RE: RE: RE: [Histonet] IHC on paraformaldehyde-fixed
Isn't petrol toxic? Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; Don’t be afraid to take a big step when one is indicated. You can’t cross a chasm in two small jumps. --Buckminster Fuller 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: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Tony Henwood Sent: 07 December 2008 22:06 To: [EMAIL PROTECTED]; Reuel Cornelia; histonet; [EMAIL PROTECTED]; Jan Shivers Subject: RE: RE: RE: [Histonet] IHC on paraformaldehyde-fixed Tf, In answer to you email: No I do not carry toxic liquid in my car. But does the PFA powder dissolve easily? My experience is that you need to make the solution alkaline then heat it. I never add methanol to my 10% neutral buffered formalin (it is buffered and diluted ie 10%). The risk of polymerisation of the formalin (since it is diluted) and formic acid formation (since it is buffered) is greatly reduced. 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: tf [mailto:[EMAIL PROTECTED] Sent: Saturday, 6 December 2008 3:01 PM To: Reuel Cornelia; Tony Henwood; histonet; [EMAIL PROTECTED]; Jan Shivers Subject: Re: RE: RE: [Histonet] IHC on paraformaldehyde-fixed you want to carry a bottle of toxic liquid on your car? or you will take a box of powder that can dissolve into useful solution easily? You have to add methanol in 10% formalin 4% formaldehyde, rather 4% paraformaldhydePFA is methanol free..it's very important. 2008-12-06 tf 发件人： Reuel Cornelia 发送时间： 2008-12-06 00:09:36 收件人： Tony Henwood; [EMAIL PROTECTED]; histonet; [EMAIL PROTECTED]; Jan Shivers 抄送： 主题： RE: RE: [Histonet] IHC on paraformaldehyde-fixed I have been curious about this discussion. we used 4% paraformaldehyde for smaller biopsies only because it has a faster penetration to tissue than 10% formalin. In all my IHC that I have done. I observe that doing an IHC with 4% paraformaldehyde does not necessarily need antigen retrieval in comparison to 10% formalin either it will be human or animal tissue but this depends on how long was it fix, our 4% paraformaldehyde we fix smaller biopsies like nerve,muscle, skin for 6 to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe you can comment on the effect on this to tissue if you say you will use 4% paraformaldehyde for storage and transportation. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 Tony Henwood [EMAIL PROTECTED] 12/04/08 9:29 PM tf wrote: I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~. I have not heard this before. Do you have a reference for this? 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: tf [mailto:[EMAIL PROTECTED] Sent: Friday, 5 December 2008 2:11 PM To: Tony Henwood; [EMAIL PROTECTED]; Jan Shivers; histonet Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed the basic principles are the same for most cross-linking fixatives and induce similar bonds the difference you observed between may due to any other variability, or the co-fixative you used. I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~.
[Histonet] thermometer calibration
Hi All, Just wondering how everyone is doing their thermometer calibrations. Inspection time is right around the corner so I want to be ready! Thanks. Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, 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 sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Marketing Position
We are currently recruiting for a marketing position in our organization. We are an Anatomic Pathology reference lab located in Lakeland, Florida and are looking for someone with both lab and marketing experience. If you are interested or know someone who is, please contact me toll-free at 866-961-2785. Thank you. Sheila Haas Laboratory Supervisor Micro Path Laboratories ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paul Monfils re: PMMA blocks
Paul, Rather than add to Jack and Gail's experience and expertise, feel free to call me if you need additional hints. I have no technical problems with large PMMA blocks. One hint is to use Pyrex beakers or crystallizing dishes, although expensive, the results are consistent. Best regards, Vicki Kalscheur Department of Surgical Sciences School of Veterinary Medicine University of Wisconsin 2015 Linden Drive Madison, WI 53706-1102 Phone: 608-262-8534 FAX: 608-263-7930 [EMAIL PROTECTED] ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PFA preparation
We routinely add paraformaldehyde to alkaline water at room temperature while stirring and wait only about 30-60 mintues for it to dissolve. Then we add a concentrated amount of PBS up to the total required volume (so that the buffer is 1x in the final volume). Then we add acid to bring the pH back down to 7. Then we filter it since not all of the PFA has dissolved (though most of it has). Merced --On Monday, December 08, 2008 9:11 AM +1100 Tony Henwood [EMAIL PROTECTED] wrote: My experience is that when you add paraformaldehyde to water all it forms is a colloidal solution (ie on standing, the paraformaldehyde settles with very little going into solution (personal experience, waited one week, then gave up). Has your experience been different? 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: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of [EMAIL PROTECTED] Sent: Sunday, 7 December 2008 7:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PFA preparation Hi, So then what is the best way to prepare formaldehyde fixative from PFA? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. I've also been taught that too much heat during preparation degrades PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it degrades. What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences ___ 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 Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 Without my flaws I'm really very boring. - random internet blog commentator ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PFA preparation: Does dissolution also mean that the PFA has depolymerized?
Hi Merced, Thanks for your reply. So then you manipulate pH rather than temperature to ensure the dissolution of PFA. Does dissolution also mean that the PFA has depolymerized? Are you saying heating to 60 degrees C is not required, or that it is not recommended? I've been told that heating 'degrades' the PFA and one wants to avoid this. But according to other sources, the heating step is required to ensure that the PFA does in indeed degrade, degrade into formaldehyde. Eric University of Calgary Medical Sciences We routinely add paraformaldehyde to alkaline water at room temperature while stirring and wait only about 30-60 mintues for it to dissolve. Then we add a concentrated amount of PBS up to the total required volume (so that the buffer is 1x in the final volume). Then we add acid to bring the pH back down to 7. Then we filter it since not all of the PFA has dissolved (though most of it has). Merced --On Monday, December 08, 2008 9:11 AM +1100 Tony Henwood [EMAIL PROTECTED] wrote: My experience is that when you add paraformaldehyde to water all it forms is a colloidal solution (ie on standing, the paraformaldehyde settles with very little going into solution (personal experience, waited one week, then gave up). Has your experience been different? 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: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of [EMAIL PROTECTED] Sent: Sunday, 7 December 2008 7:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PFA preparation Hi, So then what is the best way to prepare formaldehyde fixative from PFA? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. I've also been taught that too much heat during preparation degrades PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it degrades. What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences ___ 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 Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 Without my flaws I'm really very boring. - random internet blog commentator ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] processing times
Hello-I have just started at Roswell Park Cancer Institute and we are hoping to decrease our TAT by decreasing our processing times-having come from a facility where we hadn't changed processor times in 20 years I would appreciate it if someone could supply their schedules so I could do a comparison...we currently do a short run for bx's (5 hr), a standard surgical run (13 hr) , and a longer fatty/breast run (16 hr) Regards, Kate Kathleen Caleri, BS, MLT, HT (ASCP) Histology Lab Supervisor Pathology Roswell Park Cancer Institute (716) 845-1329 the future of mankind lies waiting for those who come to understand their lives and take up their responsibilities to help others This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] microwave processor
The Pathos can only run one run at a time. It does everything a conventional processor does. It is fully automated. On Mon, Dec 8, 2008 at 7:50 AM, Evans, Andria B. [EMAIL PROTECTED]wrote: We are currently looking at getting another microwave processor. We currently have the Histos 5 and are looking at getting another one of those or a Pathos (the big guy from milestone). But we have little information on the Pathos. I have a couple questions that maybe someone could answer Can you run more then one run at a time on the Pathos? (currently we can run 2 runs at a time on the Histos 5) Does the Pathos do the everything on the machine from fixation, to alcohol rinses, to the end product?? (We have to do all the rinses and changes manually with the Histos 5) If anyone has any other information on the microwave processors that they would like to share that would be great!! Thanks Andria B Evans, HTL(ASCP)CM CONFIDENTIALITY NOTICE: This email may contain confidential health information that is legally privileged. This information is intended for the use of the named recipient(s). The authorized recipient of this information is prohibited from disclosing this information to any party unless required to do so by law or regulation and is required to destroy the information after its stated need has been fulfilled. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or action taken in reliance on the contents of this email is strictly prohibited. If you receive this e-mail message in error, please notify the sender immediately to arrange disposition of the information. __ This e-mail has been scanned by MCI Managed Email Content Service, using Skeptic(tm) technology powered by MessageLabs. For more information on MCI's Managed Email Content Service, visit http://www.mci.com. __ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Silanized slides
The solution for making silanized slides (APES in acetone; also needs a trace of water, which does not need to be added specially) has to be used right away: it's stable for perhaps a few hours. The resulting slides, washed and dried, should be stable indefinitely if kept clean. See Microscopy Today (June 1999) pp.22-24; also available at http://publish.uwo.ca/~jkiernan/adhesivs.htm John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: Webb, Dorothy L [EMAIL PROTECTED] Date: Thursday, December 4, 2008 13:42 Subject: [Histonet] Silanized slides To: histonet@lists.utsouthwestern.edu Does anyone have any idea of how long silanized slides made in lab are good for? Would it be the same outdate as is on the solution one is using to prepare the silanized slides? Thanks, as always!! Dorothy Webb, HT Regions Histology Technical Supervisor 651-254-2962 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. ___ 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] Alias identity
Amber McKenzie [EMAIL PROTECTED] wrote: . . .We are all supposed to be professionals asking each other for advice/suggestions on the Histonet - who cares who each person is? If I post a question, I don't care if it's Jane Doe answering or John Smith ... That's true, and it certainly makes no difference if the person asking a question hides behind an alias. But what about the person answering? Can you act upon advice from an unknown source? Jane Doe might be experienced and wise, whereas John Smith might be some pimply youth who only thinks he knows all the answers. John Kiernan Anatomy, UWO London, Canada = = = ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Alias identity
While I prefer to go by my own name, I recognize that some people rightly fear to do so. An anonymous source may still have something worthwhile to say. Experience does not always teach well. The wise and experienced Linus Pauling and Erwin Chargaff were wrong about the structure of DNA; the brash young James Watson was right. -Allen A. Smith Professor of Anatomy School of Podiatric Medicine Barry University Miami Shores, FL -Original Message- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of John Kiernan Sent: Monday, December 08, 2008 3:18 PM To: Amber McKenzie Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Alias identity Amber McKenzie [EMAIL PROTECTED] wrote: . . .We are all supposed to be professionals asking each other for advice/suggestions on the Histonet - who cares who each person is? If I post a question, I don't care if it's Jane Doe answering or John Smith ... That's true, and it certainly makes no difference if the person asking a question hides behind an alias. But what about the person answering? Can you act upon advice from an unknown source? Jane Doe might be experienced and wise, whereas John Smith might be some pimply youth who only thinks he knows all the answers. John Kiernan Anatomy, UWO London, Canada = = = ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: RE: [Histonet] PFA preparation
We routinely add PFA to 0.1M PBS and heat, stir the solution for 2-3 hours. Longer heating will result in acidic pH - depolymerizing..? Then we adjust the pH to 7.4 2008-12-09 tf 发件人： Merced Leiker 发送时间： 2008-12-09 00:05:52 收件人： Tony Henwood; [EMAIL PROTECTED]; histonet@lists.utsouthwestern.edu 抄送： 主题： RE: [Histonet] PFA preparation We routinely add paraformaldehyde to alkaline water at room temperature while stirring and wait only about 30-60 mintues for it to dissolve. Then we add a concentrated amount of PBS up to the total required volume (so that the buffer is 1x in the final volume). Then we add acid to bring the pH back down to 7. Then we filter it since not all of the PFA has dissolved (though most of it has). Merced --On Monday, December 08, 2008 9:11 AM +1100 Tony Henwood [EMAIL PROTECTED] wrote: My experience is that when you add paraformaldehyde to water all it forms is a colloidal solution (ie on standing, the paraformaldehyde settles with very little going into solution (personal experience, waited one week, then gave up). Has your experience been different? 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: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of [EMAIL PROTECTED] Sent: Sunday, 7 December 2008 7:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PFA preparation Hi, So then what is the best way to prepare formaldehyde fixative from PFA? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. I've also been taught that too much heat during preparation degrades PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it degrades. What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences ___ 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 Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 Without my flaws I'm really very boring. - random internet blog commentator ___ Histonet mailing list Histonet@lists.utsouthwestern.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
