[Histonet] Cytochrome Oxidase Enzyme Histochemical Staining
Anyone out there doing COX enzyme histochemistry on fresh flash frozen EQUINE muscle...I have tried several methods and fibers are not differentiating the way they should be and are very faint ..any suggestions welcome!! Thanks - Amy Amy S. Porter, HT (ASCP) Michigan State University - Department of Physiology Investigative HistoPathology Lab - Supervisor Research Core Support Facility 567 Wilson Road - Room 2201 East Lansing, MI 48824-6458 517-884-5026 port...@msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] NK Cell markers for Mouse in Paraffin
Hi all - anyone out there have an antibody that they like for NK Cells in a mouse model - I am trying to work with a mouse monoclonal (MOM) with polymer technology. Looking for assistance from labs that have established protocols. We have tried enzymatic and heat retrieval ..not getting good results. Any comments would be appreciated - willing to change antibody vendors currently using clone PK136. Thanks - Amy Amy S. Porter, HT (ASCP) Michigan State University - Department of Physiology Investigative HistoPathology Lab - Supervisor Research Core Support Facility 567 Wilson Road - Room 2201 East Lansing, MI 48824-6458 517-884-5026 port...@msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] University Core Support Facilities
To all: I have been asked to perform a cost analysis and comparison of the rates we charge for services to our researchers here on campus and outside the university - I am looking for labs that would be will to share this information for both in house and outside clients - a lab that does both would be ideal. I would conduct these conversations off list once started to prevent overload of the list and keep your information confidential if needed. Thanks in advance - Amy Amy S. Porter, HT (ASCP) Michigan State University Investigative HistoPathology Lab Research Support Facility port...@msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Looking for assistance with Osteo-Bed Bone MM
Anyone out there using this product for undecalcified bone sections in rodents - looking for information to potentially start this process in our lab. Thanks - amy Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory port...@msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] pepsin pre-digestion before immuno staining
0.1% Trypsin + 0.1% Calcium Chloride in PBS for 10-20 minutes at 37C -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Thursday, June 27, 2013 2:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pepsin pre-digestion before immuno staining We use trypsin and protease fairly often in our lab in conjunction with immuno staining, but we have not used pepsin. An investigator wants us to use pepsin, and we do have some, but would appreciate any recommendations regarding its use - concentration, time, temperature. Thanks Paul M. ___ 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] pepsin pre-digestion before immuno staining
Disregard my previous email that was for Trypsin.duh, sorry about that!! Pepsin we use at a 0.04% in 0.2N HCl for 20 minutes at 37C -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Thursday, June 27, 2013 2:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pepsin pre-digestion before immuno staining We use trypsin and protease fairly often in our lab in conjunction with immuno staining, but we have not used pepsin. An investigator wants us to use pepsin, and we do have some, but would appreciate any recommendations regarding its use - concentration, time, temperature. Thanks Paul M. ___ 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] IHC on Gerbil Tissue
Is anyone out there performing IHC with Mouse primaries on gerbil tissue??? If so what are you using for secondaries? Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Question for Alkaline Phosphatase / Biotin System Users
We are using standard avidin / biotin staining with alkaline phosphatase labeling and sometimes, not every time and not every slide in the same run we get residual blotchy spots and streaking of the reaction on and off the tissue. We have increased to two rinses after primary and two rinses after alk phos reagent prior to substrate; still happening on some slides...anyone else have this experience or type of problem? Any experiences or suggestions are appreciated. Just can't figure out what is causing it.. Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory port...@msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Looking for US Distributor for biological dyes
Does anyone know of a U.S. distributor that carries Chroma-Gesellschaft dyes??? Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Patricia K. Senagore, M.D. - Consulting Pathologist Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: [Histology] Congo Red Stain
Sigma Aldrich makes a very nice kit for this. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sherwood, Margaret Sent: Thursday, November 08, 2012 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Re: [Histology] Congo Red Stain To all: Would people please share their method for doing Congo Red staining? (We are not interested in making up the reagents from powder form). Thanks! Peggy Peggy Sherwood Research Specialist, Photopathology Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114-2696 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-1206 (fax) msherw...@partners.org The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. ___ 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] CD31 for FFPE IHC on Mouse
This antibody was the overwhelming consensus out of all the information I received. Thanks to all for the input!! Amy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Sunday, October 21, 2012 11:20 AM To: 'Mark Elliott'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CD31 for FFPE IHC on Mouse Rat anti mouse cd31 from Dionova is the way to go it is better than all the others. Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Phone: 303-644-4538 Fax: 720-859-4110 pru...@ihctech.net -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Elliott Sent: Friday, October 19, 2012 3:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD31 for FFPE IHC on Mouse Amy Can you please share the responses you got with the rest of us Thanks Mark Message: 8 Date: Thu, 18 Oct 2012 16:21:55 -0400 From: Amy Porter port...@msu.edu Subject: RE: [Histonet] CD31 for FFPE Immunohistochemistry on Mouse Model To: 'Amy Porter' port...@msu.edu,'Histonet' histonet@lists.utsouthwestern.edu Message-ID: 003101cdad6e$3792fc80$a6b8f580$@edu Content-Type: text/plain;charset=US-ASCII Thanks to all for responses..looks like most roads lead to once place which is spectacular!! Just what I needed. Amy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Thursday, October 18, 2012 3:36 PM To: 'Histonet' Subject: [Histonet] CD31 for FFPE Immunohistochemistry on Mouse Model Anyone out there have CD31 working well on FFPE samples for Mouse Samples?? I know this might be a long shot, however I haven't looked for anything on this in quite awhile. Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Patricia K. Senagore, M.D. - Consulting Pathologist Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ***CONFIDENTIALITY NOTICE*** This electronic message and any attachments are intended only for the use of the addressee and may contain information that is privileged and confidential. Any dissemination, distribution or copying of this communication by unauthorized individuals is strictly prohibited. If you have received this communication in error, please notify the sender immediately by reply e-mail and delete the original and all copies from your system. ___ 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] Dako cd3 and cd79a
Don't know what a Flex mini kit is - however we use Dako CD3 @ 1:100 for 60 minute / Heat retrieval steamer - ph 9.0 Tris EDTA / Vector secondaries and enzyme label HRP -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stuart Beaver Sent: Monday, October 22, 2012 12:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dako cd3 and cd79a Hi all, I am looking to for approximate dilution ranges for Dako cd3 and cd79a in canine lymph node samples for T and Bcell Lymphoma. I am using the Flex mini kit at the moment with high pH antigen retrieval. Regards Stuart ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CD31 for FFPE Immunohistochemistry on Mouse Model
Thanks to all for responses..looks like most roads lead to once place which is spectacular!! Just what I needed. Amy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Thursday, October 18, 2012 3:36 PM To: 'Histonet' Subject: [Histonet] CD31 for FFPE Immunohistochemistry on Mouse Model Anyone out there have CD31 working well on FFPE samples for Mouse Samples?? I know this might be a long shot, however I haven't looked for anything on this in quite awhile. Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Patricia K. Senagore, M.D. - Consulting Pathologist Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Stain for HP
Anatech Ltd here in Michigan has a very nice stain for HP I think it is only a couple of components that can be purchased by the quart or the gallon. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Wednesday, October 17, 2012 11:04 AM To: Histonet Subject: [Histonet] Stain for HP What stain are you using for HP?--Giemsa. Warthin Starry or IHC. Do you do them routinely or only when requested? Are they done on an autostainer or with a kit? Diana ___ 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] Shelf Life of Zinc Fixative???
Does anyone out there using Zinc Fixative (prepared in lab / not commercial) have a shelf life they use successfully?? Thanks for the responses in advance. Amy Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Patricia K. Senagore, M.D. - Consulting Pathologist Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Zinc Fixative Question.....
A wonderful colleague recommended to aid in my response that I add the components I am using for preparation to assist in obtaining a more specific response, about shelf life, so here it is: 0.1 M Tris/HCl Buffer pH 7.4 - to which is added: Calcium Acetate - 0.5 gm Zinc Acetate - 5.0 gm Zinc Chloride - 5.0 gm The resulting solution was pH 5.45 According to my protocol my pH should be between 6.5 7.0 which is working. I am just starting to work on developing this protocol and my Zinc Chloride looked pretty pathetic in the bottle and I will be ordering fresh. I think this may have affected the pH - uncertain however at this point. Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Patricia K. Senagore, M.D. - Consulting Pathologist Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: zinc fixative
Thank you Gayle - I am going to order new Zinc Chloride and see where it goes from there. I have utilized the commercial vendors in the past and am just trying to save on purchasing costs if I have a client that has variable size studies - I would like to have it more readily available without having to go through university ordering system which as you know can create a variety of delays when you are trying to take care of researchers. I always make sure to inform them that a slab is not acceptable for this type of fixation. The publications which you have supported are always on my top ten reading list when working with rodent samples!! Thank you for the wealth of information you provided. Amy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of gayle callis Sent: Monday, September 10, 2012 1:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: zinc fixative Amy, You wrote: A wonderful colleague recommended to aid in my response that I add the components I am using for preparation to assist in obtaining a more specific response, about shelf life, so here it is: 0.1 M Tris/HCl Buffer pH 7.4 - to which is added: Calcium Acetate - 0.5 gm Zinc Acetate - 5.0 gm Zinc Chloride - 5.0 gm The resulting solution was pH 5.45 According to my protocol my pH should be between 6.5 7.0 which is working. I am just starting to work on developing this protocol and my Zinc Chloride looked pretty pathetic in the bottle and I will be ordering fresh. I think this may have affected the pH - uncertain however at this point. ** Invitrogen/BD Biosciences sells this fixative aka IHC Zinc Fixative (formalin free) and are only vendors in the USA as far as I know although it is sold in Europe under another name. I did not find a shelf life in either MSDS or Technical Data sheet, but you might want to contact them about this. When we tried this, we made it fresh. You may want to look at cost of buying the ready made compared to buying new chemicals or taking the time to make it up - whatever is cost effective.If you make it up, I suggest using only fresh chemicals if any of yours have been sitting around on the shelf as we ran into the same problem with zinc chloride going bad. The commercial IHC Zinc Fixative is a 10X solution (storage is RT) and diluted with distilled water just before use - very handy. If you are interested, I have many publications on file about this fixative including the original Beckstead (for human CD markers) and Nitta (for murine CD markers) articles and would be happy to send these to you privately. At least the TRIS buffer can be made up ahead of time, maybe even as a 10X solution and dilute when needed to add dry chemicals just before use. One thing is that the pH is never adjusted after adding the chemicals to the pH 7.4 TRIS buffer. This was mentioned in the original publications. Some things to think about when using this fixative.Make sure your tissues are not overly large/thick by reducing the sample size to achieve total fixation since the time of fixation is limited. BD Bioscience says up to 48 hours. I think one could perfuse nicely with this fixative too or at least inject it into lumens, fill lungs, hearts, etc. If you have incomplete fixation with ZnTRIS buffer (Beckstead's fixative), then alcohol during processing will complete the fixation which is something you do not want to happen.Nitta et al had a processing schedule in their publication but we found we had to use shorter processing schedule for murine tissues which became too dry and friable leading to poor microtomy with overly long soaking to get a section. Also the first NBF station on the processor should be replaced with this fixative. When working with formalin sensitive CD markers which is the purpose of this fixative, I wouldn't want a sniff of formalin from NBF carry over into ANY of the solvents. Others may have more suggestions on this. I am not sure what you are using zinc fixative for, but presume it is for CD markers. Good luck, Gayle Callis HTL/HT/MT(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] Loss of Matrigel material during staining
To all: I have a client that we are attempting to stain H E / as well as immuno on FFPE mouse skin with matrigel plugs. These things section like a dream and then when we attempt to stain only the matrigel part of the sample is falling off. Does anyone have any tricks with this stuff they would be willing to share? We are using charged slides and cutting at 5-6 microns per the client. Thanks - Amy S. Porter, HT(ASCP) QIHC Michigan State University Investigative HistoPathology Laboratory William S. Spielman, Ph.D. - Director Patricia K. Senagore, M.D. - Consulting Pathologist Department of Physiology / Human Pathology Biomedical Physical Sciences Building 567 Wilson Road - Room 2133 East Lansing, MI 48824-3320 Phone: 517-884-5026 Fax: 517-432-1368 port...@msu.edu www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] DRGs
I think it is strange that we are all doing similar techniques and wind up with different outcomes using the histogel. I would be curious how many of us are using the equipment sold with the histogel for warming and cooling opposed to any of us who don't. we did not purchase the equipment and I wonder sometimes if warming the histogel using other means causes some type of breakdown / and do any of you repeatedly reheat the same tube once it has been warmed and resolidified?? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Monday, February 22, 2010 9:41 AM To: HISTONET Subject: Re: [Histonet] DRGs Importance: High Hi Carol, I have used histogel for these kinds of samples and also other small, thin tissues like insect antennae and insect GI tracts and midguts. Since I get all my projects already fixed in whatever fixative the investigator chooses, rinsed and placed in 70% ETOH the histogel never touches formalin. I don't use formalin on my processor but start in 70%. I've never had a problem with the histogel. We just put the sample in the histogel flat and stand it up (turn 90°) when embedding in paraffin. I use tissue prep for embedding. If you don't want to use histogel you could try to put the drg's on GN Metricel membrane disc filters. We do this with a lot of the samples I receive, actually I have the investigators or their techs do this. The tissue sticks to the membrane and orientation is a dream. The membrane presents no problem when sectioning. You can get it from VWR. Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 happy slicing and dicing and may all your stains work perfectly - Paula Sicurello P Please consider the environment before printing this email. On Feb 18, 2010, at 11:23 AM, Barone, Carol wrote: Histonetter's...we received a boat-load of mouse DRGs that had been prepared in histogel and are cutting...well..not so good. We normally do DRGs from FS and get beautiful results. We have used histogel before with other small sample and have never had issues... not sure if it is the Histogel or DRG's (fixed in 10% NBF and then transferred to 70% before placed into the histogel).is the issue..I seem to remember that histogel requires formalin and wonder if the transfer to 70% is causing our problem ...but, obviously there is not much room for error with such tiny- tiny samples and they are already process and in paraffin? I am not quite sure how twe can improve the ones that came in histogel, and were processed to paraffin a paraffin blockany idea's? any experience? any anything? Thx- ASAP! cbar...@nemours.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PPE's embedding and cutting
Usually only if you are working with prions. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of rick.garnh...@memorialhealthsystem.com Sent: Monday, February 22, 2010 6:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PPE's embedding and cutting Anyone in histology land required to wear all PPE's to embed and cut? Rick Garnhart HT(ASCP) Memorial Health System Histology Supervisor 1400 E. Boulder St. Colorado Springs, CO 80909 Cell: 719-365-8357 Ph: 719-365-6926 Fax: 719-365-6373 rick.garnh...@memorialhealthsystem.com Mission: To provide the highest quality health care Vision: To create an outstanding health system where patients heal and people thrive Values: Compassion - Integrity - Quality - Respect - Teamwork www.memorialhealthsystem.com The information contained in or attached to this electronic message is privileged and confidential, intended only for the use of the individual(s) named above. If the reader of this message is not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please inform the sender immediately and remove any record of 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
Re: [Histonet] DRGs
We normally do CG's and DRG's in Frozen format as well and have had difficulty working with histogel and low percetages of ethanol. The histogel doesn't really bond with the samples very well when taken from an low % ethanol for us. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Barone, Carol cbar...@nemours.org To: histonet@lists.utsouthwestern.edu Sent: Thursday, February 18, 2010 1:23 PM Subject: [Histonet] DRGs Histonetter's...we received a boat-load of mouse DRGs that had been prepared in histogel and are cutting...well..not so good. We normally do DRGs from FS and get beautiful results. We have used histogel before with other small sample and have never had issues... not sure if it is the Histogel or DRG's (fixed in 10% NBF and then transferred to 70% before placed into the histogel).is the issue..I seem to remember that histogel requires formalin and wonder if the transfer to 70% is causing our problem ...but, obviously there is not much room for error with such tiny- tiny samples and they are already process and in paraffin? I am not quite sure how twe can improve the ones that came in histogel, and were processed to paraffin a paraffin blockany idea's? any experience? any anything? Thx- ASAP! cbar...@nemours.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Parvovirus Control Slides
Jennifer - could you contact me off list for what your actual need is. Thanks - Amy Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Bull, Jennifer L. jennifer.b...@northwestpathology.com To: histonet@lists.utsouthwestern.edu Sent: Thursday, February 18, 2010 2:35 PM Subject: [Histonet] Parvovirus Control Slides Does anyone out there have a resource for unstained slides or tissue infected with parvovirus? Thanks Jennifer Bull Northwest Pathology Bellingham, WA 98225 w: 360-734-2800 x503 jennifer.b...@nwpathology.commailto:jennifer.b...@nwpathology.com mailgate.hinet.org made the following annotations - NOTICE: This email message 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 email 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
Re: [Histonet] Vimentin for Mouse
I believe that Abcam makes a Rabbit anti-Vimentin, we have used in mouse with heat retrieval at ph6 and it worked really well. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Bell, Pat pat.b...@ucdenver.edu To: histonet@lists.utsouthwestern.edu Sent: Thursday, December 17, 2009 6:00 PM Subject: [Histonet] Vimentin for Mouse Thank you all very much for your help regarding the COX2 for the mouse. Now I would like to ask your help regarding Vimentin for FFPE mouse tissue. I have tried Dako and Sigma with no success. I am also not sure of what clone to use. Thanks again. Pat Pat Bell HT(ASCP) Medical Oncology; MS 8117 12801 E 17th Ave. Aurora, Co. 80045 303-724-6077 pat.b...@ucdenver.edumailto:pat.b...@ucdenver.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Autostainer service ...Dako would you respond?
I just renewed my service contract for my autostainer which is 6 years old - no problem. Just an FYI. Amy Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Horn, Hazel V hor...@archildrens.org To: McMahon, Loralee A loralee_mcma...@urmc.rochester.edu; Xipamanine Mkuze xipaman...@gmail.com; histonet@lists.utsouthwestern.edu Sent: Wednesday, December 02, 2009 9:41 AM Subject: RE: [Histonet] Autostainer service ...Dako would you respond? Dako would you respond to this issue? Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 1 Children's WaySlot 820 Little Rock, AR 72202 phone 501.364.4240 fax501.364.3155 visit us on the web at:www.archildrens.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of McMahon, Loralee A Sent: Wednesday, December 02, 2009 8:39 AM To: Xipamanine Mkuze; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Autostainer service I think that they are not supporting the older model of autostainer. They are still selling a new version of the autostainer. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Xipamanine Mkuze [xipaman...@gmail.com] Sent: Wednesday, December 02, 2009 9:35 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Autostainer service Dako will no longer support Autostainers? Why? Ninni I need to pick your collective brains here about getting a service contract for my Dako Autostainers. I remember seeing a posting awhile back with the name of a company, I think from MD, that will service the Dako Autostainers now that Dako will no longer support them. If anyone has a suggestion for a company who will service these instruments or if anyone can remember the name of that company in MD, please contact me. Happy Monday everyone, Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 ___ 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 ** The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ___ 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] GFAP IHC
If I remember correctly Dako's GFAP is a bovine immunogen (bovine spinal cord) - that is probably what is causing the problem. You may want to look for GFAP where the cow is not the immunogen / try running another brain from a different species and see if you get the same problem. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Nancy Herman nancy.her...@inspection.gc.ca To: histonet@lists.utsouthwestern.edu Sent: Wednesday, October 28, 2009 4:19 PM Subject: [Histonet] GFAP IHC We are using DAKO's ready-to-use GFAP antibody on our bovine tissue and using Envision+ HRP Rabbit detection system. When using this antibody we see a brown color on the slide itself. The staining was intense and quite uneven. We have diluted the anitbody to 1:10 but there is still the brown slide. The staining is not quite as uneven but appears more washed out. We don't see this with any of our other antibodies we are using and our no primary controls for GFAP are clean. Any advice or explanations for the slides picking up the brown color or improving GFAP IHC on bovine tissue? Thanks Nancy Herman CFIA Lethbridge Laboratory Lethbridge, Alberta ___ 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] Canine endothelial antibody
We use Dako Von Willebrand Factor VIII in Rabbit on multiple species and it works fabulously. Enzyme pretreatment with 0.03% pronase e for 10 mins at 37C / Primary at 1:200 to 1:500 depending on the species being stained. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Jackie M O'Connor Jackie.O'con...@abbott.com To: histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, October 28, 2009 4:35 PM Subject: [Histonet] Canine endothelial antibody A week or so ago I thought I saw a post about an antibody that works for canine endothelial cells, but I can't find it in the archives - was I just dreaming, because I need this so bad? I've tried Factor VIII and a CD34 on FFPE, but with poor results. Any help would be most appreciated. Thanks, Jackie O' ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] RE: reduction/elimination of red cell autofluorescence on FFPE sections for IF examination
We have been utilizing 0.25% Ammonium Hydroxide(28%) + 70% Ethanol v/v for 1 hour followed by 10 minutes in 50% Ethanol rinsing in several changes of buffer afterward and it seems to be working well for a multitude of autofluorescent problems on FFPE sections for us. We tried to Sodium Borohydride and it was a little diffiucult to work withquite hazardous. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Thurby, Christina christina.thu...@bms.com To: Thurby, Christina christina.thu...@bms.com; histonet@lists.utsouthwestern.edu Sent: Monday, August 24, 2009 11:00 AM Subject: [Histonet] RE: reduction/elimination of red cell autofluorescence on FFPE sections for IF examination Sorry I did not include my contact information. Christina Thurby christina.thu...@bms.com 812-429-8097 Thanks! -Original Message- From: Thurby, Christina Sent: Monday, August 24, 2009 9:59 AM To: 'histonet@lists.utsouthwestern.edu' Subject: reduction/elimination of red cell autofluorescence on FFPE sections for IF examination Can anyone give feedback on reagents/procedures to use for immunofluorescence to reduce/eliminate red cell autofluorescence on FFPE sections. I am using an indirect method of labeling for two antibodies (FITC and Texas Red). I have not tried 0.1% sodium borohydride. Will this help for formalin fixed specimens? I have read that it is used for gluteraldehyde autofluorescence reduction. If appropriate how long should this reagent be applied to the specimen and at what temperature. This message (including any attachments) may contain confidential, proprietary, privileged and/or private information. The information is intended to be for the use of the individual or entity designated above. If you are not the intended recipient of this message, please notify the sender immediately, and delete the message and any attachments. Any disclosure, reproduction, distribution or other use of this message or any attachments by an individual or entity other than the intended recipient is prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] CD31 working protocol
BD Biosciences has a great rodent CD31 for mouse and rat - however you need to use their Zinc Fixative. It is well worth the results!! Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Zerfas, Patricia (NIH/OD/ORS) [E] zerf...@ors.od.nih.gov To: histonet@lists.utsouthwestern.edu Sent: Monday, August 10, 2009 2:50 PM Subject: [Histonet] CD31 working protocol Dear Listservers, I have tried MANY protocols and I am unable to obtain a working protocol for the CD31 antibody. I have tried both the CD31 rat monoclonal and a CD31 rabbit polyclonal. Thanks in advance for your help. Patricia Zerfas National Institutes of Health Building 28A, Room 112 28 Library Drive Bethesda, MD 20892 ph: (301) 496-4464 fax: (301) 402-1068 ___ 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] metal embedding base molds
Try EM Sciences the distribute alot of the unique old type histology supplies. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Victoria Baker bakevicto...@gmail.com To: histonet histo...@pathology.swmed.edu Sent: Thursday, August 06, 2009 1:13 PM Subject: [Histonet] metal embedding base molds Hi- I'm trying to locate embedding molds that Sakura/Tissue Tek makes. All I'm able to find at this point are the metal molds with the 'wings' on them. Does anyone know a source? With all the mergers etc it's been a little challenging. Thanks in advance Vikki ___ 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] Clear-Rite 3
I would contact Anatech Ltd. they have a substitute Pro-Par Clearant www.anatechltdusa.com - they are all very knowledgeable and would be able to help you I am sure. I don't use substitutes in my lab at this time, however I would contact them if I were ready to make the switch. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: O'Donnell, Bill billodonn...@catholichealth.net To: Histonet@lists.utsouthwestern.edu Sent: Wednesday, August 05, 2009 9:11 AM Subject: [Histonet] Clear-Rite 3 Greetings! We have been using Clear-Rite 3 here at our lab, and we are happy with the product. Our supplier says it will be on back-order for some time now. Our crack supply folks are looking for another source. I'm taking another route to find out what products out there are comparable. Are all Xylene Substitutes pretty much the same and there for pretty much interchangable? Are there some to stay away from? Any help is appriciated. William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 ___ 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] Help with Connexin 43
Anyone out there doing FFPE IHC on rat for Connexin 43 - I am having trouble getting the titration right on this just wondering what types of dilutions anyone is using??? I am using a standard avidin/biotin complex staining system with and endogenous AV/BI block and not getting anything of any consequence in my negative control. thanks - Amy Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] NADPH-diaphorase
Try Sigma-Aldrich. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Breisch, Eric ebrei...@rchsd.org To: histonet@lists.utsouthwestern.edu Sent: Thursday, May 07, 2009 12:23 PM Subject: [Histonet] NADPH-diaphorase Hi, hoping that someone might be able to share with us where they purchase NADPH-diaphorase for the NADPH stain. Thank you. EAB Eric A. Breisch, Ph.D. Clinical Anatomist Dept. of Pathology Rady Children's Hospital and Health Center Associate Clinical Professor of Anatomy Dept. of Surgery UCSD School of Medicine ___ 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] IHC double labeling question
I would do them together making sure your end volume holds the same concentration of each reagent as you use singely. It should work fine for you and cut down on your time. You will need to find a generic protein block to utilize if your secondaries are made in two different hosts. If your antigens are co-localized you may possibly have a difficult time demonstrating them together. Good luck, Amy Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Nicole Collette collet...@mail.llnl.gov To: histonet@lists.utsouthwestern.edu Sent: Thursday, April 30, 2009 1:16 PM Subject: [Histonet] IHC double labeling question Hi, All, I am starting to do some IHC on FFPE mouse tissues, and have several antibodies working individually on my tissues (with the same retrieval protocol). The next step is to move on to double labeling, and my generic protocol calls for each label to be done sequentially (primary, secondary, followed by primary, secondary). My question is, if both of my primary antibodies are raised in different species, and are also different from my host species, can they be done together (mix the primaries for one incubation, mix the secondaries for detection)? It would save a day. I expect to see colocalization, is it better to do both primaries in one incubation so that binding of one doesn't exclude the other? I understand that I will have better control over the post-antibody washes if I do them separately, but is there another reason to do them sequentially if the retrieval is the same? Thanks for the advice! Sincerely, Nicole Collette LLNL/UCB ___ 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] Another CAP question
I use my yearly p.m. records, certified themometer maintenance records, I concentrated more on the function item and not the ongoing results. In my mind if something is broken or repaired the ongoing results have not been satisfactory. Hope this helps - Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Mary Abosso mabo...@unipathllc.com To: Histonet histonet@lists.utsouthwestern.edu Sent: Monday, April 27, 2009 1:15 PM Subject: [Histonet] Another CAP question Good Monday all - We were just talking about this reg. and wondering how everyone is answering this? Are instrument maintenance records acceptable? ANP.23075 - Is there evidence of ongoing evaluation of results of instrument maintenance and function for all devices? Mary Abosso ___ 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] Mega cassettes
You could try Surgipath Medical. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Ian Montgomery ian.montgom...@bio.gla.ac.uk To: histonet@lists.utsouthwestern.edu Sent: Friday, April 24, 2009 10:45 AM Subject: [Histonet] Mega cassettes Looking for a supplier of mega cassettes in the UK. VWR are a wee bit expensive. Ian. Dr. Ian Montgomery, Histotechnology, I.B.L.S. Support Unit, Thomson Building, University of Glasgow, Glasgow, G12 8QQ. ___ 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] CAP question
Mary - I have one generic protocol with a spreadsheet attached stating the following: Name of Antibody CD/CK # Clone Species Pretreatment Dilution Incubation time Enzyme (ABC or AP) Lot # in Use Hope this helps out it has passed in the past. take care. amy Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Mary Abosso mabo...@unipathllc.com To: Histonet histonet@lists.utsouthwestern.edu Sent: Wednesday, April 22, 2009 1:30 PM Subject: [Histonet] CAP question Hi all, I am updating procedures and am wondering if there is a better format for compliance with ANP.22250 - Does the procedure manual address all methods and antibodies currently in use? Just wondering if I need to include every antibody or a generic procedure covering most antibodies, with the exceptions only reference within the same procedure? Any help would be appreciated. Thanks, Mary Abosso ___ 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] IHC, decalcification preferences
All the mouse femurs and tibias done in our laboratory are decaled in 14% EDTA and we have a high success rate with our immunohistochemistry and Tunel staining. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu - Original Message - From: Nicole Collette collet...@mail.llnl.gov To: histonet@lists.utsouthwestern.edu Sent: Wednesday, April 01, 2009 12:44 PM Subject: [Histonet] IHC, decalcification preferences Hello, all, I am going to be doing some IHC on adult mouse bone, and since I'm new to all this antigen preservation stuff, was wondering the best way to decalcify? Normally, I use high percentage EDTA (17%) and determine end point by weight loss/weight gain. (up until now have been using this protocol for LacZ stains, works like a charm! ) We are comparing several lines of mice with bone mineral density differences, and endpoint is very phenotype and age-dependent. It takes a long time, but the results are worth the wait. I also have Cal-Rite, which is a formaldehyde/formic acid decalcifier, and obviously works much faster. I am leaning toward the EDTA, since it seems to work well for the LacZ, which is sensitive to lots of other processes, but if there's a reason not to, please let me know. Thanks for all your help and support, I have so far been given great advice from this listserv. Sincerely, Nicole Collette LLNL/UCB ___ 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] ? About fixation of excessively bloody tissues
Does anyone out there know of a reference that states or discusses tissues that a covered with a large amount of excess blood have problems with fixation. I have looked in the reference materials I have here in the lab, however I cannot find anything to support my complaints to a client that state entire mouse organs (whole liver, spleen, lung, heart...) covered in blood thrown into onecassette with to little formalin will not be well preserved. I know garbage in garbage out - I am trying to get rid of the garbage. Thanks in advance for the help. Amy Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
