RE: [Histonet] Histology
I think one of the things that I've learnt by leaving the world of Histology and dabbling (not too well) in the other disciplines is that there is no such thing as exacts. The maximum and minimum times for fixation for tissue depends on the tissue, the species, the temperature, the manufacturer of the fixative and, ah yes... The time of fixation. I think you need to do what the chemists do and control the reaction; fixatives alter proteins and the rate of reaction is dependent that which I've already said. Why concentrate on one variable? Surely the time taken to fix is just one of the variables and you need to control them all. As in all chemical reactions the time taken is dependent on all the others and you need to determine how that fixative works in your Lab, at your ambient temperature, with your manufacturer of fixative on the species of the tissue you are fixing. Multiple blocks of the same tissue, fixed for differing times and processed the way after being fixed with the same fixative; you can't go wrong. If I've been helpful then I apologise (g). Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: 07 April 2009 00:16 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology I hate to bring up histological technique for a change of pace but... Could we get some consensus of opinion as to maximum, minimum and optimal fixation times for different tissues? This is assuming that tissues will be fixed in buffered formalin at room temperature and processed to wax with a standard technique in a processor. This would also require a standard thickness for each tissue type. If there are students out there looking for projects this might seem to be suitable, as a few tissues only could be examined at one time. I know that several papers have been published about fixation in formalin but can't bring to mind any that deal with this aspect of the topic. If there are any students out there who would like a summary of fixation in general I will be happy to email it to them. Barry ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Unsubscribe
Please can you unsubscribe me from this mailing list. As an inexperienced histologist trying to find my feet I no longer feel like I could ask any questions as I'm sure I would get a torrent of abuse back instead of any helpful comments. Sophie Ainsworth Brighton and Sussex Medical School Medical Research Building Falmer East Sussex BN1 9PX Tel: #44 1273 877886 Fax: #44 1273 877884 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Large coverslips?
Hi all, I’m looking for some kind of a DIY coverslip or a tape, plastic foil… anything usable to cover some large slides (+/- 4.5cm * 15cm = 1.8inch * 5.9inch) containing botanical and zoological sections mounted in Canada Balsam. Does anyone has a (cheap…) solution for this? It doesn’t need to be pristine optical quality as the slides are primarely intended to be used on an overhead projector in the class room, but the possibility of viewing them under low power (40x - 100x) would be a real advantage. Thanks in advance four your wisdom and knowledge! Yvan. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Staying...
I am staying on the list-serve. I have learned a lot from people on this list-serve. I have been in the field since 1970 and I too am still learning things. No one has all of the answers, and sometimes there isn't just one answer. I am staying on the list-serve, and if my input can help one person, or 15 different responses can help me, then the list-serve works. There are still people out there asking questions in between the bickering. Marla Thomas, HT(ASCP) Litton Pathology Associates, PC 700 NW Hunter Dr. Blue Springs, MO 64015 Phone: 816-229-6449 Fax:816-874-4400 CONFIDENTIALITY NOTICE This message and any included attachments are from Litton Pathology Associates, P.C. 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 816-229-6449 and ask for the HIPAA/Compliance Coordinator. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Staying...
Amen! Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Marla Thomas Sent: Tuesday, April 07, 2009 7:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Staying... I am staying on the list-serve. I have learned a lot from people on this list-serve. I have been in the field since 1970 and I too am still learning things. No one has all of the answers, and sometimes there isn't just one answer. I am staying on the list-serve, and if my input can help one person, or 15 different responses can help me, then the list-serve works. There are still people out there asking questions in between the bickering. Marla Thomas, HT(ASCP) Litton Pathology Associates, PC 700 NW Hunter Dr. Blue Springs, MO 64015 Phone: 816-229-6449 Fax:816-874-4400 CONFIDENTIALITY NOTICE This message and any included attachments are from Litton Pathology Associates, P.C. 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 816-229-6449 and ask for the HIPAA/Compliance Coordinator. ___ 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] FREEZY spray
I have to agree with Jennifer on the subject of using freezing spray in cryostats containing potentially infectious agents. I have tried to promote the concept of good cryostat hygiene with everyone I train. When I started out in practice I was greeted by cryostats that have been left full of shavings. At our hospital we pathologists cut our own frozens, and it seemed to be the policy that the shavings were emptied when you could no longer close the door! One day I opened the door of the cryostat and the draft created by opening the door sent a small blizzard up and I literally inhaled shavings. From that moment on I demanded our trays be emptied and wiped clean after each use. It is tough to get some pathologists to comply as they think they are above menial cleaning tasks, and prefer there histoslaves to do there clean up. I believe these same culprits wait for their mothers to flush for them. Filthy cryostats also risk cross contamination of slides. If you drop your chuck it will come up covered in coconut. If you chunk out a precious piece of tissue in a freshly cleaned cryostat you may actually see it be able to re embed it. If you use freezing sprays in a cryostat snowstorm you will end up with a blizzard and risk not only inhalation but eye contact as well. If you must use freezing sprays, I suggest putting a dot on the chuck at 12:00, removing the chuck and spray the thing back to the ice age if you like. Return it to the chuck holder with the dot in the 12:00 position. Bring the chuck back a bit so you can start trimming over again without yet hitting the tissue in case there has been slight change in X-Y orientation. When you are done wipe it out and empty the tray. If you think this is over kill I can recommend a few polluted Beach's here in the New York metropolitan area to take your family this summer! Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 Phone and fax 201 847 7600 www.pathologyinnovations.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HT openings
I have 2 full time openings in my lab at this time. One for a supervisory level HT with IHC experience and one for a becnch level tech. Days, no weekends. If interested please call: Steve Crochiere,HT(ASCP) Histology Supervisor LifePath Partners, LLC @ Mercy Medical Center Springfield, MA 01104 413-748-9543 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 65, Issue 14
a motion is hereby offered to have the active members of histonet vote this troll off the island, or at least request that the list moderator do so. this person has proven to have absolutely nothing of value to contribute to this group, and is only intent on disrupting it. second? - Message: 16 Date: Mon, 6 Apr 2009 15:43:07 -0700 (PDT) From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] Nice one Kemlo To: histonet@lists.utsouthwestern.edu Message-ID: 453473.65848...@web43516.mail.sp1.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Instead of spitting your venom try to learn Sort of like you might consider learning correct English? I mean, its one thing if its not your first language, but despite how many people on here called you kindly and helpful, I find it perplexing that you feel compelled to call me a piece of ignorant, and TWICE, no less. I do not speak French. This list is not in French. Ergo, my awareness (or lack thereof) of proper French diction has absolutely nothing at all to do with anything... aside form providing you a vehicle to call me names and lash out. If you have something to say, say it. The simple fact that you do not like me does not give you- or anyone else- wholesale right to insult me or call me names. You should be ashamed of yourself, Rene. I never expected such antisocial behavior from you, and frankly, when you behave so regrettably, I feel no compulsion to learn anything form you! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: Histonet Digest, Vol 65, Issue 14
I second it!!! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of MKing Sent: Tuesday, April 07, 2009 6:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 65, Issue 14 a motion is hereby offered to have the active members of histonet vote this troll off the island, or at least request that the list moderator do so. this person has proven to have absolutely nothing of value to contribute to this group, and is only intent on disrupting it. second? - Message: 16 Date: Mon, 6 Apr 2009 15:43:07 -0700 (PDT) From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] Nice one Kemlo To: histonet@lists.utsouthwestern.edu Message-ID: 453473.65848...@web43516.mail.sp1.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Instead of spitting your venom try to learn Sort of like you might consider learning correct English? I mean, its one thing if its not your first language, but despite how many people on here called you kindly and helpful, I find it perplexing that you feel compelled to call me a piece of ignorant, and TWICE, no less. I do not speak French. This list is not in French. Ergo, my awareness (or lack thereof) of proper French diction has absolutely nothing at all to do with anything... aside form providing you a vehicle to call me names and lash out. If you have something to say, say it. The simple fact that you do not like me does not give you- or anyone else- wholesale right to insult me or call me names. You should be ashamed of yourself, Rene. I never expected such antisocial behavior from you, and frankly, when you behave so regrettably, I feel no compulsion to learn anything form you! ___ 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] please unsubscribe
please unsubscribe This e-mail is for the use of the intended recipient(s) only. The information contained in this communication may be confidential, privileged, or protected by copyright, and may be subject to confidentiality agreements. If you are the intended recipient and you do not wish to receive similar electronic messages from us in future then please respond to the sender to this effect. If you have received this email in error, please notify the sender immediately and then delete it. If you are not the intended recipient, you must not keep, use, disclose, copy or distribute this email without the author's prior permission. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: Histonet Digest, Vol 65, Issue 14
Can I just say that you are giving him more importance than he's worth and you are perpetuating it? Obviously he must feed off your responses so stop being manipulated; just ignore or delete or make an Outlook rule, as I have, that his e-mails go to trash!! Bernie, Bernie, where are you? I can't see you. Well I can cos people him replying to you and the craps at the bottom of the e-mail. My point so stop inflicting me on him, please! Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: 07 April 2009 14:45 To: MKing; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Histonet Digest, Vol 65, Issue 14 I second it!!! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of MKing Sent: Tuesday, April 07, 2009 6:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 65, Issue 14 a motion is hereby offered to have the active members of histonet vote this troll off the island, or at least request that the list moderator do so. this person has proven to have absolutely nothing of value to contribute to this group, and is only intent on disrupting it. second? - Message: 16 Date: Mon, 6 Apr 2009 15:43:07 -0700 (PDT) From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] Nice one Kemlo To: histonet@lists.utsouthwestern.edu Message-ID: 453473.65848...@web43516.mail.sp1.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Instead of spitting your venom try to learn Sort of like you might consider learning correct English? I mean, its one thing if its not your first language, but despite how many people on here called you kindly and helpful, I find it perplexing that you feel compelled to call me a piece of ignorant, and TWICE, no less. I do not speak French. This list is not in French. Ergo, my awareness (or lack thereof) of proper French diction has absolutely nothing at all to do with anything... aside form providing you a vehicle to call me names and lash out. If you have something to say, say it. The simple fact that you do not like me does not give you- or anyone else- wholesale right to insult me or call me names. You should be ashamed of yourself, Rene. I never expected such antisocial behavior from you, and frankly, when you behave so regrettably, I feel no compulsion to learn anything form you! ___ 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] Large coverslips?
Yvan: You could use a transparent thin polystyrene sheet cut to the size you need and cement it to your slides with the Canada balsam. I just got somewhat confused with what you wrote about low power 40x - 100x. If you are referring to an objective magnification, a 40x is always considered a high dry power and a 100x is always an oil immersion (unless it is a dry mineralogical high power objective). Any way, a polystyrene sheet will allow you to use the 40x objective. René J. --- On Tue, 4/7/09, yvan lindekens yvan_lindek...@yahoo.com wrote: From: yvan lindekens yvan_lindek...@yahoo.com Subject: [Histonet] Large coverslips? To: histonet@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 5:05 AM Hi all, I’m looking for some kind of a DIY coverslip or a tape, plastic foil… anything usable to cover some large slides (+/- 4.5cm * 15cm = 1.8inch * 5.9inch) containing botanical and zoological sections mounted in Canada Balsam. Does anyone has a (cheap…) solution for this? It doesn’t need to be pristine optical quality as the slides are primarely intended to be used on an overhead projector in the class room, but the possibility of viewing them under low power (40x - 100x) would be a real advantage. Thanks in advance four your wisdom and knowledge! Yvan. ___ 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] Please unsubscribe me
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[Histonet] Pricing of blocks
Good Morning everyone! I am opening a Histology Lab in our Dermatology practice, we routinely do approx. 10,000 biopsies a year. I would love some input from anyone out there that can assisit me with obtaining a fair price for this task. The doctors are going to be paying me per block. I will be grossing in, processing, cutting staining, maintaining CLIA, and of course maintaining quality. Thanks so much. Kathy Hicks H.T.(ASCP) DPNS Surgical Center 400 Skokie Blvd. Suite 450 Northbrook, Illinois 60062 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Job opportunity in Cambridge, MA
Hello all! We are a small pathology group within a large, Cambridge, MA based organization looking for a histologist. Our main function is to support drug development in a preclinical setting. The work would involve basic histology, IHC, and special staining on rodent tissue. Our ideal candidate is self-motivated, able to work independently but also as a team player, enthusiastic, and able to multitask. If that sounds like you, please visit our job posting at: http://www.novartis.com/careers/job-search/brassring/index.shtml and search for Job ID: 48106BR. We hope to speak with you soon! Regards, Michelle Broome Team Leader Preclinical Safety, Pathology Translational Sciences Novartis Institutes for BioMedical Research, Inc. 250 Massachusetts Avenue Cambridge, MA 02139 USA Email : michelle.bro...@novartis.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paraffin embedded mouse embryos - too dry
Dear all, I'm having trouble with my paraffin embedded mouse embryos. We use E12.5 to E15.5 embryos and we embed and cut the whole embryo. When we try to cut, the paraffin sheets are fine, but as soon as we get to the embryo, the tissue disrupts and tears. I also hear rasping or scraping sounds when cutting throught the embryo, so the problem seems to be that the tissue is too dry or too hard. Here is my current protocol: Dissect embryos from the uterus and amnion in 1x PBS, then fix overnight at 4°C in 4% PFA. Transfer to 70% Ethanol and store at 4°C. After 2 days to several weeks, we continue with the processing as follows: E12.5: 96% Ethanol for 15min, 100% Ethanol for 15min, Xylol for 15min (I also tried 10min Xylol and 8min Xylol, but with the same results). E15.5: 96% Ethanol for 30min, 100% Ethanol for 30min, Xylol for 30min (I also tried 20min Xylol and 15min Xylol, but with the same results). Then bring into Paraffin and leave overnight at 65°C, next day embedding and cooling down on-5°C cooling plate. Store at 4°C. What is the problem? Storage in 70% Ethanol for too long? Xylol incubation times too long or too short? What is the critical step - the ethanol incubation or the xylol incubation - should I try different times in Xylol or also try and change the Ethanol incubation times? Thanks in advance for any help! Best regards, Barbara -- Dipl. Biol. Barbara Schormair PhD student Institute of Human Genetics Tel.: 0049-89-3187-3953 Fax:0049-89-3187-3297 e-Mail: barbara.schorm...@helmholtz-muenchen.de Helmholtz Zentrum München German Research Center for Environmental Health (GmbH) Ingolstaedter Landstraße 1 D-85764 Neuherberg Germany Chairman of Supervisory Board: MinDir Dr. Peter Lange Board of Directors: Prof. Dr. Günther Wess and Dr. Nikolaus Blum Register of Societies: Amtsgericht München HRB 6466 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Eosinophilic granulocytes
Hi Is anyone of you aware of a marker/method to stain eosinophilic granulocytes in mouse lung tissue which is formalin fixed and paraffin embedded? Thanks Joost Bruijntjes TNO Quality of Life Zeist Holland TNO.NL http://www.tno.nl/ Joost Bruijntjes T +31 30 694 44 80 F +31 30 694 49 86 E joost.bruijnt...@tno.nl mailto:joost.bruijnt...@tno.nl Disclaimer http://www.tno.nl/tno/email/ This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Large coverslips?
Whenever I needed to prepare projection slides I coverslipped with another regular glass slide, then cleaned up the edges after the mounting medium dried. Low power microscopy was still possible providing the working distance for the objective lens was greater than 1 mm. Bryan Llewellyn - Original Message - From: yvan lindekens yvan_lindek...@yahoo.com To: histonet@lists.utsouthwestern.edu Sent: Tuesday, April 07, 2009 2:05 AM Subject: [Histonet] Large coverslips? Hi all, I’m looking for some kind of a DIY coverslip or a tape, plastic foil… anything usable to cover some large slides (+/- 4.5cm * 15cm = 1.8inch * 5.9inch) containing botanical and zoological sections mounted in Canada Balsam. Does anyone has a (cheap…) solution for this? It doesn’t need to be pristine optical quality as the slides are primarely intended to be used on an overhead projector in the class room, but the possibility of viewing them under low power (40x - 100x) would be a real advantage. Thanks in advance four your wisdom and knowledge! Yvan. ___ 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] Job opportunity in Cambridge, MA
Try: Apply this rule after message arrives with Bernie Taulin in the senders address Delete it Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of michelle.bro...@novartis.com Sent: 07 April 2009 15:01 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Job opportunity in Cambridge, MA Hello all! We are a small pathology group within a large, Cambridge, MA based organization looking for a histologist. Our main function is to support drug development in a preclinical setting. The work would involve basic histology, IHC, and special staining on rodent tissue. Our ideal candidate is self-motivated, able to work independently but also as a team player, enthusiastic, and able to multitask. If that sounds like you, please visit our job posting at: http://www.novartis.com/careers/job-search/brassring/index.shtml and search for Job ID: 48106BR. We hope to speak with you soon! Regards, Michelle Broome Team Leader Preclinical Safety, Pathology Translational Sciences Novartis Institutes for BioMedical Research, Inc. 250 Massachusetts Avenue Cambridge, MA 02139 USA Email : michelle.bro...@novartis.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
[Histonet] FW: Cervical cancer mission in Peru
From: Savaloja, Lynnette C Sent: Thursday, April 02, 2009 11:06 AM To: Webb, Dorothy L Cc: Semerad, Shelly A Subject: FW: Cervical cancer mission in Peru Importance: High Hey Dorothy, Got an old microtome laying around (see below)? Let me know... L ;) From: Fischer, Andrew [mailto:andrew.fisc...@umassmemorial.org] Sent: Thursday, April 02, 2009 10:58 AM To: memb...@lists.cytopathology.org Subject: [ASC Listserv] Cervical cancer mission in Peru Dear ASC members, Does anyone have a working microtome that could be donated to the INCCA (International Cervical Cancer Foundation) cervical cancer screening and treatment mission in Peru? This important program does not yet have a microtome to allow evaluation of the biopsies/cones. We will need it by the end of May. Please contact me if you can help. For more information on the Peru mission, see http://www.theincca.org/indextest2.html Sincerely, Andy Fischer UMASS The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material. Any review, transmission, re-transmission, dissemination or other use of, or taking of any action in reliance upon this information by persons or entities other than the intended recipient is prohibited. If you received this in error, please contact the sender and delete the material from any computer. This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Getting rid of pests
Or Bernie Taupin... Even. Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson Sent: 07 April 2009 15:16 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Job opportunity in Cambridge, MA Try: Apply this rule after message arrives with Bernie Taulin in the senders address Delete it Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of michelle.bro...@novartis.com Sent: 07 April 2009 15:01 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Job opportunity in Cambridge, MA Hello all! We are a small pathology group within a large, Cambridge, MA based organization looking for a histologist. Our main function is to support drug development in a preclinical setting. The work would involve basic histology, IHC, and special staining on rodent tissue. Our ideal candidate is self-motivated, able to work independently but also as a team player, enthusiastic, and able to multitask. If that sounds like you, please visit our job posting at: http://www.novartis.com/careers/job-search/brassring/index.shtml and search for Job ID: 48106BR. We hope to speak with you soon! Regards, Michelle Broome Team Leader Preclinical Safety, Pathology Translational Sciences Novartis Institutes for BioMedical Research, Inc. 250 Massachusetts Avenue Cambridge, MA 02139 USA Email : michelle.bro...@novartis.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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: Histonet Digest, Vol 65, Issue 14
I have tried for the past few days to stay out of this. Might I suggest that if there is a person or persons causing anyone discomfort or problems, simply add them to your blocked list in you e-mail program. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of MKing Sent: Tuesday, April 07, 2009 8:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 65, Issue 14 a motion is hereby offered to have the active members of histonet vote this troll off the island, or at least request that the list moderator do so. this person has proven to have absolutely nothing of value to contribute to this group, and is only intent on disrupting it. second? - Message: 16 Date: Mon, 6 Apr 2009 15:43:07 -0700 (PDT) From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] Nice one Kemlo To: histonet@lists.utsouthwestern.edu Message-ID: 453473.65848...@web43516.mail.sp1.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Instead of spitting your venom try to learn Sort of like you might consider learning correct English? I mean, its one thing if its not your first language, but despite how many people on here called you kindly and helpful, I find it perplexing that you feel compelled to call me a piece of ignorant, and TWICE, no less. I do not speak French. This list is not in French. Ergo, my awareness (or lack thereof) of proper French diction has absolutely nothing at all to do with anything... aside form providing you a vehicle to call me names and lash out. If you have something to say, say it. The simple fact that you do not like me does not give you- or anyone else- wholesale right to insult me or call me names. You should be ashamed of yourself, Rene. I never expected such antisocial behavior from you, and frankly, when you behave so regrettably, I feel no compulsion to learn anything form you! ___ 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] Eosinophilic granulocytes
Good morning: I have good luck by simply using haematoxylin and eosin stainingthe eosinophils pop right out by virtue of their bright pink/red granules! When used in conjunction with nuclear morphology, one can, with confidence, call these cells eosinophils. I have never performed IHC for eosinophils, but just did a quick search and found this helpful website. I am sure there are more websites like this, but this one will certainly get you started. http://www.antibodybeyond.com/reviews/cell-markers/eosinophil-marker.htm Best of luck --mtp Michele T. Pritchard, Ph.D. Research Associate Nagy Laboratory Department of Pathobiology/NE40 Lerner Research Institute Cleveland Clinic 9500 Euclid Avenue Cleveland, OH 44195 phone: 216.444.8613 fax: 216.636.1493 email: prit...@ccf.org Lab location: Lerner Research Institute NE4-214 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bruijntjes, J.P. (Joost) Sent: Tuesday, April 07, 2009 10:14 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Eosinophilic granulocytes Hi Is anyone of you aware of a marker/method to stain eosinophilic granulocytes in mouse lung tissue which is formalin fixed and paraffin embedded? Thanks Joost Bruijntjes TNO Quality of Life Zeist Holland TNO.NL http://www.tno.nl/ Joost Bruijntjes T +31 30 694 44 80 F +31 30 694 49 86 E joost.bruijnt...@tno.nl mailto:joost.bruijnt...@tno.nl Disclaimer http://www.tno.nl/tno/email/ This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet === P Please consider the environment before printing this e-mail Cleveland Clinic is ranked one of the top hospitals in America by U.S. News World Report (2008). Visit us online at http://www.clevelandclinic.org for a complete listing of our services, staff and locations. Confidentiality Note: This message is intended for use only by 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 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 contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] When sectioning small long bones in PMMA ....
... such as a mouse femur, do you prefer to orient the bone parallel to the knife edge or perpendicular to the knife edge (or diagonal)? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Entamoeba Trophozoites
I haven't checked my email for a few days! Yikes, over 150 messages! But I'll offer this anyway, however late. Entamoeba trophs can easily be demonstrated either in dried fixed smears or in paraffin sections by PAS. Their cytoplasm is loaded with glycogen so they stain very dark. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] cryoprotection possible-thanks!
Dear Histonetters, I just wanted to say a big thank you to all who responded to my cryoprotection request. We managed it to fix the samples with GA and do a sucrose step accourding to your suggestions: it worked! happy eastern to all, Frauke ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] When sectioning small long bones in PMMA ....
neither - I like it on the diagonal. Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 happy slicing and dicing and may all your stains work perfectly - Paula Sicurello On Apr 7, 2009, at 7:48 AM, Monfils, Paul wrote: ... such as a mouse femur, do you prefer to orient the bone parallel to the knife edge or perpendicular to the knife edge (or diagonal)? ___ 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] When sectioning small long bones in PMMA ....
I have always orientated the specimen in an AP view and taken longitudinal/frontal sections. As for parallel or perpendicular, I may not be completely sure of what you mean. Jack On Apr 7, 2009, at 9:48 AM, Monfils, Paul pmonf...@lifespan.org wrote: ... such as a mouse femur, do you prefer to orient the bone parallel to the knife edge or perpendicular to the knife edge (or diagonal)? ___ 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] Nice one Kemlo
I for one would be lost with out this group. I need and miss the collaboration of fellow histotechs on a daily basis. Cheri -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of alan taylor Sent: Saturday, April 04, 2009 11:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Nice one Kemlo Nice one Kemlo: /histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MAMMOGLOBIN
Has anyone used the DAKO predilute antibody, Mammoglobin, on the Ventana Benchmark XT? And if so.could you share the protocol you used? Finally looking on the Dako website it appears that they have ready to use, could you share with me which antibody should be used with the Ventana I-view detection kit? Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] unsubscribe
Cynthia Keith HT(ASCP) Histology Supervisor NorDx 102 Campus drive Scarborough, ME 04074 Tel: [207] 885-7907 Fax:[207] 885-7538 Email:kei...@mmc.org CONFIDENTIALITY NOTICE: This email message, including any attachments, is for the use of the intended recipient(s) only and may contain information that is privileged, confidential, and prohibited from unauthorized disclosure under applicable law. If you are not the intended recipient of this message, any dissemination, distribution, or copying of this message is strictly prohibited. If you received this message in error, please notify the sender by reply email and destroy all copies of the original message and attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] histonet off-topic comments
Dear Histonet members: I have removed Bernie Taupin from the list. Let's please get back to the topic of histology and related fields.Thanks Linda M Histonet administrator MKing mak...@ufl.edu 4/7/2009 8:43 AM a motion is hereby offered to have the active members of histonet vote this troll off the island, or at least request that the list moderator do so. this person has proven to have absolutely nothing of value to contribute to this group, and is only intent on disrupting it. second? - Message: 16 Date: Mon, 6 Apr 2009 15:43:07 -0700 (PDT) From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] Nice one Kemlo To: histonet@lists.utsouthwestern.edu Message-ID: 453473.65848...@web43516.mail.sp1.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Instead of spitting your venom try to learn Sort of like you might consider learning correct English? I mean, its one thing if its not your first language, but despite how many people on here called you kindly and helpful, I find it perplexing that you feel compelled to call me a piece of ignorant, and TWICE, no less. I do not speak French. This list is not in French. Ergo, my awareness (or lack thereof) of proper French diction has absolutely nothing at all to do with anything... aside form providing you a vehicle to call me names and lash out. If you have something to say, say it. The simple fact that you do not like me does not give you- or anyone else- wholesale right to insult me or call me names. You should be ashamed of yourself, Rene. I never expected such antisocial behavior from you, and frankly, when you behave so regrettably, I feel no compulsion to learn anything form you! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet /pre span style=font-weight: bold;Please consider the environment before printing this e-mail/spanbr / br / span style=font-size: 8pt;This e-mail, facsimile, or letter and any files or attachments transmitted with it containsbr / information that is confidential and privileged. This information is intended only for the use of the br / individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further br / disclosures are prohibited without proper authorization. If you are not the intended recipient, any br / disclosure, copying, printing, or use of this information is strictly prohibited and possibly a br / violation of federal or state law and regulations. If you have received this information in error, br / please notify Children's Medical Center Dallas immediately at 214-456- or via e-mail at br / priv...@childrens.com. Children's Medical Center Dallas and its affiliates hereby claim all br / applicable privileges related to this information./spanbr / br / /html ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] AE1/AE3 nuclear staining
We have recently been having dark nuclear staining with our AE1/AE3 antibody. This is a recent event and we cannot figure out what is going on. It started gradually and has been increasing in intensity. We have not changed antibody lots or any other part of our protocol. Has anyone seen this before? Any suggestions on what is happening here? Tammy Barnhart, BS, HTL(ASCP) Avera McKennan Hospital - 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. 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] Paraffin embedded mouse embryos - too dry
Good morning. A colleague of mine recently placed the 10% formalin fixed mouse embryos in 2% agarose, let them solidify, then process at 30 minutes per station under pressure/vacuum. The protocol was published, but right now I don't have that information. Hope this helps in some way! Jen Anderson The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws. If you received this in error, please contact the sender and delete/destroy this email. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Tuesday, April 07, 2009 7:40 AM Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Paraffin embedded mouse embryos - too dry I sometimes get embryos in the lab for processing and here are the protocols that I use for embryos in these stages: E12.5 - 70% Ethanol - 12 min 95% Ethanol - 12 min 100% Ethanol x 2 - 12 min each Xylene x 2 - 12 min each Paraffin - 30 min Paraffin x 2 - 1 hr each no vacuum or pressure used except for the last paraffin E15.5 - 70% Ethanol - 2 hrs (or two changes for an hour each) 95% Ethanol - 1 hr 95% Ethanol - 2 hrs 100% Ethanol x 3 - 1 hr. eash Xylene x 2 - 2 hrs each Paraffin x 4 - total of 10 hrs I use some vacuum and pressure - in the last Xylene and in the paraffins These protocols were provided to my by Gayle Callis and I have modified them a little. They work great for me and I don't have any problems with the sectioning. Good luck! Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 happy slicing and dicing and may all your stains work perfectly - Paula Sicurello On Apr 7, 2009, at 7:05 AM, Barbara Schormair wrote: Dear all, I'm having trouble with my paraffin embedded mouse embryos. We use E12.5 to E15.5 embryos and we embed and cut the whole embryo. When we try to cut, the paraffin sheets are fine, but as soon as we get to the embryo, the tissue disrupts and tears. I also hear rasping or scraping sounds when cutting throught the embryo, so the problem seems to be that the tissue is too dry or too hard. Here is my current protocol: Dissect embryos from the uterus and amnion in 1x PBS, then fix overnight at 4°C in 4% PFA. Transfer to 70% Ethanol and store at 4°C. After 2 days to several weeks, we continue with the processing as follows: E12.5: 96% Ethanol for 15min, 100% Ethanol for 15min, Xylol for 15min (I also tried 10min Xylol and 8min Xylol, but with the same results). E15.5: 96% Ethanol for 30min, 100% Ethanol for 30min, Xylol for 30min (I also tried 20min Xylol and 15min Xylol, but with the same results). Then bring into Paraffin and leave overnight at 65°C, next day embedding and cooling down on-5°C cooling plate. Store at 4°C. What is the problem? Storage in 70% Ethanol for too long? Xylol incubation times too long or too short? What is the critical step - the ethanol incubation or the xylol incubation - should I try different times in Xylol or also try and change the Ethanol incubation times? Thanks in advance for any help! Best regards, Barbara -- Dipl. Biol. Barbara Schormair PhD student Institute of Human Genetics Tel.: 0049-89-3187-3953 Fax: 0049-89-3187-3297 e-Mail: barbara.schorm...@helmholtz-muenchen.de Helmholtz Zentrum München German Research Center for Environmental Health (GmbH) Ingolstaedter Landstraße 1 D-85764 Neuherberg Germany Chairman of Supervisory Board: MinDir Dr. Peter Lange Board of Directors: Prof. Dr. Günther Wess and Dr. Nikolaus Blum Register of Societies: Amtsgericht München HRB 6466 ___ 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] Large coverslips?
Hi Yvan, Does it have to be a coverslip? There are some mounting media that harden and form a barrier with optical qualities similar to that of glass. It might not be a perfect solution, but it might work. I think the stuff I used was called Crystal Mount (?). Paula :-) Paula Sicurello VA Medical Center San Diego Veterans Medical Research Foundation (VMRF) Core Research Imaging Center 3350 La Jolla Village Dr.., MC151 San Diego, CA 92161 858-552-8585 x2397 --- On Tue, 4/7/09, yvan lindekens yvan_lindek...@yahoo.com wrote: From: yvan lindekens yvan_lindek...@yahoo.com Subject: [Histonet] Large coverslips? To: histonet@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:05 AM Hi all, I’m looking for some kind of a DIY coverslip or a tape, plastic foil… anything usable to cover some large slides (+/- 4.5cm * 15cm = 1.8inch * 5.9inch) containing botanical and zoological sections mounted in Canada Balsam. Does anyone has a (cheap…) solution for this? It doesn’t need to be pristine optical quality as the slides are primarely intended to be used on an overhead projector in the class room, but the possibility of viewing them under low power (40x - 100x) would be a real advantage. Thanks in advance four your wisdom and knowledge! Yvan. ___ 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] HI Downdraft fume extractor for sale
I have a HI downdraft fume extractor for $50 and we will provide carbon to refill the drawer. Cathy A. Mayton Wasatch Histo Consultants, Inc. 775-625-4425 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] breaking glass jar/vials for MMA embedded specimens
Yes, the scintillation vials are notorious for shattering in the freezer. However, if you put them in a plastic container with a lid on, the shattered glass will be contained and not all over your freezer. When breaking larger glass containers, again place them in the freezer, remove from the freezer after 20 minutes or so, take the lid off, wrap jar in paper towels and hold the ends closed. Wear glovesand eye protection in case shards of glass escape. Strike the jar with a hammer and unroll the paper towel over a garbage can. Rinse the block in tap water to remove any small shards of glass. This is a great way to vent frustration!! Cathy A. Mayton Wasatch Histo Consultants, Inc. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
AW: [Histonet] MAMMOGLOBIN
I tried also this antibody on Ventana Benchmark. Pity, I had no results. Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Vickroy, Jim Gesendet: Dienstag, 07. April 2009 17:54 An: 'Histonet@lists.utsouthwestern.edu' Betreff: [Histonet] MAMMOGLOBIN Has anyone used the DAKO predilute antibody, Mammoglobin, on the Ventana Benchmark XT? And if so.could you share the protocol you used? Finally looking on the Dako website it appears that they have ready to use, could you share with me which antibody should be used with the Ventana I-view detection kit? Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly 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
Re: [Histonet] AE1/AE3 nuclear staining
When was the last time you checked the pH of your retrieval solution? Or are you using an enzyme? Mark Tarango On Tue, Apr 7, 2009 at 9:53 AM, Tammy Barnhart tammy.barnh...@mckennan.orgwrote: We have recently been having dark nuclear staining with our AE1/AE3 antibody. This is a recent event and we cannot figure out what is going on. It started gradually and has been increasing in intensity. We have not changed antibody lots or any other part of our protocol. Has anyone seen this before? Any suggestions on what is happening here? Tammy Barnhart, BS, HTL(ASCP) Avera McKennan Hospital - 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. 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Tissue Capture Pen
hello, if any of you are using and/or have used the Ted Pella, Inc. Tissue Capture Pen will you be so kind as to share with me your experience (s), pros, cons and whatever else you deem necessary? Also, does it's use require special glass slides? Any info you can provide on it's use ASAP will be much appreciated. Thank you kindly, Atoska ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: I'm outta here
I can spot spam and o.t. posts and delete them in a half second each. The questions on Histonet make me aware of the extent of use of special stains, and the potential for new stains. Jjob postings are forwarded to the career service office here. The safety warnings are often very useful. HIstonet is a great source of information on antigen retrieval techniques and antibodies. I would really miss Histonet. --Allen A Smith, Ph.D. Barry University School of Podiatric Medicine ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Controls needed!
We are looking for H. Pylori control tissue and also GMS/fungus control tissue. Is there anyone out there that might have extra to share? We have good GRAM control blocks that we would be happy to exchange. Please let me know if you can help us out. Thank you! Deanne Knutson Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, N. Dak. 58506 701-530-6730 Fax 701-530-6735 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fixation question - Cerebellar granular cells
Dear Histoneters, I am doing some IHC on rat cerebellar granular cells (NOT TISSUE) and I need to fix them with PF. I have gotten 4 different answers on how to go about this, and I wanted to run this by the list to see what you think. 1) Fix in COLD, 4% PF in 1x PBS 2) Fix in COLD, 4% PF in 1x PBS with 4% Sucrose. 3) Fix in WARM (37 C) ,in 4% PF in 1x PBS. 4) Fix in WARM (37 C), in 1x PBS with 4% Sucrose. I get the fact that the PF might need to be at 37 C, since it is the temperature that the cells are in the incubator and it would probably temperature shock them. What about the sucrose? does it remove the water? I'd appreciate your thoughts... Best, Guillermo Guillermo Palchik g...@georgetown.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Recycled formalin
Is anyone using recycled formalin for primary fixation of either surgical or autopsy tissue? Thanks. Richard Richard W. Cartun, Ph.D. Director, Histology Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Unsubscribe
Sophie, I for one would never throw a torrent of abuse at any one (mmm unless they are politicians - in between football seasons!) So if my (and most others on Histonet) comments are not of some worth then we apologise. We need to try harder. (also please remember the delete key, I unfortunately have to regularly use it) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of s.j.ainswo...@bsms.ac.uk Sent: Tuesday, 7 April 2009 5:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Unsubscribe Please can you unsubscribe me from this mailing list. As an inexperienced histologist trying to find my feet I no longer feel like I could ask any questions as I'm sure I would get a torrent of abuse back instead of any helpful comments. Sophie Ainsworth Brighton and Sussex Medical School Medical Research Building Falmer East Sussex BN1 9PX Tel: #44 1273 877886 Fax: #44 1273 877884 ___ 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
RE: [Histonet] gi microwave processing
Nancy, Best advice I can offer is to ensure as much fixation as you can. We start with 30 minutes at 30oC and the ramp it up to 40oC for 30-60 minutes, rinse in 70% ethanol (5min)then continue microwave processing with isopropanol and wax. You may also consider decreasing the processing temperatures. GI biopsies being quite small do not need over-the-top processing, but in my experience, fixation is still the key. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of magod...@aol.com Sent: Tuesday, 7 April 2009 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] gi microwave processing I am starting a gi lab and using a microwave processor for my first time. Any suggestions? SHUR Wave processor. Thanks, Nancy **A Good Credit Score is 700 or Above. See yours in just 2 easy steps! (http://pr.atwola.com/promoclk/100126575x1221621488x1201450096/aol?redir =http:%2F%2Fwww.freecreditreport.com%2Fpm%2Fdefault.aspx%3Fsc%3D668072%2 6hmpgID %3D62%26bcd%3DAprilfooterNO62) ___ 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
RE: [Histonet] AE1/AE3 nuclear staining
Possibly the concentration of hydrogen peroxide in the DAB solution has been slowly increasing (maybe micropipette creep?) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tammy Barnhart Sent: Wednesday, 8 April 2009 2:54 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AE1/AE3 nuclear staining We have recently been having dark nuclear staining with our AE1/AE3 antibody. This is a recent event and we cannot figure out what is going on. It started gradually and has been increasing in intensity. We have not changed antibody lots or any other part of our protocol. Has anyone seen this before? Any suggestions on what is happening here? Tammy Barnhart, BS, HTL(ASCP) Avera McKennan Hospital - 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. 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 * 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
Re: [Histonet] Recycled formalin
Yes. ?? Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 I find that the harder I work, the more luck I seem to have. - Thomas Jefferson Richard Cartun rcar...@harthosp.org 04/07/09 7:30 PM Is anyone using recycled formalin for primary fixation of either surgical or autopsy tissue? Thanks. Richard Richard W. Cartun, Ph.D. Director, Histology Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Fixation question - Cerebellar granular cells
Hi, i normally work on tissue. I think 4% PFA in 0.1M PB is fine. so approach 3. 2009-04-08 TF 发件人： Guillermo Palchik 发送时间： 2009-04-08 05:03:13 收件人： histonet 抄送： 主题： [Histonet] Fixation question - Cerebellar granular cells Dear Histoneters, I am doing some IHC on rat cerebellar granular cells (NOT TISSUE) and I need to fix them with PF. I have gotten 4 different answers on how to go about this, and I wanted to run this by the list to see what you think. 1) Fix in COLD, 4% PF in 1x PBS 2) Fix in COLD, 4% PF in 1x PBS with 4% Sucrose. 3) Fix in WARM (37 C) ,in 4% PF in 1x PBS. 4) Fix in WARM (37 C), in 1x PBS with 4% Sucrose. I get the fact that the PF might need to be at 37 C, since it is the temperature that the cells are in the incubator and it would probably temperature shock them. What about the sucrose? does it remove the water? I'd appreciate your thoughts... Best, Guillermo Guillermo Palchik g...@georgetown.edu ___ 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] Staying...
Yes, Well-done to all of you who have decided to stay! It really separates the wheat from the chaff, as it were. Or the weenies from the real adults, more or less. And even the French-speakers from the non-French-speakers, in the case of Kemlo/Rene. Ha ha. That was a joke. Learn to laugh a little, folks. I, myself, have decided to stay. I'll try to play more nicely. Here's hoping the rest of you can be so cool about it all. SO BIG UPS TO ALL THE HISTOLOGISTS IN THE HOUSE! CAN I GET A WHUT-WHUT! From: Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov To: Marla Thomas mtho...@littonlab.com; histonet@lists.utsouthwestern.edu Sent: Tuesday, April 7, 2009 8:10:22 AM Subject: RE: [Histonet] Staying... Amen! Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Marla Thomas Sent: Tuesday, April 07, 2009 7:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Staying... I am staying on the list-serve. I have learned a lot from people on this list-serve. I have been in the field since 1970 and I too am still learning things. No one has all of the answers, and sometimes there isn't just one answer. I am staying on the list-serve, and if my input can help one person, or 15 different responses can help me, then the list-serve works. There are still people out there asking questions in between the bickering. Marla Thomas, HT(ASCP) Litton Pathology Associates, PC 700 NW Hunter Dr. Blue Springs, MO 64015 Phone: 816-229-6449 Fax:816-874-4400 CONFIDENTIALITY NOTICE This message and any included attachments are from Litton Pathology Associates, P.C. 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 816-229-6449 and ask for the HIPAA/Compliance Coordinator. ___ 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] FREEZY spray
Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE: Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned. Regards,Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ 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] FREEZY spray
I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the wayThis came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned. Regards, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ Histonet mailing list histo...@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] FREEZY spray
Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE: Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned.. Regards,Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo..com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ 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] FREEZY spray
Sorry, again, I'm confused... are you responding to me? I'm not the original poster... I never stated I had trouble with cracking, because, well, I don't. I'm Bernie Taupin, the King of Cryomicrotomy, Esq. From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:27:35 AM Subject: Re: [Histonet] FREEZY spray Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo..com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds.. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned.. Regards, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo..com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___
