Re: [Histonet] Joe Nocito's request -- oh my!
Am I the only one who thinks its funny Joe the Toe did an unsubscribe e-mail and Cheryl Kerry copied an entire digest to reply. Oh histonet pet peeves... haha On Mon, Feb 4, 2013 at 1:05 PM, Cheryl tkngfl...@yahoo.com wrote: Oh no, Joe! Say it ain't so!! Please tell us this is just to take off your military address and you aren't leaving us forever... 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 ad...@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 apn...@fullstaff.org. Please include your name and specialty in the body of the email. --- On Mon, 2/4/13, histonet-requ...@lists.utsouthwestern.edu histonet-requ...@lists.utsouthwestern.edu wrote: ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 111, Issue 5
Hi Brian, I am wondering three possibilities: 1. the PFA that you use, powder form, is rather old. ( however, you said that you have tried other powders and concoctions that didn't work as well!)... perhaps, you could try using Polysciences Formaldehyde. The sealed vials come as 16% stock that you can open and dilute immediately. 2. The procedure: For some reason, the fixative isn't reaching the brain well enough. The question to you is, do you see clear, whitish coloured brain after you dissect or would you see it reddish or pinkish in colour. The reason could be a. the heart stopped beating very early in your procedure. Perhaps, slightly lower the dose or Avertin/anaesthetic. (I have had this issue previously, but it almost always works if the brain sits in the fixative unless it is never fixed! Or, the needle is not positioned correctly in the left ventricle, and fixative did not reach circulation. 3.Fixation itself: I Have had issues with the dendrites looking bloated and nobby with swiss cheese morphology before with only PFA fixation. These, however, were slice cultures, so different from whole brain fixations. They apparently needed 4% sucrose in the fixative to take care of the However, I would definitely check that. I would suggest adding 4% sucrose to the fixative as well and see if it improves the morphology. Please let us know what worked for you when you solve the issue!!! Thanks and Best, Pow Message: 4 Date: Mon, 04 Feb 2013 15:38:51 -0800 From: Brian W. Jones bwjo...@uw.edu Subject: [Histonet] extensive troubleshooting of IF on fixed-then-frozen mouse brain To: histonet@lists.utsouthwestern.edu Message-ID: 5110468b.1050...@uw.edu Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hi All, I've used Histonet many times to answer my IF/IHC questions, but this time I'm stumped. We've been using a transcardial perfusion fixation/immunofluoresence protocol for years and it has always worked. It has worked so reliably that we would often make last-minute changes---e.g., post-fix overnight instead of 4 hours---if something else came up and it would still turn out fine. Well, obviously things aren't working now or I wouldn't be pleading for help! THE PROBLEMS: Mouse brain tissue has large holes (Swiss cheese look), lacks fine structures such as axons or dendrites (looks mushy), some of the brain regions easily separate away from others (e.g., the hippocampus separates from the thalamus), and random areas of the section are not in the same focal plane as the surrounding tissue. (I tried to post images as instructed on the web site, but I can't figure that out. Please advise as I think the images would show better than I can describe.) THE PROTOCOL: Here is a detailed description of the protocol and variations I have tried while troubleshooting. Note that I have not tried every combination of every variation, and certainly I am careful to only change one thing at a time so I can make meaningful comparisons. 1) Euthanize 3 - 5-month old mouse with Beuthansia D. Wait until no longer responsive to front paw pinch and restrain animal on dissecting board. Make incision to expose thoracic cavity and lift ribcage out of the way. Heart is still beating strongly at this point. 2) Grasp heart with blunt forceps and a) make small incision in left ventricle and insert 20-g cannula, or b) insert 20-g or 25-g needle into left ventricle. 3) Using a peristaltic pump, perfuse PBS and immediately cut right atrium for outflow. Have tried PBS at either 4 C or 35 C. Have made up PBS fresh and checked pH. Have replaced tubing and used a different pump. 4) Set flow rate to a) 1.5 mL/min, b) 3 mL/min, c) 6 mL/min, or d) 10 mL/min. (Do not have a way to measure pressure.) 5) Perfuse PBS: a) 3 mL only, b) until liver blanches (never really blanches at low flow rates), or c) 15 mL. 6) PBS is either a) PBS alone, or b) PBS with heparin. 7) Perfuse with 50 mL of fixative: 4% PFA in PBS or PLP (Nakane) fixative. Note: I have tried PFA from Sigma (two different lots) or EMS (granules or 16%). Fixative is perfused at same rate and temp as PBS. 8) For 8% PFA stock: PFA is usually dissolved in water but have tried directly in PBS. Dissolved by bringing solution to 55 C then adding PFA while stirring. A few drops of 1-4 M NaOH are added until solution clears (a few undissolved particles remain). Solution is filtered through Whatman paper or 0.45 um membrane. Always made fresh the day of the perfusion. 9) Animal's lower extremities begin to move shortly after introduction of fixative. Rarely observe perfusate exiting the nose/mouth. 10) Post-fix: after perfusion, brain is dissected from skull using scissors and forceps and placed in 4 C fixative for 4-7 hours or overnight. 11) Brain is cryoprotected in sucrose-PBS: a) 10% sucrose until brain sinks, then 20%, then 30%; or b) 30% sucrose until brain sinks; c) three changes of 25% sucrose
RE: [Histonet] Joe Nocito's request -- oh my!
You're not alone- funny! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango Sent: Tuesday, February 05, 2013 2:42 PM To: Cheryl Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Joe Nocito's request -- oh my! Am I the only one who thinks its funny Joe the Toe did an unsubscribe e-mail and Cheryl Kerry copied an entire digest to reply. Oh histonet pet peeves... haha On Mon, Feb 4, 2013 at 1:05 PM, Cheryl tkngfl...@yahoo.com wrote: Oh no, Joe! Say it ain't so!! Please tell us this is just to take off your military address and you aren't leaving us forever... 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 ad...@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 apn...@fullstaff.org. Please include your name and specialty in the body of the email. --- On Mon, 2/4/13, histonet-requ...@lists.utsouthwestern.edu histonet-requ...@lists.utsouthwestern.edu wrote: ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Joe Nocito's request -- oh my!
I figured it was a test to see who would go on a rant! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango Sent: Tuesday, February 05, 2013 2:42 PM To: Cheryl Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Joe Nocito's request -- oh my! Am I the only one who thinks its funny Joe the Toe did an unsubscribe e-mail and Cheryl Kerry copied an entire digest to reply. Oh histonet pet peeves... haha On Mon, Feb 4, 2013 at 1:05 PM, Cheryl tkngfl...@yahoo.com wrote: Oh no, Joe! Say it ain't so!! Please tell us this is just to take off your military address and you aren't leaving us forever... 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 ad...@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 apn...@fullstaff.org. Please include your name and specialty in the body of the email. --- On Mon, 2/4/13, histonet-requ...@lists.utsouthwestern.edu histonet-requ...@lists.utsouthwestern.edu wrote: ___ 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] Bronch Order Forms
Hi All, Not sure how many of you do cytologies in your departments? We do quite a bit of bronchs around here and about 5 years ago a new physician who is no longer here developed the form that we currently use. It has never been liked by the nurses in scopes etc and it is pretty complicated. It allows Drs to put up to 5 microbiology, hematology, cyto or histo samples (many of which we share in all depts) on one form and has multiple check boxes to check as to what tests they want for each sample in multiple columns. We are looking to streamline this form to hopefully make it easier for everyone involved. Any of you have any Bronch ordering forms you really like or have used some in the past. Needs to be a paper order as we are not on a Lab system in Path at this point. Thanks, Helayne Parker, H.T. (ASCP) Pathology Section Head Cox Medical Center Branson P.O. Box 650, Branson, MO 65615 Phone: 417-335-7254 Fax: 417-335-7127 Email: hpar...@skaggs.net Web: www.coxhealth.com/branson CoxHealth – ranked one of Missouri's Best Hospitals by U.S. News World Report COXHEALTH 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
Re: [Histonet] HER-2
Yes, that is sometimes a common occurrence amongst pathologists, BUT those differences have to be solved in conference before issuing the report. Difficult cases (at least at my hospital) are reviewed in conference, I agree with you: your protocol (specially if it is based on DAKO's protocol) should remain as is. The diagnosis differences should not determine a change. René J. From: Mark Tarango marktara...@gmail.com To: Wilson A wilson6...@yahoo.com Cc: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Tuesday, February 5, 2013 2:24 PM Subject: Re: [Histonet] HER-2 I'd be interested to know if all your pathologists agree that the 2+ cases are 2+. Is it possible that one of the pathologists is calling more cases as 2+ than the rest? I would have a lot of questions before modifying the staining protocol. It would be helpful you posted more info. Mark On Fri, Feb 1, 2013 at 6:52 PM, Wilson A wilson6...@yahoo.com wrote: Hi, Our pathologists are concerned we may be reporting too many 2+ HER2’s. Can someone help with this? 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PRN pay?
How much is the standard pay for PRN in Texas? IHC, specials, and routine to be included in the work. 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] Prisma HE Stainer Throughput
Hello Histonetters, I am looking for a number for throughput on Prisma stainer in your Lab. we use ovens on the stainer for baking the slides. Our throughput is less than the manufactures throughput. Any input is appreciated. thank you in advance, -Kiran ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] HER-2
Many pathologists, if they have any doubt about the score will just say that it is 2+ so that its gets HER2 by FISH which is considered the best method for determining HER2 status. Even if by the scoring criteria it is a 1+ but the intensity is a little stronger than normal (but maybe basolateral or not complete staining) they just go with 2+. Sometimes its high grade 1+ and it doesn't quite meet the 2+ staining criteria and they call it 2+ too. If these things are happening enough it could mean calling more 2+ cases. They don't always follow the scoring criteria to the letter. We use digital image analysis for HER2 IHC scoring and the computer is pretty right on (matches with FISH), but sometimes the pathologist will change the score in the report to 2+ even though it's a 1+ by the computer. There is one pathologist in particular who doesn't believe in the computer reading the HER2 score and is trying very hard to find cases that are positive by FISH but that the computer is calling the IHC 1+. He also requests FISH on some 3+ cases to try and find any over-calling by the computer. The thought of a woman getting a toxic drug needlessly really bothers him. So without Wilson posting more info there's not much help that I think anyone can offer. This is stain that has to be validated more extensively than others, so the protocol shouldn't just be tweaked. How do the controls look? Was there any lot to lot variation? Lots of questions.. Mark On Tue, Feb 5, 2013 at 12:49 PM, Rene J Buesa rjbu...@yahoo.com wrote: Yes, that is sometimes a common occurrence amongst pathologists, BUT those differences have to be solved in conference before issuing the report. Difficult cases (at least at my hospital) are reviewed in conference, I agree with you: your protocol (specially if it is based on DAKO'sprotocol) should remain as is. The diagnosis differences should not determine a change. René J. *From:* Mark Tarango marktara...@gmail.com *To:* Wilson A wilson6...@yahoo.com *Cc:* histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu *Sent:* Tuesday, February 5, 2013 2:24 PM *Subject:* Re: [Histonet] HER-2 I'd be interested to know if all your pathologists agree that the 2+ cases are 2+. Is it possible that one of the pathologists is calling more cases as 2+ than the rest? I would have a lot of questions before modifying the staining protocol. It would be helpful you posted more info. Mark T. On Fri, Feb 1, 2013 at 6:52 PM, Wilson A wilson6...@yahoo.com wrote: Hi, Our pathologists are concerned we may be reporting too many 2+ HER2’s. Can someone help with this? 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] HER-2
I would suggest that you send several of your own cases to a reference lab that reads tons of them and see what numbers they report. Then compare the two results. Joyce Weems Pathology Manager 678-843-7376 Phone 678-843-7831 Fax joyce.we...@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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Wilson A Sent: Friday, February 01, 2013 9:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HER-2 Hi, Our pathologists are concerned we may be reporting too many 2+ HER2’s. Can someone help with this? Thanks, Wilson ___ 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
[Histonet] Xylene Substitutes
Hello all, My lab is looking into xylene substitutes, and I'd love some feedback on what other labs are using. We currently use SubX, but are there other items out there more economical? Thanks, Adrienne ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cytology Question for you Cytotechs
Or a question for you HT and HTL's that manage Cytotechs in your labs Is it written ANYWHERE that a Cytotech cannot or should not review their correlations and make the call whether there is a discrepancy or miss??? Then document themselves on it? Does this make sense? Thanks, Wanda 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Frozen Section Charges
Can Frozens (88331 or 88332) be charged at a Technical AND Professional level or Professional only? Thanks ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Joe Nocito's request -- oh my!
OMGoodness. I am so sorry-- Did it from my phone...thought I'd cleaned it but didn't scroll to look. Reposting the whole thing is one of my peeves, too! Again, my apologies. I'll probably do it again some day so in advance--SO SORRY!! xoxo Cheryl ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] ER/PR and Her2 image analysis
Hey- Help! Will ya'll share what image analysis software/hardware you're using? Is anyone doing full digital quatifying analysis of ER, PR and Her2 ? Thanks! Cheryl Kerry, HT(ASCP), AP Manager Pathology Group of Louisiana cke...@pathgroupla.com (LOOK GUYS!! No digest :) ) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
