RE: [Histonet] PROBLEM GETTING GOOD SECTIONS FROM BIOPSIES AND LIVER BIOPSY
Overnight processing is usually optimal for average size tissue. Larger tissue may not get optimal processing and biopsies may be a little over processed. It is the nature of tissue processing in a general lab that does not have several processors with different programs. A solution is to rehydrate after facing in the block. I keep soapy water( any liquid soap will work) to which I have added some ammonium hydroxide and pour it on my ice. ( ammonia is great for bloody tissue) for soaking. Remember what Lee Luna said, rehydrate, rehydrate, rehydrate. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Wilson A Sent: Friday, February 24, 2012 10:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PROBLEM GETTING GOOD SECTIONS FROM BIOPSIES AND LIVER BIOPSY Hi, Please we are having problems getting great slides from tissue/biopsy specimens like gastric, esophageal, needdle biopsy and Liver Biopsy. Our pathologists are not happy at all because of this situation. I will really, really appreciate if you guys in histoland could suggest some solutions that could stop the problem. Hoping to read from you guys asap. You guys are the best. Thanks, Wilson ___ 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] anti-GFP staining rabbit polyclonal A11122
Dear all, We are trying to establish an anti-GFP staining with rabbit polyclonal A11122 from Invitrogen. We are using formalin fixed, paraffin embedded mouse tissue and want to distinguish between a GFP transgenic mouse strain and a wt mouse. The problem is, we get a strong background in the wt, especially in spleen and lymphnodes, while the staining looks fine in liver. We dilute the antibody 1:400 and use 2 min antigen retrieval in Citrate buffer. Can anyone recommend a protocol? Or any other antibody that is working better? Thanks a lot for your help, Marina Marina Pils, DVM, PhD Animal experimental unit Helmholtz Centre for Infection Research Inhoffenstr. 7 D 38124 Braunschweig Helmholtz-Zentrum f?r Infektionsforschung GmbH | Inhoffenstra?e 7 | 38124 Braunschweig | www.helmholtz-hzi.de Vorsitzende des Aufsichtsrates: MinDir'in B?rbel Brumme-Bothe, Bundesministerium f?r Bildung und Forschung Stellvertreter: R?diger Eichel, Abteilungsleiter Nieders?chsisches Ministerium f?r Wissenschaft und Kultur Gesch?ftsf?hrung: Prof. Dr. Dirk Heinz; Ulf Richter, MBA Gesellschaft mit beschr?nkter Haftung (GmbH) Sitz der Gesellschaft: Braunschweig Handelsregister: Amtsgericht Braunschweig, HRB 477 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HistoTALK Welcomes Carrie Diamond Brenda Royce
Hi NSH'ers - Program #22 of HistoTALK www.HistoTALK.com has a great interview with our Executive Director, Carrie Diamond and Brenda Royce our Membership Manager. Both ladies share with us a huge amount of information and ideas about our special day - Histotechnology Professionals Day on March 10th. Listen at home, at work (quietly) or anywhere you have an Internet connection! Yours, Dave ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Hire ASCP PA
Wish I could hit the like button for this email ;) Message: 10 Date: Fri, 24 Feb 2012 14:01:09 -0800 From: Davide Costanzo pathloc...@gmail.com Subject: RE: [Histonet] Grossing techs To: Bruce Gapinski bgapin...@pathgroup.com, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: 2522340837944687498@unknownmsgid Content-Type: text/plain; charset=ISO-8859-1 Preserve the profession - hire an ASCP Certified PA. Amy Senn, HT Holy Spirit Hospital Histology Laboratory 503 N. 21st Street, Camp Hill, PA 17011 (717) 763-2124 Attention: This Message is intended only for the use of the individual or entity to which it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination or copying of this message or the taking of any action in reliance on the contents of this message is strictly prohibited. If you have received this message in error, please notify us immediately and destroy the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Chicago Labs
Hey Chicagoland Histoneters, We (Tech One Biomedical) are in the process of updating our fliers and catalog. The designer and photographer were hoping to some shots of a real lab. If you are willing to let us do it, please let me know. We will do our best to stay out of the way of your work our could even do the work on the weekend when you are closed. Thanks for your consideration. Best Matt -- 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] anti-GFP staining rabbit polyclonal A11122
I use this exact antibody routinely at 1:1000. Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com From: Marina Pils marina.p...@helmholtz-hzi.de To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Sent: Monday, February 27, 2012 6:05 AM Subject: [Histonet] anti-GFP staining rabbit polyclonal A11122 Dear all, We are trying to establish an anti-GFP staining with rabbit polyclonal A11122 from Invitrogen. We are using formalin fixed, paraffin embedded mouse tissue and want to distinguish between a GFP transgenic mouse strain and a wt mouse. The problem is, we get a strong background in the wt, especially in spleen and lymphnodes, while the staining looks fine in liver. We dilute the antibody 1:400 and use 2 min antigen retrieval in Citrate buffer. Can anyone recommend a protocol? Or any other antibody that is working better? Thanks a lot for your help, Marina Marina Pils, DVM, PhD Animal experimental unit Helmholtz Centre for Infection Research Inhoffenstr. 7 D 38124 Braunschweig Helmholtz-Zentrum f?r Infektionsforschung GmbH | Inhoffenstra?e 7 | 38124 Braunschweig | www.helmholtz-hzi.de Vorsitzende des Aufsichtsrates: MinDir'in B?rbel Brumme-Bothe, Bundesministerium f?r Bildung und Forschung Stellvertreter: R?diger Eichel, Abteilungsleiter Nieders?chsisches Ministerium f?r Wissenschaft und Kultur Gesch?ftsf?hrung: Prof. Dr. Dirk Heinz; Ulf Richter, MBA Gesellschaft mit beschr?nkter Haftung (GmbH) Sitz der Gesellschaft: Braunschweig Handelsregister: Amtsgericht Braunschweig, HRB 477 ___ 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] egfr
Good Morning Histonetters I am looking to add EGFR to our antibody list. What clone is everyone using out there in Histoland? Any info would be greatly appreciated. Thanks in advance. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory Manager X-Cell Laboratories 716-250-9235 Ext. 232 e-mail cp...@x-celllab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] NFAT antibody
Anybody using anti-nFAT antibodies in mouse tissues or tcells? If so, could you recomend a particular antibody? Thanks! Mike ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Removing tape
I'm venturing a few guesses here, and would welcome others to do the same. My guess is using acetone might dissolve/soften the plastic from the tape but not remove the adhesive from the top of the tissue section. In thinking about removing adhesive from surfaces where it is unwanted, you sometimes cannot use solvents without compromising the integrity of the surface. Peanut butter or WD40 works well for those types of issues. So I'm thinking maybe WD40 or maybe a soak in mineral oil? I'd probably try mineral oil first, since it's a paraffinic oil. Maybe even warm it? Good luck. Let us know what works for you! Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Removing coverslipping tape
Soak the slide in acetone for 10-15 minutes; it turns to something that looks like a slice of Jell-O and slithers down the slide, leaving cells and stain and structure in perfect order. I usually dip the slide in xylene a couple times to clear the acetone and re-coverslip it. Works like a charm. LOVE the coverslipping tape! I'm down to 20 days until I retire! Not that I'm counting or anything... 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
[Histonet] fibrin IHC on FFPE mouse tissue
Is there an antibody out there that will stain for fibrin on FFPE mouse tissues; been searching but haven't found one. Does everyone just do a special stain when looking for fibrin? Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] wondering about per diem work in St. Pete, FL
Hi all, I'm wondering if anyone knows of any per diem histology jobs in St. Pete area. I am retiring from my current job and will be spending several(winter) months in St. Pete. Would I need a FL license to work per diem? Thanks for any info you can provide. Nancy Rutledge ~~ This email and any files transmitted with it are confidential, and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error contact the Help Desk for Cape Cod Healthcare. helpd...@capecodhealth.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Removing coverslipping tape
Just a note on coverslipping tape - I love it - this is the first place in 40+ years I've used it, and it's good stuff. HOWEVER, I noticed a brown cornflaking artefact on some tissues when I first arrived in this job - and we fixed it. If the last dehydrating step (100%) ethanol has the least little tiny pink from eosin - trace amounts of H20 remain on the slide even through xylene. IF you were using traditional glass coverslips and synthetic mounting medium - this trace H20 is absorbed by the mounting medium. However, the tape is not as forgiving. This trace amount of moisture shows up as brown spots on the tissue. I've proven this theory by removing the tape from affected slides, backing them through xylene to very clean 100%, clear through xylene and remounting with glass. The brown artefact disappears. In this instance it could have been mistaken for melanin on a section of rodent tongue. Jackie O' -Original Message- From: Breeden, Sara sbree...@nmda.nmsu.edu To: histonet histonet@lists.utsouthwestern.edu Sent: Mon, Feb 27, 2012 11:32 am Subject: [Histonet] Removing coverslipping tape Soak the slide in acetone for 10-15 minutes; it turns to something that ooks like a slice of Jell-O and slithers down the slide, leaving cells nd stain and structure in perfect order. I usually dip the slide in ylene a couple times to clear the acetone and re-coverslip it. Works ike a charm. LOVE the coverslipping tape! I'm down to 20 days until I retire! Not that I'm counting or nything... 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) ___ istonet mailing list isto...@lists.utsouthwestern.edu ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Removing tape - clarified
Greetings all, My response was to the post sent by Merissa which states: Dear All, I am using clear packing tape on mma embedded rabbit femurs with implants. The reasoning for the use of tape is that the implant crumbles when being cut without the tape. I have switched to a new tape that partially comes off with xylene, but does not release from the section on the slide (held with haupts). Does anyone have suggestions for removing the tape - xylene vs toluene vs acetone? At this point, the slides have been in xylene overnight and have not been released from the slides and the other tape took 45mins to lift off. Thank you so much, Merissa I'm thinking I should have provided this with my email response. My apologies. For those who are still confused, here is my response to the above dilemma: I'm venturing a few guesses here, and would welcome others to do the same. My guess is using acetone might dissolve/soften the plastic from the tape but not remove the adhesive from the top of the tissue section. In thinking about removing adhesive from surfaces where it is unwanted, you sometimes cannot use solvents without compromising the integrity of the surface. Peanut butter or WD40 works well for those types of issues. So I'm thinking maybe WD40 or maybe a soak in mineral oil? I'd probably try mineral oil first, since it's a paraffinic oil. Maybe even warm it? In addition, Diane Sterchi has a JOH article which addresses this in large paraffin block sectioning. She tried different tapes and shows which ones worked better than others. Happy Monday! Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 99, Issue 35
Bonjour, Je suis absent du laboratoire jusqu'au lundi 05 mars 2012. Je vous répondrai le plus rapidement possible. Eric Tambutté Thank you for your mail. I will be out of office till March 05th 2012. I will respond to your e-mail as soon as possible. Thank you for your understanding. Best regards Eric Tambutté ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Hydrating Frozen Tissues embedded in OCT for Sectioning
We are trouble-shooting cryosectioning mouse lung tissue. Often the lung section tears or breaks apart during sectioning. In the past, if the lung section is proving difficult to section, we take the OCT-embedded tissue and re-embed it back into OCT (basically put fresh OCT into the original mold and then place the OCT block with the tissue back into the mold such that the exposed tissue is covered back with OCT). This is then placed back in our -80°C. When sectioning the next day, the tissue is often easier to section. One person in our lab tried to resolve the problem by brushing a little bit of sterile water onto the tissue when sectioning. This appeared to hydrate the tissue and it sectioned better. However, we weren't sure if this was a good idea or not. Any feedback would be greatly appreciated. For background purposes our lung tissue are processed several ways: 1. Lungs are perfused with PBS, tissue extracted from mouse, placed in PFA/sucrose for several hours and then embedded in OCT. 2. Lungs are perfused with PBS, inflated gently with OCT/PBS (3:1), placed in PFA/sucrose for several hours and then embedded in OCT. 3. Lungs are perfused with PBS, inflated gently with OCT/PBS (3:1), embedded in OCT and frozen by immersion into liquid nitrogen (just the bottom half of the mold is lowered into LN). Much appreciation, Miki Kasai Biologist Pediatric Oncology Branch NCI, NIH CRC, 1W Rm. 1-3-888 10 Center Drive Bethesda, MD 20892 (301) 496-2318 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] conservation histotech
Well you can always run your lab in the most green way possible using recyling and reuse management of all materials in the lab. In Md there are a few state labs with coveted positions that only open up through the death or retirement of a tech because they are so rewarding. The closest thing I can thin of are labs that deal with fish and seafood as a means of testing waterways. Hope this helps and good luck in your new field. It has changed since I started in the late 70's(pan embedding, knife sharpening, eating and smoking in the lab)I can only imagine where the field will be in 30 years. Nick Madary, HT/HTL(ASCP)QIHC George Washington University Pathology Core Laboratory Ross Hall, Room 706 23rd and I Street NW Washington D.C. 20037 202.994.8916 pat...@gwumc.edu BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Joseph Madary EMAIL;WORK;PREF;NGW:pat...@gwumc.edu N:Madary;Joseph ORG:;Pathology TITLE:Senior Research Assistant TEL;PREF;FAX:202 994-5056 END:VCARD ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Hydrating Frozen Tissues embedded in OCT for Sectioning
As long as you don't need to use them for RNA analysis, the easiest thing to do is just rub your finger (without glove) across the block (very briefly) and then take the section. This will probably do the trick and hydrate it just enough to make a difference. Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From: Kasai, Miki (NIH/NCI) [E] kas...@mail.nih.gov To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Monday, February 27, 2012 1:34 PM Subject: [Histonet] Hydrating Frozen Tissues embedded in OCT for Sectioning We are trouble-shooting cryosectioning mouse lung tissue. Often the lung section tears or breaks apart during sectioning. In the past, if the lung section is proving difficult to section, we take the OCT-embedded tissue and re-embed it back into OCT (basically put fresh OCT into the original mold and then place the OCT block with the tissue back into the mold such that the exposed tissue is covered back with OCT). This is then placed back in our -80°C. When sectioning the next day, the tissue is often easier to section. One person in our lab tried to resolve the problem by brushing a little bit of sterile water onto the tissue when sectioning. This appeared to hydrate the tissue and it sectioned better. However, we weren't sure if this was a good idea or not. Any feedback would be greatly appreciated. For background purposes our lung tissue are processed several ways: 1. Lungs are perfused with PBS, tissue extracted from mouse, placed in PFA/sucrose for several hours and then embedded in OCT. 2. Lungs are perfused with PBS, inflated gently with OCT/PBS (3:1), placed in PFA/sucrose for several hours and then embedded in OCT. 3. Lungs are perfused with PBS, inflated gently with OCT/PBS (3:1), embedded in OCT and frozen by immersion into liquid nitrogen (just the bottom half of the mold is lowered into LN). Much appreciation, Miki Kasai Biologist Pediatric Oncology Branch NCI, NIH CRC, 1W Rm. 1-3-888 10 Center Drive Bethesda, MD 20892 (301) 496-2318 ___ 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] Hydrating Frozen Tissues embedded in OCT for Sectioning
Also, make sure that the block sits in the cryostat for a while to acclimate to the temperature. The cutting temp for lung (correct me if I'm wrong here) is probably about -20; the block should be the same temperature as the cryostat. Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From: Colleen Forster cfors...@umn.edu To: Kim Merriam kmerriam2...@yahoo.com Sent: Monday, February 27, 2012 1:55 PM Subject: Re: [Histonet] Hydrating Frozen Tissues embedded in OCT for Sectioning Yep, the sample is too cold. Rubbing your finger across even with a glove (just linger a bit longer) will help alot. Colleen Forster U of MN On 2/27/2012 12:47 PM, Kim Merriam wrote: As long as you don't need to use them for RNA analysis, the easiest thing to do is just rub your finger (without glove) across the block (very briefly) and then take the section. This will probably do the trick and hydrate it just enough to make a difference. Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From: Kasai, Miki (NIH/NCI) [E]kas...@mail.nih.gov To: histonet@lists.utsouthwestern.eduhistonet@lists.utsouthwestern.edu Sent: Monday, February 27, 2012 1:34 PM Subject: [Histonet] Hydrating Frozen Tissues embedded in OCT for Sectioning We are trouble-shooting cryosectioning mouse lung tissue. Often the lung section tears or breaks apart during sectioning. In the past, if the lung section is proving difficult to section, we take the OCT-embedded tissue and re-embed it back into OCT (basically put fresh OCT into the original mold and then place the OCT block with the tissue back into the mold such that the exposed tissue is covered back with OCT). This is then placed back in our -80°C. When sectioning the next day, the tissue is often easier to section. One person in our lab tried to resolve the problem by brushing a little bit of sterile water onto the tissue when sectioning. This appeared to hydrate the tissue and it sectioned better. However, we weren't sure if this was a good idea or not. Any feedback would be greatly appreciated. For background purposes our lung tissue are processed several ways: 1. Lungs are perfused with PBS, tissue extracted from mouse, placed in PFA/sucrose for several hours and then embedded in OCT. 2. Lungs are perfused with PBS, inflated gently with OCT/PBS (3:1), placed in PFA/sucrose for several hours and then embedded in OCT. 3. Lungs are perfused with PBS, inflated gently with OCT/PBS (3:1), embedded in OCT and frozen by immersion into liquid nitrogen (just the bottom half of the mold is lowered into LN). Much appreciation, Miki Kasai Biologist Pediatric Oncology Branch NCI, NIH CRC, 1W Rm. 1-3-888 10 Center Drive Bethesda, MD 20892 (301) 496-2318 ___ 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] Parameters for alcohol on Processor
Hello Histonetters, Could someone please advise me on established guidelines for changing the alcohol on the processor. We process a very small amount of blocks weekly on our ASP300S. Approximately 20 blocks per week. We have decided to use a hydrometer to test our 70% thru 100% alcohols, rather than base our changing reagents on number of blocks processed. My questions are, does a (1) % point decrease necessitate a change ? What are the parameters for changing each alcohol ? Thanking you in advance, Karen Karen E. Cruise Histologist (Research Technician II) Washington University School of Medicine Laboratory for Translational Pathology 216 S. Kingshighway Room 2332 St. Louis, MO 63110 (314) 454-8636 Phone (314) 454-5525 Fax kcru...@path.wustl.edumailto:kcru...@path.wustl.edu The materials in this email are private and may contain Protected Health Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] fibrin IHC on FFPE mouse tissue
Hi Kim, I am working on a project just like this right now and we are using Fibrinogen (AbCam ab34269) and PTAH. Drop me a line if you have any questions. Amos On Mon, Feb 27, 2012 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 20 Date: Mon, 27 Feb 2012 09:34:08 -0800 (PST) From: Kim Merriam kmerriam2...@yahoo.com Subject: [Histonet] fibrin IHC on FFPE mouse tissue To: Histonet histonet@lists.utsouthwestern.edu Message-ID: 1330364048.88489.yahoomail...@web130102.mail.mud.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Is there an antibody out there that will stain for fibrin on FFPE mouse tissues; been searching but haven't found one. Does everyone just do a special stain when looking for fibrin? Kim ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] anti-GFP staining rabbit polyclonal A11122
Marina, I agree with Paula Pierce, have used the same anti- GFP antibody on some projects but at 1:500-1:1000 area and with a much longer 8-15 minute retrieval in citrate. Also, I know you know to be aware of how much the mice are transfected . In my transfected mice (NOT GFP and in a former life) we might transfect 4 different times and get 4 different efficiencies and 4 different levels of expression in 4 different lines. And of course then in your case the variation with the type of GFP you are transfecting with might ultimately be seen at varying IHC levels since all GFP antibodies don't see all GFP variants with the same efficiency. Something I did to help ease the pain of working up a stain when working up lacZ (not GFP ) was to use a commercially available Tie2/ LacZ transgenic mouse. With that Tie2 promoter driving LacZ expression only in endothelial cells, you had neg tissue (equivalent to wt) and pos endothelial cells (your transgenic target) right in the very same section. It proved to be an exquisitely sensitive control when doing beta-gal studies. Although I've never used it I understand there is now a Tie2/ GFP transgenic available which should help optimize staining protocols as the expression levels are constant and well characterized and you need only one mouse. Don't know what other gene or transgene target you are after but any given transgene does not incorporate equally all the time as you know. Thats my take on this. Ray Koelling PhenoPath Labs Seattle, WA - Original Message - From: Marina Pils Marina. Pils @helmholtz-hzi. de To: histonet @lists. utsouthwestern . edu histonet @lists. utsouthwestern . edu Sent: Monday, February 27, 2012 4:05:56 AM Subject: [ Histonet ] anti- GFP staining rabbit polyclonal A11122 Dear all, We are trying to establish an anti- GFP staining with rabbit polyclonal A11122 from Invitrogen . We are using formalin fixed, paraffin embedded mouse tissue and want to distinguish between a GFP transgenic mouse strain and a wt mouse. The problem is, we get a strong background in the wt, especially in spleen and lymphnodes , while the staining looks fine in liver. We dilute the antibody 1:400 and use 2 min antigen retrieval in Citrate buffer. Can anyone recommend a protocol? Or any other antibody that is working better? Thanks a lot for your help, Marina Marina Pils , DVM , PhD Animal experimental unit Helmholtz Centre for Infection Research Inhoffenstr . 7 D 38124 Braunschweig Helmholtz-Zentrum f?r Infektionsforschung GmbH | Inhoffenstra ?e 7 | 38124 Braunschweig | www .helmholtz-hzi. de Vorsitzende des Aufsichtsrates : MinDir'in B? rbel Brumme-Bothe, Bundesministerium f?r Bildung und Forschung Stellvertreter : R? diger Eichel , Abteilungsleiter Nieders ? chsisches Ministerium f?r Wissenschaft und Kultur Gesch ? ftsf ? hrung : Prof. Dr. Dirk Heinz; Ulf Richter, MBA Gesellschaft mit beschr ? nkter Haftung ( GmbH ) Sitz der Gesellschaft : Braunschweig Handelsregister : Amtsgericht Braunschweig , HRB 477 ___ 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
