1342545393.60586.YahooMailNeo@web125805.mail.ne1.yahoo.com> References: <1342545393.60586.YahooMailNeo@web125805.mail.ne1.yahoo.com> Message-ID: <1342556906.2133.YahooMailNeo@web121406.mail.ne1.yahoo.com> To prevent oxidation of those 2 reagents Ren? J. ________________________________ From: Kelly Boyd To: histonet Sent: Tuesday, July 17, 2012 1:16 PM Subject: [Histonet] (no subject) Since there is?discussion of formalin and methanol, I would like to ask everyone.....Why would a vendor add methanol to their reagent alcohol and their 10% buffered formalin?? What are the advantages/disadvantages to the tissue especially for processing? Kelly? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From baderbo <@t> gmail.com Tue Jul 17 15:43:21 2012 From: baderbo <@t> gmail.com (Bader Siddiki) Date: Tue Jul 17 15:43:25 2012 Subject: [Histonet] (no subject) In-Reply-To: <1342556906.2133.YahooMailNeo@web121406.mail.ne1.yahoo.com> References: <1342545393.60586.YahooMailNeo@web125805.mail.ne1.yahoo.com> <1342556906.2133.YahooMailNeo@web121406.mail.ne1.yahoo.com> Message-ID: Hello Methanol is added to reagent alcohol (ethyl alcohol) so that people do not use for drinking purpose and for tax purposes. Alcohol (ethyl alcohol) has federal tax just like liquor. One can buy pure alcohol, but you have keep inventory and make sure it is not misused. Bader On Tue, Jul 17, 2012 at 1:28 PM, Rene J Buesa wrote: > To prevent oxidation of those 2 reagents > Ren? J. > > > ________________________________ > From: Kelly Boyd > To: histonet > Sent: Tuesday, July 17, 2012 1:16 PM > Subject: [Histonet] (no subject) > > Since there is discussion of formalin and methanol, I would like to ask > everyone.....Why would a vendor add methanol to their reagent alcohol and > their 10% buffered formalin? > > What are the advantages/disadvantages to the tissue especially for > processing? > > Kelly > _______________________________________________ > 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 > -- If any Q's please feel free to contact us Have a nice day/weekend Mit freundlichen Gr??en / With Kind Regards / avec l'aimable ce qui concerne Met vriendelijke groeten ?????? Bader Bader B Siddiki, PhD Executive director, Research and development ImmunoBioScience Corp. (IBSC) Phone: + 1 425 367 4601 Fax: + 1 425 367 4817 cell (mobile) phone: + 1 425 314 0199 e-mail address: BaderBo@Gmail.com Web site: www.ImmunoBioScience.Com Marketing: phone: + 1 650 343 IBSC (4272) E-mail: AnitaIBSC@Aol.com From Rcartun <@t> harthosp.org Tue Jul 17 18:04:39 2012 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Tue Jul 17 18:04:48 2012 Subject: [Histonet] Negative Reagent Control Message-ID: <5005B747.7400.0077.1@harthosp.org> Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing. In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen. Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists. We had a long discussion about this and he agreed to support my position within CAP. With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed. The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause". This one change stands to save the healthcare system millions of dollars. Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax From Joyce.Weems <@t> emoryhealthcare.org Tue Jul 17 18:14:32 2012 From: Joyce.Weems <@t> emoryhealthcare.org (Weems, Joyce K.) Date: Tue Jul 17 18:14:41 2012 Subject: [Histonet] Negative Reagent Control In-Reply-To: <5005B747.7400.0077.1@harthosp.org> References: <5005B747.7400.0077.1@harthosp.org> Message-ID: Wow!! Thank you, Richard et al!! I'm doing a joy, joy dance!!! May we begin eliminating now or must we wait for the official change? Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Tuesday, July 17, 2012 7:05 PM To: Histonet Subject: [Histonet] Negative Reagent Control Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing. In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen. Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists. We had a long discussion about this and he agreed to support my position within CAP. With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed. The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause". This one change stands to save the healthcare system millions of dollars. Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). From amosbrooks <@t> gmail.com Tue Jul 17 19:48:10 2012 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Tue Jul 17 19:48:13 2012 Subject: [Histonet] Golgi-Cox Sections In-Reply-To: References: Message-ID: Hi, For those that nave had the joy of performing the Golgi-Cox stain, you will likely recall that it generally calls for some thick sections. The researcher I am working with is hoping for 100 uM sections. That is way too thick for a microtome and a cryostat. I suggested a vibratome might work better, but not having used one I thought I should ask some folks that may have encountered this. I know 100 uM is *really* thick, but he *really* wants to follow the nerve tracks. Thanks, Amos From ptypea <@t> msn.com Tue Jul 17 20:53:25 2012 From: ptypea <@t> msn.com (LESLIE GARCIA) Date: Tue Jul 17 20:53:29 2012 Subject: [Histonet] Job Opportunity Message-ID: Hello Histonet, There is a wonderful job opportunity in Long Beach CA. We are a small Gastroenterology Pathology Lab seeking an experienced histotechnician. We offer competitive pay and benefits. Schedule: Days, Mon-Fri. For more information please call Linda Watson at 562-997-4540 or email lwatson@lbgastro.com or contact Leslie Garcia at 562-595-5421 extension 143 or email at lgarcia@lbgastro.com. Thank You, Leslie Garcia From AteretS <@t> migal.org.il Wed Jul 18 06:20:42 2012 From: AteretS <@t> migal.org.il (Ateret Shabtay-Yanai) Date: Wed Jul 18 06:21:00 2012 Subject: [Histonet] Image analysis for fat tissue Message-ID: Dear Histonet members I'm interested in image analysis of mice fat tissue, I would like to quantify number Of cells per area and average cell size. Does anyone know a software (+instructions/ lucid manual) which does that? Many thanks! -- Best regards -- Ateret Shabtay-Yanai Migal- Galilee Technology Center Southern Industrial Zone- kiryat Smona P.O.B 831 Kiryat Shmona 11016 Tel: +972-46955002 Mobile: +972-547946460 Email: aterets@migal.org.il From akbitting <@t> geisinger.edu Wed Jul 18 06:25:33 2012 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Wed Jul 18 06:25:47 2012 Subject: [Histonet] Negative Reagent Control In-Reply-To: <5005B747.7400.0077.1@harthosp.org> References: <5005B747.7400.0077.1@harthosp.org> Message-ID: <500664ED.2B7F.00C9.1@geisinger.edu> Our Prayers have been answered!!! >>> Richard Cartun 7/17/2012 7:04 PM >>> Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing. In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen. Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists. We had a long discussion about this and he agreed to support my position within CAP. With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed. The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause". This one change stands to save the healthcare system millions of dollars. Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. Geisinger Health System utilizes an encryption process to safeguard Protected Health Information and other confidential data contained in external e-mail messages. If email is encrypted, the recipient will receive an e-mail instructing them to sign on to the Geisinger Health System Secure E-mail Message Center to retrieve the encrypted e-mail. From mward <@t> wakehealth.edu Wed Jul 18 07:13:11 2012 From: mward <@t> wakehealth.edu (Martha Ward-Pathology) Date: Wed Jul 18 07:13:27 2012 Subject: [Histonet] Negative Reagent Control In-Reply-To: <500664ED.2B7F.00C9.1@geisinger.edu> References: <5005B747.7400.0077.1@harthosp.org> <500664ED.2B7F.00C9.1@geisinger.edu> Message-ID: What wonderful news!!!! ? Martha Ward, MT (ASCP) QIHC Manager Molecular Diagnostics Lab Medical Center Boulevard ?\? Winston-Salem, NC 27157 p 336.716.2109 ?\? f 336.716.5890 ? mward@wakehealth.edu ? ? ? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, July 18, 2012 7:26 AM To: Richard Cartun; Histonet Subject: Re: [Histonet] Negative Reagent Control Our Prayers have been answered!!! >>> Richard Cartun 7/17/2012 7:04 PM >>> Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing. In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen. Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists. We had a long discussion about this and he agreed to support my position within CAP. With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed. The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause". This one change stands to save the healthcare system millions of dollars. Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. Geisinger Health System utilizes an encryption process to safeguard Protected Health Information and other confidential data contained in external e-mail messages. If email is encrypted, the recipient will receive an e-mail instructing them to sign on to the Geisinger Health System Secure E-mail Message Center to retrieve the encrypted e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kdboydhisto <@t> yahoo.com Wed Jul 18 07:38:02 2012 From: kdboydhisto <@t> yahoo.com (Kelly Boyd) Date: Wed Jul 18 07:38:09 2012 Subject: [Histonet] Methanol Message-ID: <1342615082.82861.YahooMailNeo@web125801.mail.ne1.yahoo.com> ?Thanks for all of the replies. And I was aware of why, but this is my confusion:? Why would the MSDS and biohazard triangle be different for different venors for the same reagents? ? Kelly? From Luis.Chiriboga <@t> nyumc.org Wed Jul 18 08:01:54 2012 From: Luis.Chiriboga <@t> nyumc.org (Chiriboga, Luis) Date: Wed Jul 18 08:01:59 2012 Subject: [Histonet] IL5 Message-ID: <3E6798F00C9F494399E96B720ECD1429089054@MSGWCDCPMB22.nyumc.org> Does anyone on the net do or know of a lab that does IL-5 IHC for clinical/diagnostic purposes? TIA Luis Chiriboga Ph.D, Director OCS Experimental Pathology IHC Core Lab NYU School of Medicine Medical Science Building Room 124 646-501-6934 Luis.Chiriboga@nyumc.org From Luis.Chiriboga <@t> nyumc.org Wed Jul 18 08:07:25 2012 From: Luis.Chiriboga <@t> nyumc.org (Chiriboga, Luis) Date: Wed Jul 18 08:08:38 2012 Subject: [Histonet] Negative Reagent Control In-Reply-To: <5005B747.7400.0077.1@harthosp.org> References: <5005B747.7400.0077.1@harthosp.org> Message-ID: <3E6798F00C9F494399E96B720ECD1429089076@MSGWCDCPMB22.nyumc.org> Outstanding!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Tuesday, July 17, 2012 7:05 PM To: Histonet Subject: [Histonet] Negative Reagent Control Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing. In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen. Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists. We had a long discussion about this and he agreed to support my position within CAP. With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed. The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause". This one change stands to save the healthcare system millions of dollars. Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Wed Jul 18 08:45:25 2012 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Wed Jul 18 08:45:33 2012 Subject: [Histonet] any news on CD4, CD8 IHC on FFPE mouse tissue? Message-ID: I'm wondering if anyone has recommendations for CD4 and CD8 antibodies for FFPE mouse tissue. Looking through the archives I see there was only success on frozen tissues. Has anything changed recently?? Or should I stick with CD3. Thanks, Brett Brett M. Connolly, Ph.D. Principal Scientist, Imaging Dept. Merck & Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_connolly@merck.com T- 215-652-2501 F- 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. From kmendell <@t> goldbergmd.net Wed Jul 18 10:42:52 2012 From: kmendell <@t> goldbergmd.net (MaryK Mendell) Date: Wed Jul 18 10:42:58 2012 Subject: [Histonet] (no subject) Message-ID: <96138C8AB728814B9A576E4364EF84F9012BD954D304@EXMBX01.mmeprod.cbeyond> I work in a small lab (office bldg) that has been turned into histolab. We have a portable work station with a vent to out side. (roof) I also have my VIP processor in this room which has the filter carbon (activated) on the processor. Where we have an issue is , on very windy days there seems to be a down draft that happens causing fumes to go into other offices (not ours). I have been looking into the air filter system by leica and one mercedes sells. They have a filter that can be changed every 6 months or so. Has anyone used these with any sort of success rate? Kate Mendell Histopathology/Lab Manager HOWARD S. GOLDBERG, M.D., INC 990 Paradise Road Swampscott, MA 01907 TEL: 781.595.0151 FAX: 781.592.6780 kmendell@goldbergmd.net www.cosmesticdermcenter.com PRIVACY NOTICE: This e-mail message may contain confidential patient or other information belonging to the sender that is legally privileged. This information is intended only for the use of the individual or authorized entity named above. The authorized recipient of this patient or other confidential information is prohibited from disclosing the information to any other party. If you have received this message in error, please notify the sender immediately and delete. Please keep any information you may have viewed confidential. From relia1 <@t> earthlink.net Wed Jul 18 10:55:37 2012 From: relia1 <@t> earthlink.net (Pam Barker) Date: Wed Jul 18 10:55:42 2012 Subject: [Histonet] RELIA Histology Job Update. Summertime in the South is Sizzlin hot and it's not just the temps! Message-ID: <006201cd64fd$c6bed0f0$543c72d0$@earthlink.net> Hi Histonetters!! I hope everyone is having a great day. Are you keeping cool on these hot summer days? Summertime in the South is Sizzlin hot and it's not just the temps!! I have some great new opportunities to tell you about: I have full time permanent positions with some of the best employers in the South. They offer competitive salaries, great benefits and awesome crews to work with. Here is a list of the Sizzlin Summer Stuff! Dermpath HistoTech - Atlanta, GA Histology Tech evenings - Charlotte, NC Histology Tech nights - Charlotte, NC Histology Tech Days - Bristol, TN Histology Tech Nights - Lafayette, LA If you have any questions or know someone who might be interested please contact me. I would love to tell you about these opportunities or write another referral check like I did today if you have a friend who might be interested. I hope to hear from you soon. In the meantime grab some shade and a cool drink! Thank You! Pam Barker President/Senior Recruiting Specialist-Histology RELIA Solutions Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.facebook.com /PamBarkerRELIA www.linkedin.com/in/reliasolutions www.twitter.com/pamatrelia From mdpraet <@t> gmail.com Wed Jul 18 11:56:32 2012 From: mdpraet <@t> gmail.com (mequita praet) Date: Wed Jul 18 11:56:35 2012 Subject: [Histonet] NSH S/C Message-ID: I am flying into Bellingham, WA on Friday Sept. 28th to attend the NSH S/C (much cheaper than flying into Vancouver) at 2:38pm and am looking to share a limo to Vancouver with 5 or 6 people. If you are interested please email me. If I get enough interest, I will follow up and see if that is feasible. Mequita Praet mdpraet@gmail.com From nathanael <@t> bioquant.com Wed Jul 18 12:41:34 2012 From: nathanael <@t> bioquant.com (Nathanael Reveal) Date: Wed Jul 18 12:41:58 2012 Subject: [Histonet] Re: Image Analysis for Fat Tissue Message-ID: Hi Ateret, You've got a few choices, some of which are free and some of which are quite expensive. In many soft tissue staining protocols, adipocytes show up as white, oval, void spaces. Which is good! Software like ImageJ , or its more integrated variant FIJI (which stands for Fiji is Just Image J) is an excellent place to start. They have very good and easy to use measurement tools for area and number. Unfortunately, the topic takes a turn for the complex. Cell number and cell size (i.e. cell volume) can be subject to systematic bias due to section orientation, section thickness, and related parameters. This may cause under or over estimation of the data. This brings us to Stereology, the science of estimating data from 2d cross-sections of 3d structures. Say for example, you're sectioning at 5 microns. The 2d profiles (i.e. cross-sections) of a large adiopcyte will appear on many sections, but the 2d profile of a small adiopcyte will show up only once or twice. In this way, if you count all the adipocyte profiles you see, you could be overestimating the number of adipocytes. You could be counting some adiopocytes more than once. There are standard techniques for estimating the total number of cells in a region and the mean volume of a group of cells. The Optical Fractionator estimates the total number of cells. The Rotator estimates mean cell volume. These tools are found in most stereology systems, commercial or free. The drawback to these methods are that they require specialized commercial software and a microscope capable of monitoring real-time position in X, Y, and Z. This is commonly done with either a motorized stage or encoded stage. Some practical alternatives you could implement with ImageJ may be: 1. Measure "Adipose Volume". That is the area of adipose tissue measured over a set of serial sections. The Delesse principle(again from Stereology) says that area fractions are proportional to volume fractions in a set of serial sections. So no special hardware or random sampling is required. 2. Take care to only count nucleated adipocyte profiles. This helps minimize the problem of overcounting in serial sections. 3. Count adipocytes in sections that are not adjacent. Meaning, cut 5 micron sections but make sure the sections you count are far enough apart that you won't see the cross-sections from the same adipocyte. 4. As far as measuring the volume of individual adipocytes, you might be able use the shortest diameter of the nucleated adipocyte profiles in you see in your sections. This is a tricky one and very much subject to the shape of your adipocyte population. As always, you'll do well to mimic the methods of those that have performed similar work in the past. This will make it easier for reviewers to understand your methods and comment constructively on your paper. You might find this Google Scholar searchhelpful. It covers references to adipocyte volume and adipocyte number since 2008. Best of Luck, Nathanael -- Nathanael Reveal, CEO BIOQUANT Image Analysis Corporation 5611 Ohio Avenue, Nashville, TN 37209 phone: 615-350-7866, toll free: 800-221-0549, fax: 615-350-7282 email: nathanael@bioquant.com, web: www.bioquant.com From sdysart <@t> mirnarx.com Wed Jul 18 12:44:12 2012 From: sdysart <@t> mirnarx.com (Sarah Dysart) Date: Wed Jul 18 12:44:27 2012 Subject: [Histonet] Hold IHC overnight Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> Just found out I have to go to a meeting in 2 hours for the rest of the day (I haven't even eaten lunch yet). I do IHC manually, they just got done cooling off from HIER...I can store them overnight in buffer right? Just double checking myself as this run is an antibody validation run... -S- Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 From rosenfeldtek <@t> hotmail.com Wed Jul 18 13:17:22 2012 From: rosenfeldtek <@t> hotmail.com (Jerry Ricks) Date: Wed Jul 18 13:17:26 2012 Subject: [Histonet] RE: Image analysis for fat tissue In-Reply-To: References: Message-ID: Hi, That is a pretty common kind of measurement. You will need a digital camera attached to your microscope, and a stage micrometer. You can then use the free software "Image-J" to get area measurements. If you have some extra money "Image-Pro" software from Media Cybernetics is the deluxe way to go. Jerry Ricks Research Scientist University of Washington Department of Pathology > From: AteretS@migal.org.il > To: histonet@lists.utsouthwestern.edu > Date: Wed, 18 Jul 2012 11:20:42 +0000 > Subject: [Histonet] Image analysis for fat tissue > > Dear Histonet members > I'm interested in image analysis of mice fat tissue, I would like to quantify number > Of cells per area and average cell size. > Does anyone know a software (+instructions/ lucid manual) which does that? > Many thanks! > > -- Best regards -- > Ateret Shabtay-Yanai > Migal- Galilee Technology Center > Southern Industrial Zone- kiryat Smona > P.O.B 831 Kiryat Shmona 11016 > Tel: +972-46955002 > Mobile: +972-547946460 > Email: aterets@migal.org.il > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rlevier <@t> urologycp.com Wed Jul 18 13:26:14 2012 From: rlevier <@t> urologycp.com (Rebecca LeVier) Date: Wed Jul 18 13:26:24 2012 Subject: [Histonet] Black Biopsy Paper Message-ID: <1D89C917892F914C94DABB8AEEA1E6CC01A6492DD743@ucpaexchange> I am wondering if anyone out there is using black biopsy paper to give to offices to place the biopsy on prior to putting it into formalin? It looks kind of like construction paper. If you are using this can you please give me a list of pros and cons associated with it? Thanks, Becky ________________________________ Disclaimer Note: The information in this message with any attachment to it, may be privileged and protected from disclosure. If the reader of this message is neither the intended recipient, nor an employee or agent responsible for delivering this message to the intended recipient, then you are hereby notified that any dissemination, distribution, unauthorized use, or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ... From TGoins <@t> mt.gov Wed Jul 18 13:35:45 2012 From: TGoins <@t> mt.gov (Goins, Tresa) Date: Wed Jul 18 13:35:53 2012 Subject: [Histonet] RE: Hold IHC overnight In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: I vote No - this run would be valid only if you held all future slides overnight after HIER. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart Sent: Wednesday, July 18, 2012 11:44 AM To: histonet Subject: [Histonet] Hold IHC overnight Just found out I have to go to a meeting in 2 hours for the rest of the day (I haven't even eaten lunch yet). I do IHC manually, they just got done cooling off from HIER...I can store them overnight in buffer right? Just double checking myself as this run is an antibody validation run... -S- 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 From brett_connolly <@t> merck.com Wed Jul 18 14:08:55 2012 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Wed Jul 18 14:09:00 2012 Subject: [Histonet] RE: Black Biopsy Paper In-Reply-To: <1D89C917892F914C94DABB8AEEA1E6CC01A6492DD743@ucpaexchange> References: <1D89C917892F914C94DABB8AEEA1E6CC01A6492DD743@ucpaexchange> Message-ID: Becky, We have used this paper. What I found is that when submersed in the formalin the black dye leaches out of the paper and the portion of the tissue touching the paper turned somewhat black. These were tissues left on the paper and in formalin overnight. Since I never processed that tissue I don't know if the black washes out during processing. Brett Brett M. Connolly, Ph.D. Principal Scientist, Imaging Dept. Merck & Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_connolly@merck.com T- 215-652-2501 F- 215-993-6803 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca LeVier Sent: Wednesday, July 18, 2012 2:26 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Black Biopsy Paper I am wondering if anyone out there is using black biopsy paper to give to offices to place the biopsy on prior to putting it into formalin? It looks kind of like construction paper. If you are using this can you please give me a list of pros and cons associated with it? Thanks, Becky ________________________________ Disclaimer Note: The information in this message with any attachment to it, may be privileged and protected from disclosure. If the reader of this message is neither the intended recipient, nor an employee or agent responsible for delivering this message to the intended recipient, then you are hereby notified that any dissemination, distribution, unauthorized use, or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. From erinandjesseconley <@t> yahoo.com Wed Jul 18 14:35:13 2012 From: erinandjesseconley <@t> yahoo.com (Erin and Jesse) Date: Wed Jul 18 14:35:35 2012 Subject: [Histonet] RE: Hold IHC overnight In-Reply-To: References: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: <1D302247-DFD3-4507-BA75-427ED98453DE@yahoo.com> I agree On Jul 18, 2012, at 2:35 PM, "Goins, Tresa" wrote: > I vote No - this run would be valid only if you held all future slides overnight after HIER. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart > Sent: Wednesday, July 18, 2012 11:44 AM > To: histonet > Subject: [Histonet] Hold IHC overnight > > Just found out I have to go to a meeting in 2 hours for the rest of the day (I haven't even eaten lunch yet). I do IHC manually, they just got done cooling off from HIER...I can store them overnight in buffer right? > Just double checking myself as this run is an antibody validation run... > -S- > > 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 From lblazek <@t> digestivespecialists.com Wed Jul 18 14:40:31 2012 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Wed Jul 18 14:42:03 2012 Subject: [Histonet] RE: Hold IHC overnight In-Reply-To: <1D302247-DFD3-4507-BA75-427ED98453DE@yahoo.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> <1D302247-DFD3-4507-BA75-427ED98453DE@yahoo.com> Message-ID: <5A2BD13465E061429D6455C8D6B40E3913E9A776BC@IBMB7Exchange.digestivespecialists.com> But only because it's a validation run. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Erin and Jesse Sent: Wednesday, July 18, 2012 3:35 PM To: Goins, Tresa Cc: histonet Subject: Re: [Histonet] RE: Hold IHC overnight I agree On Jul 18, 2012, at 2:35 PM, "Goins, Tresa" wrote: > I vote No - this run would be valid only if you held all future slides overnight after HIER. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart > Sent: Wednesday, July 18, 2012 11:44 AM > To: histonet > Subject: [Histonet] Hold IHC overnight > > Just found out I have to go to a meeting in 2 hours for the rest of the day (I haven't even eaten lunch yet). I do IHC manually, they just got done cooling off from HIER...I can store them overnight in buffer right? > Just double checking myself as this run is an antibody validation run... > -S- > > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rosenfeldtek <@t> hotmail.com Wed Jul 18 14:50:52 2012 From: rosenfeldtek <@t> hotmail.com (Jerry Ricks) Date: Wed Jul 18 14:50:58 2012 Subject: [Histonet] RE: Hold IHC overnight In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D501077F4C5@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: No you can't do that. If it were just a research IHC then you could give it a whirl, write down what you did and often it would work just fine depending on antigen and antibody. But if you are validating an antibody for clinical lab you have to document you follow GLP SOP every time else you messed up. There's no getting around it. You are just going to have to run the IHC stain again. Jerry Ricks > From: sdysart@mirnarx.com > To: histonet@lists.utsouthwestern.edu > Date: Wed, 18 Jul 2012 17:44:12 +0000 > Subject: [Histonet] Hold IHC overnight > > Just found out I have to go to a meeting in 2 hours for the rest of the day (I haven't even eaten lunch yet). I do IHC manually, they just got done cooling off from HIER...I can store them overnight in buffer right? > Just double checking myself as this run is an antibody validation run... > -S- > > 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 From hardin <@t> oncology.wisc.edu Wed Jul 18 15:23:07 2012 From: hardin <@t> oncology.wisc.edu (Joe Hardin) Date: Wed Jul 18 15:23:12 2012 Subject: [Histonet] automated H+E and cover slipper Message-ID: <50071B2B.7000607@oncology.wisc.edu> Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. From Sandra.Harrison3 <@t> va.gov Wed Jul 18 15:44:12 2012 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Wed Jul 18 15:44:35 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: <50071B2B.7000607@oncology.wisc.edu> References: <50071B2B.7000607@oncology.wisc.edu> Message-ID: Leica Autostainer XL with CV5030 coverslipper and transfer station. This has been a real timesaver for us. It automatically moves the slides from the stainer to the coverslipper. It has been relatively trouble free. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 3:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. From liz <@t> premierlab.com Wed Jul 18 15:49:37 2012 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Wed Jul 18 15:49:42 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: <50071B2B.7000607@oncology.wisc.edu> Message-ID: <14E2C6176416974295479C64A11CB9AE01310A23EA8F@SBS2K8.premierlab.local> We have a Sakura Tissue Tek Prisma and Glas coverslipper we really like it. It's a workhorse. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 2:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. From BDeBrosse-Serra <@t> isisph.com Wed Jul 18 16:18:53 2012 From: BDeBrosse-Serra <@t> isisph.com (Bea DeBrosse-Serra) Date: Wed Jul 18 16:19:05 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: References: <50071B2B.7000607@oncology.wisc.edu> Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FC3B@EXCHMB01.isis.local> I second on that! Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Harrison, Sandra C. Sent: Wednesday, July 18, 2012 1:44 PM To: Joe Hardin; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] automated H+E and cover slipper Leica Autostainer XL with CV5030 coverslipper and transfer station. This has been a real timesaver for us. It automatically moves the slides from the stainer to the coverslipper. It has been relatively trouble free. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 3:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Wed Jul 18 16:24:00 2012 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Wed Jul 18 16:23:13 2012 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: It is added to formaldehyde to prevent polymerization of the formaldehyde. Bader Siddiki Sent by: histonet-bounces@lists.utsouthwestern.edu 07/17/2012 01:44 PM To Rene J Buesa cc histonet , Kelly Boyd Subject Re: [Histonet] (no subject) Hello Methanol is added to reagent alcohol (ethyl alcohol) so that people do not use for drinking purpose and for tax purposes. Alcohol (ethyl alcohol) has federal tax just like liquor. One can buy pure alcohol, but you have keep inventory and make sure it is not misused. Bader On Tue, Jul 17, 2012 at 1:28 PM, Rene J Buesa wrote: > To prevent oxidation of those 2 reagents > Ren? J. > > > ________________________________ > From: Kelly Boyd > To: histonet > Sent: Tuesday, July 17, 2012 1:16 PM > Subject: [Histonet] (no subject) > > Since there is discussion of formalin and methanol, I would like to ask > everyone.....Why would a vendor add methanol to their reagent alcohol and > their 10% buffered formalin? > > What are the advantages/disadvantages to the tissue especially for > processing? > > Kelly > _______________________________________________ > 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 > -- If any Q's please feel free to contact us Have a nice day/weekend Mit freundlichen Gr??en / With Kind Regards / avec l'aimable ce qui concerne Met vriendelijke groeten ?????? Bader Bader B Siddiki, PhD Executive director, Research and development ImmunoBioScience Corp. (IBSC) Phone: + 1 425 367 4601 Fax: + 1 425 367 4817 cell (mobile) phone: + 1 425 314 0199 e-mail address: BaderBo@Gmail.com Web site: www.ImmunoBioScience.Com Marketing: phone: + 1 650 343 IBSC (4272) E-mail: AnitaIBSC@Aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wdesalvo.cac <@t> hotmail.com Wed Jul 18 17:30:53 2012 From: wdesalvo.cac <@t> hotmail.com (WILLIAM DESALVO) Date: Wed Jul 18 17:31:03 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: <493CAA64F203E14E8823737B9EE0E25F092BA8FC3B@EXCHMB01.isis.local> References: <50071B2B.7000607@oncology.wisc.edu>, , <493CAA64F203E14E8823737B9EE0E25F092BA8FC3B@EXCHMB01.isis.local> Message-ID: Sakura Prisma w/ tape cover slip. It also is available w/ glass cover slip. Reliable, fast and a workhorse. William DeSalvo, B.S., HTL(ASCP) > From: BDeBrosse-Serra@isisph.com > To: Sandra.Harrison3@va.gov; hardin@oncology.wisc.edu; histonet@lists.utsouthwestern.edu > Date: Wed, 18 Jul 2012 14:18:53 -0700 > Subject: RE: [Histonet] automated H+E and cover slipper > CC: > > I second on that! > > Beatrice DeBrosse-Serra HT(ASCP)QIHC > Isis Pharmaceuticals > Antisense Drug Discovery > 2855 Gazelle Ct. > Carlsbad, CA 92010 > 760-603-2371 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Harrison, Sandra C. > Sent: Wednesday, July 18, 2012 1:44 PM > To: Joe Hardin; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] automated H+E and cover slipper > > Leica Autostainer XL with CV5030 coverslipper and transfer station. > > This has been a real timesaver for us. It automatically moves the slides from the stainer to the coverslipper. It has been relatively trouble free. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin > Sent: Wednesday, July 18, 2012 3:23 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] automated H+E and cover slipper > > Hi All, > I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. > > > _______________________________________________ > 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 From g.vanhaalem <@t> prosensa.nl Thu Jul 19 02:37:02 2012 From: g.vanhaalem <@t> prosensa.nl (Geert van Haalem) Date: Thu Jul 19 02:37:18 2012 Subject: [Histonet] Re: Image analysis for fat tissue Message-ID: <5F4E852313B6BC4B8C9916B69BAB4C1E30968347@pro-ex02> The software ImageJ is commonly used program for doing image analysis with lots of macros and plugins available. For your purpose I guess you need good recognition of individual cells by the program with the proper segmentation algorithm and tissue staining. PubMed ID : 21245003 -----Original Message----- Message: 10 Date: Wed, 18 Jul 2012 11:20:42 +0000 From: Ateret Shabtay-Yanai Subject: [Histonet] Image analysis for fat tissue To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="us-ascii" Dear Histonet members I'm interested in image analysis of mice fat tissue, I would like to quantify number Of cells per area and average cell size. Does anyone know a software (+instructions/ lucid manual) which does that? Many thanks! -- Best regards -- Ateret Shabtay-Yanai Migal- Galilee Technology Center Southern Industrial Zone- kiryat Smona P.O.B 831 Kiryat Shmona 11016 Tel: +972-46955002 Mobile: +972-547946460 Email: aterets@migal.org.il ------------------------------ From b-frederick <@t> northwestern.edu Thu Jul 19 07:26:01 2012 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Jul 19 07:26:06 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: References: <50071B2B.7000607@oncology.wisc.edu> Message-ID: <62C639732D3F274DACED033EBDF6ADAF1E272466@evcspmbx3.ads.northwestern.edu> I second this!!! Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-frederick@northwestern.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Harrison, Sandra C. Sent: Wednesday, July 18, 2012 3:44 PM To: Joe Hardin; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] automated H+E and cover slipper Leica Autostainer XL with CV5030 coverslipper and transfer station. This has been a real timesaver for us. It automatically moves the slides from the stainer to the coverslipper. It has been relatively trouble free. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 3:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgrobert <@t> rci.rutgers.edu Thu Jul 19 09:17:00 2012 From: kgrobert <@t> rci.rutgers.edu (kgrobert@rci.rutgers.edu) Date: Thu Jul 19 09:17:07 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: References: <50071B2B.7000607@oncology.wisc.edu> Message-ID: <79cc411b49a6556ac697985edcf896d3.squirrel@webmail.rci.rutgers.edu> I second that! I have an older model of the Leica stainer, but it's still great. Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Molecular Pathology Facility Core Dept of Pharmacology & Toxicology Rutgers, the State University of NJ 41 B Gordon Road Piscataway, NJ 08854 (732) 445-6914 FAX (732) 445-6905 > Leica Autostainer XL with CV5030 coverslipper and transfer station. > > This has been a real timesaver for us. It automatically moves the > slides from the stainer to the coverslipper. It has been relatively > trouble free. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe > Hardin > Sent: Wednesday, July 18, 2012 3:23 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] automated H+E and cover slipper > > Hi All, > I will be trying out new H+E autostainers and cover slippers soon. Does > anyone have a favorite, and why? Thanks for your responses. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Joyce.Weems <@t> emoryhealthcare.org Thu Jul 19 10:38:53 2012 From: Joyce.Weems <@t> emoryhealthcare.org (Weems, Joyce K.) Date: Thu Jul 19 10:39:03 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: <62C639732D3F274DACED033EBDF6ADAF1E272466@evcspmbx3.ads.northwestern.edu> References: <50071B2B.7000607@oncology.wisc.edu> <62C639732D3F274DACED033EBDF6ADAF1E272466@evcspmbx3.ads.northwestern.edu> Message-ID: Me too! Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Thursday, July 19, 2012 8:26 AM To: Harrison, Sandra C.; Joe Hardin; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] automated H+E and cover slipper I second this!!! Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-frederick@northwestern.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Harrison, Sandra C. Sent: Wednesday, July 18, 2012 3:44 PM To: Joe Hardin; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] automated H+E and cover slipper Leica Autostainer XL with CV5030 coverslipper and transfer station. This has been a real timesaver for us. It automatically moves the slides from the stainer to the coverslipper. It has been relatively trouble free. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 3:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. _______________________________________________ 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 ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). From mucram11 <@t> comcast.net Thu Jul 19 10:47:49 2012 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Thu Jul 19 10:48:00 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: Message-ID: <1119091903.57992.1342712869410.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> We have two Leica stainers and love them however; we are looking at the Dako Coverstainer to replace them for ease of use and reagent savings.? We don't have the transfer station on the Leicas , ?just the separate coverslipper , so it is not quite as efficient as we would like it to be.? Still Leicas are work horses and really excellent stainers. ? Pam Marcum UAMS ----- Original Message ----- From: "Joyce K. Weems " To: "Bernice Frederick" , "Sandra C. Harrison" , "Joe Hardin" < hardin @oncology. wisc . edu >, histonet @lists. utsouthwestern . edu Sent: Thursday, July 19, 2012 10:38:53 AM Subject: RE: [ Histonet ] automated H+E and cover slipper Me too! Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce . weems @ emoryhealthcare .org www . saintjosephsatlanta .org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). ?It may contain information that is privileged and confidential. ?Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet -bounces@lists. utsouthwestern . edu [ mailto : histonet -bounces@lists. utsouthwestern . edu ] On Behalf Of Bernice Frederick Sent: Thursday, July 19, 2012 8:26 AM To: Harrison, Sandra C.; Joe Hardin; histonet @lists. utsouthwestern . edu Subject: RE: [ Histonet ] automated H+E and cover slipper I second this!!! Bernice Frederick HTL ( ASCP ) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-frederick@northwestern. edu -----Original Message----- From: histonet -bounces@lists. utsouthwestern . edu [ mailto : histonet -bounces@lists. utsouthwestern . edu ] On Behalf Of Harrison, Sandra C. Sent: Wednesday, July 18, 2012 3:44 PM To: Joe Hardin; histonet @lists. utsouthwestern . edu Subject: RE: [ Histonet ] automated H+E and cover slipper Leica Autostainer XL with CV5030 coverslipper and transfer station. This has been a real timesaver for us. ?It automatically moves the slides from the stainer to the coverslipper . ?It has been relatively trouble free. -----Original Message----- From: histonet -bounces@lists. utsouthwestern . edu [ mailto : histonet -bounces@lists. utsouthwestern . edu ] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 3:23 PM To: histonet @lists. utsouthwestern . edu Subject: [ Histonet ] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. _______________________________________________ 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 ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). _______________________________________________ Histonet mailing list Histonet @lists. utsouthwestern . edu http ://lists. utsouthwestern . edu /mailman/ listinfo / histonet From HornHV <@t> archildrens.org Thu Jul 19 10:54:40 2012 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Thu Jul 19 10:54:49 2012 Subject: [Histonet] automated H+E and cover slipper In-Reply-To: References: <50071B2B.7000607@oncology.wisc.edu> Message-ID: <25A4DE08332B19499904459F00AAACB719BB4A172C@EVS1.archildrens.org> Same here for us... Hazel Horn Supervisor of Histology/Autopsy/Transcription Anatomic Pathology Arkansas Children's Hospital 1 Children's Way | Slot 820| Little Rock, AR 72202 501.364.4240 direct | 501.364.1302 office | 501.364.1241 fax hornhv@archildrens.org archildrens.org 100 YEARS YOUNG! JOIN THE PARTY AT ach100.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Harrison, Sandra C. Sent: Wednesday, July 18, 2012 3:44 PM To: Joe Hardin; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] automated H+E and cover slipper Leica Autostainer XL with CV5030 coverslipper and transfer station. This has been a real timesaver for us. It automatically moves the slides from the stainer to the coverslipper. It has been relatively trouble free. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Hardin Sent: Wednesday, July 18, 2012 3:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated H+E and cover slipper Hi All, I will be trying out new H+E autostainers and cover slippers soon. Does anyone have a favorite, and why? Thanks for your responses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ****************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************** The information contained in this message may be privileged and confidential and protected from disclosure. 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 dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. From YJiang <@t> isisph.com Thu Jul 19 13:06:36 2012 From: YJiang <@t> isisph.com (Yuhong Jiang) Date: Thu Jul 19 13:06:47 2012 Subject: [Histonet] Problem for uneven staining for using QuantiGene View RNA Assay kit for FFPE ISH from Affymetrix Message-ID: Hi Histonetter: Does anyone have the experience for using QuantiGen View RNA ISH from Affymetrix. I got the problem for uneven staining sometimes and cannot figure out why. Thank you, Yuhong Jiang Senior Research Associate ISIS Pharmaceuticals 2855 GAZELLE CT San Diego, Carlsbad CA,92010 Email: yjiang@isisph.com Phone: 760-603-3561 From kblack <@t> digestivehlth.com Thu Jul 19 14:13:14 2012 From: kblack <@t> digestivehlth.com (Konni Black) Date: Thu Jul 19 14:13:25 2012 Subject: [Histonet] New HT/HTL Positions in Bremerton, Washington Message-ID: <3E4EA095A7B79A4DB1722FF5F90CE86B53C401@BL2PRD0810MB361.namprd08.prod.outlook.com> Hi All, There are two new HT/HTL opportunities in a large multi-specialty practice in Bremerton, WA. One fulltime opening for an HT or HTL with 3 to 5 years of experience and one part-time opening for an experienced or new graduate HT or HTL. Both must meet CLIA education requirements to perform gross (minimum AA degree). Lab opening scheduled early October 2012 Bremerton is located on the Washington Peninsula across Puget Sound from Seattle (via ferry). Great location for all types of outdoor activities and great coffee! If you are interested, please contact me at kblack@digestivehlth.com. Thank you, Konni Black From cjbulmer <@t> sbcglobal.net Thu Jul 19 14:25:28 2012 From: cjbulmer <@t> sbcglobal.net (Cindy Bulmer) Date: Thu Jul 19 14:25:31 2012 Subject: [Histonet] p53, control Message-ID: <1342725928.78482.YahooMailClassic@web184504.mail.ne1.yahoo.com> What type of tissue is everyone using for a positive control for this Ab, and are you using Mouse or Rabbit Ab? Thanks, Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX From tkngflght <@t> yahoo.com Thu Jul 19 14:31:32 2012 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Jul 19 14:31:35 2012 Subject: [Histonet] Temp Travelers Needed Message-ID: <1342726292.52030.YahooMailNeo@web39406.mail.mud.yahoo.com> Hi All- ? Ever thought of temping?? In between jobs or looking for a new adventure?? ? We're always on the lookout for good flexible techs that can make a difference for facilities that need some help.? We have a couple of assignments available if you're ready now.? If you're just curious, we'll spend a little time to answer your questions and keep you posted as new opportunities come along. ? I'm a working traveler--benched two weeks ago at a favorite client--so I do know the drill.? Happy to share--give us a call! ? Cheryl Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one GREAT?Tech at a time.? 281.852.9457?Office 800.756.3309?Phone & Fax? admin@fullstaff.org Sign up for the FREE?newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please?include your name and specialty in the body of the email. From matt <@t> ka-recruiting.com Thu Jul 19 14:58:39 2012 From: matt <@t> ka-recruiting.com (Matthew Mosca) Date: Thu Jul 19 14:58:42 2012 Subject: [Histonet] KA Recruiting- Great Histology opportunities!!! Message-ID: <415918181.1342727918883.JavaMail.cfservice@sl14app4> Dear Histonet Community, I work for a nationally recognized healthcare recruiting firm, specializing in helping Lab Professionals find permanent employment. I wanted to reach out to you today and see if you are interested in taking the next step in your career. Our clients typically assist with relocation expenses. I am currently working on some great positions that you may be interested in including a Lead Histotech position with a hospital in Northwest Ohio. This is a 1st shift Histotechnologist position at one of Ohio's most highly regarded hospital networks. Recognized for superior patient satisfaction at this 113 bed facility, and a state of the art heart catheterization laboratory offers an excellent work environment. This is a position offers a terrific compensation package; including competitive hourly rate, and retirement plan. To qualify you must have an AS or BS in Histology and be either HT or HTL (ASCP) certified. Working knowledge of Tissue Processors, Micrometers, Immunohistochemical and Special Staining Instrumentation is required. If you are interested in learning more about this position, please call or e-mail me at matt@ka-recruiting.com Below is a list of some of the other opportunities we are currently working on. If you do not see an opening in a location in which you live or would like to live, please send me an email with a copy of your resume and let me know where you would be interested in a job. I will then tailor a search for you that is completely confidential. Histotech/Cytotech Openings: * PA - Philadelphia - IHC Tech * ME - Histotechnologist HT or HTL * NC - Histology Manager * KY - Histotechnologist - 3rd shift * NY - Western - Histotech * NY - NYC - Histotech * NYC - Pathology Manager (commercial background) * OH - Columbus - Histotech - 1st shift * OH - Columbus- Lead Histotech * NV - Las Vegas - Histotech, IHC * CA - Southern - Histology Manager * CA - Southern - Histolgy Supervisor * CA - Southern - Histotech Sincerely, Matt Mosca K.A. Recruiting, Inc. 10 Post Office Square, 8th Flr, Boston, MA 02109 P: (617) 692-2949 F: (617) 507-8009 Matthew@ka-recruiting.com www.ka-recruiting.com From marktarango <@t> gmail.com Thu Jul 19 16:26:51 2012 From: marktarango <@t> gmail.com (Mark Tarango) Date: Thu Jul 19 16:26:56 2012 Subject: [Histonet] p53, control In-Reply-To: <1342725928.78482.YahooMailClassic@web184504.mail.ne1.yahoo.com> References: <1342725928.78482.YahooMailClassic@web184504.mail.ne1.yahoo.com> Message-ID: Hi Cindy, The one from Dako works well at a 1:200 dilution on the Ventana platforms (catalog #M7001). We use HIGH GRADE ovarian serous carcinoma as a positive control. You should get intense nuclear staining throughout the tumor with this control. Mark On Thu, Jul 19, 2012 at 12:25 PM, Cindy Bulmer wrote: > What type of tissue is everyone using for a positive control for this Ab, > and are you using Mouse or Rabbit Ab? > > Thanks, > Cindy > > Cynthia Bulmer HT(ASCP)QIHC > > IHC Supervisor, CTPL > > Waco, TX > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From chak_bou <@t> yahoo.com Thu Jul 19 18:04:09 2012 From: chak_bou <@t> yahoo.com (Chakib Boussahmain) Date: Thu Jul 19 18:04:13 2012 Subject: [Histonet] Perilipin antibody Message-ID: <1342739049.53094.YahooMailClassic@web160801.mail.bf1.yahoo.com> Hi Histonetters, Does anyone using?Perilipin antibody? if so, can you share with me the staining protocol? Which clone are you using? Vendor? Any input will be greatly appreciated! Thank you! Chakib Boussahmain HTL(ASCP) MGH From contact <@t> histocare.com Thu Jul 19 18:11:19 2012 From: contact <@t> histocare.com (Contact HistoCare) Date: Thu Jul 19 18:11:27 2012 Subject: [Histonet] RE: automated H+E and cover slipper Message-ID: Having used several including the Leica XL and the Sakura Stainer/coverslipper, unequivocally the Sakura combo is better. The Leica stainer works without fail as it should but the coverslipper seems ancient compared to the Sakura. There are more errors with bubbles, misaligned glass coverslip. The Leica also begins with one rack but it transfers to another rack once it makes it to the coverslipper which holds about three in a vertical stack of 30 slides each. You have to watch this more and is more sensitive to slide placement in a rack. It will try to coverslip an empty slot. Sakura on the other hand maintains the same rack throughout both the stain and coverslip process. The cover tape is EASY! Comes on a roll and is precisely cut each time. There is a fast actuator-type arm that checks for slides in a rack before attempting to coverslip. Once complete, they are loaded to a carousel-style resting station above awaiting to be taken off. This resting area will hold about 10 (it may be more but can't recall) racks of 20 slides each. Hope this helps. Need reliable Histology Professionals for a temporary gig? Visit us at HistoCare.com From Milton.Gomez <@t> nyumc.org Thu Jul 19 21:07:27 2012 From: Milton.Gomez <@t> nyumc.org (Gomez, Milton) Date: Thu Jul 19 21:07:37 2012 Subject: [Histonet] collecting chemical waste Message-ID: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Hello histonetters, Do you folks enforce placing a label on a waste chemical container before collecting any waste or do you allow the label to be placed on the waste container after the waste is collected? I was taught to place the label on the waste container even before the first drop of waste is collected. MG From MSHERWOOD <@t> PARTNERS.ORG Fri Jul 20 07:52:05 2012 From: MSHERWOOD <@t> PARTNERS.ORG (Sherwood, Margaret) Date: Fri Jul 20 07:52:12 2012 Subject: [Histonet] RE: collecting chemical waste In-Reply-To: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Message-ID: <090FA56107A969459F3941DDD5585C3A1184641F@PHSX10MB10.partners.org> We have to have any container labelled (waste or reagent). We only date the container when it is full and ready for pick up. Peggy Sherwood Research Specialist, Photopathology Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114-2696 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-1206 (fax) msherwood@partners.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gomez, Milton Sent: Thursday, July 19, 2012 10:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] collecting chemical waste Hello histonetters, Do you folks enforce placing a label on a waste chemical container before collecting any waste or do you allow the label to be placed on the waste container after the waste is collected? I was taught to place the label on the waste container even before the first drop of waste is collected. MG _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. From geralddavydov <@t> gmail.com Fri Jul 20 08:17:28 2012 From: geralddavydov <@t> gmail.com (Gerald Davydov) Date: Fri Jul 20 08:17:35 2012 Subject: [Histonet] p16 Message-ID: P16 stain demonstrates week nuclear stain. Any ideas how to correct? From Joyce.Weems <@t> emoryhealthcare.org Fri Jul 20 09:36:02 2012 From: Joyce.Weems <@t> emoryhealthcare.org (Weems, Joyce K.) Date: Fri Jul 20 09:36:20 2012 Subject: [Histonet] RE: collecting chemical waste In-Reply-To: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Message-ID: Before Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 5665 Peachtree Dunwoody Road Atlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gomez, Milton Sent: Thursday, July 19, 2012 10:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] collecting chemical waste Hello histonetters, Do you folks enforce placing a label on a waste chemical container before collecting any waste or do you allow the label to be placed on the waste container after the waste is collected? I was taught to place the label on the waste container even before the first drop of waste is collected. MG _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). From b427297 <@t> aol.com Fri Jul 20 09:47:29 2012 From: b427297 <@t> aol.com (Jackie O'Connor) Date: Fri Jul 20 09:47:54 2012 Subject: [Histonet] RE: collecting chemical waste In-Reply-To: References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Message-ID: <8CF349DB88195AE-804-2134@webmail-d014.sysops.aol.com> If you don't place a label on it before filling it, how do you know what is supposed to go in it? We have separate waste containers for Xylene, Formalin and alcohol. JO'C -----Original Message----- From: Weems, Joyce K. To: 'Gomez, Milton' ; histonet Sent: Fri, Jul 20, 2012 9:36 am Subject: [Histonet] RE: collecting chemical waste Before Joyce Weems athology Manager 78-843-7376 Phone 78-843-7831 Fax oyce.weems@emoryhealthcare.org www.saintjosephsatlanta.org 665 Peachtree Dunwoody Road tlanta, GA 30342 This e-mail, including any attachments is the property of Saint Joseph's ospital and is intended for the sole use of the intended recipient(s). It may ontain information that is privileged and confidential. Any unauthorized eview, use, disclosure, or distribution is prohibited. If you are not the ntended recipient, please delete this message, and reply to the sender egarding the error in a separate email. -----Original Message----- rom: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] n Behalf Of Gomez, Milton ent: Thursday, July 19, 2012 10:07 PM o: histonet@lists.utsouthwestern.edu ubject: [Histonet] collecting chemical waste Hello histonetters, Do you folks enforce placing a label on a waste chemical container before ollecting any waste or do you allow the label to be placed on the waste ontainer after the waste is collected? I was taught to place the label on the aste container even before the first drop of waste is collected. MG ______________________________________________ istonet mailing list istonet@lists.utsouthwestern.edu ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ This e-mail message (including any attachments) is for the sole use of he intended recipient(s) and may contain confidential and privileged nformation. If the reader of this message is not the intended ecipient, you are hereby notified that any dissemination, distribution r copying of this message (including any attachments) is strictly rohibited. If you have received this message in error, please contact he sender by reply e-mail message and destroy all copies of the riginal message (including attachments). _______________________________________________ istonet mailing list istonet@lists.utsouthwestern.edu ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linda.Margraf <@t> cookchildrens.org Fri Jul 20 09:48:15 2012 From: Linda.Margraf <@t> cookchildrens.org (Linda Margraf) Date: Fri Jul 20 09:48:25 2012 Subject: [Histonet] PA job at MD Anderson Message-ID: <928719B9EBFA1C4686918B975FF845285A2534DB@CCHCSMBX04.CCHCS.LDAP> From: "Barr,Kaye H" > Date: July 19, 2012 11:37:43 AM CDT Subject: PA Position The University of Texas MD Anderson Cancer Center in Houston, Texas has a vacancy for a pathologist assistant. This is a M-F position, no weekends, no holidays. Req#5030, recruiter contact info: Nikol Blackmon, 713-745-6349. Please click on the provided link to apply. https://2xrecruit.kenexa.com/kr/cc/jsp/public/jobSearchResults.jsf Kaye Barr, HT (ASCP) | Laboratory Manager M. D. Anderson Cancer Center | Pathology/Histology Labs 1515 Holcombe Blvd, Unit 85 | Houston, TX 77030 713.792.5366 office | khbarr@mdanderson.org From Terri.Brown <@t> Northside.com Fri Jul 20 09:54:54 2012 From: Terri.Brown <@t> Northside.com (Terri Brown) Date: Fri Jul 20 09:55:05 2012 Subject: [Histonet] ThermoScientific Printmate Message-ID: <731941C266951A47BEF11E5EFAAED9C9127EA80F@nsmvexch01.northside.local> Who has the Newer versions of this product that is interfaced with CoPath V4.1 for accessioning, printing cassettes and slides? Thanks, Terri H. Brown,, HT (ASCP) Pathology Laboratory Manager Northside Hospital Atlanta terri.brown@northside.com CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. From Timothy.Morken <@t> ucsfmedctr.org Fri Jul 20 10:30:03 2012 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Timothy) Date: Fri Jul 20 10:29:30 2012 Subject: [Histonet] RE: collecting chemical waste In-Reply-To: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Message-ID: <8D7C2D242DBD45498006B21122072BF8B5D87043@MCINFRWEM003.ucsfmedicalcenter.org> Tag must be on the container once any waste is put in it. So, first use, tag it. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gomez, Milton Sent: Thursday, July 19, 2012 7:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] collecting chemical waste Hello histonetters, Do you folks enforce placing a label on a waste chemical container before collecting any waste or do you allow the label to be placed on the waste container after the waste is collected? I was taught to place the label on the waste container even before the first drop of waste is collected. MG _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LPaveli1 <@t> hurleymc.com Fri Jul 20 10:42:40 2012 From: LPaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Fri Jul 20 10:42:46 2012 Subject: [Histonet] RE: collecting chemical waste In-Reply-To: <8D7C2D242DBD45498006B21122072BF8B5D87043@MCINFRWEM003.ucsfmedicalcenter.org> References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org>, <8D7C2D242DBD45498006B21122072BF8B5D87043@MCINFRWEM003.ucsfmedicalcenter.org> Message-ID: <89F4666A496DC949A819ECC40E11C867B2A142B1@EXCHANGEMB2.hmc.hurleymc.com> >From the start. Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Morken, Timothy [Timothy.Morken@ucsfmedctr.org] Sent: Friday, July 20, 2012 11:30 AM To: Gomez, Milton; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: collecting chemical waste Tag must be on the container once any waste is put in it. So, first use, tag it. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gomez, Milton Sent: Thursday, July 19, 2012 7:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] collecting chemical waste Hello histonetters, Do you folks enforce placing a label on a waste chemical container before collecting any waste or do you allow the label to be placed on the waste container after the waste is collected? I was taught to place the label on the waste container even before the first drop of waste is collected. MG _______________________________________________ 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 From Rcartun <@t> harthosp.org Fri Jul 20 11:16:43 2012 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Jul 20 11:16:54 2012 Subject: [Histonet] p16 In-Reply-To: References: Message-ID: <50094C2A.7400.0077.1@harthosp.org> In order to troubleshoot, one would need to know the following: Test tissue Fixation Antibody clone, dilution, and incubation time Detection system Antigen retrieval Positive control tissue Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax >>> Gerald Davydov 7/20/2012 9:17 AM >>> P16 stain demonstrates week nuclear stain. Any ideas how to correct? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shive003 <@t> umn.edu Fri Jul 20 11:48:15 2012 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Jul 20 11:48:21 2012 Subject: [Histonet] Olig2 Message-ID: I'm asking for a colleague - Does anyone know a lab that does IHC staining for Olig2? Thanks in advance, Jan Shivers Univ. of Minnesota Veterinary Diagnostic Lab St. Paul, MN shive003@umn.edu From JCBRITTON <@t> Cheshire-Med.COM Fri Jul 20 11:57:31 2012 From: JCBRITTON <@t> Cheshire-Med.COM (Britton, Josette C) Date: Fri Jul 20 11:57:37 2012 Subject: [Histonet] p16 In-Reply-To: <50094C2A.7400.0077.1@harthosp.org> Message-ID: Where is everyone getting their p16 IVD antibody from? Josie Britton HT Cheshire Medical Center 580 Court Street Keene, NH 03431 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Friday, July 20, 2012 12:17 PM To: Gerald Davydov; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] p16 In order to troubleshoot, one would need to know the following: Test tissue Fixation Antibody clone, dilution, and incubation time Detection system Antigen retrieval Positive control tissue Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax >>> Gerald Davydov 7/20/2012 9:17 AM >>> P16 stain demonstrates week nuclear stain. Any ideas how to correct? _______________________________________________ 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 From Loralee_Mcmahon <@t> URMC.Rochester.edu Fri Jul 20 12:01:33 2012 From: Loralee_Mcmahon <@t> URMC.Rochester.edu (McMahon, Loralee A) Date: Fri Jul 20 12:02:18 2012 Subject: [Histonet] p16 In-Reply-To: References: <50094C2A.7400.0077.1@harthosp.org>, Message-ID: Roche/Ventana. they bought MTM labs and then raised the price. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Britton, Josette C [JCBRITTON@Cheshire-Med.COM] Sent: Friday, July 20, 2012 12:57 PM To: Richard Cartun; Gerald Davydov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] p16 Where is everyone getting their p16 IVD antibody from? Josie Britton HT Cheshire Medical Center 580 Court Street Keene, NH 03431 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Friday, July 20, 2012 12:17 PM To: Gerald Davydov; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] p16 In order to troubleshoot, one would need to know the following: Test tissue Fixation Antibody clone, dilution, and incubation time Detection system Antigen retrieval Positive control tissue Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax >>> Gerald Davydov 7/20/2012 9:17 AM >>> P16 stain demonstrates week nuclear stain. Any ideas how to correct? _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From 41dmb41 <@t> gmail.com Fri Jul 20 13:03:36 2012 From: 41dmb41 <@t> gmail.com (Drew Meyer) Date: Fri Jul 20 13:03:43 2012 Subject: [Histonet] CD200 Message-ID: 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 From patrick.lewis <@t> seattlechildrens.org Fri Jul 20 17:22:23 2012 From: patrick.lewis <@t> seattlechildrens.org (Lewis, Patrick) Date: Fri Jul 20 17:22:33 2012 Subject: [Histonet] Antibodies to Mast cells for IHC help Message-ID: <3903BE18914F4440834F0E620415FFCA0BCC7D8D@PPWEXD01B.childrens.sea.kids> Hi everyone. Would anyone be able anyone recommend a good antibody to stain mast cells in Human Tonsil for IHC? These would be in FFPE (Formalin fixed paraffin embedded sections) With secondary antibody hopefully being HRP labeled. Substrate would most likely be DAKO AEC. Thanks Patrick Lewis PS: If you have done this staining in the past and have a protocol, or antigen retrieval suggestions that would also be helpful. CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From patrick.lewis <@t> seattlechildrens.org Fri Jul 20 17:28:23 2012 From: patrick.lewis <@t> seattlechildrens.org (Lewis, Patrick) Date: Fri Jul 20 17:28:36 2012 Subject: [Histonet] Antibodies to mast cells in human tonsil Message-ID: <3903BE18914F4440834F0E620415FFCA0BCC7D99@PPWEXD01B.childrens.sea.kids> Hi everyone, That should read, Would anyone able to recommend a good antibody to stain for only mast cells in human tonsil for IHC. Thanks in advance. Patrick Lewis CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From liz <@t> premierlab.com Fri Jul 20 17:41:58 2012 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Jul 20 17:42:03 2012 Subject: [Histonet] RE: Antibodies to Mast cells for IHC help In-Reply-To: <3903BE18914F4440834F0E620415FFCA0BCC7D8D@PPWEXD01B.childrens.sea.kids> Message-ID: <14E2C6176416974295479C64A11CB9AE01310A23EAF9@SBS2K8.premierlab.local> Mast Cell Tryptase, I have used the one from Dako before, it works on human and other species. Toluidine blue will also work nicely, with air drying and coverslip in xylene. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lewis, Patrick Sent: Friday, July 20, 2012 4:22 PM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] Antibodies to Mast cells for IHC help Hi everyone. Would anyone be able anyone recommend a good antibody to stain mast cells in Human Tonsil for IHC? These would be in FFPE (Formalin fixed paraffin embedded sections) With secondary antibody hopefully being HRP labeled. Substrate would most likely be DAKO AEC. Thanks Patrick Lewis PS: If you have done this staining in the past and have a protocol, or antigen retrieval suggestions that would also be helpful. CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ashley.Troutman <@t> Vanderbilt.Edu Fri Jul 20 18:35:30 2012 From: Ashley.Troutman <@t> Vanderbilt.Edu (Troutman, Kenneth A) Date: Fri Jul 20 18:35:42 2012 Subject: [Histonet] p16 Message-ID: <7B310892042DA74CB3590053F424CFE61463BCFD94@ITS-HCWNEM06.ds.Vanderbilt.edu> The only IVD p16 available in the US is from MTM. However, Roche just bought MTM so you will have to order it through Ventana. Their Customer support number is 800-227-2155. Ashley Troutman BS, HT(ASCP) QIHC Immunohistochemistry Supervisor Vanderbilt University Histopathology 1301 Medical Center Drive TVC 4531 Nashville, TN 37232 *(Office) 615-875-3311 *(Lab) 615-343-9134 ashley.troutman@vanderbilt.edu Message: 20 Date: Fri, 20 Jul 2012 12:57:31 -0400 From: "Britton, Josette C" > Subject: To: "Richard Cartun" >, "Gerald Davydov" >, > Message-ID: > Content-Type: text/plain; charset="us-ascii" Where is everyone getting their p16 IVD antibody from? Josie Britton HT Cheshire Medical Center 580 Court Street Keene, NH 03431 From Milton.Gomez <@t> nyumc.org Fri Jul 20 20:16:26 2012 From: Milton.Gomez <@t> nyumc.org (Gomez, Milton) Date: Fri Jul 20 20:16:22 2012 Subject: [Histonet] RE: collecting chemical waste In-Reply-To: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Message-ID: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBCB5@MSGWCDCPMB27.nyumc.org> Thank you all who replied about this matter. Have a nice weekend! MG ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Gomez, Milton [Milton.Gomez@nyumc.org] Sent: Thursday, July 19, 2012 10:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] collecting chemical waste Hello histonetters, Do you folks enforce placing a label on a waste chemical container before collecting any waste or do you allow the label to be placed on the waste container after the waste is collected? I was taught to place the label on the waste container even before the first drop of waste is collected. MG _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tahseen <@t> brain.net.pk Sat Jul 21 05:57:22 2012 From: tahseen <@t> brain.net.pk (tahseen@brain.net.pk) Date: Sat Jul 21 05:57:30 2012 Subject: [Histonet] collecting chemical waste In-Reply-To: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> References: <7A7F4F00BF2D2D46A158AE61E0D2B6302A1EBC23@MSGWCDCPMB27.nyumc.org> Message-ID: <7351.203.135.35.66.1342868242.squirrel@brain.net.pk> Dear MG First when accumulate the chemical waste. Then before chemical waste can be picked up by house keeping staff, a waste tag is required. Muhammad Tahseen SKMCH&RC Lahore Pakistan Hello histonetters, > > > > Do you folks enforce placing a label on a waste chemical container before > collecting any waste or do you allow the label to be placed on the waste > container after the waste is collected? I was taught to place the label > on the waste container even before the first drop of waste is collected. > > > > MG > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pruegg <@t> ihctech.net Sun Jul 22 13:33:01 2012 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Sun Jul 22 13:33:05 2012 Subject: [Histonet] disposable blade holder Message-ID: <821D0154318D4F64AD99824B87FB2DB0@Patsyoffice> We are looking for the disposable blade holder for low profile blades that fit in microtomes like the Leica 1510 where permanent blades used to be held. We have two of these but they are both for high profile blades and we would prefer to use low profile disposable blades. Does anyone have one of these sitting around used we could buy from you or if there are vendors selling used ones please feel free to contact us. Regards, Patsy and Sherri Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net & email pruegg@flagshipbio.com web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From eca9 <@t> georgetown.edu Mon Jul 23 08:39:57 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Mon Jul 23 08:40:24 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? Message-ID: Hello, I have noticed that our biotinylated secondary antibodies on occasion cause nuclear staining in some samples. Why is this? It is not every time so I find it rather stange. Anyone know why this is happening and what I can do to avoid it? Thank you for any suggestion, Eva Permaul Georgetown University From BDeBrosse-Serra <@t> isisph.com Mon Jul 23 09:30:23 2012 From: BDeBrosse-Serra <@t> isisph.com (Bea DeBrosse-Serra) Date: Mon Jul 23 09:30:33 2012 Subject: [Histonet] disposable blade holder In-Reply-To: <821D0154318D4F64AD99824B87FB2DB0@Patsyoffice> References: <821D0154318D4F64AD99824B87FB2DB0@Patsyoffice> Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FC5B@EXCHMB01.isis.local> Check with your local Leica rep. Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Sunday, July 22, 2012 11:33 AM To: Histonet@lists.utsouthwestern.edu Cc: 'Sherri Saturley' Subject: [Histonet] disposable blade holder We are looking for the disposable blade holder for low profile blades that fit in microtomes like the Leica 1510 where permanent blades used to be held. We have two of these but they are both for high profile blades and we would prefer to use low profile disposable blades. Does anyone have one of these sitting around used we could buy from you or if there are vendors selling used ones please feel free to contact us. Regards, Patsy and Sherri Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net & email pruegg@flagshipbio.com web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Jul 23 09:57:17 2012 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Jul 23 09:57:23 2012 Subject: [Histonet] disposable blade holder In-Reply-To: <96138C8AB728814B9A576E4364EF84F9012BD954D314@EXMBX01.mmeprod.cbeyond> References: <821D0154318D4F64AD99824B87FB2DB0@Patsyoffice> <96138C8AB728814B9A576E4364EF84F9012BD954D314@EXMBX01.mmeprod.cbeyond> Message-ID: <64988B34442D48748488F3099B1D0D16@Patsyoffice> Yes it fits on the microtome where the permanent blades go for older microtomes that do not have disposable blade holding stages. Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net & email pruegg@flagshipbio.com web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: MaryK Mendell [mailto:kmendell@goldbergmd.net] Sent: Monday, July 23, 2012 6:26 AM To: Patsy Ruegg Subject: RE: [Histonet] disposable blade holder Patsy are you talking about on the microtome it self. Kate Mendell Histopathology/Lab Manager HOWARD S. GOLDBERG, M.D., INC 990 Paradise Road Swampscott, MA 01907 TEL: 781.595.0151 FAX: 781.592.6780 kmendell@goldbergmd.net www.cosmesticdermcenter.com PRIVACY NOTICE: This e-mail message may contain confidential patient or other information belonging to the sender that is legally privileged. This information is intended only for the use of the individual or authorized entity named above. The authorized recipient of this patient or other confidential information is prohibited from disclosing the information to any other party. If you have received this message in error, please notify the sender immediately and delete. Please keep any information you may have viewed confidential. ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg [pruegg@ihctech.net] Sent: Sunday, July 22, 2012 2:33 PM To: Histonet@lists.utsouthwestern.edu Cc: 'Sherri Saturley' Subject: [Histonet] disposable blade holder We are looking for the disposable blade holder for low profile blades that fit in microtomes like the Leica 1510 where permanent blades used to be held. We have two of these but they are both for high profile blades and we would prefer to use low profile disposable blades. Does anyone have one of these sitting around used we could buy from you or if there are vendors selling used ones please feel free to contact us. Regards, Patsy and Sherri Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net & email pruegg@flagshipbio.com web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Mon Jul 23 10:00:19 2012 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Mon Jul 23 10:00:25 2012 Subject: [Histonet] Negative Reagent Control In-Reply-To: <5005B747.7400.0077.1@harthosp.org> References: <5005B747.7400.0077.1@harthosp.org> Message-ID: For the last couple of years, I've thought this was true, but I didn't have the guts to say so. Thank you, Richard for bringing the truth out and getting it accepted. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Tuesday, July 17, 2012 7:05 PM To: Histonet Subject: [Histonet] Negative Reagent Control Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing. In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen. Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists. We had a long discussion about this and he agreed to support my position within CAP. With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed. The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause". This one change stands to save the healthcare system millions of dollars. Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From michelecarr10 <@t> yahoo.com Mon Jul 23 11:11:56 2012 From: michelecarr10 <@t> yahoo.com (Michele Carr) Date: Mon Jul 23 11:11:59 2012 Subject: [Histonet] counter stain for PAS for fungus Message-ID: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> Hi everyone I am having a problem with the fast green counterstain for the PAS for fungus.? The fast green stain is simply not staining anymore no matter how long I leave it on the specimen.? Do you have any suggestions on how to correct this or perhaps recommendations for a different counterstain. Our protocol is as follows: Periodic Acid 7 min quick rinse Schiff solution 20min wash for 10min Fast green stain 1 min ?Any help is appreciated. Thanks in advance, Michele Carr Medical Laboratory Services From rjbuesa <@t> yahoo.com Mon Jul 23 11:19:12 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jul 23 11:19:21 2012 Subject: [Histonet] counter stain for PAS for fungus In-Reply-To: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> References: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> Message-ID: <1343060352.20905.YahooMailNeo@web121406.mail.ne1.yahoo.com> Prepare a fresh solution Ren? J. ________________________________ From: Michele Carr To: "Histonet@lists.utsouthwestern.edu" Sent: Monday, July 23, 2012 12:11 PM Subject: [Histonet] counter stain for PAS for fungus Hi everyone I am having a problem with the fast green counterstain for the PAS for fungus.? The fast green stain is simply not staining anymore no matter how long I leave it on the specimen.? Do you have any suggestions on how to correct this or perhaps recommendations for a different counterstain. Our protocol is as follows: Periodic Acid 7 min quick rinse Schiff solution 20min wash for 10min Fast green stain 1 min ?Any help is appreciated. Thanks in advance, Michele Carr Medical Laboratory Services _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From renafail2 <@t> gmail.com Mon Jul 23 11:51:05 2012 From: renafail2 <@t> gmail.com (Rena Fail) Date: Mon Jul 23 11:51:09 2012 Subject: [Histonet] counter stain for PAS for fungus In-Reply-To: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> References: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> Message-ID: I have used a working solution light green Sf yellowish C,I. # 42095 and the only time I had problems was when someone left out the glacial acetic acid in the stock, Stock is light green SF yellowish 0.2 gm,distilled water 100.0 ml, 0.2 ml glacial acetic acid. Working is 10.0 ml of stock, 50.0 ml distilled water Rena Fail On Mon, Jul 23, 2012 at 12:11 PM, Michele Carr wrote: > Hi everyone I am having a problem with the fast green counterstain for the > PAS for fungus. The fast green stain is simply not staining anymore no > matter how long I leave it on the specimen. Do you have any suggestions on > how to correct this or perhaps recommendations for a different > counterstain. Our protocol is as follows: > Periodic Acid 7 min > quick rinse > Schiff solution 20min > wash for 10min > Fast green stain 1 min > Any help is appreciated. > Thanks in advance, > Michele Carr > Medical Laboratory Services > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From TJohnson <@t> gnf.org Mon Jul 23 12:07:24 2012 From: TJohnson <@t> gnf.org (Teri Johnson) Date: Mon Jul 23 12:07:27 2012 Subject: [Histonet] Re: counter stain for PAS for fungus Message-ID: <9F3CFEE76E51B64991C7485270890B400CDC8844@EX4.lj.gnf.org> Rena, If one uses Light Green (SF Yellowish) as a counterstain, it is important to monitor it closely. That solution will grow fungus and will show up as an artifact on stained slides. It is easy to tell the source because it is not stained, whereas your silver or PAS techniques will stain the fungus in the tissue. Some labs moved to fast green as a counterstain instead because of this issue. Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 From brendal.finlay <@t> medicalcenterclinic.com Mon Jul 23 12:57:21 2012 From: brendal.finlay <@t> medicalcenterclinic.com (Brendal Finlay) Date: Mon Jul 23 12:57:26 2012 Subject: [Histonet] (no subject) Message-ID: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> Hello all! We are budgeted to get an automated coverslipper for our department.? When we talked about it with our service rep, he asked if we wanted a glass or tape coverslipper.? I worked with a glass one many years ago when they first came out and?all I remember is breaking and sticking coverslips.? I'm sure things have improved by now and was wondering about the pros and cons of the glass versus tape coverslipper.? Which do you prefer and why? Thank you! Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com From pkarlisch <@t> hmc.psu.edu Mon Jul 23 13:13:53 2012 From: pkarlisch <@t> hmc.psu.edu (Karlisch, Patricia) Date: Mon Jul 23 13:14:02 2012 Subject: [Histonet] EM send outs Message-ID: All, We are looking for a site to send our NEURO Electron Microscopic blocks for EM ONLY (Tech only). There would be about 25-30 cases a year (muscle, nerve and possibly brain). If you have information please let me know. I would appreciate it. Patricia Karlisch Supervisor, Histology Department of Pathology and Laboratory Medicine Penn State Milton S Hershey Medical Center 500 University Drive Hershey, PA 17033-0850 Tel: 717-531-6072 Fax: 717-531-7741 Email: pkarlisch@hmc.psu.edu *****E-Mail Confidentiality Notice***** This message (including any attachments) contains information intended for a specific individual(s) and purpose that may be privileged, confidential or otherwise protected from disclosure pursuant to applicable law. Any inappropriate use, distribution or copying of the message is strictly prohibited and may subject you to criminal or civil penalty. If you have received this transmission in error, please reply to the sender indicating this error and delete the transmission from your system immediately. From wbenton <@t> cua.md Mon Jul 23 13:32:26 2012 From: wbenton <@t> cua.md (Walter Benton) Date: Mon Jul 23 13:32:47 2012 Subject: [Histonet] RE: EM send outs In-Reply-To: References: Message-ID: <0B8979A204680A42B93A52B486088CD92CCB563CC1@CUAEXH1.GCU-MD.local> Try Hopkins or Univ of Maryland Medical Center. Walter Benton HT(ASCP)QIHC Histology Supervisor Chesapeake Urology Associates 806 Landmark Drive, Suite 126 (All Deliveries to Suite 127) Glen Burnie, MD 21061 443-471-5850 (Direct) 410-768-5961 (Lab) 410-768-5965 (Fax) wbenton@cua.md ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Karlisch, Patricia [pkarlisch@hmc.psu.edu] Sent: Monday, July 23, 2012 2:13 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] EM send outs All, We are looking for a site to send our NEURO Electron Microscopic blocks for EM ONLY (Tech only). There would be about 25-30 cases a year (muscle, nerve and possibly brain). If you have information please let me know. I would appreciate it. Patricia Karlisch Supervisor, Histology Department of Pathology and Laboratory Medicine Penn State Milton S Hershey Medical Center 500 University Drive Hershey, PA 17033-0850 Tel: 717-531-6072 Fax: 717-531-7741 Email: pkarlisch@hmc.psu.edu *****E-Mail Confidentiality Notice***** This message (including any attachments) contains information intended for a specific individual(s) and purpose that may be privileged, confidential or otherwise protected from disclosure pursuant to applicable law. Any inappropriate use, distribution or copying of the message is strictly prohibited and may subject you to criminal or civil penalty. If you have received this transmission in error, please reply to the sender indicating this error and delete the transmission from your system immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: The information contained in this electronic message is intended solely for the personal and confidential use of the designated recipient(s) named above and may contain information that is protected from disclosure under applicable law. If you are not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this transmission is strictly prohibited. If you have received this transmission in error, please notify the transmitting person/department immediately by email or telephone (410) 581-5881 and delete the message without making a copy. From CIngles <@t> uwhealth.org Mon Jul 23 14:31:11 2012 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Mon Jul 23 14:31:45 2012 Subject: [Histonet] (no subject) References: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A876@UWHC-MAIL2.uwhis.hosp.wisc.edu> I'd love to know too. We also use a xylene substitute. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Brendal Finlay Sent: Mon 7/23/2012 12:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hello all! We are budgeted to get an automated coverslipper for our department. When we talked about it with our service rep, he asked if we wanted a glass or tape coverslipper. I worked with a glass one many years ago when they first came out and all I remember is breaking and sticking coverslips. I'm sure things have improved by now and was wondering about the pros and cons of the glass versus tape coverslipper. Which do you prefer and why? Thank you! Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shernandez <@t> st-joseph.org Mon Jul 23 14:38:34 2012 From: shernandez <@t> st-joseph.org (shernandez@st-joseph.org) Date: Mon Jul 23 14:38:40 2012 Subject: [Histonet] CAP and expiration dates Message-ID: Hi All, I have a PA that uses acetic alcohol for lymph nodes. I have tried all of the premade ones and he doesn't like any of them, so we are back to making it again. How do you determine expiration dates of solutions made in house? Thanks, Susan Baker ________________________________ The documents accompanying this transmission may contain confidential health information that is legally privileged. This information is intended only for the use of the individual or entity named on this sheet. The authorized recipient of this information is prohibited from disclosing this information to any party unless required to do so by law or regulation and is required to destroy the information after its stated need has been fulfilled. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or action taken in reliance on the contents of these documents is strictly prohibited. Violators will be prosecuted. If you have received this information in error, please notify the sender immediately and arrange for the destruction of these documents. From BDeBrosse-Serra <@t> isisph.com Mon Jul 23 14:52:25 2012 From: BDeBrosse-Serra <@t> isisph.com (Bea DeBrosse-Serra) Date: Mon Jul 23 14:52:30 2012 Subject: [Histonet] (no subject) In-Reply-To: <064F1ACBAE8A78469AE2E41D533D87E505A876@UWHC-MAIL2.uwhis.hosp.wisc.edu> References: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> <064F1ACBAE8A78469AE2E41D533D87E505A876@UWHC-MAIL2.uwhis.hosp.wisc.edu> Message-ID: <493CAA64F203E14E8823737B9EE0E25F092BA8FC60@EXCHMB01.isis.local> It all depends on how long you want to store the slides. The tape has a tendency to come off, whereas the glass slides, if covered properly, can be stored "indefinitely". The glass coverslippers have improved quite a bit, and there are several vendors out there who have them. We have one here, and we love it! As far as xylene substitutes, you may want to check about this with the vendor. Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Monday, July 23, 2012 12:31 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) I'd love to know too. We also use a xylene substitute. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Brendal Finlay Sent: Mon 7/23/2012 12:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hello all! We are budgeted to get an automated coverslipper for our department. When we talked about it with our service rep, he asked if we wanted a glass or tape coverslipper. I worked with a glass one many years ago when they first came out and all I remember is breaking and sticking coverslips. I'm sure things have improved by now and was wondering about the pros and cons of the glass versus tape coverslipper. Which do you prefer and why? Thank you! Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com _______________________________________________ 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 From eca9 <@t> georgetown.edu Mon Jul 23 15:07:10 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Mon Jul 23 15:07:33 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: References: Message-ID: We believe it has to be the secondary as we have nuclear staining in the absence of primary antibody. Eva On Mon, Jul 23, 2012 at 12:58 PM, ihcs-wojcieszyn wrote: > Hello, > Please explain how you know that it is the secondary and not the > primary? We Have confirmed that in canine lymphomas, nuclear > CD79a is a real and frequent observation. > > john > IHC Services > > On Monday, July 23, 2012, at 08:39 AM, Eva Permaul wrote: > > Hello, >> >> I have noticed that our biotinylated secondary antibodies on occasion >> cause >> nuclear staining in some samples. Why is this? It is not every time so I >> find it rather stange. Anyone know why this is happening and what I can do >> to avoid it? >> >> Thank you for any suggestion, >> Eva Permaul >> Georgetown University >> ______________________________**_________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.**edu >> http://lists.utsouthwestern.**edu/mailman/listinfo/histonet >> >> > From jmacdonald <@t> mtsac.edu Mon Jul 23 15:08:11 2012 From: jmacdonald <@t> mtsac.edu (Jennifer MacDonald) Date: Mon Jul 23 15:08:54 2012 Subject: [Histonet] counter stain for PAS for fungus In-Reply-To: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> References: <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> Message-ID: Add some acetic acid to the fast green. On Jul 23, 2012, at 9:11 AM, Michele Carr wrote: > Hi everyone I am having a problem with the fast green counterstain for the PAS for fungus. The fast green stain is simply not staining anymore no matter how long I leave it on the specimen. Do you have any suggestions on how to correct this or perhaps recommendations for a different counterstain. Our protocol is as follows: > Periodic Acid 7 min > quick rinse > Schiff solution 20min > wash for 10min > Fast green stain 1 min > Any help is appreciated. > Thanks in advance, > Michele Carr > Medical Laboratory Services > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ccrowder <@t> vetmed.lsu.edu Mon Jul 23 17:14:48 2012 From: ccrowder <@t> vetmed.lsu.edu (Cheryl Crowder) Date: Mon Jul 23 17:14:49 2012 Subject: [Histonet] Light green for PAS Message-ID: Michelle - I agree that the light green is "dead" and you should make some new or buy a commercially made solution. However, if you want to try a different counterstain try a light hematoylin (usually used for glycogen) or metanil yellow. Either will give good contrast to the pink of the PAS. Cheryl Crowder From tony.henwood <@t> health.nsw.gov.au Mon Jul 23 17:59:30 2012 From: tony.henwood <@t> health.nsw.gov.au (Tony Henwood (SCHN)) Date: Mon Jul 23 17:59:44 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: References: Message-ID: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> 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 H&E 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-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@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. Why is this? It is not every time so I find it rather stange. Anyone know why this is happening and what I can do to avoid it? Thank you for any suggestion, Eva Permaul Georgetown University _______________________________________________ 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. ********************************************************************************* From tony.henwood <@t> health.nsw.gov.au Mon Jul 23 18:16:44 2012 From: tony.henwood <@t> health.nsw.gov.au (Tony Henwood (SCHN)) Date: Mon Jul 23 18:17:01 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> Message-ID: <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> 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-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@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. Why is this? It is not every time so I find it rather stange. Anyone know why this is happening and what I can do to avoid it? Thank you for any suggestion, Eva Permaul Georgetown University _______________________________________________ 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. ********************************************************************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From naveedafahim <@t> yahoo.ca Mon Jul 23 19:32:26 2012 From: naveedafahim <@t> yahoo.ca (naveeda arshad) Date: Mon Jul 23 21:32:39 2012 Subject: [Histonet] Re: ZN control making procedure inhouse Message-ID: <1343089946.31512.YahooMailClassic@web161704.mail.bf1.yahoo.com> Hi All?does any one has procedure for making inhouse ZN control if they can share with me . I will?appreciate itthanksNaveeda? --- On Mon, 7/23/12, histonet-request@lists.utsouthwestern.edu wrote: From: histonet-request@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 104, Issue 26 To: histonet@lists.utsouthwestern.edu Received: Monday, July 23, 2012, 8:30 PM 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-request@lists.utsouthwestern.edu You can reach the person managing the list at ??? histonet-owner@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. disposable blade holder (Patsy Ruegg) ???2. Secondary antibody causing nuclear staining? (Eva Permaul) ???3. RE: disposable blade holder (Bea DeBrosse-Serra) ???4. RE: disposable blade holder (Patsy Ruegg) ???5. RE: Negative Reagent Control (Smith, Allen) ???6. counter stain for PAS for fungus (Michele Carr) ???7. Re: counter stain for PAS for fungus (Rene J Buesa) ???8. Re: counter stain for PAS for fungus (Rena Fail) ---------------------------------------------------------------------- Message: 1 Date: Sun, 22 Jul 2012 12:33:01 -0600 From: "Patsy Ruegg" Subject: [Histonet] disposable blade holder To: Cc: 'Sherri Saturley' Message-ID: <821D0154318D4F64AD99824B87FB2DB0@Patsyoffice> Content-Type: text/plain;??? charset="us-ascii" We are looking for the disposable blade holder for low profile blades that fit in microtomes like the Leica 1510 where permanent blades used to be held.? We have two of these but they are both for high profile blades and we would prefer to use low profile disposable blades.? Does anyone have one of these sitting around used we could buy from you or if there are vendors selling used ones please feel free to contact us. Regards, Patsy and Sherri Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email? pruegg@ihctech.net & email? pruegg@flagshipbio.com web site? www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. ------------------------------ Message: 2 Date: Mon, 23 Jul 2012 09:39:57 -0400 From: Eva Permaul Subject: [Histonet] Secondary antibody causing nuclear staining? To: histonet@lists.utsouthwestern.edu Message-ID: ??? Content-Type: text/plain; charset=ISO-8859-1 Hello, I have noticed that our biotinylated secondary antibodies on occasion cause nuclear staining in some samples. Why is this? It is not every time so I find it rather stange. Anyone know why this is happening and what I can do to avoid it? Thank you for any suggestion, Eva Permaul Georgetown University ------------------------------ Message: 3 Date: Mon, 23 Jul 2012 07:30:23 -0700 From: Bea DeBrosse-Serra Subject: RE: [Histonet] disposable blade holder To: "'Patsy Ruegg'" , ??? "Histonet@lists.utsouthwestern.edu" ??? Cc: 'Sherri Saturley' Message-ID: ??? <493CAA64F203E14E8823737B9EE0E25F092BA8FC5B@EXCHMB01.isis.local> Content-Type: text/plain; charset="us-ascii" Check with your local Leica rep. Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Sunday, July 22, 2012 11:33 AM To: Histonet@lists.utsouthwestern.edu Cc: 'Sherri Saturley' Subject: [Histonet] disposable blade holder We are looking for the disposable blade holder for low profile blades that fit in microtomes like the Leica 1510 where permanent blades used to be held.? We have two of these but they are both for high profile blades and we would prefer to use low profile disposable blades.? Does anyone have one of these sitting around used we could buy from you or if there are vendors selling used ones please feel free to contact us. Regards, Patsy and Sherri Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email? pruegg@ihctech.net & email? pruegg@flagshipbio.com web site? www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 23 Jul 2012 08:57:17 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] disposable blade holder To: "'MaryK Mendell'" Cc: Histonet@lists.utsouthwestern.edu Message-ID: <64988B34442D48748488F3099B1D0D16@Patsyoffice> Content-Type: text/plain;??? charset="us-ascii" Yes it fits on the microtome where the permanent blades go for older microtomes that do not have disposable blade holding stages. Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net & email pruegg@flagshipbio.com web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: MaryK Mendell [mailto:kmendell@goldbergmd.net] Sent: Monday, July 23, 2012 6:26 AM To: Patsy Ruegg Subject: RE: [Histonet] disposable blade holder Patsy are you talking about on the microtome it self.? Kate Mendell Histopathology/Lab Manager HOWARD S. GOLDBERG, M.D., INC 990 Paradise Road Swampscott, MA? 01907 TEL:? 781.595.0151 FAX:? 781.592.6780 kmendell@goldbergmd.net www.cosmesticdermcenter.com PRIVACY NOTICE: This e-mail message may contain confidential patient or other information belonging to the sender that is legally privileged.? This information is intended only for the use of the individual or authorized entity named above. The authorized recipient of this patient or other confidential information is prohibited from disclosing the information to any other party.? If you have received this message in error, please notify the sender immediately and delete.? Please keep any information you may have viewed confidential. ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg [pruegg@ihctech.net] Sent: Sunday, July 22, 2012 2:33 PM To: Histonet@lists.utsouthwestern.edu Cc: 'Sherri Saturley' Subject: [Histonet] disposable blade holder We are looking for the disposable blade holder for low profile blades that fit in microtomes like the Leica 1510 where permanent blades used to be held.? We have two of these but they are both for high profile blades and we would prefer to use low profile disposable blades.? Does anyone have one of these sitting around used we could buy from you or if there are vendors selling used ones please feel free to contact us. Regards, Patsy and Sherri Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email? pruegg@ihctech.net & email? pruegg@flagshipbio.com web site? www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 23 Jul 2012 15:00:19 +0000 From: "Smith, Allen" Subject: RE: [Histonet] Negative Reagent Control To: Richard Cartun Cc: "histonet@lists.utsouthwestern.edu" ??? Message-ID: ??? ??? Content-Type: text/plain; charset="us-ascii" For the last couple of years, I've thought this was true, but I didn't have the guts to say so. Thank you, Richard for bringing the truth out and getting it accepted. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Tuesday, July 17, 2012 7:05 PM To: Histonet Subject: [Histonet] Negative Reagent Control Most of you know that I have been advocating for the elimination of the "Negative Reagent Control" when using a non-avidin-biotin detection system (polymer) in immunohistochemical testing.? In my opinion, it is a waste of healthcare dollars and, more importantly, precious patient specimen.? Last year at the NSH IHC Forum in Denver I met James Dvorak, MT(ASCP) from the College of American Pathologists.? We had a long discussion about this and he agreed to support my position within CAP.? With the support of James, and the help of Dr. Regan Fulton (from the CAP IHC Committee), the wording on the CAP Anatomic Pathology checklist for question ANP.22570 will be changed.? The new wording includes the following sentence: "Immunohistochemical tests using polymer-based detection systems (biotin-free) are sufficiently free of background reactivity to obviated the need for a negative reagent control and such controls may be omitted at the discretion of the laboratory director." I announced this change at the 2012 NSH IHC/ISH Forum this past weekend in Windsor, CT to "loud applause".? This one change stands to save the healthcare system millions of dollars.? Thank you Jim and Dr. Fulton for making this happen! Richard Richard W. Cartun, MS, PhD Director, Histology & Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT? 06102 (860) 545-1596 Office (860) 545-2204 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 23 Jul 2012 09:11:56 -0700 (PDT) From: Michele Carr Subject: [Histonet] counter stain for PAS for fungus To: "Histonet@lists.utsouthwestern.edu" ??? Message-ID: ??? <1343059916.21147.YahooMailNeo@web120704.mail.ne1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi everyone I am having a problem with the fast green counterstain for the PAS for fungus.? The fast green stain is simply not staining anymore no matter how long I leave it on the specimen.? Do you have any suggestions on how to correct this or perhaps recommendations for a different counterstain. Our protocol is as follows: Periodic Acid 7 min quick rinse Schiff solution 20min wash for 10min Fast green stain 1 min ?Any help is appreciated. Thanks in advance, Michele Carr Medical Laboratory Services ------------------------------ Message: 7 Date: Mon, 23 Jul 2012 09:19:12 -0700 (PDT) From: Rene J Buesa Subject: Re: [Histonet] counter stain for PAS for fungus To: Michele Carr , ??? "Histonet@lists.utsouthwestern.edu" ??? Message-ID: ??? <1343060352.20905.YahooMailNeo@web121406.mail.ne1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Prepare a fresh solution Ren? J. ________________________________ From: Michele Carr To: "Histonet@lists.utsouthwestern.edu" Sent: Monday, July 23, 2012 12:11 PM Subject: [Histonet] counter stain for PAS for fungus Hi everyone I am having a problem with the fast green counterstain for the PAS for fungus.? The fast green stain is simply not staining anymore no matter how long I leave it on the specimen.? Do you have any suggestions on how to correct this or perhaps recommendations for a different counterstain. Our protocol is as follows: Periodic Acid 7 min quick rinse Schiff solution 20min wash for 10min Fast green stain 1 min ?Any help is appreciated. Thanks in advance, Michele Carr Medical Laboratory Services _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 23 Jul 2012 12:51:05 -0400 From: Rena Fail Subject: Re: [Histonet] counter stain for PAS for fungus To: Michele Carr Cc: "Histonet@lists.utsouthwestern.edu" ??? Message-ID: ??? Content-Type: text/plain; charset=ISO-8859-1 I have used a working solution light green Sf yellowish C,I. # 42095 and the only time I had problems was when someone left out the glacial acetic acid in the stock, Stock is light green SF yellowish 0.2 gm,distilled water 100.0 ml, 0.2 ml glacial acetic acid. Working? is 10.0 ml of stock, 50.0 ml? distilled water Rena Fail On Mon, Jul 23, 2012 at 12:11 PM, Michele Carr wrote: > Hi everyone I am having a problem with the fast green counterstain for the > PAS for fungus.? The fast green stain is simply not staining anymore no > matter how long I leave it on the specimen.? Do you have any suggestions on > how to correct this or perhaps recommendations for a different > counterstain. Our protocol is as follows: > Periodic Acid 7 min > quick rinse > Schiff solution 20min > wash for 10min > Fast green stain 1 min >? Any help is appreciated. > Thanks in advance, > Michele Carr > Medical Laboratory Services > _______________________________________________ > 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 End of Histonet Digest, Vol 104, Issue 26 ***************************************** From betterpath10 <@t> yahoo.com Mon Jul 23 21:44:24 2012 From: betterpath10 <@t> yahoo.com (betterpath10@yahoo.com) Date: Mon Jul 23 21:45:06 2012 Subject: [Histonet] (no subject) In-Reply-To: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> References: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> Message-ID: Hi - all hands down the film. Faster, less daily issues. I was fortunate to be one if the first to try this in the Boston area in the early 90's. I have done numerous studies including long term storage. You need to ensure adequate 'clean' xylene drips 3-5 per slide. GOOD LUCK!!!! Candace Connected by DROID on Verizon Wireless -----Original message----- From: Brendal Finlay To: histonet@lists.utsouthwestern.edu Sent: Mon, Jul 23, 2012 17:58:08 GMT+00:00 Subject: [Histonet] (no subject) Hello all! We are budgeted to get an automated coverslipper for our department.? When we talked about it with our service rep, he asked if we wanted a glass or tape coverslipper.? I worked with a glass one many years ago when they first came out and?all I remember is breaking and sticking coverslips.? I'm sure things have improved by now and was wondering about the pros and cons of the glass versus tape coverslipper.? Which do you prefer and why? Thank you! Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From louise.renton <@t> gmail.com Tue Jul 24 06:14:37 2012 From: louise.renton <@t> gmail.com (Louise Renton) Date: Tue Jul 24 06:14:50 2012 Subject: [Histonet] techniques for archaeological bone Message-ID: 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? From ADuddey <@t> firsthealth.org Tue Jul 24 06:43:12 2012 From: ADuddey <@t> firsthealth.org (Duddey, Aimee) Date: Tue Jul 24 06:43:49 2012 Subject: [Histonet] (no subject) In-Reply-To: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> References: <044756548c9f715026ef579f70384b58@medicalcenterclinic.com> Message-ID: Hi Brenda, We used a tape coverslipper for years with little issue day to day. After storing, though, many of the old coverslips became yellowed and bubbled and came off the slides. We switched to the glass coverslips with the stainer coverslipper combo unit. We found that you have to use a very specific coverslip for them not to stick together (double coverslips). There is no issue with breaking routinely. There have been occasions when it gets jammed or has some residual goo which resulted in a broken slide from time to time but nothing regularly enough to outweigh its benefits. Hope this helps! Aimee -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Brendal Finlay Sent: Monday, July 23, 2012 1:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hello all! We are budgeted to get an automated coverslipper for our department.? When we talked about it with our service rep, he asked if we wanted a glass or tape coverslipper.? I worked with a glass one many years ago when they first came out and?all I remember is breaking and sticking coverslips.? I'm sure things have improved by now and was wondering about the pros and cons of the glass versus tape coverslipper.? Which do you prefer and why? Thank you! Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Tue Jul 24 07:13:36 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue Jul 24 07:14:04 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> Message-ID: 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: > histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto: > histonet-bounces@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. Why is this? It is not every time > so I find it rather stange. Anyone know why this is happening and what I > can do to avoid it? > > Thank you for any suggestion, > Eva Permaul > Georgetown University > _______________________________________________ > 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. > > ********************************************************************************* > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From one_angel_secret <@t> yahoo.com Tue Jul 24 07:18:48 2012 From: one_angel_secret <@t> yahoo.com (Kim Donadio) Date: Tue Jul 24 07:23:02 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> Message-ID: <6A8688F1-DF64-464A-97BD-98D6399955BC@yahoo.com> 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 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@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. Why is this? It is not every time >> so I find it rather stange. Anyone know why this is happening and what I >> can do to avoid it? >> >> Thank you for any suggestion, >> Eva Permaul >> Georgetown University >> _______________________________________________ >> 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. >> >> ********************************************************************************* >> >> _______________________________________________ >> 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 From eca9 <@t> georgetown.edu Tue Jul 24 07:41:55 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue Jul 24 07:42:24 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: <9C33FEBF8553D44884B4093C2AB256761B1E4346@VMW-EXCMBD2S05.dhe.duke.edu> References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> <9C33FEBF8553D44884B4093C2AB256761B1E4346@VMW-EXCMBD2S05.dhe.duke.edu> Message-ID: 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.burchette@duke.edu> wrote: > What is your heat retrieval process? > > Jim Burchette, HT(ASCP) QIHC > Histologist and Fly Fishing Bum > Orlando, Florida > > ________________________________________ > From: histonet-bounces@lists.utsouthwestern.edu [ > histonet-bounces@lists.utsouthwestern.edu] on behalf of Eva Permaul [ > eca9@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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: > > histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto: > > histonet-bounces@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. Why is this? It is not every time > > so I find it rather stange. Anyone know why this is happening and what I > > can do to avoid it? > > > > Thank you for any suggestion, > > Eva Permaul > > Georgetown University > > _______________________________________________ > > 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. > > > > > ********************************************************************************* > > > > _______________________________________________ > > 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 > From eca9 <@t> georgetown.edu Tue Jul 24 08:17:59 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue Jul 24 08:18:28 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: <9C33FEBF8553D44884B4093C2AB256761B1E436F@VMW-EXCMBD2S05.dhe.duke.edu> References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> <9C33FEBF8553D44884B4093C2AB256761B1E4346@VMW-EXCMBD2S05.dhe.duke.edu> <9C33FEBF8553D44884B4093C2AB256761B1E436F@VMW-EXCMBD2S05.dhe.duke.edu> Message-ID: 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.burchette@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-bounces@lists.utsouthwestern.edu [ > histonet-bounces@lists.utsouthwestern.edu] on behalf of Eva Permaul [ > eca9@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.burchette@duke.edu> wrote: > > > What is your heat retrieval process? > > > > Jim Burchette, HT(ASCP) QIHC > > Histologist and Fly Fishing Bum > > Orlando, Florida > > > > ________________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu [ > > histonet-bounces@lists.utsouthwestern.edu] on behalf of Eva Permaul [ > > eca9@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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: > > > histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto: > > > histonet-bounces@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. Why is this? It is not every > time > > > so I find it rather stange. Anyone know why this is happening and what > I > > > can do to avoid it? > > > > > > Thank you for any suggestion, > > > Eva Permaul > > > Georgetown University > > > _______________________________________________ > > > 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. > > > > > > > > > ********************************************************************************* > > > > > > _______________________________________________ > > > 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 mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From donna_suresch <@t> merck.com Tue Jul 24 10:16:21 2012 From: donna_suresch <@t> merck.com (Suresch, Donna L.) Date: Tue Jul 24 10:16:42 2012 Subject: [Histonet] Cyclin D1 IHC Message-ID: <2C9A1D9608959940943F357E0A470FF8A8457310B9@USCTMXP51005.merck.com> 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: WP44K Office: 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. From eca9 <@t> georgetown.edu Tue Jul 24 10:26:35 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue Jul 24 10:27:03 2012 Subject: [Histonet] Cyclin D1 IHC In-Reply-To: <2C9A1D9608959940943F357E0A470FF8A8457310B9@USCTMXP51005.merck.com> References: <2C9A1D9608959940943F357E0A470FF8A8457310B9@USCTMXP51005.merck.com> Message-ID: 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. 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: WP44K Office: 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 > > From marktarango <@t> gmail.com Tue Jul 24 10:33:25 2012 From: marktarango <@t> gmail.com (Mark Tarango) Date: Tue Jul 24 10:33:33 2012 Subject: [Histonet] Cyclin D1 IHC In-Reply-To: <2C9A1D9608959940943F357E0A470FF8A8457310B9@USCTMXP51005.merck.com> References: <2C9A1D9608959940943F357E0A470FF8A8457310B9@USCTMXP51005.merck.com> Message-ID: 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. 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: WP44K Office: 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 > > From screnshaw <@t> oregonmed.net Tue Jul 24 10:51:37 2012 From: screnshaw <@t> oregonmed.net (Crenshaw, Stacey) Date: Tue Jul 24 10:51:46 2012 Subject: [Histonet] Unsubscribe Message-ID: 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. From LBUSTAMANTE <@t> cvm.tamu.edu Tue Jul 24 11:02:07 2012 From: LBUSTAMANTE <@t> cvm.tamu.edu (Bustamante, Lin) Date: Tue Jul 24 11:02:16 2012 Subject: [Histonet] Question about waste storage temperature Message-ID: <94B6DC15AAF2F046BF847D4C1CA9AAC973096845@CVMMB02.cvm.tamu.edu> 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. From histotalk <@t> yahoo.com Tue Jul 24 11:18:40 2012 From: histotalk <@t> yahoo.com (David Kemler) Date: Tue Jul 24 11:18:42 2012 Subject: [Histonet] HistoTALK Show Message-ID: <1343146720.95915.YahooMailNeo@web121506.mail.ne1.yahoo.com> 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 From nmhisto <@t> comcast.net Tue Jul 24 11:36:49 2012 From: nmhisto <@t> comcast.net (nmhisto@comcast.net) Date: Tue Jul 24 11:37:45 2012 Subject: [Histonet] Unsubscribe In-Reply-To: Message-ID: <1656138725.85282.1343147809231.JavaMail.root@sz0075a.emeryville.ca.mail.comcast.net> OH NO!? They're BACK!? Saints preserve us! ----- Original Message ----- From: "Stacey Crenshaw" To: "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 From ihcman2010 <@t> hotmail.com Tue Jul 24 12:13:00 2012 From: ihcman2010 <@t> hotmail.com (Glen Dawson) Date: Tue Jul 24 12:13:07 2012 Subject: [Histonet] CD200 In-Reply-To: References: Message-ID: I would be interested in this as well... Thank-you, Glen Dawson BS, HT(ASCP), QIHC Histology Technical Specialist Janesville, WI > From: 41dmb41@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 From sdysart <@t> mirnarx.com Tue Jul 24 13:35:43 2012 From: sdysart <@t> mirnarx.com (Sarah Dysart) Date: Tue Jul 24 13:36:01 2012 Subject: [Histonet] Non-Histo. question Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D50107809C5@BL2PRD0710MB363.namprd07.prod.outlook.com> 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 From jqb7 <@t> cdc.gov Tue Jul 24 13:41:27 2012 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/OID/NCEZID)) Date: Tue Jul 24 13:41:38 2012 Subject: [Histonet] RE: Non-Histo. question In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D50107809C5@BL2PRD0710MB363.namprd07.prod.outlook.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D50107809C5@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: This tells you how to temporarily unsubscribe: Welcome to the Histonet@lists.utsouthwestern.edu mailing list! Welcome to HISTONET, the World's largest electronic mail list for the Histology profession, which has been in continuous operation since January 1996. Histonet currently has more than 3200 members from throughout the world. The Histonet listserver is managed as a service to the field of Histology by Dr. Linda Margraf and the University of Texas Southwestern Medical Center. HOW THE LIST WORKS: To subscribe to the list or change your subscription to the daily digest mode etc. you need to go to http://lists.utsouthwestern.edu/mailman/listinfo/histonet You will need to request a password if you do not have one. Once you are on the list you will begin to receive email messages from the list members. If you wish to send a message to the list members, please send your email message to histonet@lists.utsouthwestern.edu The server will only circulate messages sent from current list members. If your email address changes, even slightly, from the address on the membership list, you will need to update your address before you can send a message to the list. WHAT ARE THE RULES ABOUT CONTENT? You may post any questions you wish pertaining to histology, pathology, in-situ hybridization, immunohistochemistry etc. Equipment and reagent evaluations, laboratory management issues, government regulations, and job opportunities are all appropriate topics. The University asks that we restrict the use of its hardware and software to business purposes only (occasional jokes do slip through but PLEASE use restraint). Vendors and those with commercial interests in histology products are welcome contributors however,we ask that blatant advertisements be avoided at all times. It is fine to refer to product that your company produces if it is pertinent to a topic being discussed on the list. Unsolicited advertisements are poorly tolerated by the members and you will likely receive a number of negative comments if you overstep the boundaries. Please contact Linda Margraf at linda.margraf@childrens.com if you are not sure about the appropriateness of a message you wish to post. BASIC HISTONET "NETIQUETTE" It is most helpful to the list members if you post your responses to queries to everyone on the list and not just as a personal reply to the person asking the question. That way duplicate messages are minimized and we all learn from each other's comments. Likewise, if you post a question and get a number of responses back directly to you, it is helpful to everyone if you could send out a summary of the replies you got to Histonet. Please avoid abbreviations unless they are explained in your message. For example: immunohistochemistry (IHC). This list circulates to a wide variety of individuals and what seems obvious to you may have no meaning on the other side of the world. Please sign your letter and include your institution or affiliation and location. Not all email systems have headers which identify the sender. Do use the subject line to indicate the topic of your message. DON'T USE ONLY CAPITAL LETTERS -it is considered shouting. IMPORTANT INFORMATION: Please do not send images or any other attachments with your message. The server will block all attachments to protect the list against viruses. Images can be posted at the Histonet web site (www.histonet.org) Please see the web site for instructions. Histosearch archives the messages from the Histonet listserver and provides a search engine to search the messages by keyword(s). There are currently over 30,000 messages in the archives beginning in October, 1998. These archives are managed by Marvin Hanna. The archives are available at www.histosearch.com/histonet.html. To post to this list, send your email to: histonet@lists.utsouthwestern.edu General information about the mailing list is at: http://lists.utsouthwestern.edu/mailman/listinfo/histonet If you ever want to unsubscribe or change your options (eg, switch to or from digest mode, change your password, etc.), visit your subscription page at: http://lists.utsouthwestern.edu/mailman/options/histonet/jqb7%40cdc.gov You can also make such adjustments via email by sending a message to: Histonet-request@lists.utsouthwestern.edu with the word `help' in the subject or body (don't include the quotes), and you will get back a message with instructions. You must know your password to change your options (including changing the password, itself) or to unsubscribe. Normally, Mailman will remind you of your lists.utsouthwestern.edu mailing list passwords once every month, although you can disable this if you prefer. This reminder will also include instructions on how to unsubscribe or change your account options. There is also a button on your options page that will email your current password to you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart Sent: Tuesday, July 24, 2012 2:36 PM To: histonet Subject: [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 From anne.lewin <@t> bms.com Tue Jul 24 13:42:37 2012 From: anne.lewin <@t> bms.com (Lewin, Anne) Date: Tue Jul 24 13:42:44 2012 Subject: [Histonet] unsubscribe Message-ID: <4895A1696F956D4CB56011A8C61312820C8A4E2E22@ushpwbmsmmp008.one.ads.bms.com> 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. From eca9 <@t> georgetown.edu Tue Jul 24 13:50:32 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue Jul 24 13:51:00 2012 Subject: [Histonet] Non-Histo. question In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D50107809C5@BL2PRD0710MB363.namprd07.prod.outlook.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D50107809C5@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: 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 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 > From eca9 <@t> georgetown.edu Tue Jul 24 14:32:58 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue Jul 24 14:33:24 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: <6A8688F1-DF64-464A-97BD-98D6399955BC@yahoo.com> References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> <6A8688F1-DF64-464A-97BD-98D6399955BC@yahoo.com> Message-ID: 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 wrote: > 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 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: > >> histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto: > >> histonet-bounces@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. Why is this? It is not every > time > >> so I find it rather stange. Anyone know why this is happening and what I > >> can do to avoid it? > >> > >> Thank you for any suggestion, > >> Eva Permaul > >> Georgetown University > >> _______________________________________________ > >> 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. > >> > >> > ********************************************************************************* > >> > >> _______________________________________________ > >> 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 > From twheelock <@t> mclean.harvard.edu Tue Jul 24 14:36:11 2012 From: twheelock <@t> mclean.harvard.edu (Tim Wheelock) Date: Tue Jul 24 14:36:18 2012 Subject: [Histonet] Bielschowsky questions, Luxol Fast Blue answers, and Protocol Drift. Message-ID: <500EF92B.3040407@mclean.harvard.edu> 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 From renafail2 <@t> gmail.com Tue Jul 24 14:41:48 2012 From: renafail2 <@t> gmail.com (Rena Fail) Date: Tue Jul 24 14:41:50 2012 Subject: [Histonet] Tennessee B&B Message-ID: 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 From ratliffjack <@t> hotmail.com Tue Jul 24 15:46:28 2012 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Tue Jul 24 15:46:32 2012 Subject: [Histonet] 2012 Hard Tissue Forum Event in Bethesda, MD (August 18th!) Message-ID: 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 (Aubrey@nsh.org) or myself (ratliffjack@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 From monettes <@t> mskcc.org Tue Jul 24 15:56:27 2012 From: monettes <@t> mskcc.org (monettes@mskcc.org) Date: Tue Jul 24 15:56:48 2012 Subject: [Histonet] Job Opportunity at Weill Cornell Medical College, NYC Message-ID: 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 S?bastien 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: monettes@mskcc.org Email WCMC: sem2010@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. From SteveM <@t> mcclainlab.com Tue Jul 24 16:29:35 2012 From: SteveM <@t> mcclainlab.com (Steve McClain) Date: Tue Jul 24 16:29:47 2012 Subject: [Histonet] counter stain for PAS for fungus Message-ID: <012ADA4B5CC00F4AB5E4BAA399E0A5DF11B40436@ML1.McClainLabs.local> 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 From CIngles <@t> uwhealth.org Tue Jul 24 17:24:51 2012 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Tue Jul 24 17:25:33 2012 Subject: [Histonet] Unsubscribe References: <1656138725.85282.1343147809231.JavaMail.root@sz0075a.emeryville.ca.mail.comcast.net> Message-ID: <064F1ACBAE8A78469AE2E41D533D87E505A87B@UWHC-MAIL2.uwhis.hosp.wisc.edu> I vote for Raspberry! Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of nmhisto@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! From Tony_Reilly <@t> health.qld.gov.au Tue Jul 24 18:23:58 2012 From: Tony_Reilly <@t> health.qld.gov.au (Tony Reilly) Date: Tue Jul 24 18:25:04 2012 Subject: [Histonet] Secondary antibody causing nuclear staining? In-Reply-To: <6A8688F1-DF64-464A-97BD-98D6399955BC@yahoo.com> References: <6D6BD1DE8A5571489398B392A38A71579D1C9B96@xmdb02.nch.kids> <6D6BD1DE8A5571489398B392A38A71579D1C9C1E@xmdb02.nch.kids> <6A8688F1-DF64-464A-97BD-98D6399955BC@yahoo.com> Message-ID: <500FBB2D.411C.0039.0@health.qld.gov.au> 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_reilly@health.qld.gov.au Web: www.health.qld.gov.au/qhcss/ >>> Kim Donadio 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 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@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 H&E 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-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@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. Why is this? It is not every time >> so I find it rather stange. Anyone know why this is happening and what I >> can do to avoid it? >> >> Thank you for any suggestion, >> Eva Permaul >> Georgetown University >> _______________________________________________ >> 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. >> >> ********************************************************************************* >> >> _______________________________________________ >> 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ******************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. 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Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. ********************************************************************************** From lpwenk <@t> sbcglobal.net Tue Jul 24 18:57:14 2012 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Jul 24 18:57:17 2012 Subject: [Histonet] How to UNSUBSCRIBE In-Reply-To: <4895A1696F956D4CB56011A8C61312820C8A4E2E22@ushpwbmsmmp008.one.ads.bms.com> References: <4895A1696F956D4CB56011A8C61312820C8A4E2E22@ushpwbmsmmp008.one.ads.bms.com> Message-ID: >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 From phass <@t> ethz.ch Wed Jul 25 01:54:55 2012 From: phass <@t> ethz.ch (Hass Philipp) Date: Wed Jul 25 01:55:02 2012 Subject: AW: [Histonet] Unsubscribe In-Reply-To: <064F1ACBAE8A78469AE2E41D533D87E505A87B@UWHC-MAIL2.uwhis.hosp.wisc.edu> References: <1656138725.85282.1343147809231.JavaMail.root@sz0075a.emeryville.ca.mail.comcast.net> <064F1ACBAE8A78469AE2E41D533D87E505A87B@UWHC-MAIL2.uwhis.hosp.wisc.edu> Message-ID: <0124CE54B5793C4787431313856B527A28C9DA06@MBX20.d.ethz.ch> Please remove me from the list... From louise.renton <@t> gmail.com Wed Jul 25 01:58:06 2012 From: louise.renton <@t> gmail.com (Louise Renton) Date: Wed Jul 25 01:58:09 2012 Subject: [Histonet] old bones Message-ID: Thanks all for suggestions ... as always you guys rock! Sadly, this may almost be my last post - after close to 30 years in the lab, I'm moving into another arena all together - environmental health! So this may well be farewell to all you great guys and gals who have provided help and advice over the years -- 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? From JGarrigan <@t> cbiolabs.com Wed Jul 25 07:41:11 2012 From: JGarrigan <@t> cbiolabs.com (Jennifer Garrigan) Date: Wed Jul 25 07:42:56 2012 Subject: [Histonet] Markers for Rat Samples Message-ID: <91BD8268D893C44A823BE2653A60CE53CC432B57@cbiolabs05.CBiolabs.local> Hi All, I am in search of an IF marker to use in the detection of rat natural killer cells. From what I have found, I have seen only antibodies that were made using rat tissue which would not work for us. Any suggestions? We are also looking for markers for rat macrophage, rat B-cells, and rat T-cells (analogs of CD-3, CD-4, and CD-8 , of mouse and human ) Any help is appreciated. Thank you, Jennifer Cleveland BioLbas Inc This communication may contain privileged information. It is intended solely for the use of the addressee. If you are not the intended recipient, you are strictly prohibited from disclosing, copying, distributing or using any of this information. If you received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. This communication may contain nonpublic information about individuals and businesses subject to the restrictions of the Gramm-Leach-Bliley Act. You may not directly or indirectly reuse or redisclose such information for any purpose other than to provide the services for which you are receiving the information. From LPaveli1 <@t> hurleymc.com Wed Jul 25 07:59:17 2012 From: LPaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Wed Jul 25 07:59:26 2012 Subject: [Histonet] Bielschowsky questions, Luxol Fast Blue answers, and Protocol Drift. In-Reply-To: <500EF92B.3040407@mclean.harvard.edu> References: <500EF92B.3040407@mclean.harvard.edu> Message-ID: <89F4666A496DC949A819ECC40E11C867B2A177ED@EXCHANGEMB2.hmc.hurleymc.com> Hi Tim, In our lab, we use positive charged slides for all special stains. We clean our glassware using a "low-sudsing" soap with bleach (~1 cup) added and then rinsed 3 times using distilled water. When we make up the formaldehyde solution, we consider the 40% to be 100% in making the dilution. hope this helped, Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Tim Wheelock [twheelock@mclean.harvard.edu] Sent: Tuesday, July 24, 2012 3:36 PM To: histonet@lists.utsouthwestern.edu Subject: [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 From fpearsa <@t> clemson.edu Wed Jul 25 08:06:49 2012 From: fpearsa <@t> clemson.edu (Fran Pearsall) Date: Wed Jul 25 08:06:59 2012 Subject: [Histonet] GMS stain on blood smear Message-ID: <5.0.0.25.2.20120725085453.00bc4ac8@mail.clemson.edu> I have an aspirate from a mass on an animal. It was very bloody, I have spun down a portion of the sample. Does it need to be fixed with alcohol before I make slide smears? I need to do a GMS on a direct smear. What are the staining steps I need to do for these smears? Any info would be greatly appreciated! Thanks. From dreynold <@t> mdanderson.org Wed Jul 25 09:04:46 2012 From: dreynold <@t> mdanderson.org (Reynolds,Donna M) Date: Wed Jul 25 09:04:59 2012 Subject: [Histonet] RE: Secondary antibody causing nuclear staining Message-ID: <785BBF0C5F49CE41BA74460A43A08F02322D1B7A48@DCPWVMBXC0VS3.mdanderson.edu> If you are running a negative control (no primary)with your ABC staining wouldn't you see the same nuclear labeling in the NC ? Thus alerting you to false staining and indicating that you should try a HRP conjugated secondary or use a polymer system. Good discussion thank Tony. Donna Reynolds HT(ASCP) Chief Histologist IHC Lab -----Original Message----- > 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@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 H&E 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 >> From paintedsplashes <@t> yahoo.com Wed Jul 25 09:14:06 2012 From: paintedsplashes <@t> yahoo.com (Jeanne Clark) Date: Wed Jul 25 09:14:14 2012 Subject: [Histonet] Decalcifying bone Message-ID: <1343225646.39594.YahooMailNeo@web161703.mail.bf1.yahoo.com> In looking through old procedures, I have found several different 'solutions' that have been used for fixation and decalcification of bone (particularly bone marrow cores). ?I would very much appreciate hearing what people are using today for optimal fixation and decalcification of bone for routine pathology and IHC testing. Thank you, Jeanne Clark Histology/IHC Stanford Hospital and Clinics From eca9 <@t> georgetown.edu Wed Jul 25 09:59:58 2012 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Wed Jul 25 10:00:22 2012 Subject: [Histonet] RE: Secondary antibody causing nuclear staining In-Reply-To: <785BBF0C5F49CE41BA74460A43A08F02322D1B7A48@DCPWVMBXC0VS3.mdanderson.edu> References: <785BBF0C5F49CE41BA74460A43A08F02322D1B7A48@DCPWVMBXC0VS3.mdanderson.edu> Message-ID: I do see positive nuclei in the NC. That is what I am asking about. I know I could switch methods but my question is also why if it is happening is it not as strong all the time? Why are the cells very light one day and dark the next? What is causing them to stain? Just curious is all. Eva On Wed, Jul 25, 2012 at 10:04 AM, Reynolds,Donna M wrote: > > If you are running a negative control (no primary)with your ABC staining > wouldn't you see the same nuclear labeling in the NC ? Thus alerting you to > false staining and indicating that you should try a HRP conjugated > secondary or use a polymer system. > Good discussion thank Tony. > Donna Reynolds HT(ASCP) Chief Histologist IHC Lab > > -----Original Message----- > > 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: > >> histonet-bounces@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 H&E 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 > >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From brendal.finlay <@t> medicalcenterclinic.com Wed Jul 25 10:12:34 2012 From: brendal.finlay <@t> medicalcenterclinic.com (Brendal Finlay) Date: Wed Jul 25 10:12:46 2012 Subject: [Histonet] Thank you for coverslipper responses Message-ID: <929594e776a7d135c058e3f19d1aa73f@medicalcenterclinic.com> Thank you everyone for your input on the different types of coverslippers.? I'll be sharing the information with the others that I work with so we can make a decision.? I'll be happy to share my thoughts after using whichever we choose for a while. Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com phone - 850.474.8758 fax - 850.474.8584 From jgoldsmi <@t> bidmc.harvard.edu Wed Jul 25 10:33:00 2012 From: jgoldsmi <@t> bidmc.harvard.edu (jgoldsmi@bidmc.harvard.edu) Date: Wed Jul 25 10:33:12 2012 Subject: [Histonet] CAP Sample Exchange Registry Message-ID: All, I am the current chair of the CAP's Immunohistochemistry Committee. One of the things we've been grappling with is how to help IHC laboratories evaluate difficult-to-validate antibodies (e.g. HSV, spirochetes, etc.). Some years ago, our molecular pathology colleagues at the CAP instituted a 'Sample Exchange Registry' that is designed to facilitate interlaboratory validation. The College has now expanded this service to all laboratory disciplines, including IHC laboratories. This service is free of charge. Additional information can be found below, and on the referenced website. Please direct any questions to me or Ms. P. Vasalos (email: pvasalo@cap.org). Jeff The College of American Pathologists (CAP) has announced the expansion of its current Sample Exchange Registry for Alternative Assessment. This service now includes all clinical laboratory disciplines. The Sample Exchange Registry is an Internet-based service designed to connect laboratories performing testing where no formal proficiency testing (PT) is available. Laboratories can participate in the registry service at any time. When at least three laboratories are identified as testing for the same analyte, the CAP will facilitate the sample exchange. Website: http://www.cap.org/apps/cap.portal?_nfpb=true&cntvwrPtlt_actionOverride=%2Fportlets%2FcontentViewer%2Fshow&_windowLabel=cntvwrPtlt&cntvwrPtlt%7BactionForm.contentReference%7D=laboratory_resources%2Fexc.html&_state=maximized&_pageLabel=cntvwr Jeffrey Goldsmith, MD Director, Surgical Pathology Laboratory and Gastrointestinal Pathology Fellowship Department of Pathology and Laboratory Medicine Beth Israel Deaconess Medical Center Assistant Professor of Pathology, Harvard Medical School 330 Brookline Avenue Boston, MA 02215 Consultant in Gastrointestinal Pathology Children's Hospital Boston 300 Longwood Avenue Boston, MA 02215 BIDMC Office: 617 667 2586 BIDMC Fax: 617 975 5620 CHB Pathology Department (for CHB adminstrative matters only): 617 355 7431 From carl.hobbs <@t> kcl.ac.uk Wed Jul 25 11:32:22 2012 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Wed Jul 25 11:33:11 2012 Subject: [Histonet] Re: Markers for Rat Samples Message-ID: <11D9615B89C10747B1C985966A63D7CA386298F0B6@KCL-MAIL04.kclad.ds.kcl.ac.uk> You could have a look here, in the image gallery- top left link under Immunhistochemistry . Then scroll down to individual protein/ab hyperlinks. Maybe you will find some suitable Abs. NB: Sure you can use rat primaries on rat tissue, it just depends on whether you are looking for NK cells within B-cell/plasma cell population. Include a no -primary, just to see.. Sure, ...happy to be proved Good luck. Best wishes. Carl From louise.renton <@t> gmail.com Wed Jul 25 11:53:45 2012 From: louise.renton <@t> gmail.com (Louise Renton) Date: Wed Jul 25 11:53:53 2012 Subject: [Histonet] Thanks for all the good wishes Message-ID: Also....I'll try to get the "UNSUSCIBE" "UNSCRIBE" "UNSUSCRIBE" thingy right when the time comes! -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa +27 11 717 2298 (tel & fax) 073 5574456 (emergencies only) A good head and a good heart are always a formidable combination. Nelson Mandela From LPaveli1 <@t> hurleymc.com Wed Jul 25 12:14:13 2012 From: LPaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Wed Jul 25 12:14:21 2012 Subject: [Histonet] Decalcifying bone In-Reply-To: <1343225646.39594.YahooMailNeo@web161703.mail.bf1.yahoo.com> References: <1343225646.39594.YahooMailNeo@web161703.mail.bf1.yahoo.com> Message-ID: <89F4666A496DC949A819ECC40E11C867B2A178A1@EXCHANGEMB2.hmc.hurleymc.com> For fixation, we use Z-fix from Anatech, LTD. It gets the great sharp nuclear detail that's so imperative to diagnosis but without the environmental issues of the old B-5 fixative. After fixation, we decalcify in a formic acid solution called Immunocal from Decal Chemical Corp. Our IHC works well with this combo. regards, Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] on behalf of Jeanne Clark [paintedsplashes@yahoo.com] Sent: Wednesday, July 25, 2012 10:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Decalcifying bone In looking through old procedures, I have found several different 'solutions' that have been used for fixation and decalcification of bone (particularly bone marrow cores). I would very much appreciate hearing what people are using today for optimal fixation and decalcification of bone for routine pathology and IHC testing. Thank you, Jeanne Clark Histology/IHC Stanford Hospital and Clinics _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rsrichmond <@t> gmail.com Wed Jul 25 12:31:19 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Wed Jul 25 12:31:25 2012 Subject: [Histonet] Re: Decalcifying bone Message-ID: Jeanne Clark, Histology/IHC, Stanford Hospital and Clinics asks: >>In looking through old procedures, I have found several different 'solutions' that have been used for fixation and decalcification of bone (particularly bone marrow cores). I would very much appreciate hearing what people are using today for optimal fixation and decalcification of bone for routine pathology and IHC testing.<< I don't think that any of the present fixatives are significantly better for bone marrow (and other surgical bone) than neutral buffered formalin, and the ordinary commercial decalcifiers suffice. More important is meticulous technique. Specimens must be adequately fixed, and that often means overnight fixation before decalcifying; do not use mixtures that combine fixative and decalcifier. Large clots must be cut into thin slices so that they fix promptly. Femoral heads and the like need to be slabbed with a saw, and the slab fixed overnight. Decalcification must not be prolonged. Constant feedback among histotechnologist, pathologist, oncologist, and the person assisting the oncologist is essential, though rarely achieved. Of course, if the oncologist has given you crappy material to start out with, then none of this is going to help! Bob Richmond Samurai Pathologist Knoxville TN From Mark.Elliott <@t> hli.ubc.ca Wed Jul 25 12:36:30 2012 From: Mark.Elliott <@t> hli.ubc.ca (Mark Elliott) Date: Wed Jul 25 12:36:38 2012 Subject: [Histonet] Rabbit M1 and M2 Markers In-Reply-To: <91E7A005.809@mail.mrl.ubc.ca> References: <91E7A005.809@mail.mrl.ubc.ca> Message-ID: <500FCC2E020000D60004F7E7@mail.mrl.ubc.ca> What are people using to differentiate between M1 and M2 cells in rabbit tissues?? Mark ***CONFIDENTIALITY NOTICE*** This electronic message and any attachments are intended only for the use of the addressee and may contain information that is privileged and confidential. Any dissemination, distribution or copying of this communication by unauthorized individuals is strictly prohibited. If you have received this communication in error, please notify the sender immediately by reply e-mail and delete the original and all copies from your system. From thomas.huynh <@t> mdanderson.org Wed Jul 25 12:44:52 2012 From: thomas.huynh <@t> mdanderson.org (Huynh,Thomas) Date: Wed Jul 25 12:44:59 2012 Subject: [Histonet] RE: Decalcifying bone In-Reply-To: <0eb54a92-b37f-4550-a76d-920ef4e39633@DCPWPRTR02.mdanderson.edu> References: <0eb54a92-b37f-4550-a76d-920ef4e39633@DCPWPRTR02.mdanderson.edu> Message-ID: Ms. Clark We are currently use 10% formic Acid in a microwave oven to decal ours for about 2 and 1/2 hour after they were fix in 10% neutral buffer Formalin. We were told that they are Ok for immuno-stains. Thomas Huynh, HT( ASCP) Clinical Histology Technician Pathology/ Histology/ Bone Lab thuynh@mdanderson.org T 713-745-4759 F 713-792-2046 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, July 25, 2012 12:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 104, Issue 30 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. Decalcifying bone (Jeanne Clark) 2. Re: RE: Secondary antibody causing nuclear staining (Eva Permaul) 3. Thank you for coverslipper responses (Brendal Finlay) 4. CAP Sample Exchange Registry (jgoldsmi@bidmc.harvard.edu) 5. Re: Markers for Rat Samples (Hobbs, Carl) 6. Thanks for all the good wishes (Louise Renton) ---------------------------------------------------------------------- Message: 1 Date: Wed, 25 Jul 2012 07:14:06 -0700 (PDT) From: Jeanne Clark Subject: [Histonet] Decalcifying bone To: "histonet@lists.utsouthwestern.edu" Message-ID: <1343225646.39594.YahooMailNeo@web161703.mail.bf1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 In looking through old procedures, I have found several different 'solutions' that have been used for fixation and decalcification of bone (particularly bone marrow cores). ?I would very much appreciate hearing what people are using today for optimal fixation and decalcification of bone for routine pathology and IHC testing. Thank you, Jeanne Clark Histology/IHC Stanford Hospital and Clinics ------------------------------ Message: 2 Date: Wed, 25 Jul 2012 10:59:58 -0400 From: Eva Permaul Subject: Re: [Histonet] RE: Secondary antibody causing nuclear staining To: "Reynolds, Donna M" , histonet Message-ID: Content-Type: text/plain; charset=ISO-8859-1 I do see positive nuclei in the NC. That is what I am asking about. I know I could switch methods but my question is also why if it is happening is it not as strong all the time? Why are the cells very light one day and dark the next? What is causing them to stain? Just curious is all. Eva On Wed, Jul 25, 2012 at 10:04 AM, Reynolds,Donna M wrote: > > If you are running a negative control (no primary)with your ABC staining > wouldn't you see the same nuclear labeling in the NC ? Thus alerting you to > false staining and indicating that you should try a HRP conjugated > secondary or use a polymer system. > Good discussion thank Tony. > Donna Reynolds HT(ASCP) Chief Histologist IHC Lab > > -----Original Message----- > > 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.henwood@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-bounces@lists.utsouthwestern.edu [mailto: > >> histonet-bounces@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 H&E 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 > >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 3 Date: Wed, 25 Jul 2012 10:12:34 -0500 From: "Brendal Finlay" Subject: [Histonet] Thank you for coverslipper responses To: histonet@lists.utsouthwestern.edu Message-ID: <929594e776a7d135c058e3f19d1aa73f@medicalcenterclinic.com> Content-Type: text/plain; charset="utf-8" Thank you everyone for your input on the different types of coverslippers.?? I'll be sharing the information with the others that I work with so we can make a decision.?? I'll be happy to share my thoughts after using whichever we choose for a while. Brendal C. Finlay, HT (ASCP) West Florida Medical Center Clinic brendal.finlay@medicalcenterclinic.com phone - 850.474.8758 fax - 850.474.8584 ------------------------------ Message: 4 Date: Wed, 25 Jul 2012 11:33:00 -0400 From: Subject: [Histonet] CAP Sample Exchange Registry To: Cc: pvasalo@cap.org Message-ID: Content-Type: text/plain; charset="us-ascii" All, I am the current chair of the CAP's Immunohistochemistry Committee. One of the things we've been grappling with is how to help IHC laboratories evaluate difficult-to-validate antibodies (e.g. HSV, spirochetes, etc.). Some years ago, our molecular pathology colleagues at the CAP instituted a 'Sample Exchange Registry' that is designed to facilitate interlaboratory validation. The College has now expanded this service to all laboratory disciplines, including IHC laboratories. This service is free of charge. Additional information can be found below, and on the referenced website. Please direct any questions to me or Ms. P. Vasalos (email: pvasalo@cap.org). Jeff The College of American Pathologists (CAP) has announced the expansion of its current Sample Exchange Registry for Alternative Assessment. This service now includes all clinical laboratory disciplines. The Sample Exchange Registry is an Internet-based service designed to connect laboratories performing testing where no formal proficiency testing (PT) is available. Laboratories can participate in the registry service at any time. When at least three laboratories are identified as testing for the same analyte, the CAP will facilitate the sample exchange. Website: http://www.cap.org/apps/cap.portal?_nfpb=true&cntvwrPtlt_actionOverride=%2Fportlets%2FcontentViewer%2Fshow&_windowLabel=cntvwrPtlt&cntvwrPtlt%7BactionForm.contentReference%7D=laboratory_resources%2Fexc.html&_state=maximized&_pageLabel=cntvwr Jeffrey Goldsmith, MD Director, Surgical Pathology Laboratory and Gastrointestinal Pathology Fellowship Department of Pathology and Laboratory Medicine Beth Israel Deaconess Medical Center Assistant Professor of Pathology, Harvard Medical School 330 Brookline Avenue Boston, MA 02215 Consultant in Gastrointestinal Pathology Children's Hospital Boston 300 Longwood Avenue Boston, MA 02215 BIDMC Office: 617 667 2586 BIDMC Fax: 617 975 5620 CHB Pathology Department (for CHB adminstrative matters only): 617 355 7431 ------------------------------ Message: 5 Date: Wed, 25 Jul 2012 17:32:22 +0100 From: "Hobbs, Carl" Subject: [Histonet] Re: Markers for Rat Samples To: "histonet@lists.utsouthwestern.edu" Message-ID: <11D9615B89C10747B1C985966A63D7CA386298F0B6@KCL-MAIL04.kclad.ds.kcl.ac.uk> Content-Type: text/plain; charset="us-ascii" You could have a look here, in the image gallery- top left link under Immunhistochemistry . Then scroll down to individual protein/ab hyperlinks. Maybe you will find some suitable Abs. NB: Sure you can use rat primaries on rat tissue, it just depends on whether you are looking for NK cells within B-cell/plasma cell population. Include a no -primary, just to see.. Sure, ...happy to be proved Good luck. Best wishes. Carl ------------------------------ Message: 6 Date: Wed, 25 Jul 2012 18:53:45 +0200 From: Louise Renton Subject: [Histonet] Thanks for all the good wishes To: Histonet Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Also....I'll try to get the "UNSUSCIBE" "UNSCRIBE" "UNSUSCRIBE" thingy right when the time comes! -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa +27 11 717 2298 (tel & fax) 073 5574456 (emergencies only) A good head and a good heart are always a formidable combination. Nelson Mandela ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 104, Issue 30 ***************************************** From rceades <@t> gmail.com Wed Jul 25 15:15:45 2012 From: rceades <@t> gmail.com (Eric Eades) Date: Wed Jul 25 15:15:48 2012 Subject: [Histonet] Bielschowsky questions, Luxol Fast Blue answers, and Protocol Drift. In-Reply-To: <500EF92B.3040407@mclean.harvard.edu> References: <500EF92B.3040407@mclean.harvard.edu> Message-ID: Hi Tim, It is safe to consider "40% formaldehyde" to be a 100% solution, because that is the maximum amount of formaldehyde that will dissolve into an aqueous solution. Happy Fixation, -Eric Eades Phenopath Laboratories Seattle, WA On Tue, Jul 24, 2012 at 12:36 PM, Tim Wheelock wrote: > 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 > From sdysart <@t> mirnarx.com Thu Jul 26 10:25:53 2012 From: sdysart <@t> mirnarx.com (Sarah Dysart) Date: Thu Jul 26 10:26:04 2012 Subject: [Histonet] RRAS IHC Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D5018D4A418@BL2PRD0710MB363.namprd07.prod.outlook.com> Has anyone ever done any staining with RRAS. I tried doing this about a year ago and thought it had been swept under the rug...nope...I'm trying to find a good positive control...and possibly a picture of it? Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 From robyn.rhoden <@t> narmc.com Thu Jul 26 12:20:10 2012 From: robyn.rhoden <@t> narmc.com (Robyn Rhoden) Date: Thu Jul 26 12:20:16 2012 Subject: [Histonet] Bond Max for sale....in excellent shape Message-ID: <1FEB17E5D46F284CA6EC9E5315BF5B2FBF17DB4CC2@mercury.NARMC.com> We have a Bond Max for sale. It just had an $18,000 PM done on it and it's in excellent shape. The Bond Max is the best immunostainer I've ever used, and I've used them all. We just got a new one. If you're interested give us a call at 870-414-4088, ask for Bev. Robyn Rhoden From hfedor <@t> jhmi.edu Thu Jul 26 12:40:48 2012 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Thu Jul 26 12:40:58 2012 Subject: [Histonet] Bliss Imaging system Message-ID: Hello, We have a Bliss imaging system that we are no longer using. I has a great Zeiss axiophot microscope with high quality lenses. I believe that Olympus is now the company that has taken over the Bacus Labs Imaging system. Please contact me if you have a desire to acquire this system from us or know of anyone who might be interested in it.. Thanks for your consideration, Helen L. Fedor Prostate Tissue Bank, Manager TMA Core Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ From SteveM <@t> mcclainlab.com Thu Jul 26 15:31:49 2012 From: SteveM <@t> mcclainlab.com (Steve McClain) Date: Thu Jul 26 15:32:10 2012 Subject: [Histonet] Counter stain for PAS for fungus (Steve McClain) Message-ID: <012ADA4B5CC00F4AB5E4BAA399E0A5DF11B40792@ML1.McClainLabs.local> Change all your reagents. Change periodic acid every week at least. PAS unlike many other stains is a chemical reaction and is sensitive to oxidizer 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, by 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, either PAS+AB+H or PAS+AB 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. AB also demonstrates conidia and microconidia and internal structure in many fungi. AlcianBlue also stains bacteria, making it useful where polymicrobial (fungal and bacterial) infections are common. Consider it your biofilm assay. Many fungi produce mucin making PAS-AB with hema 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 more difficult for pathologists. Steedman wrote the original description , in 1951 and it remains the best paper. Steve Steve A. McClain, MD McClain Labs, LLC 45 Manor Road, Smithtown, NY 11787 631 361 4000 From susan <@t> labpath360.com Thu Jul 26 16:14:25 2012 From: susan <@t> labpath360.com (Susan Dowling) Date: Thu Jul 26 16:14:29 2012 Subject: [Histonet] PA Needed - Finder's Fee Message-ID: <001f01cd6b73$a2fec8b0$e8fc5a10$@labpath360.com> July 26, 2012 To whom it may concern: I am a recruiter in the laboratory / diagnostic industry, and we are seeking a PA for a large group in the southeast. This is an excellent opportunity for financial and career growth with excellent benefits in a very desirable area of the U.S. Our primary focus is pathology and techs, so our database is not very large in the PA arena. If you or someone you know that is from an NAACLES-Accredited Pathologists? Assistant program and is ASCP certified with a minimum of one year experience, please forward them my contact information. We also offer a "Finder's Fee", so your recommendation doesn't go unnoticed. Please advise them to mention your name as a referral source. We keep an updated database of referrals. Furthermore, we are always looking for new HT's and HTL's, so feel free to send us your resume or contact information. Our practice is based on utmost integrity - We do not share your information without your prior permission. As always, Kind regards, Susan Dowling, President 3106 Bethel Road, Suite 66 Simpsonville, SC 29681 864-236-5094 (office) 864-915-2698 (cell) 864-642-3696 (fax) www.labpath360.com susan@labpath360.com ?Finding Tomorrow?s Success Today? Confidential: This e-mail and any attachments are confidential and are intended solely for the addressee. If you are not the intended recipient of this transmission, you are hereby notified that distribution or copying this information, and any unauthorized use of the information in this transmission, are strictly prohibited and may be unlawful. If you have received this information in error, please notify the original sender and destroy its contents. The contents in this email are those of the individual sender and do not necessarily reflect LabPath360, LLC. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steve McClain Sent: Thursday, July 26, 2012 4:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counter stain for PAS for fungus (Steve McClain) Change all your reagents. Change periodic acid every week at least. PAS unlike many other stains is a chemical reaction and is sensitive to oxidizer 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, by 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, either PAS+AB+H or PAS+AB 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. AB also demonstrates conidia and microconidia and internal structure in many fungi. AlcianBlue also stains bacteria, making it useful where polymicrobial (fungal and bacterial) infections are common. Consider it your biofilm assay. Many fungi produce mucin making PAS-AB with hema 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 more difficult for pathologists. Steedman wrote the original description , in 1951 and it remains the best paper. Steve Steve A. McClain, MD McClain Labs, LLC 45 Manor Road, Smithtown, NY 11787 631 361 4000 From nmhisto <@t> comcast.net Thu Jul 26 16:23:44 2012 From: nmhisto <@t> comcast.net (nmhisto@comcast.net) Date: Thu Jul 26 16:23:51 2012 Subject: [Histonet] New Mexico Delegate to 2012 NSH Symposium? Message-ID: <1977832344.183343.1343337824716.JavaMail.root@sz0075a.emeryville.ca.mail.comcast.net> Anyone out there, from NEW MEXICO, attending NSH in Vancouver this year?? If so, please contact me as soon as possible.? This relates to New Mexico's representation at the NSH House of Delegates.? Thank you! From renafail2 <@t> gmail.com Thu Jul 26 16:54:55 2012 From: renafail2 <@t> gmail.com (Rena Fail) Date: Thu Jul 26 16:55:00 2012 Subject: [Histonet] Counter stain for PAS for fungus (Steve McClain) In-Reply-To: <012ADA4B5CC00F4AB5E4BAA399E0A5DF11B40792@ML1.McClainLabs.local> References: <012ADA4B5CC00F4AB5E4BAA399E0A5DF11B40792@ML1.McClainLabs.local> Message-ID: earlier I recommended using light green yellowish as a counterstain for Pas for fungus. Some Pathlogists prefer the green counterstain when staining with PAS for fungus. Using a 5% chromic acid solution as the oxidizer and controlling the time and temperature" will eliminate reactive aldehydes in all but the structures that have the greatest concentration of carbohydrate " Using chromic acid makes it much easier to detect fungus Histologic Dec.2005 XXXVIII(2):35 Rena Fail the first pathologist I worked for had AB-Pas stains done every day and you are right they are wickedly good. On Thu, Jul 26, 2012 at 4:31 PM, Steve McClain wrote: > Change all your reagents. > Change periodic acid every week at least. > PAS unlike many other stains is a chemical reaction and is sensitive to > oxidizer > > 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, > by 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, either PAS+AB+H or PAS+AB > 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. > AB also demonstrates conidia and microconidia and internal structure in > many fungi. > > AlcianBlue also stains bacteria, making it useful where polymicrobial > (fungal and bacterial) infections are common. > Consider it your biofilm assay. > > Many fungi produce mucin making PAS-AB with hema 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 more difficult for > pathologists. > > Steedman wrote the original description , in 1951 and it remains the best > paper. > > 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 > > From cindy38017 <@t> yahoo.com Fri Jul 27 04:53:54 2012 From: cindy38017 <@t> yahoo.com (cindy38017@yahoo.com) Date: Fri Jul 27 04:55:02 2012 Subject: [Histonet] (no subject) Message-ID: <1343382834.65140.YahooMailNeo@web37103.mail.mud.yahoo.com> http://escortozlem.net/ekle/template.php?vast223.html From christiegowan <@t> msn.com Fri Jul 27 14:25:18 2012 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Fri Jul 27 14:25:23 2012 Subject: [Histonet] Inking Skin Message-ID: Dear Colleagues, My derm pathologist has requested that we ink the epithelium on MOHs derm cases (new pathologist). We process many derm specimens a day but it seems we are having trouble getting full epithelium on our cases that come over from the MOHs clinic. I am not sure that inking the skin will help with the problem but let's just say that if we do decide to ink the surface of these specimens, does anyone know of an ink that will actually stick to skin? I would appreciate anyones input. Thanks and have a great weekend. Christie Gowan UAB Hospital Birmingham, Alabama From kstoll <@t> mcw.edu Fri Jul 27 14:47:40 2012 From: kstoll <@t> mcw.edu (Stoll, Kathryn) Date: Fri Jul 27 14:47:47 2012 Subject: [Histonet] Sections washing off slides Message-ID: I have some old H&E slides that I would like to try IHC staining on. I am having trouble keeping the tissue on the slide. I put a slide in xylene to remove the cover slip and eventually the section floated off the slide when I tried to rehydrate. The second time I tried Mount-Quick from Newcomer Supply and was able to put the section on a charged slide. The section stayed on the slide but was loose after hydration. It was just hanging on after retrieval and did not stain well. I have a limited number of slides. If anyone has experience in this area and knows the best way to keep tissue on a slide I would really appreciate your advice. Thanks in advance. Kathryn Stoll, HT(ASCP) Department of Pathology Medical College of Wisconsin 9200 W Wisconsin Ave Milwaukee WI 53226 414.805.1525 kstoll@mcw.edu From christiegowan <@t> msn.com Fri Jul 27 14:52:47 2012 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Fri Jul 27 14:52:50 2012 Subject: [Histonet] Inking Skin In-Reply-To: <5012A7F9020000AF00009BD3@nodcdmg2.no.trinity-health.org> References: , <5012A7F9020000AF00009BD3@nodcdmg2.no.trinity-health.org> Message-ID: Just to clarify: We do ink our excisions as described below in Cindi's response howerver, our MOHs specimens come to us mounted on a pieces of sturdy paper and are processed in place. We only describe them at gross and then directly place the paper and skin into a cassette for processing. The specemins are skin side up and then placed on edge when embedded. The clinician has stated that he will ink the skin for us so we have a better success rate at getting a full epithelial section at microtomy. Thanks Cindi for your input. We actually use a diluted acetic acid solution to set our inks which is the same thing as vinegar. Christie > Date: Fri, 27 Jul 2012 15:38:49 -0400 > From: robinsoc@mercyhealth.com > To: christiegowan@msn.com > Subject: Re: [Histonet] Inking Skin > > We use cotton swabs dipped in india ink in a variety of colors (purchased from the local Hobby Lobby) and set it with vinegar. We ink the resected margin not the skin. If there is no orientation we use black. If we have orientation we use at least three colors. The ellipse is inked with one color from tip to tip on one half. The other half is divided between two colors. We cut perpendicular to the side with the solid color so each cross section has two colors on it. > > > Cindi Robinson HT(ASCP) > Mercy Medical Center > Dunes Medical Laboratories > 350 W Anchor Dr > Dakota Dunes SD 57049 > phone-712-279-2768 > robinsoc@mercyhealth.com > > > >>> CHRISTIE GOWAN 7/27/2012 2:25 PM >>> > > Dear Colleagues, > My derm pathologist has requested that we ink the epithelium on MOHs derm cases (new pathologist). We process many derm specimens a day but it seems we are having trouble getting full epithelium on our cases that come over from the MOHs clinic. I am not sure that inking the skin will help with the problem but let's just say that if we do decide to ink the surface of these specimens, does anyone know of an ink that will actually stick to skin? I would appreciate anyones input. Thanks and have a great weekend. > Christie Gowan > UAB Hospital > Birmingham, Alabama _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From evzmedia <@t> yahoo.com Fri Jul 27 19:04:55 2012 From: evzmedia <@t> yahoo.com (Carl) Date: Fri Jul 27 19:05:05 2012 Subject: [Histonet] Opinions regarding Avantik RVG1 Message-ID: Hello Everyone, What has been your experiences with the company Avantik? Have you used their processor the RVG1 and what has been your experiences? Any input would be great. Regards, Carl Sent from my iPad From mehlikafaire <@t> hotmail.com Fri Jul 27 19:54:23 2012 From: mehlikafaire <@t> hotmail.com (Mehlika Faire) Date: Fri Jul 27 19:54:27 2012 Subject: [Histonet] Sections washing off slides In-Reply-To: References: Message-ID: Have you tried baking or placing your slide on a slide warmer for a few hrs or overnight prior to staining? That seems to help make the sections stick.I would do this step after transferring the section to a charged slide. -Mehlika > From: kstoll@mcw.edu > To: histonet@lists.utsouthwestern.edu > Date: Fri, 27 Jul 2012 19:47:40 +0000 > Subject: [Histonet] Sections washing off slides > > I have some old H&E slides that I would like to try IHC staining on. I am having trouble keeping the tissue on the slide. > I put a slide in xylene to remove the cover slip and eventually the section floated off the slide when I tried to rehydrate. > The second time I tried Mount-Quick from Newcomer Supply and was able to put the section on a charged slide. The section stayed on the slide but was loose after hydration. It was just hanging on after retrieval and did not stain well. > I have a limited number of slides. If anyone has experience in this area and knows the best way to keep tissue on a slide I would really appreciate your advice. Thanks in advance. > > Kathryn Stoll, HT(ASCP) > Department of Pathology > Medical College of Wisconsin > 9200 W Wisconsin Ave > Milwaukee WI 53226 > 414.805.1525 > kstoll@mcw.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Sat Jul 28 05:07:41 2012 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Jul 28 05:07:47 2012 Subject: AW: [Histonet] Sections washing off slides In-Reply-To: References: Message-ID: <002401cd6ca8$d39ade60$7ad09b20$@gmx.at> Although I have no experience in this special case, I would try to "soften down" the retrieval step. Eg. prolonged incubation time in buffer at lower temperature. If you have to remount the section, be careful, that any water is removed under the section. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Stoll, Kathryn Gesendet: Freitag, 27. Juli 2012 21:48 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Sections washing off slides I have some old H&E slides that I would like to try IHC staining on. I am having trouble keeping the tissue on the slide. I put a slide in xylene to remove the cover slip and eventually the section floated off the slide when I tried to rehydrate. The second time I tried Mount-Quick from Newcomer Supply and was able to put the section on a charged slide. The section stayed on the slide but was loose after hydration. It was just hanging on after retrieval and did not stain well. I have a limited number of slides. If anyone has experience in this area and knows the best way to keep tissue on a slide I would really appreciate your advice. Thanks in advance. Kathryn Stoll, HT(ASCP) Department of Pathology Medical College of Wisconsin 9200 W Wisconsin Ave Milwaukee WI 53226 414.805.1525 kstoll@mcw.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Sat Jul 28 05:10:45 2012 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Jul 28 05:10:50 2012 Subject: AW: [Histonet] Inking Skin In-Reply-To: References: , <5012A7F9020000AF00009BD3@nodcdmg2.no.trinity-health.org> Message-ID: <002501cd6ca9$41157310$c3405930$@gmx.at> In my mind comes the pen that is used to mark patient's skin for surgery. Does it survive processing? Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von CHRISTIE GOWAN Gesendet: Freitag, 27. Juli 2012 21:53 An: robinsoc@mercyhealth.com; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] Inking Skin Just to clarify: We do ink our excisions as described below in Cindi's response howerver, our MOHs specimens come to us mounted on a pieces of sturdy paper and are processed in place. We only describe them at gross and then directly place the paper and skin into a cassette for processing. The specemins are skin side up and then placed on edge when embedded. The clinician has stated that he will ink the skin for us so we have a better success rate at getting a full epithelial section at microtomy. Thanks Cindi for your input. We actually use a diluted acetic acid solution to set our inks which is the same thing as vinegar. Christie > Date: Fri, 27 Jul 2012 15:38:49 -0400 > From: robinsoc@mercyhealth.com > To: christiegowan@msn.com > Subject: Re: [Histonet] Inking Skin > > We use cotton swabs dipped in india ink in a variety of colors (purchased from the local Hobby Lobby) and set it with vinegar. We ink the resected margin not the skin. If there is no orientation we use black. If we have orientation we use at least three colors. The ellipse is inked with one color from tip to tip on one half. The other half is divided between two colors. We cut perpendicular to the side with the solid color so each cross section has two colors on it. > > > Cindi Robinson HT(ASCP) > Mercy Medical Center > Dunes Medical Laboratories > 350 W Anchor Dr > Dakota Dunes SD 57049 > phone-712-279-2768 > robinsoc@mercyhealth.com > > > >>> CHRISTIE GOWAN 7/27/2012 2:25 PM >>> > > Dear Colleagues, > My derm pathologist has requested that we ink the epithelium on MOHs derm cases (new pathologist). We process many derm specimens a day but it seems we are having trouble getting full epithelium on our cases that come over from the MOHs clinic. I am not sure that inking the skin will help with the problem but let's just say that if we do decide to ink the surface of these specimens, does anyone know of an ink that will actually stick to skin? I would appreciate anyones input. Thanks and have a great weekend. > Christie Gowan > UAB Hospital > Birmingham, Alabama _______________________________________________ > 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 From rjbuesa <@t> yahoo.com Sat Jul 28 10:36:41 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Jul 28 10:36:45 2012 Subject: [Histonet] Sections washing off slides In-Reply-To: References: Message-ID: <1343489801.47555.YahooMailNeo@web121406.mail.ne1.yahoo.com> How old the sections are may give you some clue about if they were initially mounted on (+) slides or not. In any event, I would try to soften the mounting medium first with heat. Try to displace the coverslip with your finger when the mounting medium softens. It usually does. If not, use warm xylene to remove the coverslip in the least amount of time possible and instead of HIER try to use protease and all steps as gently as possible. Ren? J. ________________________________ From: "Stoll, Kathryn" To: "histonet@lists.utsouthwestern.edu" Sent: Friday, July 27, 2012 3:47 PM Subject: [Histonet] Sections washing off slides I have some old H&E slides that I would like to try IHC staining on.? I am having trouble keeping the tissue on the slide. I put a slide in xylene to remove the cover slip and eventually the section floated off the slide when I tried to rehydrate. The second time I tried Mount-Quick from Newcomer Supply and was able to put the section on a charged slide.? The section stayed on the slide but was loose after hydration.? It was just hanging on after retrieval and did not stain well. I have a limited number of slides.? If anyone has experience in this area and knows the best way to keep tissue on a slide I would really appreciate your advice.? Thanks in advance. Kathryn Stoll, HT(ASCP) Department of Pathology Medical College of Wisconsin 9200 W Wisconsin Ave Milwaukee WI 53226 414.805.1525 kstoll@mcw.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mdpraet <@t> gmail.com Sat Jul 28 12:31:53 2012 From: mdpraet <@t> gmail.com (mequita praet) Date: Sat Jul 28 12:31:57 2012 Subject: [Histonet] Re: Inking skins Message-ID: Christie, The problem you are having might be more with the embedding. Puttting the epithelium on edge can be real tricky. Lots of times you think it is on edge when it is not. Try placing the skin with the epithelium on edge and then rolling it with the forceps so that it is not slightly leaning over. Make sure you have only a very thin layer of paraffin beneath it all along the edge. You have to be very quick. Then when you look at the paraffin block if you can't see the epithelium all along the edge melt it and re-embed it. I know that takes time but better to do it over than not have the epithelium. Hope this helps. Wish I was closer, I would come by to see if I could help. Mequita Praet From rsrichmond <@t> gmail.com Sat Jul 28 12:49:23 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Sat Jul 28 12:49:27 2012 Subject: [Histonet] Re: Inking Skin Message-ID: Christie Gowan at UAB Hospital. Birmingham, Alabama asks: >>My derm pathologist has requested that we ink the epithelium on MOHs derm cases (new pathologist). We process many derm specimens a day but it seems we are having trouble getting full epithelium on our cases that come over from the MOHs clinic. I am not sure that inking the skin will help with the problem but let's just say that if we do decide to ink the surface of these specimens, does anyone know of an ink that will actually stick to skin? Your new pathologist should be able to specify what he wants. About time they started teaching pathology residents where slides come from. Mohs is somebody's name, not an acronym. Specifically Frederic Mohs, an American surgeon (1910-2002): http://en.wikipedia.org/wiki/Frederic_E._Mohs You need a particulate ink that will stick to the specimen, withstand frozen section or paraffin processing, and be visible on the slide. Particulate marking inks for surgical pathology are made by a variety of manufacturers. I prefer the Davidson marking inks, but there are several other brands. They come in at least seven colors. A wooden case that keeps the tall narrow bottles from tipping over is usually included. Tattoo inks are cheap and come in an almost unlimited variety of colors, and in the one lab I've used them in work extremely well. The downside is you have to read some pretty appalling catalogs if you don't like tattoos and piercings (gimme a break, I'm 73 years old.) You can get ordinary India ink (which by definition is black) very easily. I didn't know that craft stores like Hobby Lobby offer a variety of colored particulate inks - I'd want to know exactly what to buy, to make sure I got a suitable colored ink that would show up on the slide. All of these inks have a good shelf life if you keep them tightly capped except when you're actually using them. If they become excessively viscous or solidify, they need to be thrown out. The specimen must be blotted thoroughly dry before inking, and the ink blotted off afterwards. If you do this you won't need to use anything to "set" the ink. I never use these fixatives. If you do want to use such a fixing solution, 2 or 3% acetic acid (diluted from glacial acetic acid) or half strength ordinary grocery store white vinegar is supposed to work as well as anything. It's unnecessary to use Bouin's fixative or acetone, both of which are serious hazmats. Bob Richmond Samurai Pathologist Knoxville TN From christiegowan <@t> msn.com Sat Jul 28 19:44:54 2012 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Sat Jul 28 19:44:58 2012 Subject: [Histonet] Re: Inking Skin In-Reply-To: References: Message-ID: Thanks for all the info everyone. The tattoo ink sounds intriguing. We don't seem to have a problem getting full sections on any other derm specimens so I think it is specimen specific or technique specific meaning that the epithelial layer of these samples vary in thickness so I will keep you all posted as to the outcome as I know you are all waiting with anticipation.... Thanks again Christie > Date: Sat, 28 Jul 2012 13:49:23 -0400 > From: rsrichmond@gmail.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Inking Skin > > Christie Gowan at UAB Hospital. Birmingham, Alabama asks: > > >>My derm pathologist has requested that we ink the epithelium on MOHs derm cases (new pathologist). We process many derm specimens a day but it seems we are having trouble getting full epithelium on our cases that come over from the MOHs clinic. I am not sure that inking the skin will help with the problem but let's just say that if we do decide to ink the surface of these specimens, does anyone know of an ink that will actually stick to skin? > > Your new pathologist should be able to specify what he wants. About > time they started teaching pathology residents where slides come from. > > Mohs is somebody's name, not an acronym. Specifically Frederic Mohs, > an American surgeon (1910-2002): > http://en.wikipedia.org/wiki/Frederic_E._Mohs > > You need a particulate ink that will stick to the specimen, withstand > frozen section or paraffin processing, and be visible on the slide. > > Particulate marking inks for surgical pathology are made by a variety > of manufacturers. I prefer the Davidson marking inks, but there are > several other brands. They come in at least seven colors. A wooden > case that keeps the tall narrow bottles from tipping over is usually > included. > > Tattoo inks are cheap and come in an almost unlimited variety of > colors, and in the one lab I've used them in work extremely well. The > downside is you have to read some pretty appalling catalogs if you > don't like tattoos and piercings (gimme a break, I'm 73 years old.) > > You can get ordinary India ink (which by definition is black) very > easily. I didn't know that craft stores like Hobby Lobby offer a > variety of colored particulate inks - I'd want to know exactly what to > buy, to make sure I got a suitable colored ink that would show up on > the slide. > > All of these inks have a good shelf life if you keep them tightly > capped except when you're actually using them. If they become > excessively viscous or solidify, they need to be thrown out. > > The specimen must be blotted thoroughly dry before inking, and the ink > blotted off afterwards. If you do this you won't need to use anything > to "set" the ink. I never use these fixatives. If you do want to use > such a fixing solution, 2 or 3% acetic acid (diluted from glacial > acetic acid) or half strength ordinary grocery store white vinegar is > supposed to work as well as anything. It's unnecessary to use Bouin's > fixative or acetone, both of which are serious hazmats. > > Bob Richmond > Samurai Pathologist > Knoxville TN > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wilson6848 <@t> yahoo.com Sun Jul 29 07:44:47 2012 From: wilson6848 <@t> yahoo.com (Wilson A) Date: Sun Jul 29 07:44:50 2012 Subject: [Histonet] GROSS PROCEDURE Message-ID: <1343565887.10906.YahooMailNeo@web125401.mail.ne1.yahoo.com> ?Hi, ??? Please I am trying to review my gross procedure( i.e preparing for gross and assisting with gross). I will appreciate it, if you guys could share some of your procedures with me. ? Thank you very much for your usual cooperation. ? ?? Wilson. From rsrichmond <@t> gmail.com Sun Jul 29 16:43:25 2012 From: rsrichmond <@t> gmail.com (Bob Richmond) Date: Sun Jul 29 16:43:29 2012 Subject: [Histonet] Re: Gross Procedure Message-ID: Someone identified only as Wilson asks: >>Please I am trying to review my gross procedure (i.e. preparing for gross and assisting with gross). I will appreciate it, if you guys could share some of your procedures with me.<< Any edition (the current one is the tenth, published last year) of Rosai and Ackerman's Surgical Pathology has an extensive and very widely used appendix on gross examination of complex surgical specimens. This is a book one of your pathologists is very likely to have. http://www.amazon.com/Rosai-Ackermans-Surgical-Pathology-Volume/dp/0323013422 Bob Richmond Samurai Pathologist Knoxville TN From wilson6848 <@t> yahoo.com Sun Jul 29 18:34:35 2012 From: wilson6848 <@t> yahoo.com (Wilson A) Date: Sun Jul 29 18:34:37 2012 Subject: [Histonet] Fw: GROSSING PROCEDURE In-Reply-To: <1343565887.10906.YahooMailNeo@web125401.mail.ne1.yahoo.com> References: <1343565887.10906.YahooMailNeo@web125401.mail.ne1.yahoo.com> Message-ID: <1343604875.29763.YahooMailNeo@web125405.mail.ne1.yahoo.com> ? ?Hi, ??? Please I am trying to review my grossing procedure( i.e preparing for gross and assisting with gross). I will appreciate it, if you guys could share some of your procedures with me. ? Thank you very much for your usual cooperation. ? ?? Wilson. From SteveM <@t> mcclainlab.com Sun Jul 29 21:11:02 2012 From: SteveM <@t> mcclainlab.com (Steve McClain) Date: Sun Jul 29 21:11:08 2012 Subject: [Histonet] Inking skins Message-ID: <012ADA4B5CC00F4AB5E4BAA399E0A5DF11B40E8F@ML1.McClainLabs.local> Many apologies for being cantankerous, BUT NEVER INK TISSUE WITH A COTTON TIPPED APPLICATOR Just to be clear- NEVER Cotton fibers stick to tissue and remain in block. Fibers hang on microtome blade during cutting, causing tears and artifacts in final section. I've heard it doesn't matter, we don't see a problem, etc. True, it matters less when slicing Boorshead bologna. And more when making museum-quality slides. Pathologists can see past minor imperfections. But why deliberately add to the noise or distract the pathologist? Examples: When photographing slides, time is wasted getting to the perfect section/area/ region of interest. (pathologists time is valued at zero) -repeating IHC stains due to sections falling off (double reagent cost for same fee; tech time) -holes and lost sections in tissue microarray. (those blocks are worth thousands $ and some many thousands) You get my point. The studies we do are too valuable to waste on cotton fluff. Use plain wood end of stick. Use small pipette. Steve A, McClain, MD McClain Laboratories, LLC Smithtown NY 631 361 4000 From mw <@t> personifysearch.com Mon Jul 30 08:00:55 2012 From: mw <@t> personifysearch.com (Matt Ward) Date: Mon Jul 30 08:01:02 2012 Subject: [Histonet] IHC Specialist Opportunity in FL Message-ID: Good Morning Histonet, We are currently searching for an experienced Histotech who has a background in IHC that would be interested in working with a global leader in a field support role. The IHC Field Specialist will be the expert on the company?s IHC products and will interact with clients on a daily bases. Our client will consider qualified candidates who are based in Central and Northern FL. The position offers a competitive base salary + bonus and a full benefit package including: Monthly Car Allowance, Gas Card, Paid Expenses, Cell Phone, Laptop, Health/Dental/Vision, and company matching 401k. If you or someone you may know are interested in learning more please contact me directly at mw@personifysearch.com with an updated resume, or call me at 800.875.6188 ext. 103. Have a great week! Regards, Matt Ward *Account Executive* *Personify* 5020 Weston Parkway Suite 315 Cary NC 27513 (Tel) 800.875.6188 direct ext 103 (Fax) 919.460.0642 www.personifysearch.com From LLoss <@t> dermwisconsin.com Mon Jul 30 09:06:44 2012 From: LLoss <@t> dermwisconsin.com (Lee Loss) Date: Mon Jul 30 09:06:53 2012 Subject: [Histonet] Defrost -80 Message-ID: <2B99CB40EC5CC7409888A2961CBF9688CDABF7@EX2010.DERM.LOCAL> We need to defrost our -80 freezer in a derm lab. We do DIF procedures requiring the controls to be stored in the -80. We don't have immediate access to another -80 freezer. We do have a chest freezer that will maintain -20. Can we hold those DIF controls at -20 while the -80 is down? How long can those controls be stored that way without losing their IF? Lee Loss ________________________________ The materials and information in this e-mail are confidential and may contain Protected Health Information covered under the HIPAA Privacy Rule. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution, or action taken in reliance on the contents of this information is strictly forbidden by law. If you have received this e-mail in error, please notify me by reply e-mail and then delete this message. Do not pass any of this information to anyone else. Thank you for your cooperation. From Jonathan.Cremer <@t> med.kuleuven.be Mon Jul 30 09:17:23 2012 From: Jonathan.Cremer <@t> med.kuleuven.be (Jonathan Cremer) Date: Mon Jul 30 09:17:38 2012 Subject: [Histonet] RE: Defrost -80 In-Reply-To: <2B99CB40EC5CC7409888A2961CBF9688CDABF7@EX2010.DERM.LOCAL> References: <2B99CB40EC5CC7409888A2961CBF9688CDABF7@EX2010.DERM.LOCAL> Message-ID: Can't you obtain a replacement unit from your manufacturer for a week? Otherwise, depending on the size and amount of the material to be stored, you could use dry ice in insulating boxes. That's not very user friendly though, and I wouldn't do it overnight unless someone stays to watch the amount of ice left. --- Jonathan Cremer Laboratory Technician TARGID - KU Leuven ________________________________________ Van: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] namens Lee Loss [LLoss@dermwisconsin.com] Verzonden: maandag 30 juli 2012 16:06 To: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Defrost -80 We need to defrost our -80 freezer in a derm lab. We do DIF procedures requiring the controls to be stored in the -80. We don't have immediate access to another -80 freezer. We do have a chest freezer that will maintain -20. Can we hold those DIF controls at -20 while the -80 is down? How long can those controls be stored that way without losing their IF? Lee Loss ________________________________ The materials and information in this e-mail are confidential and may contain Protected Health Information covered under the HIPAA Privacy Rule. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution, or action taken in reliance on the contents of this information is strictly forbidden by law. If you have received this e-mail in error, please notify me by reply e-mail and then delete this message. Do not pass any of this information to anyone else. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Jul 30 09:22:50 2012 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jul 30 09:22:54 2012 Subject: [Histonet] Defrost -80 In-Reply-To: <2B99CB40EC5CC7409888A2961CBF9688CDABF7@EX2010.DERM.LOCAL> References: <2B99CB40EC5CC7409888A2961CBF9688CDABF7@EX2010.DERM.LOCAL> Message-ID: <1343658170.50516.YahooMailNeo@web121406.mail.ne1.yahoo.com> Temporarily store your DIF controls in a plastic chest with dry ice (it is a much less than -80?C) and try to do everything as fast as you can. Ren? J.? ________________________________ From: Lee Loss To: "histonet@lists.utsouthwestern.edu" Sent: Monday, July 30, 2012 10:06 AM Subject: [Histonet] Defrost -80 We need to defrost our -80 freezer in a derm lab.? We do DIF procedures requiring the controls to be stored in the -80.? We don't have immediate access to another -80 freezer.? We do have a chest freezer that will maintain -20.? Can we hold those DIF controls at -20 while the -80 is down?? How long can those controls be stored that way without losing their IF? Lee Loss ________________________________ The materials and information in this e-mail are confidential and may contain Protected Health Information covered under the HIPAA Privacy Rule. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution, or action taken in reliance on the contents of this information is strictly forbidden by law. If you have received this e-mail in error, please notify me by reply e-mail and then delete this message. Do not pass any of this information to anyone else. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From madelinegi <@t> yahoo.com Mon Jul 30 09:59:56 2012 From: madelinegi <@t> yahoo.com (Madeline Gi) Date: Mon Jul 30 10:00:03 2012 Subject: [Histonet] Histology Postion Post Message-ID: <1343660396.28789.YahooMailClassic@web162302.mail.bf1.yahoo.com> Hello everyone in Histonet I was just wondering if its ok to post a postion on this site I work in a small GI lab and need some help, Just wanted to make sure that its ok to post before post the information? Thank you all Madeline Rotger Milanese H.T. BSHCS 500 New Hempstead Rd. New City N.Y. 10965 845-362-3200 Ext 129 madelinegi@yahoo.com From keva.brannock <@t> wallawallaclinic.com Mon Jul 30 10:40:44 2012 From: keva.brannock <@t> wallawallaclinic.com (Keva Brannock) Date: Mon Jul 30 10:40:51 2012 Subject: [Histonet] How to UNSUBSCRIBE In-Reply-To: References: <4895A1696F956D4CB56011A8C61312820C8A4E2E22@ushpwbmsmmp008.one.ads.bms.com> Message-ID: <006901cd6e69$aea36e20$0bea4a60$@wallawallaclinic.com> How long before the unsubscribe actually takes place? I requested it over a week ago and am still receiving messages? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 24, 2012 4:57 PM To: Lewin, Anne; histonet@lists.utsouthwestern.edu Subject: [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 ----- No virus found in this message. Checked by AVG - www.avg.com Version: 2012.0.2197 / Virus Database: 2437/5163 - Release Date: 07/29/12 ----- No virus found in this message. Checked by AVG - www.avg.com Version: 2012.0.2197 / Virus Database: 2437/5163 - Release Date: 07/29/12 From Norm.Burnham <@t> propath.com Mon Jul 30 11:23:18 2012 From: Norm.Burnham <@t> propath.com (Norm Burnham) Date: Mon Jul 30 11:23:25 2012 Subject: [Histonet] Immediate Opening for a Pathologists' Assistant Message-ID: <82C7248978CB50469FD6BA68EBBEFE67083F4317@exchange.propathlab.com> PATHOLOGISTS' ASSISTANT ProPath, a high volume, privately owned pathology practice, is seeking a Pathologists' Assistant. In this position you will be responsible for accurate description of specimens, correct dissection for microscopic diagnosis and dictation of findings for pathologist's review. The ideal candidate will have a minimum of 3 years' experience. Prefer a degree from a NACCL-accredited Pathologist's Assistant program. AAPA Fellowship/ASCP also preferred. Applicants without a degree must be able to provide documentation showing eligibility under CLIA requirements for high complexity testing. The hours for this position are 6pm to 2:30am Monday - Friday. Benefits include medical, dental, Short and Long Term Disability insurance, a matched 401K plan and more! Don't Follow the Leader! Join the Leader! EOE For consideration send resume to: ProPath, Human Resources 1355 River Bend Drive Dallas, TX 75247 www.propath.com. Accessibility Accommodations If you require an accommodation to navigate our careers site, please send your request to accessibility@propath.com ______________________________________________ Norm Burnham, MBA, MT(ASCP) Director, Anatomic Laboratory Operations ProPath - The Leader in Pathology Services 1355 River Bend Drive Dallas, TX 75247 norm.burnham@propath.com 214.237.1602 Office 214.237.1802 Fax 214.709.7127 Cell To learn more about ProPath, please visit www.propath.com This electronic message is intended to be for the use only of the named recipient and may contain information that is confidential or privileged. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this message in error or are not the named recipient, please notify us immediately by contacting the sender at the electronic mail address noted above, and delete and destroy all copies of this message. From SJMccabe <@t> drmc.org Mon Jul 30 13:40:44 2012 From: SJMccabe <@t> drmc.org (McCabe, Sara) Date: Mon Jul 30 13:42:32 2012 Subject: [Histonet] RE: Histonet Digest, Vol 104, Issue 28 In-Reply-To: <5308a0d6-8d93-4185-b731-443ef1cc6da0@EX07.drmc.org> References: <5308a0d6-8d93-4185-b731-443ef1cc6da0@EX07.drmc.org> Message-ID: <02AE2390303AAB43A823930EAD6B63AE4D738649@ex07> ________________________________________ Hi Donna, We use Novocastra/Leica Cyclin D1 (CYCLIND1-GM-L-CE) 1:50. We use a mantle cell lymphoma as control. Sara McCabe, HT (ASCP) DuBois Regional Medical Center This email and any attached files are confidential and intended solely for the intended recipient(s). If you are not the named recipient you should not read, distribute, copy or alter this email. Any views or opinions expressed in this email are those of the author and do not represent those of the company. Warning: Although precautions have been taken to make sure no viruses are present in this email, the company cannot accept responsibility for any loss or damage that arise from the use of this email or attachments. From sdysart <@t> mirnarx.com Tue Jul 31 09:38:40 2012 From: sdysart <@t> mirnarx.com (Sarah Dysart) Date: Tue Jul 31 09:38:57 2012 Subject: [Histonet] Contract rate Message-ID: <8A70A9B2ECDD084DACFE6C59FCF86D5018D4CA61@BL2PRD0710MB363.namprd07.prod.outlook.com> Hola peeps!! So does anyone know what the going rate for after hours contact HT work is. We are an urban research lab and it would be after hours work. We are in Austin Texas if that helps any =) 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 From mtighe <@t> trudeauinstitute.org Tue Jul 31 10:49:22 2012 From: mtighe <@t> trudeauinstitute.org (Mike Tighe) Date: Tue Jul 31 10:49:39 2012 Subject: [Histonet] Equipment service and repair Message-ID: <108215434A378A4E8246C567E79DD481138B34A8@CH1PRD0710MB356.namprd07.prod.outlook.com> I am wondering if there are any service company's that will repair and service Leica equipment in northern New York (Saranac Lake)? I have a Leica VT 1000s that needs some attention and would need some occasional PM visits. Thanks for any suggestions! Mike From NSEARCY <@t> swmail.sw.org Tue Jul 31 11:05:41 2012 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Tue Jul 31 11:05:55 2012 Subject: [Histonet] Symphony Issues Message-ID: <5017BC05.5D38.00EF.0@swmail.sw.org> This inquiry is from the lead histotech inour lab: Is anyone having problems with moisture on the slide, bubbles and misaligned coverslips or multiple coverslips. If so, what are they being told from Ventana? What have they done to resolve issues? Can you respond to her directly @ Pwebster@swmail.sw.org Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas, 76502 nsearcy@swmail.sw.org 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:254-724-2438 ORG:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division TEL;PAGER:633-2370 END:VCARD BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:254-724-2438 ORG:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division TEL;PAGER:633-2370 END:VCARD From melissa <@t> alliedsearchpartners.com Tue Jul 31 11:32:45 2012 From: melissa <@t> alliedsearchpartners.com (Melissa Phelan) Date: Tue Jul 31 11:32:53 2012 Subject: [Histonet] Histotech/IHC/Grossing Jobs In NY/NJ/CT areas (Permanent) Message-ID: Allied Search Partners has been retained for the following searches. We have openings in Immunohistochemistry, Grossing, and Histology. Please forward this along to anyone who you know that would be interested in any of the following positions. We do offer generous referral bonuses. 1. Please email a copy of updated resume to me or fax to 888-388-7572 for a full job description. Please note: No other information about the location or organization will be given prior to receiving an updated copy of your resume. Thank you! We have the following positions available: Immunohistochemistry (IHC) Histotech LOCATION: East White Plains/Near Port Chester, NY area DEPARTMENT & SCHEDULE: Tuesday-Saturday 2PM-10:20PM OR Monday-Friday 9:30pm-5pm Immunohistochemistry (IHC) Histotech LOCATION: Clifton, NJ area (Northern NJ) DEPARTMENT & SCHEDULE: Monday-Friday, Day Shift Histotech LOCATION: East White Plains/Near Port Chester, NY area DEPARTMENT & SCHEDULE: Tuesday-Saturday 7am-3:30PM Histotech LOCATION: Binghamton, NY (East of Ithaca) DEPARTMENT & SCHEDULE: Monday-Friday Day Shift 8AM-4:30PM Grossing Histotech LOCATION: Port Chester, NY area/East White Plains DEPARTMENT & SCHEDULE: Monday-Friday 7pm-12am To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php Melissa Phelan LinkedIn: http://www.linkedin.com/in/melissaphelan President, Laboratory Staffing Allied Search Partners P: 888.388.7571 F: 888.388.7572 M: 407.697.1175 www.alliedsearchpartners.com From Charlotte.Kopczynski <@t> baycare.org Tue Jul 31 12:16:24 2012 From: Charlotte.Kopczynski <@t> baycare.org (Kopczynski, Charlotte) Date: Tue Jul 31 12:16:28 2012 Subject: [Histonet] Ventana Symphony In-Reply-To: <9a0a62c7-a1b6-4b24-bcfe-c0fa988869a5@BCEXHT02.BCAD.BAYCARE.ORG> References: <9a0a62c7-a1b6-4b24-bcfe-c0fa988869a5@BCEXHT02.BCAD.BAYCARE.ORG> Message-ID: <16323DA35673C448BA6824C1C12426E631E45C@BCEXMBX03.BCAD.BAYCARE.ORG> We had a Symphony instrument for 6 months and had that problem - variable staining due to the humidity, bubbles, etc. Ventana engineers came to our lab and worked on the issue. Then Ventana brought a new instrument in but was never able to resolve the problem for us. We do not have a Symphony in our lab currently. It is really too bad because we really wanted it to work. The concept is the most patient safe and lean of any of the dip and dunk stain instruments. Thanks, Charlotte Kopczynski, HTL (ASCP) Pathology Manager Morton Plant Mease Health System BayCare Health System Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. From gwen <@t> gmi-inc.com Tue Jul 31 12:45:15 2012 From: gwen <@t> gmi-inc.com (Gwen Vaughn) Date: Tue Jul 31 12:45:25 2012 Subject: [Histonet] RE: Equipment service and repair In-Reply-To: <108215434A378A4E8246C567E79DD481138B34A8@CH1PRD0710MB356.namprd07.prod.outlook.com> References: <108215434A378A4E8246C567E79DD481138B34A8@CH1PRD0710MB356.namprd07.prod.outlook.com> Message-ID: Speaking of Leica VT1000S Vibrating microtome - I am looking for one, for a research facility. Let me know if you have a used one available. Thank you, Gwen Vaughn GMI, Inc. ?...Instrumental to Science Buyer http://www.gmi-inc.com/ T: 1-800-745-2710 | F: 763-712-8724 | 1-800-745-2710 Sign up to GMI for deals on New & Recertified Laboratory Instrumentation - Download Product GMI Catalogs -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike Tighe Sent: Tuesday, July 31, 2012 10:49 AM To: histonet@lists.utsouthwestern.edu (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Equipment service and repair I am wondering if there are any service company's that will repair and service Leica equipment in northern New York (Saranac Lake)? I have a Leica VT 1000s that needs some attention and would need some occasional PM visits. Thanks for any suggestions! Mike _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jaylundgren <@t> gmail.com Tue Jul 31 12:47:49 2012 From: jaylundgren <@t> gmail.com (Jay Lundgren) Date: Tue Jul 31 12:47:53 2012 Subject: [Histonet] Contract rate In-Reply-To: <8A70A9B2ECDD084DACFE6C59FCF86D5018D4CA61@BL2PRD0710MB363.namprd07.prod.outlook.com> References: <8A70A9B2ECDD084DACFE6C59FCF86D5018D4CA61@BL2PRD0710MB363.namprd07.prod.outlook.com> Message-ID: Is that full contact, or just sparring? Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) From contact <@t> excaliburpathology.com Tue Jul 31 13:23:02 2012 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Tue Jul 31 13:23:06 2012 Subject: [Histonet] (no subject) Message-ID: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> Greetings, does anyone know of a less expensive substitute for Kim-wipes? ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com From lblazek <@t> digestivespecialists.com Tue Jul 31 14:05:23 2012 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Tue Jul 31 14:07:20 2012 Subject: [Histonet] (no subject) In-Reply-To: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> References: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> Message-ID: <5A2BD13465E061429D6455C8D6B40E39163A794B79@IBMB7Exchange.digestivespecialists.com> Old telephone book pages! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, July 31, 2012 2:23 PM To: Histonet Subject: [Histonet] (no subject) Greetings, does anyone know of a less expensive substitute for Kim-wipes? ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Tue Jul 31 14:17:13 2012 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Tue Jul 31 14:17:20 2012 Subject: [Histonet] (no subject) In-Reply-To: <5A2BD13465E061429D6455C8D6B40E39163A794B79@IBMB7Exchange.digestivespecialists.com> References: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> <5A2BD13465E061429D6455C8D6B40E39163A794B79@IBMB7Exchange.digestivespecialists.com> Message-ID: Second that - learned that trick for cleaning the water bath surface from Pete Emanuele at AFIP a looooooong time ago. Brett M. Connolly, Ph.D. Principal Scientist, Imaging Dept. Merck & Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_connolly@merck.com T- 215-652-2501 F- 215-993-6803 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Blazek, Linda Sent: Tuesday, July 31, 2012 3:05 PM To: 'Paula Pierce'; Histonet Subject: RE: [Histonet] (no subject) Old telephone book pages! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, July 31, 2012 2:23 PM To: Histonet Subject: [Histonet] (no subject) Greetings, does anyone know of a less expensive substitute for Kim-wipes? ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com _______________________________________________ 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 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. From contact <@t> histocare.com Tue Jul 31 14:23:28 2012 From: contact <@t> histocare.com (Contact HistoCare) Date: Tue Jul 31 14:23:36 2012 Subject: [Histonet] Expert Level Cutting and Embedding Message-ID: Hi, We all know there are many labs in a transitional stage or are having to deal with employee turnover at any given point in time. It takes time to find the right candidate who is the best fit for your lab and its goals. HistoCare helps to alleviate workload stress while providing embedding and microtomy support for your lab with excellent and detailed results. Our interest is in short term contract partnerships (1-90 days) and are available on short notice in most states. We provide punctual and reliable assistance to your lab. Embedding experience is top notch and we haven't met a fatty or hard bony block we couldn't cut yet. We can definitely back up these claims in person! Visit www.HistoCare.com to see photos of our work. Videos demonstrating our skills are available upon request. ****Please pass this info on to anyone who's having a hard time finding great team members but still need their laboratory to run efficiently and continue to meet turn around time. From nmhisto <@t> comcast.net Tue Jul 31 14:42:16 2012 From: nmhisto <@t> comcast.net (nmhisto@comcast.net) Date: Tue Jul 31 14:42:24 2012 Subject: [Histonet] (no subject) In-Reply-To: <5A2BD13465E061429D6455C8D6B40E39163A794B79@IBMB7Exchange.digestivespecialists.com> Message-ID: <1766160803.349608.1343763736599.JavaMail.root@sz0075a.emeryville.ca.mail.comcast.net> Back in the Olde Days, when we gathered wax for embedding from the beehives in the back of the hospital and used the eggs from the hospital's chicken coops to make albumin as a slide adhesive (I'm being facetious here, I hope you know...), we used ScotTowels, torn in half, for cleaning water bath surfaces while cutting.? I've sent this idea to Paula Pierce but it bears repeating here as I'm sure one could find very inexpensive paper towels in the "pick-a-size" package.? I've always thought Kimwipes were too wimpy for cleaning the water surface but I apparently was spoiled by ScotTowels!? And, yes, I am enjoying retirement but I rather miss the routine.? I'll get over it! ----- Original Message ----- From: "Linda Blazek" To: "Paula Pierce" , "Histonet" Sent: Tuesday, July 31, 2012 1:05:23 PM Subject: RE: [Histonet] (no subject) Old telephone book pages! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, July 31, 2012 2:23 PM To: Histonet Subject: [Histonet] (no subject) Greetings, does anyone know of a less expensive substitute for Kim-wipes? ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com _______________________________________________ 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 From Wanda.Smith <@t> HCAhealthcare.com Tue Jul 31 14:48:19 2012 From: Wanda.Smith <@t> HCAhealthcare.com (Wanda.Smith@HCAhealthcare.com) Date: Tue Jul 31 14:48:41 2012 Subject: [Histonet] (no subject) In-Reply-To: <5A2BD13465E061429D6455C8D6B40E39163A794B79@IBMB7Exchange.digestivespecialists.com> References: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> <5A2BD13465E061429D6455C8D6B40E39163A794B79@IBMB7Exchange.digestivespecialists.com> Message-ID: <9E2D36CE2D7CBA4A94D9B22E8328A3BA275B00D634@NADCWPMSGCMS03.hca.corpad.net> Done that!!! And old shiny magazines. WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC? 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Blazek, Linda Sent: Tuesday, July 31, 2012 3:05 PM To: 'Paula Pierce'; Histonet Subject: RE: [Histonet] (no subject) Old telephone book pages! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, July 31, 2012 2:23 PM To: Histonet Subject: [Histonet] (no subject) Greetings, does anyone know of a less expensive substitute for Kim-wipes? ? Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com _______________________________________________ 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 From christiegowan <@t> msn.com Tue Jul 31 15:14:25 2012 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Tue Jul 31 15:14:30 2012 Subject: [Histonet] Symphony Issues In-Reply-To: <5017BC05.5D38.00EF.0@swmail.sw.org> References: <5017BC05.5D38.00EF.0@swmail.sw.org> Message-ID: Hi Nita, We had a similar problem and our Technical Service rep narrowed it down to where the coverslippers were actually manufactured. After changing out the coverslipper 3 times, he ordered a new one directly from Tuscon and we have not had the problem again. So far as humidity goes, there could be a number of issues. The first thing you need to check is with your engineers to make sure they are not pumping moisture into the air. Many hospitals do this so that is where I would start. Also, keep that filter changed regularly. I wish you luck and hope you can make it work for you. Christie Date: Tue, 31 Jul 2012 11:05:41 -0500 From: NSEARCY@swmail.sw.org To: histonet@lists.utsouthwestern.edu; PWEBSTER@swmail.sw.org CC: Subject: [Histonet] Symphony Issues This inquiry is from the lead histotech inour lab: Is anyone having problems with moisture on the slide, bubbles and misaligned coverslips or multiple coverslips. If so, what are they being told from Ventana? What have they done to resolve issues? Can you respond to her directly @ Pwebster@swmail.sw.org Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas, 76502 nsearcy@swmail.sw.org 254-724-2438 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From we3smitty <@t> yahoo.com Tue Jul 31 16:19:25 2012 From: we3smitty <@t> yahoo.com (angela smith) Date: Tue Jul 31 16:19:33 2012 Subject: [Histonet] Symphony Issues In-Reply-To: <5017BC05.5D38.00EF.0@swmail.sw.org> Message-ID: <1343769565.13222.YahooMailClassic@web125403.mail.ne1.yahoo.com> Report this to the tech support. We had to have an adjustment and also got rid of a bad lot of coverglass. they replaced it at no cost to us. --- On Tue, 7/31/12, Nita Searcy wrote: From: Nita Searcy Subject: [Histonet] Symphony Issues To: histonet@lists.utsouthwestern.edu, "Patricia Webster" Date: Tuesday, July 31, 2012, 12:05 PM This inquiry is from the lead histotech inour lab: Is anyone having problems with moisture on the slide, bubbles and misaligned coverslips or multiple coverslips. If so, what are they being told from Ventana? What have they done to resolve issues? Can you respond to her directly @ Pwebster@swmail.sw.org Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas, 76502 nsearcy@swmail.sw.org 254-724-2438 -----Inline Attachment Follows----- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From renafail2 <@t> gmail.com Tue Jul 31 16:32:34 2012 From: renafail2 <@t> gmail.com (Rena Fail) Date: Tue Jul 31 16:32:38 2012 Subject: [Histonet] (no subject) In-Reply-To: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> References: <1343758982.12170.YahooMailNeo@web5716.biz.mail.ne1.yahoo.com> Message-ID: Can't beat phone books. Cheap, torn in half they are wider than a kimwipe, you're recycling, and providing therapy in the form of stress relief obtained tearing up phone books. Rena On Tue, Jul 31, 2012 at 2:23 PM, Paula Pierce < contact@excaliburpathology.com> wrote: > Greetings, > > does anyone know of a less expensive substitute for Kim-wipes? > > > Paula K. Pierce, HTL(ASCP)HT > President > Excalibur Pathology, Inc. > 8901 S. Santa Fe, Suite G > Oklahoma City, OK 73139 > 405-759-3953 Lab > 405-759-7513 Fax > www.excaliburpathology.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From k84as <@t> yahoo.com Tue Jul 31 16:51:13 2012 From: k84as <@t> yahoo.com (mohamed abd el razik) Date: Tue Jul 31 16:51:17 2012 Subject: [Histonet] our first histology conference Message-ID: <1343771473.60746.YahooMailNeo@web140402.mail.bf1.yahoo.com> Dear all we are now arrage for the first conference of our department at Faculty of veterinary medicine- Cairo University- Egypt . Within the range of 6 monthes.....the details well be specified and tell you about....it well be an amazing opportunity for you to take a?trip and?visit Egypt and enjoy the sunny?weather....cruize the River Nile....visit Khan al- Khalili market the largiest market in the world for torrists to buy exotic perfume bottels, arab clothes and amazing sauvenirs.......Also visiting the Pyramids the only Ancient Wonder of the world that haven't been destroid by time and many many other interesting places ? So we want to enrich our new conference by 2-3 speakers who are going to visit Egypt and participate in our conference as speaker - lecturer for any topic related to our amazing feild......and realy a good opportunity to made a channel of connection between us in Egypt and you......Please don't hisitat to?replay to?me whatever your position and the favorite topic you would like to share us with it.......and don't miss the opprtunity to introduce yourself to us and enjoing the trip in Egypt......Also for companies which would like to share us email me for arrangment.... all advice from you well be appreciated ....thanks ? Dr. Mohamed abd El Razik Ass. Lec. of Vet. Histology Faculty Of Veterinary Medicine Cairo University - Egypt http://www.facebook.com/mohamed.abdelrazik.9 From Young.Kwun <@t> sswahs.nsw.gov.au Tue Jul 31 19:41:12 2012 From: Young.Kwun <@t> sswahs.nsw.gov.au (Young Kwun) Date: Tue Jul 31 19:43:16 2012 Subject: [Histonet] Proteolytic enzyme pretreatment before immunostaining Message-ID: <28587D1EA667F84BBAF99926FE94F5F40108C453@livma07.intra.swsahs.nsw.gov.au> Dear Histonetters, Could anyone explain about the difference between proteolytic enzymes for immuostaining? We use enzyme pretreatment rarely nowadays, and apart from some ready-to-use one (Dako's Proteinase K), I have used Protease (Sigma) previously when I did manual staining. At the moment I am using Leica's BondMax autostainer and their enzyme pretreatment kit (Trypsin ??, usually one drop per 7 ml vial). I know the pretreatment condition would be affected by the concentration of enzymes, pH, temperature, incubation time etc. My question is that do they have different mode of action on tissues? I am helping a research project and our antibody includes various clones of Integrin 6 subunit and uPAR. I have tried enzyme pretreatment with autostainer and manual staining with Proteinase K (Dako). It seems that some antibodies work better with certain enzymes. I mean that some antibodies work well after BondMax enzyme treatment, but some antibody works better with proteinase K pretreatment manually. I am using the same polymer detection system (Leica Microsystem) for both methods. I would like to find an enzyme which works for both of our antibodies at the same time. Thank you. Young Kwun Senior Hospital Scientist Immunohistochemistry Dept. of Anatomical Pathology Concord Repatriation General Hospital Concord NSW 2139 Australia 02-9767-6075 (Tel) 02-9767-8427 (Fax) Young.Kwun@sswahs.nsw.gov.au From chapcl <@t> yahoo.com Tue Jul 31 20:01:57 2012 From: chapcl <@t> yahoo.com (William Chappell) Date: Tue Jul 31 20:02:01 2012 Subject: [Histonet] Proteolytic enzyme pretreatment before immunostaining In-Reply-To: <28587D1EA667F84BBAF99926FE94F5F40108C453@livma07.intra.swsahs.nsw.gov.au> References: <28587D1EA667F84BBAF99926FE94F5F40108C453@livma07.intra.swsahs.nsw.gov.au> Message-ID: <4A3708CF-6DA7-4F63-9837-9EC6493502E1@yahoo.com> Enzyme pretreatment, and all steps in epitope retrieval, should follow the same quality control steps as antibody incubation and antibody concentration. Enzyme pretreatment is not magic, however the kinetics involved are very difficult to predict. Epitopes are typically chemical "shapes" within tertiary protein structure, however they can involve secondary structure and quartinary structure. The purpose of enzyme pretreatment is to get the right epitope in the right configuration so it can be recognized by the antibody. Heat retrieval is meant to break formalin cross linkages, but enzyme pretreatment actually eats away at part of the protein (depending on the protease). Too little protease treatment and it does nothing, too much and the epitope is destroyed. The bottom line, novel antibodies need to be validated with numerous retrieval methods. If it is deemed that a protease is better, numerous times and numerous concentrations should be tried -- even at different temperatures. Finally, there is no reason to believe that different novel antibodies will require the same pretreatment, however, a common pretreatment may be sufficient (though possibly not optimal) for each antibody. One last thing, if you know more about the nature of the antigen used to create the antibody, you may be able to predict the required pretreatment -- talk to the primary investigator. Will Chappell, HTL(ASCP)QIHC Anatomic Pathology Supervisor Children's Hospital of Orange County On Jul 31, 2012, at 8:41 PM, Young Kwun wrote: > Dear Histonetters, > > Could anyone explain about the difference between proteolytic enzymes > for immuostaining? > > We use enzyme pretreatment rarely nowadays, and apart from some > ready-to-use one (Dako's Proteinase K), I have used Protease (Sigma) > previously when I did manual staining. > > At the moment I am using Leica's BondMax autostainer and their enzyme > pretreatment kit (Trypsin ??, usually one drop per 7 ml vial). I know > the pretreatment condition would be affected by the concentration of > enzymes, pH, temperature, incubation time etc. > > > > My question is that do they have different mode of action on tissues? I > am helping a research project and our antibody includes various clones > of Integrin 6 subunit and uPAR. > > I have tried enzyme pretreatment with autostainer and manual staining > with Proteinase K (Dako). It seems that some antibodies work better with > certain enzymes. > > I mean that some antibodies work well after BondMax enzyme treatment, > but some antibody works better with proteinase K pretreatment manually. > I am using the same polymer detection system (Leica Microsystem) for > both methods. > > I would like to find an enzyme which works for both of our antibodies at > the same time. > > Thank you. > > > > > > > > Young Kwun > > Senior Hospital Scientist > > Immunohistochemistry > > Dept. of Anatomical Pathology > > Concord Repatriation General Hospital > > Concord NSW 2139 Australia > > > > 02-9767-6075 (Tel) > > 02-9767-8427 (Fax) > > Young.Kwun@sswahs.nsw.gov.au > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jaylundgren <@t> gmail.com Tue Jul 31 23:41:24 2012 From: jaylundgren <@t> gmail.com (Jay Lundgren) Date: Tue Jul 31 23:41:29 2012 Subject: [Histonet] Expert Level Cutting and Embedding In-Reply-To: References: Message-ID: Um, isn't blatant advertising frowned upon in this forum? Sincerely, Jay A. Lundgren, M.S., HTL (ASCP)