Re: [Histonet] OT: How do you call...
hahaha that's pretty clever, actually! On 6/14/2011 6:56 AM, Paula Sicurello wrote: This reminds me of a joke I learned as a kid. How to use analyze and anatomy in a sentence (song actually). My analyze over the ocean, my analyze over the sea, my analyze over the ocean, so bring back my anatomy. :-) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] OT: How do you call...
I'm guessing, then, that anatomal and pathalogal aren't words...? On 6/13/2011 2:51 PM, Jan Shivers wrote: When taking coursework eons ago, one of my professors addressed this very issue. He said that '-ic' and '-al' were BOTH adjective suffixes... and there was no need to include both at the end of a noun to turn it into an adjective (redundancy). So 'anatomic' it is. And it follows that another correct word spelling is 'pathologic', not 'pathological' though the latter is commonly used. Jan Shivers UMN - Original Message - From: Breeden, Sara sbree...@nmda.nmsu.edu To: histonet@lists.utsouthwestern.edu Sent: Monday, June 13, 2011 1:33 PM Subject: [Histonet] OT: How do you call... I've figured out how to fund my approaching retirement! A poll! So... is the correct term Anatomical Pathology or Anatomic Pathology? Send $5.00 with your email reply and I'll have the down payment for that tropical island with internet. Seriously - what is the consensus and/or the correct usage? Figure I better get all these Deep Questions answered while I have the chance. (P.S., I'm only kidding about the $5.00 but I do take PayPal). Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services 1101 Camino de Salud NE Albuquerque, NM 87102 505-383-9278 (Histology Lab) ___ 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] Unsubscribe
**Listserv master: Some people are reporting problems with managing their subscription as they have submitted their unsubscription multiple times in the prescribed manner and are still getting Histonet emails. This was reported to me by one such (failed) unsubscriber.** --On Thursday, April 21, 2011 10:25 AM -0400 Emily Sours talulahg...@gmail.com wrote: Jupiter's thunder two people in a row? really?!!?!? LOOK AT THE BOTTOM OF THE EMAIL. YOU UNSUBSCRIBE ON THE SAME PAGE THAT YOU SUBSCRIBED ON. Both of you are officially banned from the internet. Forever. Don't use it again. And don't EVER join a mailing list. EVER. Emily, who would love to unsubscribe you so I didn't have to get your ridiculous emails about how you can't do it your damn self. A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6118 Fx: (716) 829-2665 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PFA preparation
I concur with the others. For about 6 years I've made it from powder but the toxicity risk bothered me until I realized how safely and cheaply you can just buy the same stuff. We get a discount through our stockroom on a 4L bottle of 10% Buffered Formalin from Fisher (SF100-4) for $28. I started getting neutral buffered formalin instead from Fisher. --On Monday, March 21, 2011 11:25 AM -0500 Montina Van Meter montina.vanme...@pbrc.edu wrote: Nick, I too have tried to limit my exposure after making it from scratch for 30+ years. I currently purchase it from USB Corp. located in Cleveland Ohio. They have been acquired by Affymetrix and you will be directed to that website when you type in www.usbweb.com. Cat.# 19943 1LT Paraformaldehyde Solution, 4% in PBS, 1L $48.00 Regards, Tina Montina J. Van Meter Lab Manager Autonomic Neuroscience 225-763-2564 vanme...@pbrc.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Monday, March 21, 2011 10:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PFA preparation Anyone have a decent method to prepare paraformaldehyde. I have the powder and the NOAH, hood, heat source but forgot the method. Nick Madary, HT/HTL(ASCP)QIHC Histology Mgr, Medimmune 301.398.6360(lab), 4745(vm),9745(fax) To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary. This communication is expected to be read and/or used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. ___ 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 Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Vectashield mounting media problem?
We had the EXACT same problem about a year ago! We don't believe we had storage issues, either. We've been using SlowFade Gold (Molecular Probes, Invitrogen) for about a year now and it's been a great aqueous mounting medium. We add our own DAPI (2ul of a 1mg/ml DAPI solution per 500ul SlowFade) and it works fine. Merced --On Tuesday, April 21, 2009 10:51 AM -0400 Suhyoung Jeong suhyoung.je...@gmail.com wrote: Hello everyone, Our lab has been using 'Vectashield mounting media w/ DAPI' for many years and had no problem of fading. Recently, two bottles with different lot numbers in a row gone bad in around four months (the signal disappears in front of your eyes, tested on a couple of different microscopes). 1) Has anyone experienced same problem? Vector lab only says that this is a very stable product and should not be happening when stored at 4C. 2) Can anyone recommend a good aqueous mounting media with DAPI for immunofluorescence? Thank you in advance, Regards, Suh ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] low profile blades and cutting angle
AccuEdge by Sakura Finetek are the best (avail. through VWR) Not only have I tried a few other brands, but our former knowledgeable and experienced histotech here at UB recommends AccuEdge. Merced --On Thursday, April 02, 2009 9:55 AM -0700 Stacey Barrick barricksta...@yahoo.com wrote: Hi everyone, What brand low profile blades does everyone prefer and where do you order them? Also, what blade angle does everyone prefer for cryosectioning? Thanks Stacey ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] looking for a MAC1 antibody (aka CD11b/CD18, or Integrin alpha-M beta-2)
I am looking for a good antibody for MAC-1 (CD11b/CD18, Integrin alpha-M beta-2) that is targeted to rodent tissue. Any ideas where I can find one? Thanks! Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Please help! In dire need of user manuals
I always find it interesting to discover the vast sea of temperaments out there that is elucidated so clearly in the light of email responses. The same email can provoke anger, inflict hurt, or elicit laughter from different people. :-) With that in mind studies have shown (so I'm told by a friend who works in IT here at UB - I don't have the source) that 90% of communication is lost in email. When you don't see facial expression, gesturing, and, most importantly, tone of voice, you may never appreciate the sender's true intent. Especially if the person is someone you've never met, and, most importantly for this listserv, may be coming from another culture or even subculture. I have been caught up in several arguments over personal email (not on this listserv) with people I KNEW that were never meant to escalate as they did, simply because of miscommunication on one or both ends. :-) I am not saying that Mr. Royer was not in an argumentative (or satirical, or sarcastic) state, as key words and phrases from his email that others pointed out indicate he may not have been in the best of moods at that time. And there are people who, if in an especially sensitive mood and are in dire need of help, can certainly take very personally what has been said by someone they've never met before (both my hands are raised). And there are people who become highly offended for the sake of the offended person. And there are those who see it all quite comically (as I actually did at first until I started re-reading the emails from different perspectives). :-) Just my thoughts! Merced --On Saturday, March 28, 2009 8:16 AM -0700 Mark Tarango marktara...@gmail.com wrote: It was pretty funny alright. One for the Histonet Posts of Shame. There are a few of us that could be runners-up with Ford. haha Mark On Fri, Mar 27, 2009 at 7:13 PM, Emily Sours talulahg...@gmail.com wrote: I thought the rant was pretty funny. Just repeat to yourself it's an email list, I should really just relax. Emily -- prometheus, thief of light, giver of light, bound by the gods, must have been a book. -mark danielewski, house of leaves ___ 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 Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IF
We don't let our unfixed sections air-dry (room temp) long before fixing them - a few minutes? For some people here I let them sit a few minutes then just store them in the -80 for up to a weeks weeks before they fix them and commence with their staining procedure. If the animals were perfusion-fixed maybe they can air-dry longer. I've heard everything from 5 min to overnight. At room temperature. Just my input. --On Tuesday, March 24, 2009 12:17 PM -0400 Vanessa J. Phelan vjp2...@columbia.edu wrote: Hi everyone, When carrying out IF on frozen sections how long after the cutting the sections should you leave the slides to dry and where before starting the staining procedure? Thanks a mill, V ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IF
sorry, that should read for up to a few weeks. --On Tuesday, March 24, 2009 1:00 PM -0400 Merced Leiker lei...@buffalo.edu wrote: We don't let our unfixed sections air-dry (room temp) long before fixing them - a few minutes? For some people here I let them sit a few minutes then just store them in the -80 for up to a weeks weeks before they fix them and commence with their staining procedure. If the animals were perfusion-fixed maybe they can air-dry longer. I've heard everything from 5 min to overnight. At room temperature. Just my input. --On Tuesday, March 24, 2009 12:17 PM -0400 Vanessa J. Phelan vjp2...@columbia.edu wrote: Hi everyone, When carrying out IF on frozen sections how long after the cutting the sections should you leave the slides to dry and where before starting the staining procedure? Thanks a mill, V ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] questions re: fixing in general and Histochoice in particular
1. Generally 24 hrs for 2-3 mm thick piece. At least a 10:1 formalin:tissue volume. 2. Room temp or 4oC - I've heard debates, and I've heard it doesn't matter, but we do it at 4oC. --On Tuesday, March 24, 2009 4:54 PM -0400 Jacqui Detmar det...@lunenfeld.ca wrote: Hi all. Having a bit of an internal debate here, so I would like to get the opinions of some of you in Histoland, please. Here are the questions: 1.When fixing with 10% NBF, for how long should you fix and what volume ratio of fixative:tissue should you use? 2. At what temperature should one be fixing tissues? Regarding Histochoice: 1.For how long should you fix the tissue? 2. What volume ratio of fixative:tissue should you use? 3. How long can you store Histochoice-fixed tissues in 70% ethanol? I think that's about it. Thanks in advance, Jacqui Jacqui Detmar, Post-doctoral Fellow Samuel Lunenfeld Research Institute Mount Sinai Hospital 25 Orde Street, room 6-1001AJ Toronto, ON M5T 3H7 Email:det...@lunenfeld.ca Phone: 416-586-4800 x5607 Fax:416-586-5993 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Paraffin Block trimming
Now I'm interested in how you embed so as to not have to scrape...if i don't add enough wax (enough to end up rising around the edges of the cassette, hence the scraping later), i don't get a secure hold of the block to the cassette... ML --On Thursday, March 19, 2009 7:37 AM -0700 Martin, Gary gmar...@marshallmedical.org wrote: No you're not the only one ... I was wondering the same thing ... why all the scraping. It seems to me that clean embedding does the trick with a few exceptions. G -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine (CDC/CCID/NCZVED) Sent: Thursday, March 19, 2009 6:58 AM To: Paula Pierce; Histonet Subject: RE: [Histonet] Paraffin Block triming I was wondering if I was the only one out there that rarely has to scrape a block. 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 Paula Pierce Sent: Thursday, March 19, 2009 9:57 AM To: Histonet Subject: [Histonet] Paraffin Block triming I try to embed so as to have a minimal amount of paraffin to scrape from the blocks. ;) But, I do scrape using the handle end of the same forceps I use to pick up the ribbon and tease the sections. No sharp edge. No electricity. PKP From: Peter Carroll carro...@umdnj.edu To: histonet@lists.utsouthwestern.edu Sent: Thursday, March 19, 2009 8:30:34 AM Subject: Re: [Histonet] Paraffin Block triming Does anybody use a paraffin block dewaxer ? Yep, it's called my own two hands and a metal spatula, ha ha :) I find that it's not only very quick, but quite accurate... Scott wrote: Hi, Does anybody use a paraffin block dewaxer ? If so does it save any time, how well does it work? Thanks, Scott Hendricksen HT (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Paraffin Block trimming
hmmm...I do too...Fisher 15-182-505B HistoPrep Stainless-Steel Base Molds...maybe it's the cassette design or type of wax used? (VWR 15147-839 POLYFIN, VWR 87002-362 HISTOSETTE Cassettes, Simport Plastics Biopsy Cassettes) --On Thursday, March 19, 2009 10:57 AM -0400 Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov wrote: Don't know what to say.I fill mine up and as long as I don't get to rough with it (bump it or something where it sloshes) I don't have any excess paraffin around the edges. I use metal base molds. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: Merced Leiker [mailto:lei...@buffalo.edu] Sent: Thursday, March 19, 2009 10:56 AM To: Martin, Gary; Bartlett, Jeanine (CDC/CCID/NCZVED); Paula Pierce; Histonet Subject: RE: [Histonet] Paraffin Block trimming Now I'm interested in how you embed so as to not have to scrape...if i don't add enough wax (enough to end up rising around the edges of the cassette, hence the scraping later), i don't get a secure hold of the block to the cassette... ML --On Thursday, March 19, 2009 7:37 AM -0700 Martin, Gary gmar...@marshallmedical.org wrote: No you're not the only one ... I was wondering the same thing ... why all the scraping. It seems to me that clean embedding does the trick with a few exceptions. G -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine (CDC/CCID/NCZVED) Sent: Thursday, March 19, 2009 6:58 AM To: Paula Pierce; Histonet Subject: RE: [Histonet] Paraffin Block triming I was wondering if I was the only one out there that rarely has to scrape a block. 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 Paula Pierce Sent: Thursday, March 19, 2009 9:57 AM To: Histonet Subject: [Histonet] Paraffin Block triming I try to embed so as to have a minimal amount of paraffin to scrape from the blocks. ;) But, I do scrape using the handle end of the same forceps I use to pick up the ribbon and tease the sections. No sharp edge. No electricity. PKP From: Peter Carroll carro...@umdnj.edu To: histonet@lists.utsouthwestern.edu Sent: Thursday, March 19, 2009 8:30:34 AM Subject: Re: [Histonet] Paraffin Block triming Does anybody use a paraffin block dewaxer ? Yep, it's called my own two hands and a metal spatula, ha ha :) I find that it's not only very quick, but quite accurate... Scott wrote: Hi, Does anybody use a paraffin block dewaxer ? If so does it save any time, how well does it work? Thanks, Scott Hendricksen HT (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Paraffin Block trimming
I guess one has to go to histology school to learn how to do it, then (sent under separate cover from another histonetter...) I 'm too researchy. ;-) --On Thursday, March 19, 2009 11:45 AM -0400 Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov wrote: We have used a variety of cassettes and paraffin over the years...same result. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: Merced Leiker [mailto:lei...@buffalo.edu] Sent: Thursday, March 19, 2009 11:16 AM To: Bartlett, Jeanine (CDC/CCID/NCZVED); Martin, Gary; Paula Pierce; Histonet Subject: RE: [Histonet] Paraffin Block trimming hmmm...I do too...Fisher 15-182-505B HistoPrep Stainless-Steel Base Molds...maybe it's the cassette design or type of wax used? (VWR 15147-839 POLYFIN, VWR 87002-362 HISTOSETTE Cassettes, Simport Plastics Biopsy Cassettes) --On Thursday, March 19, 2009 10:57 AM -0400 Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov wrote: Don't know what to say.I fill mine up and as long as I don't get to rough with it (bump it or something where it sloshes) I don't have any excess paraffin around the edges. I use metal base molds. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: Merced Leiker [mailto:lei...@buffalo.edu] Sent: Thursday, March 19, 2009 10:56 AM To: Martin, Gary; Bartlett, Jeanine (CDC/CCID/NCZVED); Paula Pierce; Histonet Subject: RE: [Histonet] Paraffin Block trimming Now I'm interested in how you embed so as to not have to scrape...if i don't add enough wax (enough to end up rising around the edges of the cassette, hence the scraping later), i don't get a secure hold of the block to the cassette... ML --On Thursday, March 19, 2009 7:37 AM -0700 Martin, Gary gmar...@marshallmedical.org wrote: No you're not the only one ... I was wondering the same thing ... why all the scraping. It seems to me that clean embedding does the trick with a few exceptions. G -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine (CDC/CCID/NCZVED) Sent: Thursday, March 19, 2009 6:58 AM To: Paula Pierce; Histonet Subject: RE: [Histonet] Paraffin Block triming I was wondering if I was the only one out there that rarely has to scrape a block. 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 Paula Pierce Sent: Thursday, March 19, 2009 9:57 AM To: Histonet Subject: [Histonet] Paraffin Block triming I try to embed so as to have a minimal amount of paraffin to scrape from the blocks. ;) But, I do scrape using the handle end of the same forceps I use to pick up the ribbon and tease the sections. No sharp edge. No electricity. PKP From: Peter Carroll carro...@umdnj.edu To: histonet@lists.utsouthwestern.edu Sent: Thursday, March 19, 2009 8:30:34 AM Subject: Re: [Histonet] Paraffin Block triming Does anybody use a paraffin block dewaxer ? Yep, it's called my own two hands and a metal spatula, ha ha :) I find that it's not only very quick, but quite accurate... Scott wrote: Hi, Does anybody use a paraffin block dewaxer ? If so does it save any time, how well does it work? Thanks, Scott Hendricksen HT (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending
Re: [Histonet] Mosquito GI Tract
lol...I wanted to say the same thing...and how in the world can you even see it when you section it? How do you stain it without losing it from the slide? Is there really enough surface area for it to adhere and withstand washing? ...and it's ALMOST Friday...! --On Thursday, March 19, 2009 12:09 PM -0600 Breeden, Sara sbree...@nmda.nmsu.edu wrote: And here I thought I had an unusual project today, finding out as I did that my boss is thinking of examining up to 1000 cattle for TB (at 10 lymph nodes/beast) using AFB and asking me to cost it for him. I think the Mosquito GI Tract Processing and Embedding question has my project beat! How would you even KNOW you had the darned GI tract to begin with? Are these Texas Mosquitos? I'm just sayin'... Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cutting fresh-frozen brains from 1 week old rat pups
Definitely do at least that one sucrose cryopreservation step you mentioned. Even step up to it in 10% and 20% sucrose. --On Wednesday, March 18, 2009 4:04 PM +1100 Adam Galle adam.ga...@student.unsw.edu.au wrote: Hi all, Currently I am working with brains from 7 day old rat pups, that undergo an hypoxic-ischemic injury (Levine/Vannucci technique). These brains are unfixed, frozen in isopentane and cut at 20um on a croystat. These brains are not cutting very well compared to an adult brain (potenially due to unfinished myelination?), they are very 'crumbly' for want of a better term and always have cracks or generally poor preservation of morphology. I have tried all the standard tricks of different temperatures, section thickness and knife angle to no avail. I am going to perfuse fix my next cohort of animals with PFA and then a 30% sucrose step to see if that helps, but I was hoping that someone out there would have some tips on cutting these immature brains. Thanks, Adam. Adam Galle, Neuropharmacology and Brain Injury Lab Department of Pharmacology School of Medical Sciences UNSW Sydney ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] cryoprotection possible?
I agree with Frances and anh2006. Cryospreservation is important for brains especially and must be done after fixation, before freezing, and the glutaraldeyde can irreversibly mask antigens. Merced --On Wednesday, March 18, 2009 8:49 AM -0500 Swain, Frances L swainfranc...@uams.edu wrote: You are so correct, I forgot you do not rinse the sucrose out you blot it and then freeze. Sorry about that. I do not cut frozen sections very often and I forget. Frances L. Swain HT(ASCP) A. A. S. Special Procedures Technician Department of Orthopaedic Surgery Center for Orthopaedic Research Barton Research Building 2R28 4301 West Markham Street Little Rock AR 72205 (501) 686-8739 PHONE (501) 686-8987 FAX swainfranc...@uams.edu email -Original Message- From: anh2...@med.cornell.edu [mailto:anh2...@med.cornell.edu] Sent: Wednesday, March 18, 2009 8:41 AM To: Swain, Frances L; Dr. med. Frauke Neff; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] cryoprotection possible? Good advice indeed. However, I don't recommend you rinse in water after sucrose. Sort of defeats the purpose. Instead if you need to remove excess sucrose rinse in either 50/50 sucrose/OCT or just OCT. Also, glutaraldehyde fixation usually renders tissue difficult to immunostain. You have to consider whether your fixation is appropriate for your antigen/antibody. -Original Message- From: Swain, Frances L swainfranc...@uams.edu Date: Wed, 18 Mar 2009 08:12:45 To: Dr. med. Frauke Neffne...@staff.uni-marburg.de; histonet@lists.utsouthwestern.eduhistonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cryoprotection possible? Usually the cryoprotection is carried out after the specimens are fixed and before they are frozen. If you have a sample you can spare you might try making up some 20% Sucrose placing the frozen sample in it. putting it in the refrigerator and letting it stay in the 20% sucrose until it drops to the bottom of the container. You might have to change the 20% Sucrose a couple of times as you know sucrose can grow bacteria easily. When the specimen has dropped to the bottom of the container, remove it, rinse in a couple of changes of distilled water and quick freeze the sample. Try cutting and staining the sample and see if the morphology is good if it is then you can do all of your samples to correct the problem. There should be no thawing between removal of the sample from the storage to the cryostat. Hope this helps Frances Swain From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dr. med. Frauke Neff [ne...@staff.uni-marburg.de] Sent: Wednesday, March 18, 2009 3:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cryoprotection possible? Dear Histonetters, I'm supposed to do a noradrenaline ihc on rat brains that have been snap frozen and afterwards glutaraldehyd fixed. While the morphology is okay in the test-tissue we used to establish the protocol, it was very poor in the tissue of the trial animals. It appeared mooth eaten and disrupted. I assume the brains were thawed and refrosted due to moving the samples between several cities. Does anyone know a method /a tip how I can save some proper morphology? I checked the archive and found sucrose as cryoprotectant, but this is supposed to do it while the tissue is frozen or is it possible to use it while the tissue thaws? Thank you all in advance for your patience and help, Frauke -- Dr. med. Frauke Neff AG Neurologische Therapieforschung Neurologie Philipps-Universität Marburg Rudolph-Bultmann Str. 8 35039 Marburg 06421/5866304 This message was sent using IMP, the Internet Messaging Program. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 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
Re: [Histonet] curling on cryostat brain sections and other issues
I make 8um-thick sections of various tissues and sometimes they do curl. I do not use an anti-roll plate. So what I do while making the section is to cut just enough of the section so that it is attached by a very small amount to the top of the block, (so that most of the block has gone down below the blade and won't interfere when you pick up the section with your slide). The free end of the section will still curl; however, you only have this end to deal with since the other is fixed! I use my brushes to flatten out this edge as much as possible, then quickly place the slide on it before it starts curling up again. It usually will curl up rather slowly after having its edges brushed, giving me plenty of time. Sometimes it stays flat and doesn't curl at all. But remember this is 8-um sections, not 15um. Merced --On Tuesday, March 17, 2009 12:18 PM -0400 Guillermo Palchik g...@georgetown.edu wrote: Hi there, I am having problems with my brain sections (15 um - flash-frozen) curling up once I remove the antiroll plate on my cryostat... does anyone know how i can prevent this? Also - does anybody have a quick guide on what to improve while cutting, based on the damage seen on the brain slice: for example I heard that if there are vertical lines along the slice, it might mean a nick in the blade, that cracking of the tissue might be cutting with the blade too perpendicular, and so on... i am looking for a general troubleshooting guide for cryostat cutting... thanks Gil -- There are people who fight one day and are good... There are those who fight one year and are better... There are people who fight many years and are very good... But there are those who fight their entire lives: they are indispensable... Bertolt Brecht (1898-1956) How can it be that mathematics, being after all a product of human thought which is independent of experience, is so admirably appropriate to the objects of reality? Is human reason, then, without experience, merely by taking thought, able to fathom the properties of real things? Albert Einstein (1879-1955) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Disposal of Formaldehyde
Ugh. We are not allowed to dispose any chemical, including formaldehyde, down our drains here at UB (Buffalo, NY). Everything goes into appropriately-labeled waste bottles for Hazardous Waste to dispose of, however they do it. --On Saturday, March 14, 2009 12:05 AM +0800 TF ti...@foxmail.com wrote: Hard to say... we perfuse sooo many animals everyday...several litres for one lab into the sea 2009-03-14 TF Jessica Piche ? 2009-03-13 23:25:02 histonet ??? ??? [Histonet] Disposal of Formaldehyde ? Hi All, We have a question regarding the disposal of formaldehyde. We were told at our hospital that a consultant said it was okay to dump formaldehyde down the drain. I believe they said it was okay to dump 15 gallons or so a day! We are not to fond of this idea and would like to know what everyone else is doing. How is everyone disposing of their formaldehyde? We would be especially interested in what other hospitals in CT are doing. Thanks, Jessica Piche-Grocki, HT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Dako, Thermo and others
LOL! It's ALMOST Friday, people, ALMOST... :-) --On Thursday, March 12, 2009 11:46 AM -0500 Jackie M O'Connor Jackie.O'con...@abbott.com wrote: Not Dako - Leica purchased Surgipath. Burton, Lynn lynn.bur...@illinois.gov Sent by: histonet-boun...@lists.utsouthwestern.edu 03/12/2009 10:46 AM To Pamela Marcum pmar...@vet.upenn.edu, Victor Tobias vic...@pathology.washington.edu, jeff jlhow...@yrmc.org cc histonet@lists.utsouthwestern.edu Subject RE: [Histonet] Dako, Thermo and others Dako did acquire Surgipath recently. The Surgipath rep said it has been a good marriage for them so far. Lynn Burton Lab Assoc. I Animal Disease Lab Galesburg, Il 61401 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Pamela Marcum Sent: Thu 3/12/2009 10:24 AM To: 'Victor Tobias'; 'jeff' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dako, Thermo and others AMEN Pamela A Marcum University of Pennsylvania School of Veterinary Medicine Comparative Orthopedic Laboratory (CORL) 382 W Street Rd Kennett Square PA 19438 610-925-6278 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Victor Tobias Sent: Thursday, March 12, 2009 11:02 AM To: jeff Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Dako, Thermo and others Jeff, It is not just Dako, but many vendors have jacked up the cost of goods. Let's take the PSLIM from Accuplace. About a month ago you could purchase this for $5K. Now that Thermo has acquired distribution rights it costs over $10K and nothing has changed. To me, it is better to sell 10 at $5K than 5 at $10K. We were ready to purchase several more of these, but have to reconsider our options now. No wonder Healthcare costs are out of control. Victor Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 vic...@pathology.washington.edu 206-598-2792 206-598-7659 Fax = Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. jeff wrote: I was wondering if anyone else is being blackmailed by Dako to sign a contract with them. We have enjoyed our relationship a number of years buying there product as we need it. We also have changed detection kits as we where advised. Now they are telling us that if we do not go with price per slide they will raise our costs of purchasing our supplies. Example EnVision + dual link system last year cost 1,580.70 this year 3,300 and that this will go in effect March 1st ( just talked to the rep about this Tues) Just 1 example. S. Has anyone else had this happen to them and what did you do? Thanks Jeff 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 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 Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cutting angle?
I have a refurbished Reichert-Jung 2030, but with a knife holder from a newer model (so I'm told by the refurbisher). I use 0-1 degree angle. Merced --On Tuesday, March 10, 2009 4:15 PM -0700 Va Paula Sicurello vapat...@yahoo.com wrote: Hello Listers, I have inherited a Reichert-Jung 2030 microtome. The knife holder is a bit funky and hard to set the clearance angle. What angle do y'all use? I'm trying to get 4 or 5 degrees. Thanks, 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] slide disposal
We have a broken glass waste container we put ours in for pick up and disposal (don't deal with non-animal patients, so not an issue). -M --On Wednesday, March 11, 2009 4:56 AM -0400 Sharon Campbell shar...@celligent.net wrote: Hi everyone! Thank you for all the great responses to my last question about metal vs. plastic molds. I have another question being debated however, How to dispose of slides once the required time is up (10 years for us). We have put the slide in sharps containers and then into biohazard, but are they really bioharzard, the slides only contain the patients accession number - no personal ID on the slides? How do you guys dispose of your slides when space and money are an issue? Thank you Sharon Campbell, HTL(ASCP)CM, BSBM Histology Supervisor Celligent Diagnostics, LLC Formerly Pathology Associates Services 101 East W.T. Harris Blvd, Suite 1212 Charlotte, NC 28262 800-524-6779 ext. 104 704-970-3304 Direct Line shar...@celligent.net ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Bad sections
What animal tissues are you using? If rodent, may need less time on the steps - like 10-20 min. --On Wednesday, March 11, 2009 11:29 AM -0700 Va Paula Sicurello vapat...@yahoo.com wrote: Hi Histo netters, I am am my wit's end. I know how to section but my sections are compressing like crazy. I've altered my processing protocol thinking that I was overprocessing my samples. I've tried different knife angles, different brands of razor blade knives. It's either me or the microtome. Info- I process animal tissues with between 30-60 minute steps. Vacuum paraffins for 30 minutes. Section with Reichert-Jung 2030 set at 4-ish degrees (a little below the middle line on the knife holder). Soak blocks on Downey ice blocks, water bath at 44 degrees. Suggestions? Waa, 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] metal molds vs. disposable molds
I like my metal molds. They are neat, you get even heat transfer, and are easy to clean when they need it. I clean them by tossing them in some boiling water with an excess of cleaning powder (Alconox or Sparkleen) or any detergent you have on hand (make sure it doesn't get too bubbly and froth over though). Then I rinse them under hot top water, spread on paper towels to dry, spray with mold release, and stack in the oven. Merced --On Tuesday, March 10, 2009 11:55 AM -0400 Sharon Campbell shar...@celligent.net wrote: Hello everyone, We have a debate going on about purchasing metal molds vs. disposable plastic molds. Which is better? Is the cost better in the long run to buy metal? Also, how does everyone clean their metal molds? Thanks Sharon Campbell, HTL(ASCP)CM, BSBM Histology Supervisor Celligent Diagnostics, LLC Formerly Pathology Associates Services 101 East W.T. Harris Blvd, Suite 1212 Charlotte, NC 28262 800-524-6779 ext. 104 704-970-3304 Direct Line shar...@celligent.net ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] metal molds vs. disposable molds
...which might be the better thing to do then...since it'll all be floating on the surface anyway and we don't want to risk clogging drain pipes with the wax once it cools... --On Tuesday, March 10, 2009 12:59 PM -0400 Weems, Joyce jwe...@sjha.org wrote: And if you let the water cool, the paraffin will harden and you discard it without pouring down the drain... -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Merced Leiker Sent: Tuesday, March 10, 2009 12:22 PM To: Sharon Campbell; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] metal molds vs. disposable molds I like my metal molds. They are neat, you get even heat transfer, and are easy to clean when they need it. I clean them by tossing them in some boiling water with an excess of cleaning powder (Alconox or Sparkleen) or any detergent you have on hand (make sure it doesn't get too bubbly and froth over though). Then I rinse them under hot top water, spread on paper towels to dry, spray with mold release, and stack in the oven. Merced --On Tuesday, March 10, 2009 11:55 AM -0400 Sharon Campbell shar...@celligent.net wrote: Hello everyone, We have a debate going on about purchasing metal molds vs. disposable plastic molds. Which is better? Is the cost better in the long run to buy metal? Also, how does everyone clean their metal molds? Thanks Sharon Campbell, HTL(ASCP)CM, BSBM Histology Supervisor Celligent Diagnostics, LLC Formerly Pathology Associates Services 101 East W.T. Harris Blvd, Suite 1212 Charlotte, NC 28262 800-524-6779 ext. 104 704-970-3304 Direct Line shar...@celligent.net ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Water bubble on slides
Hi Vanessa, I have read in at least one online protocol and have also been advised by our excellent and experienced (former) histotech who used to be here at UB NOT to warm the slides without air-drying first. We typically air-dry the slides, yes, standing up, overnight, before warming them. Merced --On Wednesday, March 04, 2009 4:35 PM -0500 Vanessa J. Phelan vjp2...@columbia.edu wrote: Hi everyone, Quick question, I am finding after cutting a lot of slides for HEs onto VWR Superfrost Plus slides (these are the only slides we have at the moment) that when I lift sections from the water bath ( of distilled water) a water bubble stays under the sections on the slide. When I lie them flat on the hot plate to dry the water bubble is tending to distort the tissue by escaping out through he tissue, eventually. I have not experienced this before, the water has ran off the slide no problem. Would it be the slides that are the problem or anything I might need to add to the water? I have just ordered a histology oven, this may be the solution...drying the slides standing up! Vanessa ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Section thickness
I've read that 4-5um is average, and that's what I typically go by for paraffins, but then it could depend on tissue type as Rene recommended. I end up having to cut frozens at 8-10um because for some reason it's very difficult to cut any thinner on the cyrostat I use (an old Vibratome model). But doesn't seem to matter for our applications - we do mostly immunofluorescence some HE. Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. --On Tuesday, March 03, 2009 9:14 AM -0500 Vanessa J. Phelan vjp2...@columbia.edu wrote: Hi Guys, Just wondering what thickness you cut sections at? I was always used to cutting at 2-3 microns in my last lab, however in my new place they are cutting at 6 microns (for both H Es and IHC), which seems to me as really quite thick! What would be the average cutting thickness? Thanks a mill, Vanessa ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvienienced! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] distilled water
ummm...distilled water works best, unless you like cleaning out mineral deposit build-up and such... --On Monday, March 02, 2009 11:18 AM -0800 kristen arvidson arvidsonkris...@yahoo.com wrote: Are most people using distilled water in the water baths? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvienienced! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] HIER
In our experience, the heat is definitely more important...we retrieve epitopes ranging from cytoskeletal markers (Troponins, Myosin Heavy Chain,...) to nucler (PCNA) and others (c-kit, beta-catenin, vonWillebrand Factor-vWF). We use a steamer set to 30', just place the slides in flat on the bottom of the basket: encircle the sections with Pap pen first, then pipette some buffer on. I always test pH 6 and pH 9 but haven't seen a difference yet between the two with the epitopes we're unmasking.) We have this wicked vWF antibody from Calbiochem (PC313) that unmasks in PBS (pH7) in the steamer and binds very well on both paraffins and frozens. (And it does need retrieval - without retrieval it is gives very little stain). Ok, I digress!! But you get the point...that's from our experience, anyway! ~Merced --On Monday, March 02, 2009 7:52 PM +0100 Casper Hempel casperhem...@gmail.com wrote: Hi histonetters We have been talking a lot about how to retrieve your epitopes using a microwave in the best possible way. We haven't come to an agreement and people in my lab both argue that temperature and pH are important issues. Without a doubt both factors are important for a proper retrieval, but if you have to focus on one of the factors, which would you consider the most important? Temperature or pH? The issue is mainly longer incubations of the slides in boiling buffer. The buffer is evaporating and the solution/buffer gets less and less pH neutral and you need to top up with dH2O. However, if you boil with reduced intensity, less evaporation will occur and the pH will remain more stable. Do you have any suggestions or comments to this issue? Looking forward to your replies. Cheers Casper ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building 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 No trees were harmed in the sending of this email. However, many electrons were severely inconvienienced! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] looking for CD45 antibody
Thank you everyone who replied with a suggestion!!! --On Tuesday, February 17, 2009 3:17 PM -0500 Merced Leiker lei...@buffalo.edu wrote: Hello Histonetters, I am looking for a good CD45 (CLA) antibody for use in paraffin immunofluorescence that meets the following criteria: 1. unconjugated 2. non-mouse isotype 3. reacts with hamster OR greatest likelihood of reacting with hamster (e.g., reacts with many other rodent species...) Can anyone refer one? Thank you! 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 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] looking for CD45 antibody
(I'm going to try asking this again, as I only got 1 response and it wasn't what I was looking for. Thank you!) Hello Histonetters, I am looking for a good CD45 (LCA) antibody for use in paraffin immunofluorescence (or ANY application at this point) that is of a NON-mouse isotype (that is, it was raised in goat, rabbit, rat, or other species) Can anyone refer one? Thank you! 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Starfrost slides
I have used them almost daily for a year or more now and have had no problems with the slides giving off any autofluorescence in immunofluorescence applications. --On Thursday, January 22, 2009 2:56 PM -0500 Nancy Lemke ns...@neuro.hfh.edu wrote: Has anyone using Starfrost slides (7255) from Mercedes had any problems with autofluoresence when doing IF? Thanks in advance. Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit = = CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by reply email, delete the email from your computer system and shred any paper copies. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. See our Privacy Policy and Henry Ford My Health at www.henryford.com for more detailed information. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. = = ___ 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] pH strips
LOL, Jackie!!! I've used 10-year old strips that seemed to have worked fine. --On Wednesday, December 31, 2008 12:57 PM -0600 Jackie M O'Connor Jackie.O'con...@abbott.com wrote: I heard of some that went on a graffiti rampage in downtown Chicago last year . . . . . Happy New Year. Amber McKenzie amber.mcken...@gastrodocs.net Sent by: histonet-boun...@lists.utsouthwestern.edu 12/31/2008 12:43 PM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] pH strips Do pH strips go bad after a certain time period? ___ 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 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] Dark vs. Light Tissue?
Hey all, While embedding rodent tissues I have found that some specimen within the SAME muscle group (such as quadriceps) appear very dark in color while others are very pale. I just recently started noticing this while I've been embedding. (I embed manually; I'm a research tech who does a lot of histological work). Any insights as to why the color difference? 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] Dark vs. Light Tissue?
Just to clarify, this is the same rodent muscle. We take out the whole quadriceps from one rodent, the whole quadriceps from another rodent, and both will look different colors while embedding. --On Friday, December 19, 2008 3:03 PM -0500 Merced Leiker lei...@buffalo.edu wrote: Hey all, While embedding rodent tissues I have found that some specimen within the SAME muscle group (such as quadriceps) appear very dark in color while others are very pale. I just recently started noticing this while I've been embedding. (I embed manually; I'm a research tech who does a lot of histological work). Any insights as to why the color difference? 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 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] RE: Recruiting Techs at work
This indeed sounds like the most honest and integrous approach as well as a way to keep positions competitive. I like it. --On Monday, December 15, 2008 11:00 AM -0500 Terri Braud tbr...@holyredeemer.com wrote: I agree that if you have a great work environment with competitive pay your staff will stay put, but how many of us at the department supervisory/manager level can actually make the final decision concerning salary and pay scales for a position. I suspect that most of us, including myself, can only make well documented pleas for salary increases. When it comes to spending money, most institutions are reactive rather than proactive. If salaries are low, then you may have to lose a valuable tech before you can make the point that salaries need to be increased. It may be a tough pill to swallow, but I am more than willing to let my folk listen to any pitchman that is trying to get their ear. Any tech worth his/her salt, knows that these calls are out there, so why try to hide it. If a tech can better themselves by a job change, then who am I to stand in their way. We generally discuss these in departmental meetings and I tell techs that I get frequent calls from recruiters and if they would like to correspond with them, may I give the recruiter their personal contact info. I ask in return, that if they do go elsewhere for a better salary, that they please give me the info, so I can use it to help increase the salary of those who remain. This keeps communication up front, honest, and open and I think in the long run, helps to quell murmuring about low salaries which are beyond my control as a supervisor. Just another viewpoint. Terri Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Laboratory Holy Redeemer Hospital and Medical Center 1648 Huntingdon Pike Meadowbrook, PA 19046 (215) 938-3676 phone (215) 938-3689 fax Message: 18 Date: Sat, 13 Dec 2008 13:46:57 + (UTC) From: lee2...@comcast.net Subject: Re: [Histonet] Offended I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. - CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. ___ 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] (reply) silly questions.---PFA
Oh, I'd like to know that, too, please! --On Friday, December 12, 2008 7:48 AM -0600 Walters, Katherine S katherine-walt...@uiowa.edu wrote: This may be another silly question, but how does one test the concentration of formaldehyde in solution? Thanks, Kathy Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of tf Sent: Friday, December 12, 2008 2:35 AM To: Tony Henwood; Pat Flannery; histo...@lists.utsouthwestern.ed Subject: [Histonet] (reply) silly questions.---PFA I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the fixation seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!! Tony: Do you think this is because of inproper preparation of PFA in his lab, or the common problem in all researchers using PFA? I do think most biomedical labs currently are using PFA to prepare the fixatives! So, anyone has the idea on a correction preparation procedure of 4% PFA? I noticed some of you dissolve PFA powder in NaOH-conditioned alkaline water, then add concentrated PB solution. We here dissolve PFA in concentrated PB solution directly (heat stir for 2-3 hours), then adjust pH to 7.4. We dont have big problem in tissue quailityexcept when one want to cut the brain in a cryostat rather sliding microtome. Many times the brain sections from the cryostat have cheese like holes/cavities, which almost never appear on sliding microtome-prepared sections. 2008-12-12 tf ·¢¼þÈË£º Tony Henwood ·¢ËÍʱ¼ä£º 2008-12-12 06:18:47 ÊÕ¼þÈË£º Pat Flannery; histonet@lists.utsouthwestern.edu ³ËÍ£º Ö÷Ì⣺ RE: [Histonet] Silly Question? Pat, I agree with you. In a routine diagnostic histopathology laboratory, it makes little difference what you use. Around the world for over 100 years most labs use 10% neutral buffered formalin made from concentrated 38%(or there abouts) formalin (or formaldehyde). Researchers, though, are a different kettle of fish. They will tend to hang on to misinformed, mystical methods believing they are being scientific. Funny, you would think that they, as a group, would be the ones pushing the boundaries and critically assessing each step of their research, ensuring that they understand what and why they are doing it. (Disclaimer - not all researchers are like this, thank heavens!!) Using a formaldehyde solution made from polyformaldehyde can cause problems. One researcher used it and wondered why their morphology was sub-optimal and their p53 immunohistochemistry was negative. He assured me that he had fixed small samples of tissue for 6 hours in 4% formaldehyde and then processed them using ethanol, xylene and wax. I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the fixation seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!! This also explains the loss of p53 staining. I gave him some of our routine 10% phosphate buffered fomalin, asked him to fix overnight, and try agin. Low and behold problem solved. How's that for a Friday Flamming!!! 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 Pat Flannery Sent: Friday, 12 December 2008 3:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in
RE: [Histonet] Silly Question?
So I have a question about the cross-linking aspect of PFA...while I agree I need it to keep my epitope in place, is there such a thing as OVER-crosslinking (i.e., tissue spending TOO much time in formalin - weeks? months?) that would make my epitope difficult to near-impossible to retrieve? Loving the formaldehyde soap-boxes histonetters get onto... --On Friday, December 12, 2008 10:36 AM -0700 pru...@ihctech.net wrote: i have had this argument with the researchers at the University for 30 years, somewhere back in the day they were told that commercially made formalin had methanol in it (it does but just a little and does not hurt anything in my experience) and that methanol would damage their tissue for IHC, so they think they must use paraformaldehyde and make it fresh themselves. Since new people make it all the time it often does not get made up correctly and their stress over this issue is miss placed as they should be using something commercial for consistancy and paying more attention to adequate time for fixation in reg formalin. Another anoying myth that is difficult to combat with them is that we should limit the fixation time in aldehyde fixatives because it will cross link the proteins masking them for IHC, there fore i am always getting tissue that has not been fixed long enough (at least 24 hrs. to protect it from paraffin processing, because if the proteins are not cross linked they can be alcohol fixed and/or washed away forever), the people in research know about the cross linking fo aldehydes but do not know that cross linking of proteins is a good thing and they also do not know that we have advanced methods HIER or EIER for unmasking the proteins, but we have no way of getting a protein back that has been lost in processing because the sample was not adequately fixed. there i will get off my Friday soap box.. Happy Holidays to all! Patsy Original Message Subject: RE: [Histonet] Silly Question? From: Merced Leiker lei...@buffalo.edu Date: Fri, December 12, 2008 8:12 am To: Edwards, R.E. r...@leicester.ac.uk, 'Pat Flannery' pjfne...@duke.edu Cc: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu In research lab situations particularly, one does not have the time or technique for nailing down the ways of making each of the buffers, reagents, and procedures work the right way or the most optimum way...a lot of times it's students or postdocs just focused on getting their project done and not caring how their fixative is made as long as it works to some degree and, alas, it's up to us already over-booked technicians to figure out the best way to make the PFAand we usually don't have a whole day (week, or year) to spend researching the back-and-forth arguments, either! ;-) Merced --On Friday, December 12, 2008 2:04 PM + Edwards, R.E. r...@leicester.ac.uk wrote: You hit the nail on the head That's what we always use, fear of change is a common human condition. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: 11 December 2008 16:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, That's what we always use. Thanks. -Pat Flannery (not a real histologist - I just play one in the lab) ___ 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 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 Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State
RE: [Histonet] Silly Question? - Need help quickly!
So...is a polymer of paraformaldehyde considered depolymerized if it remains somewhere between 1-50 molecules long once it's been dissolved in solution, however it's dissolved? (the dissolving being a separate topic of debate on Histonet). Does it matter for tissue fixation purposes if there are formaldehyde chain lengths of 50 molecules present in solution - not long enough to precipitate out, but perhaps long enough to affect its penetration and fixing of tissues? Any ideas? Merced --On Thursday, December 11, 2008 9:58 AM -0800 Rene J Buesa rjbu...@yahoo.com wrote: Joyce: Methanal, which is the chemical name of formaldehyde, polymerizes. If it forms a polymer of at least 50 molecules or more, it gets solid = para-formaldehyde. Formalin (a trade name as formol is also another trade name)is the 37-50% aqueous solution of formaldehyde (with some additiveses to prevent polymerization). You can prepare BNF using the formalin solution or dissolving the amount of solid para-formaldehydede to get to the concentrationon you desire. The chemical in both solutions is the same = methanal or formaldehyde.René J. --- On Thu, 12/11/08, Weems, Joyce jwe...@sjha.org wrote: From: Weems, Joyce jwe...@sjha.org Subject: RE: [Histonet] Silly Question? - Need help quickly! To: Pat Flannery pjfne...@duke.edu, histonet@lists.utsouthwestern.edu Date: Thursday, December 11, 2008, 12:12 PM I was just going to post a question regarding paraformaldhyde myself! Just last week I believe I remember someone saying that paraformaldehyde and formalin are the same and they had put the same solution in two different containers for one of their researchers because they were so insistent to have two different solutions. Are they the same? Well, today I have a request to put tissue for a researcher in formalin and paraformaldehyde. So Without percentage required, do I use 10% NBF? Do I call somewhere and get paraformaldehyde and make 4% paraformaldehyde? I have asked the surgeon twice for the number for the lab so I can find out - don't have it yet. I have two fresh adrenals in the fridge. Help!! Thanks in advance... Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Thursday, December 11, 2008 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, That's what we always use. Thanks. -Pat Flannery (not a real histologist - I just play one in the lab) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this 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 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] PCNA staining on paraffin
Does anyone have any suggestions for staining PCNA on paraffin? We are using Santa Cruz's PCNA (FL261) SC-7907. We have already stained with it successfully on frozen sections, but it does not appear to be working on paraffin. Thank you. 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
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] JCAHO
Sounds like you found out the hard way? --On Wednesday, December 03, 2008 6:07 PM -0600 Ingles Claire [EMAIL PROTECTED] wrote: FYI Everyone: If you are still waiting for your inspection, don't bother putting up Christmas decorations. Claire ___ 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] 200 proof alcohol?
At our university in Buffalo, NY, we have to sign out the alcohol we purchase from the university (we could buy it from Sigma or other places, I suppose, but generally want to avoid the SH fee). I was told the State of NY required the signature...why? So if we're caught driving drunk they can link it to the alcohol we purchased through the university and diluted to have a party?? And the university would be held liable?? ;-) Merced --On Friday, November 07, 2008 8:56 AM -0500 Emily Sours [EMAIL PROTECTED] wrote: Our alcohol (95 to 100%) is sold by the university only, we don't get a choice. Hopefully if I ever need to use molecular biology grade, the stuff they sell will be good enough. Is it regulated in other universities? I have no idea why they do this, unless they think we're going to have parties. Actually, in PA you can't buy grain liquor, so maybe that's why. Emily -- You would know her for all the things she was...a woman who knew her way in and out of every new book without being singed, pinched, bumped or tickled by any line or chapter. John O'Hara, Appointment in Samarra ___ 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] 200 proof alcohol?
Ahhh...they are federally regulated... http://www.sigmaaldrich.com/chemistry/solvents/products.html?TablePage=14577624 --On Friday, November 07, 2008 9:29 AM -0500 Merced Leiker [EMAIL PROTECTED] wrote: At our university in Buffalo, NY, we have to sign out the alcohol we purchase from the university (we could buy it from Sigma or other places, I suppose, but generally want to avoid the SH fee). I was told the State of NY required the signature...why? So if we're caught driving drunk they can link it to the alcohol we purchased through the university and diluted to have a party?? And the university would be held liable?? ;-) Merced --On Friday, November 07, 2008 8:56 AM -0500 Emily Sours [EMAIL PROTECTED] wrote: Our alcohol (95 to 100%) is sold by the university only, we don't get a choice. Hopefully if I ever need to use molecular biology grade, the stuff they sell will be good enough. Is it regulated in other universities? I have no idea why they do this, unless they think we're going to have parties. Actually, in PA you can't buy grain liquor, so maybe that's why. Emily -- You would know her for all the things she was...a woman who knew her way in and out of every new book without being singed, pinched, bumped or tickled by any line or chapter. John O'Hara, Appointment in Samarra ___ 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 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] RE (Histonet) Bones
This cleaning-bones discussion keeps getting better and better! What an excellent topic for this Halloween week! --On Monday, October 27, 2008 10:19 AM +1100 Farish, Craig [EMAIL PROTECTED] wrote: Hi Ian, Dermestid beetles are definitely the least smelly option (although they are not perfect). You'll find that there will usually be a residual smell in the short term. They can, however, damage small or delicate bones and the best way to deal with these is by bacterial maceration. This smells beyond belief. The anatomy dept here uses flow hoods but you can still smell it from the other side of the building. I've read about people putting them in tubs and leaving them outside(a very long way away) but that's probably not an option in the West End of Glasgow. When they are done, do not use bleach to whiten the bones as that makes them brittle - use dilute hydrogen peroxide (around 3 - 5%). It's worth trying some of the taxidermy forums as well - taxidermy.net is a good place to start. Have fun, Craig Craig (Joe) Farish Senior Technical Officer Veterinary Diagnostics School of Animal and Veterinary Sciences Charles Sturt University Wagga Wagga NSW 2678 Australia ' I don't want to achieve immortality through my work, I want to achieve immortality through not dying' - Woody Allen ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 BRB (packages) / 140 Farber Hall (mail) 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] Histotechnology Society of Delaware - Fall Open Meeting and Hayride/Bonfire
We need this kind of Society in Buffalo, NY. This sounds like too much fun. --On Thursday, October 23, 2008 4:10 PM -0400 Kristen Yaros [EMAIL PROTECTED] wrote: This is a reminder to all Delaware members and non-members that we will be holding our open meeting and Hayride/Bonfire at Carousel Park this Saturday, October, 25th from 3:00 to 6:00 pm. 3:00 Food and Open Meeting 4:00 Hayride 5:00 Bonfire and Smore's HSD is providing the main food. Everyone is asked to bring a covered side dish and fold up chair. Please RSVP ASAP to me! -- Kristen Yaros, HT (ASCP)CM Histotechnology Society of Delaware Correspondence Secretary [EMAIL PROTECTED] ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 BRB (packages) / 140 Farber Hall (mail) 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] RE: [IHCRG] MAOB IHC on either frozen or FFPE sections
3ug/ml sounds awful low for a stock concentration... --On Friday, October 24, 2008 11:51 AM -0500 Swain, Frances L [EMAIL PROTECTED] wrote: Hi bert ifyou will look at the image for the antibody for IHC of MAOB you will see at the bottom that the antibody demonstrated was at a concentration of 3 ug/ml Frances L. Swain HT(ASCP) A. A. S. Special Procedures Technician Department of Orthopaedic Surgery Center for Orthopaedic Research Barton Research Building 2R28 4301 West Markham Street Little Rock AR 72205 (501) 686-8739 PHONE (501) 686-8987 FAX [EMAIL PROTECTED] email From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Connolly, Brett M Sent: Friday, October 24, 2008 10:41 AM To: [EMAIL PROTECTED]; histonet@lists.utsouthwestern.edu Subject: [IHCRG] MAOB IHC on either frozen or FFPE sections Has anyone done it? I have an antibody from Abnova. They illustrate staining of FFPE kidney at 3ug/mL, but when I asked what the stock Ab concentration is they say they never determined that. Go figure? I'm looking for a starting point dilution. Thanks, Brett Brett M. Connolly, Ph.D. Research Fellow, Imaging Research Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. [EMAIL PROTECTED] Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. --~--~-~--~~~---~--~~ You received this message because you are subscribed to the Google Groups ihcrg group. To post to this group, send email to [EMAIL PROTECTED] To unsubscribe from this group, send email to [EMAIL PROTECTED] For more options, visit this group at http://groups.google.com/group/ihcrg?hl=en -~--~~~~--~~--~--~--- 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 Merced M Leiker Research Technician II 354 BRB (packages) / 140 Farber Hall (mail) 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] RE: [IHCRG] MAOB IHC on either frozen or FFPE sections
...considering the stock Ab concentration isn't even known, as Brett mentioned. --On Friday, October 24, 2008 1:09 PM -0400 Merced Leiker [EMAIL PROTECTED] wrote: 3ug/ml sounds awful low for a stock concentration... --On Friday, October 24, 2008 11:51 AM -0500 Swain, Frances L [EMAIL PROTECTED] wrote: Hi bert ifyou will look at the image for the antibody for IHC of MAOB you will see at the bottom that the antibody demonstrated was at a concentration of 3 ug/ml Frances L. Swain HT(ASCP) A. A. S. Special Procedures Technician Department of Orthopaedic Surgery Center for Orthopaedic Research Barton Research Building 2R28 4301 West Markham Street Little Rock AR 72205 (501) 686-8739 PHONE (501) 686-8987 FAX [EMAIL PROTECTED] email From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Connolly, Brett M Sent: Friday, October 24, 2008 10:41 AM To: [EMAIL PROTECTED]; histonet@lists.utsouthwestern.edu Subject: [IHCRG] MAOB IHC on either frozen or FFPE sections Has anyone done it? I have an antibody from Abnova. They illustrate staining of FFPE kidney at 3ug/mL, but when I asked what the stock Ab concentration is they say they never determined that. Go figure? I'm looking for a starting point dilution. Thanks, Brett Brett M. Connolly, Ph.D. Research Fellow, Imaging Research Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. [EMAIL PROTECTED] Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. --~--~-~--~~~---~--~~ You received this message because you are subscribed to the Google Groups ihcrg group. To post to this group, send email to [EMAIL PROTECTED] To unsubscribe from this group, send email to [EMAIL PROTECTED] For more options, visit this group at http://groups.google.com/group/ihcrg?hl=en -~--~~~~--~~--~--~--- 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 Merced M Leiker Research Technician II 354 BRB (packages) / 140 Farber Hall (mail) 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 Merced M Leiker Research Technician II 354 BRB (packages) / 140 Farber Hall (mail) 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] Medical Equipment Source
Hi Ann, Our lab purchased a Leica 2030 microtome (using disposable blades)through Medical Equipment Source. I'd looked at several companies that sell refurbished microtomes, and went with this company because of the cost ($3000) that included a warranty and a newer knife holder than the original one (at my request). They were also very pleasant to deal with. I talked to Luigi Mascio and he was good at keeping in touch by phone and email while the unit was being refurbished for us. We purchased the microtome back in May of this year (5 months ago) and have had no problems with it thus far - it works beautifully. Merced --On Monday, October 20, 2008 3:44 PM -0400 [EMAIL PROTECTED] wrote: Has anyone had any dealings with a company located in Mars, PA called Medical Equipment Source?? I visited their booth at the NSH and am comtemplating purchasing equipment (new and used) from them.? I was wondering if anyone has used them and, if so, what comments can you share about your experience. Annj ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 BRB (Lee Lab) / 140 Farber Hall (mail) 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 In order to put yourself in someone else's shoes, you must first take off yours. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Manual Microwave Method of Tissue Processing
Thanks to everyone who replied to my query! I received many insightful responses. Merced --On Thursday, October 09, 2008 5:19 PM -0400 Merced Leiker [EMAIL PROTECTED] wrote: What are people's exerience with manual processing of FFPE tissues using a microwave? My boss has me looking into a manual method of processing tissues to cut costs - small lab with small budget getting smaller all the time... Merced M Leiker Research Technician II 354 BRB (Lee Lab) / 140 Farber Hall (mail) 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 In order to put yourself in someone else's shoes, you must first take off yours. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 BRB (Lee Lab) / 140 Farber Hall (mail) 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 In order to put yourself in someone else's shoes, you must first take off yours. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] paraffin sections
Hi James, While I know that others with more experience are going to reply and have very good insights to add, in my few years of experience I've stored the paraffin sections at room temp. for up to several months. Sections are stored only after baking them in a 60 C for 1 hour. This baking is done after the cut slides have air dried overnight. I have not had problems with the sections falling off. I know that optimum paraffin section storage is a topic of debate - ambient air vs. nitrogen vs. covered in paraffin; 4 C vs. room temp., as well as the length of storage. While I haven't seen any report of sections falling off due to how they are stored, maybe others on this list have. There may be other issues, like what type of slides or paraffin blend you use. In our lab we use Superfrost Adhesive Slides Platinum Line (Mercedes Medical) and Polyfin (paraffin blended with plasticizers). Also, epitope and tissue type may be a factor. I stain for VWF, Beta-catenin, and cytoskeletal markers such as Myosin Heavy Chain and Tropopin I, all on rodent skeletal muscle and heart tissues, and so far haven't had any problems that I can tell are due to how the sections are stored. Hope this helps in some way! Merced --On Wednesday, October 08, 2008 9:31 AM -0700 James Dooley [EMAIL PROTECTED] wrote: I am a beginning to do paraffin section. I need to know the following as I have heard many things and I have received my best advice here. As of this moment I was storing them at 4 degrees following sections. The problem I am having is many of my section have been coming off when I deparaffinize the tissue. I have been baking the tissue for 60 minutes prior to the deparaffiniztion step. 1. How to store them? 2. How long they can be stored? 3. Under what conditions should they be stored? 4. When melting the paraffin how long should this be done for and at what temperature prior to deparaffinization? Thank you, James ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 BRB (Lee Lab) / 140 Farber Hall (mail) 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 In order to put yourself in someone else's shoes, you must first take off yours. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
