Re: [Histonet] Aqueous mounting media
The un-coupled Naphthol salts (from the phosphatase reaction, dissolved in the lipids of the tissue) degrade and cause N2 bubbles . We solved this problem (in the 60ties!)with a post treatment after the reaction: After the reaction, rinse in dist water, post fix in formalin 1:4 parts water (just dist. water) for 15 min. (re-usable), Rinse in dist water and post-treat in a solution of 1% Sulfanilic acid in 10% Acetic acid (re-usable) for 15 min., rinse in dist water 4x and mount. (we used at that time glycerin jelly and/or Aquamount) Regards, Markus F. Meyenhofer Microscopy Labs Box 338 Red Bank, NJ 07701 732 747 6228 mi...@superlink.net - Original Message - From: "Kevin Bennett" To: Sent: Tuesday, August 16, 2011 2:48 PM Subject: [Histonet] Aqueous mounting media All, Can anyone recommend a quality aqueous mounting media? I was using Aquatex from EMD chemicals but it's been discontinued, tried Aqua-Mount from Thermo but show air bubbles a few days after coverslipping. Thanks, Kevin Bennett 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] Aqueous mounting media
Sorry let me send that again with a link *http://tinyurl.com/3uw5lcm *I have no idea why we use this particular medium. * * A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron On Tue, Aug 16, 2011 at 9:40 PM, Emily Sours wrote: > We use Gel Mount > > > A great book should leave you with many experiences, and slightly > exhausted. You should live several lives while reading it. > -William Styron > > > > On Tue, Aug 16, 2011 at 2:48 PM, Kevin Bennett wrote: > >> All, >> Can anyone recommend a quality aqueous mounting media? I was using Aquatex >> from EMD chemicals but it's been discontinued, tried Aqua-Mount from >> Thermo >> but show air bubbles a few days after coverslipping. >> >> Thanks, >> Kevin Bennett 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] Aqueous mounting media
We use Gel Mount A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron On Tue, Aug 16, 2011 at 2:48 PM, Kevin Bennett wrote: > All, > Can anyone recommend a quality aqueous mounting media? I was using Aquatex > from EMD chemicals but it's been discontinued, tried Aqua-Mount from Thermo > but show air bubbles a few days after coverslipping. > > Thanks, > Kevin Bennett 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: H&E on Frozen
I concur on this. If my boss will let me, I can send you our elaborate protocol for H&E staining, It involves many dishes and many washes. But your H&E will be perfect. I don't have the protocol with me right now,as I'm at home but tomorrow I will ask about sending you the protocol. I don't see why we couldn't send you ours, but I'm going to ask just in case. A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron On Tue, Aug 16, 2011 at 2:53 PM, Rathborne, Toni < trathbo...@somerset-healthcare.com> wrote: > Our protocol is to: > > Use a positively charged slide > Fix in 10% formalin for 1 minute > Rinse in tap (a few dips) > Gill 3 for 1 minute > Rinse in tap (a few dips) > 3-5 dips in bluing > Rinse in tap (a few dips) > Eosin 2-3 dips > 100% ETOH (a few dips) > 100% ETOH (a few dips) > Xylene (dip until clear) > Mount with Richard- Allan Mounting Medium > > After the Gill3, Eosin, and last alcohol, the back of the slide is wiped > clean of excess solution to prevent carryover. > > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu [mailto: > histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Reynolds,Donna M > Sent: Tuesday, August 16, 2011 10:13 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] H&E on Frozen > > We do this frequently. > Fix slides in Methanol (45ml) + formaldehyde (5ml) for 10 minutes. > Wash several changes of tap water > Stain with Harris Hematoxylin for 3 minutes Proceed as you would for > paraffin sections Get great staining. > I am sure if you are using a different Hematoxylin you could use it the > same way you use it for paraffin > > Donna Reynolds HT(ASCP) > Chief Histology Technician, Core Immuno Lab U. T. M.D. Anderson Cancer > Center, Houston, TX > 713-792-8106 > e-mail dreyn...@mdanderson.org > > Message: 2 > Date: Fri, 12 Aug 2011 08:52:11 -0700 (PDT) > From: Jennifer Sipes > Subject: [Histonet] H&E for Frozen Tissue > To: "histonet@lists.utsouthwestern.edu" > > Message-ID: ><1313164331.88113.yahoomail...@web125406.mail.ne1.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Does anyone have a good protocol for staining frozen sections with H&E?? > Also, any mounting medium suggestions?? This is the first time I've been > asked to do this for frozen. > ? > Thanks a bunch everyone! > Jen > ? > Jennifer K. Sipes, ALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 933 > 720 Rutland Avenue > Baltimore, MD? 21205 > phone: 410-614-0131 > fax: 410-955-9677 > cell: 443-631-6361 > e-mail:? jsip...@jhmi.edu > > -- > > ** > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > CONFIDENTIALITY NOTICE > This message and any included attachments are from Somerset Medical Center > and are intended only for the addressee. The information contained in this > message is confidential and may contain privileged, confidential, > proprietary and/or trade secret information entitled to protection and/or > exemption from disclosure under applicable law. Unauthorized forwarding, > printing, copying, distribution, or use of such information is strictly > prohibited and may be unlawful. If you are not the addressee, please > promptly delete this message and notify the sender of the delivery error > by e-mail or you may call Somerset Medical Center's computer Help Desk > at 908-685-2200, ext. 4050. > > Be sure to visit Somerset Medical Center's Web site - > www.somersetmedicalcenter.com - for the most up-to-date news, > event listings, health information and more. > > ___ > 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] Art Challenge
Hey All, I have been working on updating the look of our website and booth for NSH. The graphic design people I am working with don't seem to be getting the look I am going for. This gave me an idea. I would like to challenge all of the crafty Histoneters to draw, paint or Photoshop (or whatever your preferred medium is) a cartoon, painting or caricature of histology equipment. The winner will receive a gift and my unending gratitude. Thanks -- Matthew Mincer Tech One Biomedical Services 159 N Marion Street, PMB163 Oak Park, IL 60301 (708) 383-6040 X 10 fax (708) 383-6045 cell (708) 822-3738 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Aqueous mounting media
Probe mount aqueous based mounting media from Innovex. My personal favorite!! Loralei Dewe On Tue, Aug 16, 2011 at 12:05 PM, Montina Van Meter < montina.vanme...@pbrc.edu> wrote: > ProLong Gold (Invitrogen) > > > > > > > Montina J. Van Meter, HT (ASCP) > Lab Manager > Dept. of Autonomic Neuroscience > Pennington Biomedical Research Center > 6400 Perkins Rd. > Baton Rouge, LA 70791 > > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kevin > Bennett > Sent: Tuesday, August 16, 2011 1:49 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Aqueous mounting media > > All, > Can anyone recommend a quality aqueous mounting media? I was using > Aquatex from EMD chemicals but it's been discontinued, tried Aqua-Mount > from Thermo but show air bubbles a few days after coverslipping. > > Thanks, > Kevin Bennett 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 > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Aqueous mounting media
ProLong Gold (Invitrogen) Montina J. Van Meter, HT (ASCP) Lab Manager Dept. of Autonomic Neuroscience Pennington Biomedical Research Center 6400 Perkins Rd. Baton Rouge, LA 70791 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kevin Bennett Sent: Tuesday, August 16, 2011 1:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Aqueous mounting media All, Can anyone recommend a quality aqueous mounting media? I was using Aquatex from EMD chemicals but it's been discontinued, tried Aqua-Mount from Thermo but show air bubbles a few days after coverslipping. Thanks, Kevin Bennett 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] Aqueous mounting media
We have been using an aqueous mounting media from Diagnostic Biosystems. http://dbiosys.com/ 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 Kevin Bennett [bennett...@gmail.com] Sent: Tuesday, August 16, 2011 2:48 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Aqueous mounting media All, Can anyone recommend a quality aqueous mounting media? I was using Aquatex from EMD chemicals but it's been discontinued, tried Aqua-Mount from Thermo but show air bubbles a few days after coverslipping. Thanks, Kevin Bennett 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
[Histonet] RE: H&E on Frozen
Our protocol is to: Use a positively charged slide Fix in 10% formalin for 1 minute Rinse in tap (a few dips) Gill 3 for 1 minute Rinse in tap (a few dips) 3-5 dips in bluing Rinse in tap (a few dips) Eosin 2-3 dips 100% ETOH (a few dips) 100% ETOH (a few dips) Xylene (dip until clear) Mount with Richard- Allan Mounting Medium After the Gill3, Eosin, and last alcohol, the back of the slide is wiped clean of excess solution to prevent carryover. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Reynolds,Donna M Sent: Tuesday, August 16, 2011 10:13 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] H&E on Frozen We do this frequently. Fix slides in Methanol (45ml) + formaldehyde (5ml) for 10 minutes. Wash several changes of tap water Stain with Harris Hematoxylin for 3 minutes Proceed as you would for paraffin sections Get great staining. I am sure if you are using a different Hematoxylin you could use it the same way you use it for paraffin Donna Reynolds HT(ASCP) Chief Histology Technician, Core Immuno Lab U. T. M.D. Anderson Cancer Center, Houston, TX 713-792-8106 e-mail dreyn...@mdanderson.org Message: 2 Date: Fri, 12 Aug 2011 08:52:11 -0700 (PDT) From: Jennifer Sipes Subject: [Histonet] H&E for Frozen Tissue To: "histonet@lists.utsouthwestern.edu" Message-ID: <1313164331.88113.yahoomail...@web125406.mail.ne1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone have a good protocol for staining frozen sections with H&E?? Also, any mounting medium suggestions?? This is the first time I've been asked to do this for frozen. ? Thanks a bunch everyone! Jen ? Jennifer K. Sipes, ALAT Sr. Laboratory Technician Johns Hopkins University Ross 933 720 Rutland Avenue Baltimore, MD? 21205 phone: 410-614-0131 fax: 410-955-9677 cell: 443-631-6361 e-mail:? jsip...@jhmi.edu -- ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Aqueous mounting media
All, Can anyone recommend a quality aqueous mounting media? I was using Aquatex from EMD chemicals but it's been discontinued, tried Aqua-Mount from Thermo but show air bubbles a few days after coverslipping. Thanks, Kevin Bennett HT ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Details about Masson Trichrome Stain !
http://stainsfile.info/StainsFile/dyes/26905.htm This says 66... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of ? ? Sent: Tuesday, August 16, 2011 13:14 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Details about Masson Trichrome Stain ! Details about Masson Trichrome Stain ! Biebrich Scarlet: Biebrich scarlet 2.7 gm Acid fuchsin 0.3 gm Distilled water 300.0 ml Glacial acetic acid 3.0 ml I am not sure what the Biebrich Scarlet here is ? Acid Red 66 or acid Red 73 ? Acid Red 73 http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=49823|FLUKA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC Acid Red 66 http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=B6008|SIGMA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC What is the main difference between these two? May I choose 66, 73 or anyone? Thanks ! Wang Lian, MD Medical School of Nanjing University Hankou Road 22# Nanjing, Jiangsu Province, China e-mail: wangliane...@yahoo.com.cn ___ 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] Scorched tissue
Hi, I am wondering if anyone can help me out with some processing issues that our lab has been having. We recently have been having reagent contamination issues with alcohol and xylene, where our tissues are soft and cloudy after processing. It seems that the reagent valves on the processor were not operating properly. We changed all reagents after each processing run (we were waiting for replacement parts and needed to continue processing) this seemed to work but then we noticed an additional artifact. The outer cell layer (all tissue types) appears scorched, this does not appear throughout the tissue only on the outer edge. I am wondering if anyone else has seen this artifact and how they resolved the issue? Any comments or advice are greatly appreciated! Tammy ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Details about Masson Trichrome Stain !
Details about Masson Trichrome Stain ! Biebrich Scarlet: Biebrich scarlet 2.7 gm Acid fuchsin 0.3 gm Distilled water 300.0 ml Glacial acetic acid 3.0 ml I am not sure what the Biebrich Scarlet here is ? Acid Red 66 or acid Red 73 ? Acid Red 73 http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=49823|FLUKA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC Acid Red 66 http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=B6008|SIGMA&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC What is the main difference between these two? May I choose 66, 73 or anyone? Thanks ! Wang Lian, MD Medical School of Nanjing University Hankou Road 22# Nanjing, Jiangsu Province, China e-mail: wangliane...@yahoo.com.cn ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] use of isopentane substitutes for tissue freezing
Hello all, I am considering purchasing a freezing unit through Thermo that can be used with either isopentane of a new solution, HFE-7000 that is sold with the unit. I need to be able to have frozen samples that allow high quality DNA and RNA extracted. Does anyone have any experience with this fluid and molecular analytes? Thanks very much, Nancy W Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital rm 3115 E&R Bldg 2799 W Grand Blvd Detroit, MI 48202 (313) 916-8648 phone (313) 916-9855 fax == 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] Toxoplasma Control
Hello everyone, Thanks for the response regarding the Toxoplasma control. It's nice to have such a supportive network available for these types of needs. Margaret Margaret G. Coppin, HT(ASCP) Technical Supervisor--Immunohistochemistry ARUP Laboratories 500 Chipeta Way Salt Lake City, UT 84108 (801)583-2787 X3869 copp...@aruplab.com - -- The information transmitted by this e-mail and any included attachments are from ARUP Laboratories and are intended only for the recipient. The information contained in this message is confidential and may constitute inside or non-public information under international, federal, or state securities laws, or protected health information and is intended only for the use of the recipient. Unauthorized forwarding, printing, copying, distributing, or use of such information is strictly prohibited and may be unlawful. If you are not the intended recipient, please promptly delete this e-mail and notify the sender of the delivery error or you may call ARUP Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 (800) 522-2787 ext. 2100 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Peloris Rapid tissue processor
Dear Colleagues: I have just received the following e-mail from Leica Microsystems. It is evident that I was wrong and that now the tissues in the Peloris instrument go directly from the 2-propanol to the parafin wax and are not air dried. I just wanted to share this correction with you. René J. --- On Mon, 8/15/11, mary.che...@leica-microsystems.com wrote: From: mary.che...@leica-microsystems.com Subject: Re: [Histonet] Peloris Rapid tissue processor To: "Rene J Buesa" Date: Monday, August 15, 2011, 7:03 PM Dear Rene, I read your statement on Peloris today. It is certainly not our intent to try to censor the information placed on Histonet. However, I would like to point out that there is some inaccuracy in your assessment. The inaccuracy is in the statement that tissues are subjected to "DRY HOT evaporation.". This is simply not true. Within the xylene-free protocols of the instrument, isopropyl does act as a clearant instead of xylene. Therefore, tissue transitions from isopropyl directly into paraffin. The temperature of the 1st two paraffins are elevated to "evaporate" the isopropyl but the tissue is physically in paraffin undergoing infiltration during this simultaneous process. If you have any questions, we would be happy to answer them. I am hopeful that you will see fit to write in to Histonet with a clarifying statement. We would be happy to send you white papers on the Peloris technology if you are interested in receiving them. Best Regards, Mary Cheles, MPH, HTL, DLM (ASCP) Director, Technical Services & Education Programs Leica Microsystems Biosystems Division 1700 Leider Lane Buffalo Grove, IL 60089 847.317.5916 Office 847.236.3058 Fax www.leica-microsystems.com Rene J Buesa To Sent by: histonet@lists.utsouthwestern.edu, histonet-bounces@ tahs...@brain.net.pk lists.utsouthwest cc ern.edu histol...@skm.org.pk Subject Re: [Histonet] Peloris Rapid tissue 08/13/2011 09:19 processor AM Muhammad: The only thing I dislike about the Peloris technology (it is a technology in itself) is that after the dehydration with 2-propanol, the tissues are subjected to a DRY HOT evaporation of the 2-propanol in vacuum before the infiltration step with melted paraffin. That step of drying out the tissues to eliminate the 2-propanol to "facilitate" the infiltration is the one I do not like because the tissues are exposed to a very high gradient. If you end buying the instrument I think you should run a large series of validation tests to find out if the results you obtain with the Peloris compare with what you are used to, not referring to the sectioning quality of the blocks, but to their microscopic appearance.. Peloris was developed in Australia by VisionBioSystems and later bought by Leica Microsystems. Although there were tests published by VisionBioSystems, all referred to animal tissues, and the instrument was never independently validated. Leica did not make known (published) independent validations. I for one would never subject the tissues to a hot dry desiccation before infiltration. I hope this will help you in your decision. René J. --- On Sat, 8/13/11, tahs...@brain.net.pk wrote: From: tahs...@brain.net.pk Subject: [Histonet] Peloris Rapid tissue processor To: histonet@lists.utsouthwestern.edu Cc: histol...@skm.org.pk Date: Saturday, August 13, 2011, 6:18 AM Dear All, Our lab is in the process of purchasing our automated tissue processor (Peloris Rapid tissue processor). I would really appreciate comments from anyone who really likes, or dislikes the processor that they are using. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology <@t> skm.org.pk ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http
[Histonet] H&E on Frozen
We do this frequently. Fix slides in Methanol (45ml) + formaldehyde (5ml) for 10 minutes. Wash several changes of tap water Stain with Harris Hematoxylin for 3 minutes Proceed as you would for paraffin sections Get great staining. I am sure if you are using a different Hematoxylin you could use it the same way you use it for paraffin Donna Reynolds HT(ASCP) Chief Histology Technician, Core Immuno Lab U. T. M.D. Anderson Cancer Center, Houston, TX 713-792-8106 e-mail dreyn...@mdanderson.org Message: 2 Date: Fri, 12 Aug 2011 08:52:11 -0700 (PDT) From: Jennifer Sipes Subject: [Histonet] H&E for Frozen Tissue To: "histonet@lists.utsouthwestern.edu" Message-ID: <1313164331.88113.yahoomail...@web125406.mail.ne1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone have a good protocol for staining frozen sections with H&E?? Also, any mounting medium suggestions?? This is the first time I've been asked to do this for frozen. ? Thanks a bunch everyone! Jen ? Jennifer K. Sipes, ALAT Sr. Laboratory Technician Johns Hopkins University Ross 933 720 Rutland Avenue Baltimore, MD? 21205 phone: 410-614-0131 fax: 410-955-9677 cell: 443-631-6361 e-mail:? jsip...@jhmi.edu -- ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] protocol for in situ RT_PCR
I've used 10% zinc buffered formalin from Anatech (no alcohol) on mouse bones with good results. It's possible that the alcohol is denaturing your antigens of interest. Is there a reason you need to use alcoholic formalin? Adam On Tue, Aug 16, 2011 at 7:45 AM, Liang, Frank wrote: > Hello, > > Does anyone have used alcoholic zinc-formalin to fix mouse tissues? We > tried a few times with this fixative for immunohistochemical staining, it > did not work, but when switched back to 4% paraformaldehyde, the > immunostaining worked. We like alcoholic zinc-formalin as it gives better > morphology. Any suggestions? > > Thanks, > > Frank > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu [mailto: > histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Louise Renton > Sent: Tuesday, August 16, 2011 4:17 AM > To: Barone, Carol; Histonet > Subject: Re: [Histonet] protocol for in situ RT_PCR > > hi could you add me on too pelase? > > On Mon, Aug 15, 2011 at 11:09 PM, Barone, Carol > wrote: > > > Anyone got a hot protocol for In situ RT-PCR? Haven't done one in 5 > > years (on my Perk and Elmer)...Know there must be some great new > > reagents and kits out there...opinions? Send to cbar...@nemours.org. Thx > > CB > > ___ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > +27 11 717 2298 (tel & fax) > 073 5574456 (emergencies only) > "There are nights when the wolves are silent and only the moon howls". > George Carlin > No trees were killed in the sending of this message. > However, many electrons were terribly inconvenienced. > ___ > 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] IHC on Frozen Tissue
Jan, Thanks, I really appreciate the help. This is my first attempt, so we'll see how it turns out. :) Sheila -Original Message- From: Jan Shivers [mailto:shive...@umn.edu] Sent: Tuesday, August 16, 2011 9:21 AM To: Sheila Fonner Subject: Re: [Histonet] IHC on Frozen Tissue Sheila, On frozens, there should be no need to do tissue conditioning/pretreatment, since there are no fixation-caused aldehyde bonds to break. Bring your frozens to room temperature slowly, immerse in your rinse buffer for a few minutes, then proceed with your IHC staining protocol. With frozens, since the tissue can at times be more fragile, I always do the H2O2 step AFTER the primary antibody (but before the HRP-conjugated linking Ab; you don't want to quench your substrate enzyme) and I use 0.3% H2O2 instead of 3%. Jan Shivers IHC Section Head UMN Vet Diag Lab - Original Message - From: "Sheila Fonner" To: "'Richard Yeo'" ; Sent: Tuesday, August 16, 2011 7:24 AM Subject: RE: [Histonet] IHC on Frozen Tissue > Rich, > > > > Isn't the cell conditioning (retrieval) mostly done for tissues that have > been in formalin? If the tissue is fresh (frozen), it has not seen > formalin. That is the reason I questioned the cell conditioning step. > > > > Thank you for your help. > > Sheila > > > > > > > > From: Richard Yeo [mailto:r...@wchosp.org] > Sent: Tuesday, August 16, 2011 7:54 AM > To: Sheila Fonner > Subject: RE: [Histonet] IHC on Frozen Tissue > > > > Sheila, > > > > The cell conditioning step is for retrieval. It will depend on what > antibody > you are going to run. If you use it for Depar. you will also use it > wiith-out. > > > > > > Rich Y > > > > _ > > From: histonet-boun...@lists.utsouthwestern.edu on behalf of Sheila Fonner > Sent: Tue 8/16/2011 7:44 AM > To: 'Yahoo'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] IHC on Frozen Tissue > > Does anyone else leave out the cell conditioning step? > > > -Original Message- > From: Yahoo [mailto:kim.tourn...@yahoo.com] > Sent: Tuesday, August 16, 2011 7:41 AM > To: Sheila Fonner > Subject: Re: [Histonet] IHC on Frozen Tissue > > I would leave out the pretreatment step as well i.e. Enzyme, CC1/CC2. > > Sent from the iPhone of Kim Tournear > > On Aug 16, 2011, at 6:03 AM, "Sheila Fonner" wrote: > >> Thank you. >> >> >> -Original Message- >> From: Mierow, Brett T. [mailto:brett.mie...@essentiahealth.org] >> Sent: Tuesday, August 16, 2011 7:02 AM >> To: Sheila Fonner >> Subject: RE: [Histonet] IHC on Frozen Tissue >> >> You are correct, leave out the depar. step. >> >> >> Brett Mierow, HT (ASCP) >> Histology specialist >> Essentia Health >> SMDC Laboratory >> 407 East 3rd Street, Duluth, MN 54880 >> (218) 786-4510 | brett.mie...@essentiahealth.org >> >> -Original Message- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila >> Fonner >> Sent: Tuesday, August 16, 2011 5:41 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] IHC on Frozen Tissue >> >> Good morning everyone, >> >> >> >> I was wondering if anyone could help me with a protocol change for the >> Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, >> and Zoster Virus on frozen tissue that was received for IF studies. I >> know it's not Monday, but it is early, and I was hoping for some >> information about how to change the protocol for frozen vs. paraffin >> tissue. Do I just need to delete the depar. step and leave everything >> else the same? I appreciate any help you could give. >> >> >> >> Thanks, >> >> Sheila >> >> >> >> ___ >> 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] protocol for in situ RT_PCR
Hello, Does anyone have used alcoholic zinc-formalin to fix mouse tissues? We tried a few times with this fixative for immunohistochemical staining, it did not work, but when switched back to 4% paraformaldehyde, the immunostaining worked. We like alcoholic zinc-formalin as it gives better morphology. Any suggestions? Thanks, Frank -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Louise Renton Sent: Tuesday, August 16, 2011 4:17 AM To: Barone, Carol; Histonet Subject: Re: [Histonet] protocol for in situ RT_PCR hi could you add me on too pelase? On Mon, Aug 15, 2011 at 11:09 PM, Barone, Carol wrote: > Anyone got a hot protocol for In situ RT-PCR? Haven't done one in 5 > years (on my Perk and Elmer)...Know there must be some great new > reagents and kits out there...opinions? Send to cbar...@nemours.org. Thx > CB > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa +27 11 717 2298 (tel & fax) 073 5574456 (emergencies only) "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. ___ 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 on Frozen Tissue
Rich, Isn't the cell conditioning (retrieval) mostly done for tissues that have been in formalin? If the tissue is fresh (frozen), it has not seen formalin. That is the reason I questioned the cell conditioning step. Thank you for your help. Sheila From: Richard Yeo [mailto:r...@wchosp.org] Sent: Tuesday, August 16, 2011 7:54 AM To: Sheila Fonner Subject: RE: [Histonet] IHC on Frozen Tissue Sheila, The cell conditioning step is for retrieval. It will depend on what antibody you are going to run. If you use it for Depar. you will also use it wiith-out. Rich Y _ From: histonet-boun...@lists.utsouthwestern.edu on behalf of Sheila Fonner Sent: Tue 8/16/2011 7:44 AM To: 'Yahoo'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC on Frozen Tissue Does anyone else leave out the cell conditioning step? -Original Message- From: Yahoo [mailto:kim.tourn...@yahoo.com] Sent: Tuesday, August 16, 2011 7:41 AM To: Sheila Fonner Subject: Re: [Histonet] IHC on Frozen Tissue I would leave out the pretreatment step as well i.e. Enzyme, CC1/CC2. Sent from the iPhone of Kim Tournear On Aug 16, 2011, at 6:03 AM, "Sheila Fonner" wrote: > Thank you. > > > -Original Message- > From: Mierow, Brett T. [mailto:brett.mie...@essentiahealth.org] > Sent: Tuesday, August 16, 2011 7:02 AM > To: Sheila Fonner > Subject: RE: [Histonet] IHC on Frozen Tissue > > You are correct, leave out the depar. step. > > > Brett Mierow, HT (ASCP) > Histology specialist > Essentia Health > SMDC Laboratory > 407 East 3rd Street, Duluth, MN 54880 > (218) 786-4510 | brett.mie...@essentiahealth.org > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila > Fonner > Sent: Tuesday, August 16, 2011 5:41 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC on Frozen Tissue > > Good morning everyone, > > > > I was wondering if anyone could help me with a protocol change for the > Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, > and Zoster Virus on frozen tissue that was received for IF studies. I > know it's not Monday, but it is early, and I was hoping for some > information about how to change the protocol for frozen vs. paraffin > tissue. Do I just need to delete the depar. step and leave everything > else the same? I appreciate any help you could give. > > > > Thanks, > > Sheila > > > > ___ > 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] IHC on Frozen Tissue
I leave out the cell conditioning step as well. Sandra Sandra Esparza HT(ASCP)QIHC Lead Technologist Dell Children's Medical Center of Central Texas 512-324- x87061 sespa...@seton.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila Fonner Sent: Tuesday, August 16, 2011 6:44 AM To: 'Yahoo'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC on Frozen Tissue Does anyone else leave out the cell conditioning step? -Original Message- From: Yahoo [mailto:kim.tourn...@yahoo.com] Sent: Tuesday, August 16, 2011 7:41 AM To: Sheila Fonner Subject: Re: [Histonet] IHC on Frozen Tissue I would leave out the pretreatment step as well i.e. Enzyme, CC1/CC2. Sent from the iPhone of Kim Tournear On Aug 16, 2011, at 6:03 AM, "Sheila Fonner" wrote: > Thank you. > > > -Original Message- > From: Mierow, Brett T. [mailto:brett.mie...@essentiahealth.org] > Sent: Tuesday, August 16, 2011 7:02 AM > To: Sheila Fonner > Subject: RE: [Histonet] IHC on Frozen Tissue > > You are correct, leave out the depar. step. > > > Brett Mierow, HT (ASCP) > Histology specialist > Essentia Health > SMDC Laboratory > 407 East 3rd Street, Duluth, MN 54880 > (218) 786-4510 | brett.mie...@essentiahealth.org > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila > Fonner > Sent: Tuesday, August 16, 2011 5:41 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC on Frozen Tissue > > Good morning everyone, > > > > I was wondering if anyone could help me with a protocol change for the > Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, > and Zoster Virus on frozen tissue that was received for IF studies. I > know it's not Monday, but it is early, and I was hoping for some > information about how to change the protocol for frozen vs. paraffin > tissue. Do I just need to delete the depar. step and leave everything > else the same? I appreciate any help you could give. > > > > Thanks, > > Sheila > > > > ___ > 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] IHC on Frozen Tissue
Does anyone else leave out the cell conditioning step? -Original Message- From: Yahoo [mailto:kim.tourn...@yahoo.com] Sent: Tuesday, August 16, 2011 7:41 AM To: Sheila Fonner Subject: Re: [Histonet] IHC on Frozen Tissue I would leave out the pretreatment step as well i.e. Enzyme, CC1/CC2. Sent from the iPhone of Kim Tournear On Aug 16, 2011, at 6:03 AM, "Sheila Fonner" wrote: > Thank you. > > > -Original Message- > From: Mierow, Brett T. [mailto:brett.mie...@essentiahealth.org] > Sent: Tuesday, August 16, 2011 7:02 AM > To: Sheila Fonner > Subject: RE: [Histonet] IHC on Frozen Tissue > > You are correct, leave out the depar. step. > > > Brett Mierow, HT (ASCP) > Histology specialist > Essentia Health > SMDC Laboratory > 407 East 3rd Street, Duluth, MN 54880 > (218) 786-4510 | brett.mie...@essentiahealth.org > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila > Fonner > Sent: Tuesday, August 16, 2011 5:41 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC on Frozen Tissue > > Good morning everyone, > > > > I was wondering if anyone could help me with a protocol change for the > Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, > and Zoster Virus on frozen tissue that was received for IF studies. I > know it's not Monday, but it is early, and I was hoping for some > information about how to change the protocol for frozen vs. paraffin > tissue. Do I just need to delete the depar. step and leave everything > else the same? I appreciate any help you could give. > > > > Thanks, > > Sheila > > > > ___ > 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] Re:
I want you to look throw this site! It�s the most interesting stuff I�ve seen last time!... http://listadoweb.com/com.page.php?jnohot=49ek4 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC on Frozen Tissue
Thank you. -Original Message- From: Mierow, Brett T. [mailto:brett.mie...@essentiahealth.org] Sent: Tuesday, August 16, 2011 7:02 AM To: Sheila Fonner Subject: RE: [Histonet] IHC on Frozen Tissue You are correct, leave out the depar. step. Brett Mierow, HT (ASCP) Histology specialist Essentia Health SMDC Laboratory 407 East 3rd Street, Duluth, MN 54880 (218) 786-4510 | brett.mie...@essentiahealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila Fonner Sent: Tuesday, August 16, 2011 5:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on Frozen Tissue Good morning everyone, I was wondering if anyone could help me with a protocol change for the Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, and Zoster Virus on frozen tissue that was received for IF studies. I know it's not Monday, but it is early, and I was hoping for some information about how to change the protocol for frozen vs. paraffin tissue. Do I just need to delete the depar. step and leave everything else the same? I appreciate any help you could give. Thanks, Sheila ___ 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 Frozen Tissue
Good morning everyone, I was wondering if anyone could help me with a protocol change for the Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, and Zoster Virus on frozen tissue that was received for IF studies. I know it's not Monday, but it is early, and I was hoping for some information about how to change the protocol for frozen vs. paraffin tissue. Do I just need to delete the depar. step and leave everything else the same? I appreciate any help you could give. Thanks, Sheila ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] protocol for in situ RT_PCR
hi could you add me on too pelase? On Mon, Aug 15, 2011 at 11:09 PM, Barone, Carol wrote: > Anyone got a hot protocol for In situ RT-PCR? Haven't done one in 5 > years (on my Perk and Elmer)...Know there must be some great new > reagents and kits out there...opinions? Send to cbar...@nemours.org. Thx > CB > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa +27 11 717 2298 (tel & fax) 073 5574456 (emergencies only) "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
