Re: [Histonet] RE: B-cell Ab to work on rat tissue
Carl's choice is excellent. The classical B-cell marker might be CD20, and does work great, but as with all antibodies, you have to be aware of exactly what you are after. CD20 comes on line after CD19 induction so CD20 might not be seen on very early b-cells. Like wise CD79a is accompanied by chaperone proteins prior to it's assemblage with the B cell receptor (during the pro-B stage). Won't mark these very early cells well. The assemblage is complete at pre-B stage and continues expression even till plasma cell stage. So true for almost all b cells, CD79a is great and you can see rat tissue IHC of CD79A out on the web, have used several of them, along with protocols (HIER as Carl pointed out works well). The complex is found on virtually no other cell as it is the signalling complex for the b cell receptor. Ray Ray Koelling PhenoPath Labs Seattle, WA - Original Message - From: "Carl Hobbs" To: histonet@lists.utsouthwestern.edu Sent: Sunday, January 15, 2012 8:24:59 AM Subject: [Histonet] RE: B-cell Ab to work on rat tissue It may be that CD79a Ab is appropriate? It works well on Pwax sections, after HIER. I do not know the specificity of this Ag for B cells, so would be interested in feedback Check here for images : http://www.immunoportal.com/modules.php?name=gallery2 Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ___ 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: Formalin Neutralizing
Thanks Rob for reminding me. It was John A. Kiernan who originally posted the message that I refer to below on Histonet back in 2002. As everyone will agree John is a great teacher and one of our "National Treasures" in Histotechnology. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 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 Bob Richmond Sent: Saturday, 14 January 2012 4:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Formalin Neutralizing Three years ago Tony Henwood in Australia posted a formula for neutralizing formaldehyde with ammonia. He also alludes to using sodium bisulfite (a.k.a. metabisulfite), but I don't know the proportions or how the reaction works. - I've posted a copy of his note, below. A lot of commercial formaldehyde neutralizers are pure mumbo-jumbo. Of course, this has become an issue where what managers and regulators think is a great deal more important than what actually happens at the chemical level where MBA's fear to tread. Bob Richmond Samurai Pathologist Knoxville TN * Date: Wed, 25 Mar 2009 09:45:43 +1100 From: "Tony Henwood" Subject: RE: [Histonet] FW: formalin neutralizers To: "Burton, Lynn" , This is what we use: Neutralization and Disposal of Formalin Fixative 10% Formalin can be neutralised with sodium bisulfite or concentrated ammonia. The reaction with ammonia results in the formation of hexamethylenetetramine (commonly known as hexamine or methenamine). This can then be safely disposed of either as a liquid fertiliser or via the sewage system (check with your local authority). The reaction proceeds as follows: 6 CH2O + 4 NH3 ---> C6H12N4 (Hexamethylenetetramine) + 6 H2O Procedure: 1. Before beginning, personnel must have the following safety equipment readily available in the event of an accidental spill: sorbent material (spill pillows, bulk sorbent) formaldehyde rated respirator. 2. Personnel must wear a lab coat or apron, safety goggles and neoprene gloves. 3. A pH meter or pH paper 4. To 1000 ml of 10% formalin (= 4% formaldehyde) add 56 ml of strong ammonia solution (27%). This will generate 31 g of hexamine (approximately a 3% solution). 5. Stir well. Reaction may produce heat. 6. Initially, the pH of the formaldehyde solution will be about 6. As ammonia is added and stirred, a fluffy white precipitate will result. Addition of sufficient ammonia will raise the pH to about 8. Because the neutralization of the formaldehyde requires less molecules of ammonia than the apparent acid-base reaction supplies hydronium ions, the pH change from acid to base is used as an indicator that an excess of ammonia has been added. 7. Let set overnight (12 hours). 8. The smell of formalin is greatly reduced or replaced by a faint whiff of ammonia. 9. Schiff's reagent is perhaps the best, most sensitive and available reagent in any lab to test for the presence of aldehydes. If the "neutralized" formalin turns purplish with the addition of Schiff's reagent, it is not totally neutralized and you will need to add more ammonia. 10. Dispose of appropriately. I am not sure how the bisulphite method works. I picked it up from a reference on formalin neutralisation but have never tried it. And would you believe that after some searching I can't even find that reference (I probably have it on my home computer). The notes come from my "Infamous" text book I have been writing for the last 20 years. As my staff call it, the book that will probably never be published. But then the chapters are quite usefull for teaching so they are of some use. 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 ___ 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
[Histonet] Cryostat cutting?
Hello, We need to cut clean and consistent 50 micron sections of OCT embedded tissue on our leica cryostat (CM-1850). Does anyone on the list have any tips or tricks for doing this when compared to cutting the usual <10 micron sections that we usually deal with in our lab. -- Regards, Francis ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Gary Steinke is out of the office
I will be out of the office starting 01/15/2012 and will not return until 01/19/2012. I will be out of the office starting 01/15/2012 and will not return until 01/19/2012. I will be attending our National Sales Meeting, and may not be able to respond in a very timely manner. For immediate assistance, please contact Healthcare Customer Service at 877-881-1192. Otherwise leave a message and I will return it as soon as possible. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] B-cell Ab to work on rat tissue
Are you asking about pancreata B-cells? ___ James E. Staruk HT(ASCP) www.masshistology.com www.nehorselabs.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Louro, Pedro Sent: Friday, January 13, 2012 3:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] B-cell Ab to work on rat tissue Hello out there in Histoland, I'm looking for a B-cell antibody will stain rat paraffin embedded tissues. Anyone out there that has had any luck with this? Thanks in advance for your input. Pedro Our Values: Excellence, Drive, Ownership, Challenge, Teamwork, Respect LEGAL NOTICE This message is confidential and contains information which may be legally privileged. It is intended for the stated addressee(s) only. Access to this email by anyone else is unauthorised. If you are not the intended addressee, any disclosure or copying of the contents of this email or any action taken (or not taken) in reliance on it is unauthorised and is unlawful. If you are not the addressee, please inform the sender immediately and destroy the original message. The data in this email will have been screened for the presence of computer viruses known by the Company at the time the email was produced. The Company cannot guarantee, however, that the email or any attachments are virus free. Delivery of the email will be at the Client's risk. Registered in England No. 1815730 VAT Reg. No. GB425507072 Registered Office: Woolley Road, Alconbury, Huntingdon, Cambridgeshire PE28 4HS ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - No virus found in this message. Checked by AVG - www.avg.com Version: 2012.0.1901 / Virus Database: 2109/4740 - Release Date: 01/13/12 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: B-cell Ab to work on rat tissue
It may be that CD79a Ab is appropriate? It works well on Pwax sections, after HIER. I do not know the specificity of this Ag for B cells, so would be interested in feedback Check here for images : http://www.immunoportal.com/modules.php?name=gallery2 Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re:
..I can give you a good advice: visit this drugstore and your problem will be solved. http://test.toshibapod.bi-int.com/new-year.link.php?cyzlink_friend_id=09ag1 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
