[Histonet] mouse testis in Bouins
Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] mouse testis in Bouins
Fix for a week or two, then rinse out most of the picrates with 50-70% ethanol. Fixation preserves cell morphology, it does not distort it. Geoff On 9/14/2012 9:06 AM, Margaret Horne wrote: Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] mouse testis in Bouins
Hi, I always fix them for 48h and move them to 70% ethanol. Regards Jeanne Jeanne Estabel, PhD Scientific Manager Histology Operations Manager Mouse Genetics Project Wellcome Trust Sanger Institute Cambridge, UK Tel:+44 (0)1223 834244 ext 8306 Find Sanger Mouse Genetics Project phenotyping data on http://www.sanger.ac.uk/mouseportal/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margaret Horne Sent: 14 September 2012 14:07 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] mouse testis in Bouins Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret -- The Wellcome Trust Sanger Institute is operated by Genome Research Limited, a charity registered in England with number 1021457 and a company registered in England with number 2742969, whose registered office is 215 Euston Road, London, NW1 2BE. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] mouse testis in Bouins- more info
Thanks for the incoming info; it's a big help. Sorry I need a little bit more detail so to explain a little more: Some of the testis are showing gaps between the tubules, more towards the centre of the testis and the researcher though it might be the protocol but we saw the gap in subsequent testis when we were cutting them in half after being fixed in Bouins for 24 hours-48hrs. Is this normal? Half the testis was processed but the other half was left in Bouins in case he wants to use it for something somewhere down the line. Some have been there a month- too long or ok? 70% EtOH better for long term ( months to years) storage? Thanks everyone, Margaret Margaret Horne mho...@upei.ca 14/09/2012 10:06 AM Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] mouse testis in Bouins- more info
Fixation is not long enough and/or the piece of tissue is too large, that is why you see problems at the center. The connective tissue between the tubules is delicate and the normal shrinkage due to paraffin processing may show some gaps Fix longer, 24-48 hours is not sufficient. A month in Bouin's is probably better than 1-2 days. Long term months-years in rarely a good idea, why not just embed the tissue? Geoff On 9/14/2012 9:58 AM, Margaret Horne wrote: Thanks for the incoming info; it's a big help. Sorry I need a little bit more detail so to explain a little more: Some of the testis are showing gaps between the tubules, more towards the centre of the testis and the researcher though it might be the protocol but we saw the gap in subsequent testis when we were cutting them in half after being fixed in Bouins for 24 hours-48hrs. Is this normal? Half the testis was processed but the other half was left in Bouins in case he wants to use it for something somewhere down the line. Some have been there a month- too long or ok? 70% EtOH better for long term ( months to years) storage? Thanks everyone, Margaret Margaret Horne mho...@upei.ca 14/09/2012 10:06 AM Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] cell membrane
Patsy Not sure if this will work, but I remember a project we did on fixed vibratome sections of mouse liver. We did a bunch of IF markers I think we used Phalloidin and that seemed to outline the liver cells nicely. We used it already conjugated. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Thursday, September 13, 2012 12:06 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cell membrane Dear Histochemist, Does anyone know of a stain or histochemical reaction which will exclusively stain cell membrane and no other parts of the cell or other tissue components? Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email mailto:pru...@ihctech.net pru...@ihctech.net email mailto:pru...@flagshipbio.com pru...@flagshipbio.com web site http://www.ihctech.net www.ihctech.net ___ 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] Re: mouse testis in Bouins
Haven't seen mouse balls in years. My laptop has a trackpad. But seriously, folks, I concur - excessive time in Bouin's fixative will harden the tissue - move it to 70% alcohol. I've used Davidson's fixative (3 parts water, 3 parts alcohol, 2 parts 37% formaldehyde, 1 part acetic acid) with human testis as an alternative to Bouin's fixative. Bob Richmond Samurai Pathologist Maryville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: mouse testis in Bouins
Hi Margaret, Our protocol for whole mouse embryos E14.5-E18.5 was to fix in Bouin's for 5-7 days at room temp (I have gone longer, but it isn't exactly recommended). Most of the length of time, however, was to compensate for the large tissue size and need for good penetration. I'm not sure how that converts to your particular tissue of interest. For long term storage, John Shelton at UT Southwestern (who did our vacuum processing for large embryos) told me that it was preferred to put them in 1% neutral buffered formalin and store them at room temp. We had previously been storing them in 70% EtOH, but John said that the long exposure to EtOH leads to excessive drying of the tissue and ultimately brittleness if used later. I'm assuming this thought could be applied to any tissue piece, but I don't have enough experience to really know. We have successfully gotten beautiful paraffin sections from 3mo-1year samples that have been stored this way. I'm hoping this will be of some help to you, and perhaps others in the list can comment. Best of luck, ~Francie *** Francie Barron, Ph.D. Postdoctoral Fellow, Joseph Wu Lab Stanford University School of Medicine Lorry I. Lokey Stem Cell Research Building 265 Campus Drive, Room G1105 Stanford, CA 94305-5454 Phone: (650) 724-5564 or (650) 724-9240 Fax: (650) 736-0234 *** Message: 7 Date: Fri, 14 Sep 2012 10:06:33 -0300 From: Margaret Horne mho...@upei.ca Subject: [Histonet] mouse testis in Bouins To: histonet@lists.utsouthwestern.edu Message-ID: 505301a902d100018...@oes-grpwise.novell.upei.ca Content-Type: text/plain; charset=us-ascii Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: m endothelial cell in FFPE
I recently tested a mCD34 from GeneTex cat GTX28158 that gave very nice labeling of mouse endothelial cells on FFPE. Used EDTA pH 8 antigen retrieval and no amplification. For those of you using the Dianovo mCD31 antibody what are you using for antigen retrieval? And are you using amplification? I tried this antibody a year or two ago and was not very happy with the results. I have also used the VonWillebrand. It give very nice label of large established vessels but not the new or smaller vessels. Donna Reynolds HT(ASCP Chief Histology Tech, Cancer Biology IHC Research Lab M.D. Anderson Cancer Center Houston, TX 713-792-8106 Message: 1 Date: Thu, 13 Sep 2012 13:05:37 -0600 From: Patsy Ruegg pru...@ihctech.net Subject: RE: [Histonet] endothelial cell marker To: 'Amos Brooks' amosbro...@gmail.com, histonet@lists.utsouthwestern.edu, joost.bruijnt...@tno.triskelion.nl Message-ID: D9F8AB2BD9D14EF783339ABC4BD9A0CF@DESKTOP3 Content-Type: text/plain; charset=us-ascii By far the best ab for endothelial cells in mouse tissue is rat anti CD31 from Dianovo, it picks up the really early forming vessels better than any other cd31 or F8 or SMA I have ever used. We like it in AP/red. Regards, Patsy Hi, My favorite by far is VonWillebrandt (spelling?) factor VIII. Dako has a really good one (Cat# A0082) that works great in mouse tissue. CD31 is OK but it is really finicky. CD34 works but F8 is easier and more reliable. Amos On Wed, Sep 5, 2012 at 1:01 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 5 Date: Wed, 5 Sep 2012 10:04:58 + From: Bruijntjes, J.P. (Joost) joost.bruijnt...@tno.triskelion.nl Subject: [Histonet] endothelial cell marker To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu Message-ID: a3a9d8b3f110d44fa1022b49913668668835b...@exc-mbx03.tsn.tno.nl Content-Type: text/plain; charset=us-ascii Hi all Is anyone of you familiar with an antibody directed against endothelial cells which can be applied on FFPE mouse tissues? Best regards Joost Bruijntjes ___ 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: air drying special stain slides rather than
Yes Rene :) US Virgin Islands is still going as green as we can! Thank you for this gift! We have been de-waxing slides off line for over 14 months now ;TJC and CMS inspections 100% success during this transition. I was able to report to our PI department that we have experienced a cost reduction from $83.01/100 slides down to 4 pennies! My tech's are breathing in much deeper :) and excited to continue becoming as green as possible! Best Regards- Michelle Moore Histopathology Supervisor/Medical Examiner Ofc Schneider Regional Medical Center St. Thomas, USVI, 00802 From: Rene J Buesa rjbu...@yahoo.com To: E. Wayne Johnson e...@pigsqq.org Cc: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu; Mayer, Toysha N tnma...@mdanderson.org Sent: Wednesday, September 12, 2012 10:13 AM Subject: Re: [Histonet] RE: air drying special stain slides rather than EWayne (et al): So, there you have it! He (or she) who still uses xylene in the histology lab is just because he (or she) has decided to do so! At this moment what you describe is standard procedure for several private labs in the US and the US Virgin Islands, Canada, Russia and Spain. Besides dewaxing with dishwasher soap and air drying before cover-slipping, you can also eliminate xylene from tissue processing by just following the instructions outlinedin the articles I sent. you. Try to contact as many colleagues as you can and spread the word: xylene is out of our lives, as long as we want to. Thank you for the information René J. From: E. Wayne Johnson e...@pigsqq.org To: Rene J Buesa rjbu...@yahoo.com Cc: Mayer,Toysha N tnma...@mdanderson.org; 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Sent: Wednesday, September 12, 2012 9:49 AM Subject: Re: [Histonet] RE: air drying special stain slides rather than Dishwashing machines are not at all common in China even in Beijing so we could not find dishwasher detergent nearby. But we took a bus and subway ride toward the city center to Zhongguancun (the computer district) and found a Carrefour's (Jia Le Fu 家乐福) that had one brand of powdered detergent Finish (Reckett Benckiser). Finish was formerly called Electrasol. Actually I was a bit afraid of Finish. If I had known it was the same thing as the familiar Electrasol, I would not have had any concerns. Anyway, we took the Finish powder back to the lab where we had a covered water bath waiting at 90C. I mixed 40 grams in with 2L of tap water and we followed the protocol Rene' sent with some test tissues from some pigs we examined a few days ago (lung, liver, heart, spleen, kidney, gut). We heated the detergent solution on an induction stove and poured in into some square glass jars in the water bath. The procedure took the paraffin right off. We did an HE and dried the slide in the 60C oven after a water wash to clean up after Eosin. Ver-r-ry nice result. Jane tried the technique then by herself with 3 more slides including one slide with some honking big pieces of pig cerebellum. The sections all stayed put on the slides. Sometimes we can lose most of the cerebellum in processing, so we think it is a good demonstration that section loss is not going to be much of a problem. The stain was a Harris hematoxylin regressed with 1% HCl. We blued some with tap water and some with Scott's. I sort of prefer the plain tap water bluing. Our Eosin is an Eosin/Biebrich Scarlet (Acid Red 66)/Orange G that we make up. We usually use a treatment in alcoholic Picric Acid to get rid of formalin pigment but we omitted that step today and the slides turned out well. Indeed these pigs were a field necropsy and the formalin was simple 10% unbuffered formalin in plain local farm tap water, but there were only some traces of formalin pigment. We are wondering if perhaps the detergent is taking out some of the formalin pigment for us. We got a few white paraffin spots on one slide but even that would not be an issue in reading or photographing the slide. Jane thinks she can tweak the procedure to eliminate the paraffin spots. Jane's opinion on the procedure? She will be bottling up all of the xylenes and alcohols and storing them away first thing tomorrow morning. This fixes a big problem for us because the histolab is on the first floor along with some offices. We do our work under a fume hood and we are careful, but we had an incident where students left containers of xylene uncapped outside the hood overnight in hot weather vaporizing a large amount of xylene into the hallway. Not cool. We moved the tissue processor and autostainer to a remoter spot on the 4th floor but the water quality issues there made our autostainer a problem. We can now bring our autostainer back and set it up for special and routine stains. The procedure with detergent from beginning to end is significantly shorter than the xylene
[Histonet] Position North of Boston, MA-Histotechs
Hello and Happy Friday, I have two positions available just North of Boston, MA for full time/permanent bench level job openings. Please message for details. Thank you, To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php Melissa Phelan LinkedIn: http://www.linkedin.com/in/melissaphelan President, Laboratory Staffing Allied Search Partners P: 888.388.7571 F: 888.388.7572 M: 407.697.1175 www.alliedsearchpartners.com http://www.alliedsearchpartners.com/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] cell membrane
Liz, It is possible to play with different catenins (gamma etc.) to get rid of nuclear staining. But if the target is epithelium I would try to use EpCAM first. And in case of different epithelial tumors EpCAM usually is up-regulated. Anatoli Gleiberman, PhD Director of Histopathology Cleveland Biolabs, Inc 73 High Street Buffalo, NY 14203 phone:716-849-6810 ext.354 fax:716-849-6817 e-mail: agleiber...@cbiolabs.com -Original Message- From: Elizabeth Chlipala [mailto:l...@premierlab.com] Sent: Friday, September 14, 2012 4:08 PM To: Anatoli Gleiberman; pru...@ihctech.net Subject: RE: [Histonet] cell membrane Thanks Anatoli We have worked with beta-catenin before (4 different sources of antibody) and in the tissue that we looked at (tumor) we saw a combination of membrane, cytoplasmic and nuclear staining. I went back to some of the images from the phalloidin staining and cell membrane staining was variable, some cells were outlined nicely others were not. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: Anatoli Gleiberman [mailto:agleiber...@cbiolabs.com] Sent: Friday, September 14, 2012 12:41 PM To: Elizabeth Chlipala; pru...@ihctech.net Subject: RE: [Histonet] cell membrane Liz, Patsy, Phalloidin will definitely stain under membrane actin in liver cells - but for all other cell types it will stain all intra-cytoplasmic actin stress fibrils and cytoplasmic actin network. For many epithelial cells with exclusion of keratinocytes and hepatocytes the best membrane marker is EpCAM. There are rat anti-mouse EpCAM antibody and mouse anti-human EpCAM antibody commercially available. It is possible to use anti-beta-catenin antibody for most of epithelia. So, combination of EpCAM and beta-catenin will stain on sections cell membranes of all epithelia. However, there are no satisfactory staining for cell membranes of connective tissue and smooth muscle cells and I am not sure about neurons and skeletal muscle cells. I am afraid, there are no such things as one reagent staining exclusively cell membranes of all cell types. Anatoli Gleiberman, PhD Director of Histopathology Cleveland Biolabs, Inc 73 High Street Buffalo, NY 14203 phone:716-849-6810 ext.354 fax:716-849-6817 e-mail: agleiber...@cbiolabs.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Friday, September 14, 2012 10:43 AM To: 'Patsy Ruegg'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cell membrane Patsy Not sure if this will work, but I remember a project we did on fixed vibratome sections of mouse liver. We did a bunch of IF markers I think we used Phalloidin and that seemed to outline the liver cells nicely. We used it already conjugated. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Thursday, September 13, 2012 12:06 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cell membrane Dear Histochemist, Does anyone know of a stain or histochemical reaction which will exclusively stain cell membrane and no other parts of the cell or other tissue components? Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC Director of Histology and IHC IHCtech a subsidiary of Flagship Bio-Sciences, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80045 P-720-859-4060 F-720-859-4110 email mailto:pru...@ihctech.net pru...@ihctech.net email mailto:pru...@flagshipbio.com pru...@flagshipbio.com web site http://www.ihctech.net www.ihctech.net ___ 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: mouse testis in Bouins
As a GLP tox lab, we have done away with using Bouin's altogether - there is literature out there (somewhere - not handy now) that indicates Modified Davidson's fixative provides the same testicular detail of bouins, without the picric acid danger. We switched about 3-4 years ago, and our testicle experts are happy. I believe most labs are getting away from Bouins. Jackie O' -Original Message- From: Frances Elizabeth Barron fbar...@stanford.edu To: histonet histonet@lists.utsouthwestern.edu Sent: Fri, Sep 14, 2012 12:21 pm Subject: [Histonet] RE: mouse testis in Bouins Hi Margaret, Our protocol for whole mouse embryos E14.5-E18.5 was to fix in Bouin's for 5-7 days at room temp (I have gone longer, but it isn't exactly recommended). Most of the length of time, however, was to compensate for the large tissue size and need for good penetration. I'm not sure how that converts to your particular tissue of interest. For long term storage, John Shelton at UT Southwestern (who did our vacuum processing for large embryos) told me that it was preferred to put them in 1% neutral buffered formalin and store them at room temp. We had previously been storing them in 70% EtOH, but John said that the long exposure to EtOH leads to excessive drying of the tissue and ultimately brittleness if used later. I'm assuming this thought could be applied to any tissue piece, but I don't have enough experience to really know. We have successfully gotten beautiful paraffin sections from 3mo-1year samples that have been stored this way. I'm hoping this will be of some help to you, and perhaps others in the list can comment. Best of luck, ~Francie *** Francie Barron, Ph.D. Postdoctoral Fellow, Joseph Wu Lab Stanford University School of Medicine Lorry I. Lokey Stem Cell Research Building 265 Campus Drive, Room G1105 Stanford, CA 94305-5454 Phone: (650) 724-5564 or (650) 724-9240 Fax: (650) 736-0234 *** Message: 7 Date: Fri, 14 Sep 2012 10:06:33 -0300 From: Margaret Horne mho...@upei.ca Subject: [Histonet] mouse testis in Bouins To: histonet@lists.utsouthwestern.edu Message-ID: 505301a902d100018...@oes-grpwise.novell.upei.ca Content-Type: text/plain; charset=us-ascii Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ 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: mouse testis in Bouins
What danger of Picric Acid are you concerned with? Surely its not the hyped explosion hazards. We use picric acid and as inquisitive boys we have tried very hard to ignite it thinking it would be fun. We dried some down and wrapped it in aluminum foil and with appropriate protection outdoors beat it with a hammer. So very disappointing. We only made it flat. We tried heating some. It does burn pretty good but not really dramatically. We tried purifying and recrystallizing it and it still didnt do anything spectacular. Our conclusion that as fireworks, pure picric acid is pretty much a dud. I have done some reading about picric acid and it seems that in lab conditions a picric acid explosion is very unlikely maybe impossible even if the stuff is very dry indeed. We do keep our picric acid wet in a safe spot for storage. Some metal salts of picric acid are said to be much more sensitive. We havent made any lead picrate to play with since we are worried about aerosolizing the lead when it does explode or flash. There are some youtube movies about how to make explosive derivatives of picric acid. it seems that picric acid is just not a very good explosive, and that small amounts in free open air are unlikely to explode. I have been unable to find any reference to any lab accidents with picric acid. Does anyone have any information to the contrary? On 9/15/2012 7:55 AM, Jackie O'Connor wrote: As a GLP tox lab, we have done away with using Bouin's altogether - there is literature out there (somewhere - not handy now) that indicates Modified Davidson's fixative provides the same testicular detail of bouins, without the picric acid danger. We switched about 3-4 years ago, and our testicle experts are happy. I believe most labs are getting away from Bouins. Jackie O' -Original Message- From: Frances Elizabeth Barronfbar...@stanford.edu To: histonethistonet@lists.utsouthwestern.edu Sent: Fri, Sep 14, 2012 12:21 pm Subject: [Histonet] RE: mouse testis in Bouins Hi Margaret, Our protocol for whole mouse embryos E14.5-E18.5 was to fix in Bouin's for 5-7 days at room temp (I have gone longer, but it isn't exactly recommended). Most of the length of time, however, was to compensate for the large tissue size and need for good penetration. I'm not sure how that converts to your particular tissue of interest. For long term storage, John Shelton at UT Southwestern (who did our vacuum processing for large embryos) told me that it was preferred to put them in 1% neutral buffered formalin and store them at room temp. We had previously been storing them in 70% EtOH, but John said that the long exposure to EtOH leads to excessive drying of the tissue and ultimately brittleness if used later. I'm assuming this thought could be applied to any tissue piece, but I don't have enough experience to really know. We have successfully gotten beautiful paraffin sections from 3mo-1year samples that have been stored this way. I'm hoping this will be of some help to you, and perhaps others in the list can comment. Best of luck, ~Francie *** Francie Barron, Ph.D. Postdoctoral Fellow, Joseph Wu Lab Stanford University School of Medicine Lorry I. Lokey Stem Cell Research Building 265 Campus Drive, Room G1105 Stanford, CA 94305-5454 Phone: (650) 724-5564 or (650) 724-9240 Fax: (650) 736-0234 *** Message: 7 Date: Fri, 14 Sep 2012 10:06:33 -0300 From: Margaret Hornemho...@upei.ca Subject: [Histonet] mouse testis in Bouins To:histonet@lists.utsouthwestern.edu Message-ID:505301a902d100018...@oes-grpwise.novell.upei.ca Content-Type: text/plain; charset=us-ascii Hello Everyone, I am asking this for a friend. How long can mouse testis be kept in Bouins without distortion of cell morphology? Days? weeks? months? years? I noticed in the Archives that many people fix in Bouins , rinse, then store in 70% EtOH. This is preferable I assume. Again, how long is ok? Thanks in advance for the sharing of your accumulated wisdom, Margaret ___ 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
