RE: [Histonet] Leaving sample over night in ETOH 70% during fixation
Storing your samples overnight in 70% ethanol after fixation won't harm your samples, but they should be adequately fixed prior to this. I do not like xylene for mouse tissues, it makes them too brittle. Instead I used histoclear 11 from National Diagnostics. Histoclear 11 is less expensive than the original histoclear, but performs well in my experience. I found that for most of the tissues I processed (I never tried diaphragm) a shorter schedule worked well and in fact the process you describe may be too long for some tissues. The Society of histotechnologists produces a booklet with suggested processing schedules for a wide range of animals and tissues, I found this invaluable in designing my protocols. My process for mouse tissues took aroung 6 hours using a processing machine equipped with vacuum on all stations. Unfortunately as I have retired I don't have access to my SOP's any more. However, if you obtain the booklet from the Society, you will be able to devise a good protocol. At the end of the day, if your tissues section and stain well, then your process is satisfactory. It's always helpful to compare your sections with other people's if you can then you have a benchmark of quality. This can be difficult and frustrating in a small research lab. You may find that postfixing your liver samples in formol alcohol (90ml absolute ethanol to 10ml 37% formaldehyde or use Pen fix from thermo shandon) will improve the morphology. I used to do this for a couple of hours; you may need to experiment, especially if you are doing immunohistochemistry which is presumably why you are using paraformaldehyde. Good luck Margaret From: histonet-boun...@lists.utsouthwestern.edu on behalf of Itai Moshe Sent: Thu 07/07/2011 09:22 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leaving sample over night in ETOH 70% during fixation Dear All, Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C during the fixation process, will be better for the tissue fixation, and does not harm the sample ? Does the fixation process should be done straight forward from the first step to the last one without any over night stops (except from the PFA, Bouin's step) ? My fixation protocol is like this: 1) immediately after killing the mouse i'm putting the sections in a fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml DDW - pH 7, Or bouin's solution over night at 4C. 2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT 3) Xylen x2 - each for 1Hr at RT. 4) Paraffin x3 - each at 60C for 1Hr. Thank you all very much in advance Itai M Doe ___ 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] MSDS info
Your suppliers are required by law to supply msds and they would no doubt be please to do so. However the Sigma website is great as you can search all their reagents and click on MSDS and download a PDF file of the MSDS. http://www.sigmaaldrich.com/ This hyperlink should take you to their site. Regards Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dianne E. Holmes Sent: 27 May 2011 17:48 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] MSDS info HELP !!! I am having to list all my chemicals in the lab for the Safety dept. Several have no MSDS sheet with them? (been here longer than I have!) Where can I obtain this info to put in my records? Individuals who have received this information in error or are not authorized to receive it must promptly return or dispose of the information and notify the sender. Those individuals are hereby notified that they are strictly prohibited from reviewing, forwarding, printing, copying, distributing or using this information in any way. ___ 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] a Perfect Red
Dear All, Thank you all for your contributions to my request for information about A Perfect Red, Yes I should have searched it on the Net. Just didn't think! Anyway I shall be ordering it and will enjoy reading it. Thank you all a million. Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] off topic/on topic
Hi Andi, Would you kindly let me have the reference for A perfect red, please? Thanks Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Grantham, Andrea L - (algranth) Sent: 26 May 2011 20:29 To: HISTONET Subject: [Histonet] off topic/on topic This is mostly for the girls - I dropped into the Ulta store yesterday to pick up a few things and the sales person wanted me to try this somewhat new product by Benefit. It was a lip and cheek tint - came in a little bottle - looked like mucicarmine working solution. I looked at the box that it came in and sure enough, one of the first ingredients was CARMINE! So I had to relate a few of the stories about beetles and Carmine and pirates that I learned from reading A Perfect Red recommended by Dick Dapson...wonder if I bored her? Maybe she won't bother me the next time I'm rushing through the store trying to get my stuff and get out of there. Andi ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thank you
Hello everyone! Thank you all for your good wishes on my retirement. I knew you were all great people: you have just proved it big time! Thank you all from the bottom of my heart. Keep smiling! Much love Margaret ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] This is goodbye
To everyone in histoland, This is my last full week at work. From next Tuesday I will be retiring from my job at Cambridge University. I have thoroughly enjoyed my chosen profession as a research technician specialising in histology. I realise that because I have always worked in research my experience is somewhat limited: I haven't been asked for the same range of techniques that you find in a routine clinical lab. However, I have enjoyed my work and have learnt and devised some techniques for a range of interesting samples. Now it's time to learn something new! I hope to spend more time with my lace pillow and learn to do it properly, maybe learn to do Northamptonshire lace which I believe is a variation on Bucks point. (I come originally from Northants) I have had a go at Bedfordshire and will learn more, it is very pretty. I also plan to travel; my next trip is to New Zealand in November. I won't go on. I just want to say this: You are all great guys and girls; you share your experience and friendship with great generosity, which I have always felt is what a scientist should do. Thank you for your comradeship and help over the years. Maybe I'll get to meet some of you by chance during my travels. Best wishes to you all and a great big Thank You, From Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] No patient ID: Ink dissolved from Cassettes duringprocessing.
Pencil does not dissolve in processing reagents and is safe. I have also found the marker pens from Surgipath to be the best of those I have tried, but with the proviso that the labels should be left to dry for a couple of hours before being immersed in ethanol. I routinely use one of these pens for labelling my slides. The old fashioned way was to put a labelled slip of card or paper into the cassette with the tissue. I don't know how this would fit in with US regulations... Margaret -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kiranjit Grewal Sent: 29 April 2011 22:44 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] No patient ID: Ink dissolved from Cassettes duringprocessing. Hi All, What is the standard practice out in histology world if hand written cassette id washed away during processing? Please share if you had any experience and how did you resolve this and what is your current practice. Thank you so much! -Kiranjit ___ 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] science for kids
Me too, even in the UK! Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pam Barker Sent: 05 April 2011 20:25 To: 'Emily Sours' Cc: 'Histonet' Subject: RE: [Histonet] science for kids Me TOO How do we get them? Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: rel...@earthlink.net www.facebook.com search Pam Barker RELIA www.linkedin.com/reliasolutions www.myspace.com/pamatrelia www.twitter.com/pamatrelia -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Emily Sours Sent: Tuesday, April 05, 2011 3:19 PM To: Grantham, Andrea L - (algranth); histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] science for kids A coloring book?! I want that for myself!!! A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron On Tue, Apr 5, 2011 at 1:49 PM, Grantham, Andrea L - (algranth) algra...@email.arizona.edu wrote: Emily, NSH has a coloring book that explains histology. Andi Grantham On Apr 5, 2011, at 7:14 AM, Emily Sours wrote: I'm going to visit my nieces this summer and they want to know what I do at work. Does anyone know of a good book or website that explains DNA and/or histology for 4 to 6 year olds? Just thought I'd throw it out there, since googling it would yield way more than I can deal with. Emily 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 ___ 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
RE: [Histonet] RE: OT: April Fool's Nonsense
I like that one! But I have to say I would have been feeling very, very squeamish had I been a fly on the wall. Sally here's to a happy retirement for you. If you come over to the UK, look me up in Cambridge. Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lemke, Nancy Sent: 29 March 2011 15:32 To: Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: OT: April Fool's Nonsense The absolute best one I ever witnessed was when I worked in a histo lab that was next to the microbiology lab. To set up the director, some one came in with a stool specimen cup that was used in parasitology. It had a very official patient sticker on the side and was shown to the director. He was told that a test had been ordered that the tech did not recognize. The director was also unfamiliar with the requested test, so he asked what the exact nature of the specimen was, something that must have happened often, so the tech pulled the lid off of the container, whipped out a tongue depressor and stirred up the tan, gloppy contents, all the while looking like he was going to retch. He then scooped up a large dollop of the contents and ate it!!! The director came very close to apoplexy as did everyone else in the lab. At that point the tech shouted April Fools! and revealed that the container had butterscotch pudding in it! It took quite a while to calm down the lab, but the joke was gleefully told throughout the hospital! For strictly histo, it is always good to have a tray of slides of odd items, such as ffpe hotdogs, etc and see how far the diagnosis goes! Nancy W Lemke Research Coordinator Hermelin Brain Tumor Center Neurosurgery Research Henry Ford Hospital Detroit, MI (313) 916-8648 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara [sbree...@nmda.nmsu.edu] Sent: Tuesday, March 29, 2011 9:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] OT: April Fool's Nonsense I like what I've gotten so far but I'm obviously a lot more evil than some of you... I'm thinking about how I could temporarily glue the radio station dial on the boss's radio to classical music in order to keep him from listening to Rush Limbaugh and Glen Beck. Or Saran Wrap over the urinal. Vaseline on office door handles. Filling one of their offices with bubble wrap. Gluing their desk drawer shut (the one with the food in it). Let's get SERIOUS here - I need more! I want people to be glad I've retired! Heh...heh... No pathologists were harmed in the making of this Tomfoolery. Darn it. And, yes, I do have work to do today... so far, anyway. 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 == 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Stupid Rabbit primaries!
Having read all the other comments, here's my 2.5 pence worth! Have you tried a background reducing antibody diluent? You can get these from DAKO or Menarini Diagnostics: http://www.dako.com/uk/ar38/p107410/prod_products.htm. Or MP-905-25 / MP-905-100, depending on the volume you require, from Menarini Diagnostics, website http://www.dako.com/uk/ar38/p107410/prod_products.htm. I found these useful for rabbit polyclonals. Good luck Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@mirnarx.com Sent: 22 March 2011 21:04 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stupid Rabbit primaries! So I haven't had to deal with rabbit polyclonal primaries in a long time because I remember how much the background sucks with them. Unfortunately the only available antibody is a rabbit polyclonal. Does anyone have any suggestions for how to eliminate the background? I have diluted almost to the point of the antigens not showing! Thanks guys and gals!! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ 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] Fat tissue
Has anyone experience of cryostat sectioning of fat tissue? This would be rodent fat pads, particularly the gonadal fat pad which usually consists largely of adipocytes. What temperature would you cut at? Is there anything that could be used to infiltrate the tissue prior to sectioning to provide more support? Regards Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Formalin-fixed paraffin embedded question
I agree with all the comments so far, and I think all the steps are far too long, particularly the overnight in molten wax. You are effectively cooking your samples. I would avoid overnight in 100% ethanol too; it will tend to harden the tissue. Get hold of the manual for animal tissues available from the Society for histotechnology. I found this invaluable and now most of my tissues can be sectioned without soaking at all. I just chill them slightly, if necessary. Good luck with your samples. Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Noel Gray Sent: 21 February 2011 20:55 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Formalin-fixed paraffin embedded question I am having an issue with formalin-fixed, paraffin embedded tissue that I am sectioning. I am using a microtome to cut the tissue (mouse brain, cerebellum and spinal cord) in 10 um sections which I will stain using cressyl violet. Sometimes, the tissue in a block will splinter once it hits the blade. Usually all samples from the same animal splinter but this is not always the case. If I put the block into the water bath (sectioning surface exposed to water or not) this seems to stop the splintering for 1-200 um worth of tissue. However, I am afraid this may bring error into the histological analysis of my tissue. I assume it has something to do with the protocol I am using to prepare the tissue. I guessed that maybe the dehydration, alcohol clearing, or paraffin infiltration are not complete, resulting in the problem I have. However, I looked at various FFPE protocols and each of my wash steps are longer, which may be the problem? I was wondering if anyone has encountered this before, or if anyone knows exactly what is going on with my tissue and how I can fix it? Thank you. Here is my protocol: -Anesthetize mouse followed by a system flush of 30 ml of PBS and then slow perfusion of 50 ml of 4% PFA. -Brain and spinal cord are removed as a single, in tact unit and placed into 70% ethanol for 4 hours -80% EtOH for 4 hours -90% EtOH over night -100% EtOH #1 for 4 hours -100% EtOH #2 for 4 hours -100% EtOH #3 over night, forebrain cerebellum and spinal cord are separated -Xylene wash #1 for 4 hours -Xylene wash #2 for 4 hours -Xylene wash #3 over night -Moulton paraffin wash #1 for 4 hours -Moulton paraffin wash #2 for 4 hours -Moulton paraffin wash #3 over night -Tissue is embedded and then sectioned into 10 um sections Again thank you for your time. Noel W Gray Neuroscience Graduate Program SUNY Upstate Medical University 3219 Weiskotten Hall 766 Irving Ave Syracuse, NY 13210-1630 (315) 464-8144 gr...@upstate.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] FFPE tissue - sucrose fixing necessary?
This sounds like something you would do before freezing not processing to paraffin. Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Abbey Mortimer Sent: 10 February 2011 02:22 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] FFPE tissue - sucrose fixing necessary? Hello helfpul Histonetters! I am curious about your formalin-fixed paraffin embedded (FFPE) tissue protocols. If the tissue has been fixed with saline and paraformaldehyde during perfusion, then further fixed in paraformaldehyde for 24 hours, do you still utilise a sucrose wash before running it through the automatic tissue processor? Or do you not need this step? I have heard mixed responses about this and wonder if any of you have had experience with this! Thanks so much, Abbey ___ 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] fixation question for IHC
It is possible to freeze paraformaldehyde in aliquots, so you don't have to keep on making it up repeatedly as we did in the old days. I have also seen that you can buy Pure formaldehyde, but I don't remember who the supplier was. It was in sealed ampoules and was 16% so you could dilute it in your preferred buffer. I have to say that this was about 10 years ago! Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jan Berry Sent: 28 January 2011 18:15 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixation question for IHC Is there a difference between using paraformaldehyde and neutral buffered formalin when choosing a fixative for IHC? I would prefer to use formalin because of easier preparation, but am willing to put in the extra time to make fresh paraformaldehyde solution if there is a compelling reason. Jan Berry University of Michigan ___ 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] Beta Galactosidase antibody
Hi Everyone, Does anyone have a favourite antibody for beta Galactosidase that they would be confident to recommend? Thanks a lot. Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Animal tissue processing
Generally rodent tissues require shorter times for processing. See the Animal Processing Manual published by the NSH for a range of tried and tested protocols. Below I have pasted a copy of my schedule which works well for mouse tissues. As you will see I designed it in the first instance for pancreas which tends to harden on longer processes, now I use it for all my mouse tissues, with the exception of intact heads (these require much longer, i.e. 2 hours per station.) All my processes are developed using the methods in the above mentioned manual as a guide. My processor is a dunk and dip type of machine, the Leica TP1020. I hope this helps. SOP 3 PROGRAMME 4: FOR MOUSE PANCREAS COSHH CBH001 STATION REAGENT DURATION VACUUM 1 Formalin/70% ethanol 30 mins Y 2 80% Ethanol 30 mins Y 3 90% Ethanol 20 mins Y 4 Absolute ethanol/IMS 20 mins Y 5 Absolute ethanol/IMS 30 mins Y 6 Absolute ethanol/IMS 30 mins Y 7 Histoclear II 20 mins Y 8 Histoclear II 30 mins Y 9 Histoclear II 30 mins Y 10 Wax 30 mins Y 11 Wax 45 mins Y 12 Wax 45 mins Y This programme can be run during the day as long as it is started early enough; if not, a delay must be set in order that the samples are not left in hot wax for extended times - refer to instrument manual. Good luck Margaret Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Gill, Caula A. Sent: 20 January 2011 16:49 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Animal tissue processing Hi All, I work in a hospital where we process human tissue. As a favor to a friend the pathologist would like us to process animal tissue. My questions are could we process the animal tissue on the same processor with the human tissue? And Are there different processing times and reagents for animal tissue? Thanks for any help you can give Caula (HT) ___ 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: Static issues
I live in moist old UK and have always breathed on my ribbons, can't get out of the habit, but do remember to breath and not too heavily! On the cryostat front, I used to suffer from static terribly and tried all sorts of things, but since using disposable blades have had fewer problems. I just move the blade along or replace it and it often cures the problem. Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: 06 January 2011 21:50 To: Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Static issues I was going to say the same thing... Just hadn't taken time. So may of us are full of hot air.. J:) -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Thursday, January 06, 2011 16:45 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Static issues This is New Mexico where humidity is a rumor. The humidity in the lab here - as I write - is 18% and that's on a really wet day! If I have static issues with my ribbons, I just lean a little bit toward the block and breathe on it and the ribbons just float (in a good way) off the knife. I do that so often that when I use my sewing machine, I find myself breathing on the material. That's just sad! But try the Breathing Thing. Or not. 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 Confidentiality Notice: This e-mail, 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 delete this message, and reply to the sender regarding the error in a separate email. ___ 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
