[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 -

[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

[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

[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

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

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:

[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

[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

[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

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:

RE: [Histonet] CD31 for FFPE IHC on Mouse

] 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

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]

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

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

[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. -

[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

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

[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

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

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:

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

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:

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:

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

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.

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 -

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

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

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:

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.

[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

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

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

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

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:

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

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

[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