[Histonet] RE: Re: Methanol in H2O2 explanation
Hi Carl, What do you mean by "Why do people rehydrate after dewaxing?" Do you really mean that slides do not require rehydration or do you mean that slides can be left to dry after de-waxing prior to staining. Re-hydration is necessary, otherwise xylene will prevent aqueous stains from doing their thing efficiently. I was lead to believe that as H2O2 was catalysed by endogenous peroxidase, the reactive oxygen reacted with the methanol to then degrade the enzyme, but I need to look closer at this chemistry. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl Sent: Wednesday, 11 July 2012 1:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Methanol in H2O2 explanation Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Methanol in H2O2 explanation
Why do some people use methanolic H2O2? Why do people rehydrate after dewaxing? Both are unnecessary, under usual conditions. Methanol was used in the early days as a peroxidase blocker by itself. The combination was devised as a "belt and bracer" method. As you stated, you use aq H2O2 effectively. So do I and many others. However, for unfixed frozen sections, I would never use aq H2O2, if I wanted sections remaining on my slides After dewaxing, rinse sections in 4x 100% alcohol, rinse in water, endog. Px block while you make up you AR solutions Carl Hobbs Histology and Imaging Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Another Microscope Question
Hi Laurie, How are you? The idea of "pushing out of Koehler" as you described gives you a "poor-man's", pseudo-phase effect. I use it for unstained frozen sections of renal and skin biopsies that are destined for immunofluorescence. And then I get mean and request the students who I have demonstrated this technique to, to return the microscope to "Koehler perfection". I suppose it teaches the student to be familiar with the microscopy results and to recognise when a better image is achievable. I am such a mean person. A useful reference?: Head ES. Tschen EH. (1982) Unstained negative image patterns. A checkpoint for quality slides. American Journal of Dermatopathology. 4(2):143-8. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Reilly, Laurie Sent: Wednesday, 11 July 2012 9:22 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Another Microscope Question Dear All, My microscope training has been mainly on brightfield microscopes using Koehler illumination. Recently, in discussions with Veterinary Pathologists, they have been advocating winding the condenser way down to the field diaphragm when they are examining urine samples. This goes against the microscopy principles that I have been taught and now teach to our students. Can anyone enlighten me on the value of this practice? Could the same effect be obtained by closing the aperture diaphragm? Thanks in advance for your wisdom. Regards, Laurie. Mr. Laurie REILLY Histopathology School of Veterinary and Biomedical Sciences James Cook University Townsville Qld. 4811 Australia. Phone 07 4781 4468 * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] direct IF on "normal" animal tissue
Hi Liette, I didnt see if you got help yet. Ill give it a try. First I would go with the pig kidney,. and this antibody looks like it should work . http://www.abcam.com/Pig-IgG-secondary-antibody-H-L-ab6911.html This is my best guess off the top of my head. If you try it and it works, let us know, oh and you can look on that site for procedures too. and i bet you could call and get some good info. hope this helps, good luck :) Kim D From: Liette Tougas To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 2:57 PM Subject: [Histonet] direct IF on "normal" animal tissue I teach immunology and histology to biomedical laboratory technology students and I would like to perform a simple direct immunofluorescence technique on "normal" animal tissue obtained from the butcher. I believe I could easily obtain either kidney, heart or liver either from pig or beef. Therefore, I was wondering which fluorescent antibody to purchase in order to demonstrate an antigen normally present in such animal tissues and that would not produce too much background, considering an expectable certain degree of autolysis. I appreciate any suggestion and detailed information if possible. Thank you in advance, Liette Tougas, RT, B.Sc., M.Sc. Biomedical Laboratory Technology Department Dawson College, Montreal, Qc ___ 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] Another Microscope Question
Dear All, My microscope training has been mainly on brightfield microscopes using Koehler illumination. Recently, in discussions with Veterinary Pathologists, they have been advocating winding the condenser way down to the field diaphragm when they are examining urine samples. This goes against the microscopy principles that I have been taught and now teach to our students. Can anyone enlighten me on the value of this practice? Could the same effect be obtained by closing the aperture diaphragm? Thanks in advance for your wisdom. Regards, Laurie. Mr. Laurie REILLY Histopathology School of Veterinary and Biomedical Sciences James Cook University Townsville Qld. 4811 Australia. Phone 07 4781 4468 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] microscope ocular questions
Tim, The duel-adjustable eyepiece vs the single adjustable one, I believe has to do with the type of prism that is used. Mis-matched eyepieces can be a problem, especially if from different manufacturers or even magnification. Magnification differences would be somewhat obvious as you will never get them to focus for two eyes. In order to use a binocular telescope (really cool instrument, once had the chance to use one of those) you need, not only matched eyepieces, but ideally from the same lot #. However, focusing two ten-inch telescopes to the same focal plane is a bit touchier than a microscope. Its one of the reasons you don't see too many of them. Have you tried simply swapping them (left to right/right to left)? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy Sent: Tuesday, July 10, 2012 1:49 PM To: Lee & Peggy Wenk; Histonet Subject: RE: [Histonet] microscope ocular questions Thanks Peggy, That is clear. I used microscopes for years with one fixed ocular and one focusable ocular. I was wondering about why now both oculars are "focusable" yet one has more usability than the other. Maybe to accomodate greater variation? Or maybe is due to the advent of parfocal microscopes I found some instructions on parfocal adjustment that refers to setting both oculars to zero when doing the initial focus at high magnification, then setting the ocular adjustment for each eye at low magnification. So that makes sense for individualistic adjustment. However, I was asked why one ocular has easier use and more graduations that the other and I didn't have a good answer to that...The person thought the oculars were not the same so there was some problem with the microscope. Tim -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 10, 2012 11:27 AM To: Histonet Subject: RE: [Histonet] microscope ocular questions Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -Original Message- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.mor...@ucsfmedctr.org ___ 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 This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/h
Fw: [Histonet] microscope ocular questions
Tim, You are welcome, but I'm not Peggy. I'm Lee, her husband. I'm very familiar with binoculars, so I only assumed that they are the same as microscopes. Lee Wenk -Original Message- From: Morken, Timothy Sent: Tuesday, July 10, 2012 2:49 PM To: Lee & Peggy Wenk ; Histonet Subject: RE: [Histonet] microscope ocular questions Thanks Peggy, That is clear. I used microscopes for years with one fixed ocular and one focusable ocular. I was wondering about why now both oculars are "focusable" yet one has more usability than the other. Maybe to accomodate greater variation? Or maybe is due to the advent of parfocal microscopes I found some instructions on parfocal adjustment that refers to setting both oculars to zero when doing the initial focus at high magnification, then setting the ocular adjustment for each eye at low magnification. So that makes sense for individualistic adjustment. However, I was asked why one ocular has easier use and more graduations that the other and I didn't have a good answer to that...The person thought the oculars were not the same so there was some problem with the microscope. Tim -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 10, 2012 11:27 AM To: Histonet Subject: RE: [Histonet] microscope ocular questions Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -Original Message- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.mor...@ucsfmedctr.org ___ 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] direct IF on "normal" animal tissue
I teach immunology and histology to biomedical laboratory technology students and I would like to perform a simple direct immunofluorescence technique on "normal" animal tissue obtained from the butcher. I believe I could easily obtain either kidney, heart or liver either from pig or beef. Therefore, I was wondering which fluorescent antibody to purchase in order to demonstrate an antigen normally present in such animal tissues and that would not produce too much background, considering an expectable certain degree of autolysis. I appreciate any suggestion and detailed information if possible. Thank you in advance, Liette Tougas, RT, B.Sc., M.Sc. Biomedical Laboratory Technology Department Dawson College, Montreal, Qc ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] microscope ocular questions
Thanks Peggy, That is clear. I used microscopes for years with one fixed ocular and one focusable ocular. I was wondering about why now both oculars are "focusable" yet one has more usability than the other. Maybe to accomodate greater variation? Or maybe is due to the advent of parfocal microscopes I found some instructions on parfocal adjustment that refers to setting both oculars to zero when doing the initial focus at high magnification, then setting the ocular adjustment for each eye at low magnification. So that makes sense for individualistic adjustment. However, I was asked why one ocular has easier use and more graduations that the other and I didn't have a good answer to that...The person thought the oculars were not the same so there was some problem with the microscope. Tim -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Tuesday, July 10, 2012 11:27 AM To: Histonet Subject: RE: [Histonet] microscope ocular questions Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -Original Message- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.mor...@ucsfmedctr.org ___ 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] microscope ocular questions
Tim, etal: This is easily understood: focusing and setup work the same on binoculars, one eyepiece is focused with the main focusing system, the other is used to match focusing with both eyes. First focus the scope (binoculars or microscope) thru the simple (non focusing) eyepiece, then use the focusing eyepiece to fine tune focus for the other eye. Once you've determined the setting on the focusing eyepiece, you can return the scope to this setting with ease and you should be able to use the scope for hours at a time without fatigue. Each microscope or binoculars is different. The setting for each person will be different (everybody's eyes are different). Each of our eyes are different, thus the need for independent focusing for one eye. Try defocusing the focusing eyepiece and using scope for a period. Your eyes will have to work overtime to keep the image in focus (if you are young you might last longer than I would at 65) and you could get a headache or suffer fatigue. Lee Wenk (Peggy's husband) -Original Message- From: Morken, Timothy Sent: Tuesday, July 10, 2012 1:39 PM To: Histonet Subject: [Histonet] microscope ocular questions Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.mor...@ucsfmedctr.org ___ 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] IVD P63
Yes, it is true. I just talked with Biogenex Laboratories and they will be carrying both the 4A4 and 4B1E12 clones in the foreseeable future. Other places I have talked to are depleting their stock and not carrying P63 anymore. Ventana wants $800+ dollars for 50 tests worth and do not carry concentrates. What a racket! Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IVD P63
Clare, Just got off the phone with my Biocare rep, Lissa Wall. I'm afraid that it IS true. Long story short, legal issues going on, and Biocare will stop selling P63 TODAY, 7/10/12. Please talk to your rep ASAP to get those last minute orders in today. I well remember when the last (no name) company received sole rights to an antibody and the pricing quadrupled overnight! HURRY!! Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of Clare Thornton [cthorn...@dahlchase.com] Sent: Tuesday, July 10, 2012 12:48 PM To: 'Kim Donadio'; Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IVD P63 I just spoke with my Biocare rep. He said it is untrue at this time, and they are still selling p63. Clare J. Thornton, HTL(ASCP),QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Tuesday, July 10, 2012 12:03 PM To: Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IVD P63 Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 ___ 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] microscope ocular questions
Histonet gurus, Why is each microscope ocular marked and operated differently? For instance the right one has a knurled focusing ring, is easily focused and has detailed graduations while the left one is not really set up to focus quickly and has only minimal graduations? Always wondered about this but can't find anything about it! Thanks for your insights! Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center 505 Parnassus Ave, Box 1656 Room S570 San Francisco, CA 94143 (415) 353-1266 (ph) (415) 514-3403 (fax) tim.mor...@ucsfmedctr.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Formalin Fixation issues (Bob Richmond)
If you have large specimens, it is best to "open" them as soon as possible. If it is a breast, at least do your prelim measurements and slice it open, leaving it still attached, so that the formain can penetrate the tissue. If you go ahead and slice it up, at least wrap it, in order with paper towels, to keep orientation while it fixes. You can always block it and then let it fix too. You just need to be sure to slice it thin so that it fixes; and make sure you have adequate spacing between cassettes. You just need to figure out what way is best for your facility. Amanda Amador, AAS, ASCP(CM) Sr. Histotechnician Pathology Consultants of New Mexico 600 N. Richardson Roswell, NM 88202 575-622-5600, ext 218 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of histonet-requ...@lists.utsouthwestern.edu [histonet-requ...@lists.utsouthwestern.edu] Sent: Tuesday, July 10, 2012 11:02 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 104, Issue 11 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Formalin Fixation issues (Bob Richmond) 2. RE: Need help with CD40 on frozen sections (Teri Johnson) 3. Re: Formalin Fixation issues (Bob Richmond) 4. Day Shift Histotech Job Near Ithaca, NY (Melissa Phelan) 5. Re: Re: Formalin Fixation issues (Jill Cox) 6. RE: Re: Formalin Fixation issues (Weems, Joyce K.) 7. RE: Re: Formalin Fixation issues (Elizabeth Chlipala) 8. GBS IHC (Richard Cartun) 9. AUTO: Ramona Nelson is out of the office. (returning 07/16/2012) (ramona_nel...@bd.com) 10. Subject: Re: [PATHO-L] refreshing perspective on formaldehyde (White, Lisa M.) 11. Beecher tissue array 1 mm. punch set. (Mohammad Sayeeduddin) 12. CD40 (Reynolds,Donna M) 13. IVD P63 (Cathy Crumpton) 14. human vimentin in mouse tissue (Kim Merriam) 15. Re: IVD P63 (Kim Donadio) 16. RE: IVD P63 (Clare Thornton) -- Message: 1 Date: Mon, 9 Jul 2012 13:25:05 -0400 From: Bob Richmond Subject: [Histonet] Re: Formalin Fixation issues To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Jill Cox asks: >>Is anyone having problems with breast fixation prior to processing in formalin? For a couple of months now our breast specimens aren't fixing very well before gross. Our pathologist thinks they have changed something in the formalin itself. We utilize 15/1 ratio and in some cases let fix over the weekend. This has only been happening over the last couple of months and can't seem to figure this out. Any advice or similar problems, would love to hear from you.<< Breast specimens shouldn't be expected to fix before they're grossed, or at least before they've been cut into thin slices. Delaying fixation compromises immunostaining, to say nothing of H & E. They should be grossed as promptly as possible after they're received, and should never sit over the weekend without dissection. I seriously doubt that anything in the formalin has changed. It's your technique that needs to change. Unfortunately, this is a difficult task to delegate. Bob Richmond Samurai Pathologist Knoxville TN -- Message: 2 Date: Mon, 9 Jul 2012 17:46:25 + From: Teri Johnson Subject: [Histonet] RE: Need help with CD40 on frozen sections To: "histonet@lists.utsouthwestern.edu" Cc: "Bea DeBrosse-Serra \(bdebrosse-se...@isisph.com\)" Message-ID: <9f3cfee76e51b64991c7485270890b400cdc4...@ex4.lj.gnf.org> Content-Type: text/plain; charset="us-ascii" Hi Bea! I just left you a voicemail message. I looked up this antibody data sheet and to say the least the staining on the image is underwhelming! I would try fixing the cryosections with Beckstead's zinc prior to staining, and you can even consider fixing the spleens with Zinc and then cryoprotecting with sucrose prior to freezing. It appears they recommend using a biotinylated secondary and then streptavidin-HRP for staining. You might see if that helps as well. Teri Teri Johnson, HT(ASCP)QIHC GNF Histology Lab Manager Genomics Institute of the Novartis Research Foundation 858-332-4752 -- Message: 3 Date: Mon, 9 Jul 2012 13:51:10 -0400 From: Bob Richmond Subject: [Histonet] Re: Formalin Fixation issues To: histonet@lists.utsouthwestern.edu M
[Histonet] Methanol in H2O2 explanation
Hi everyone, Can someone refresh/enlighten me as to why Methanol is used in H2O2 blocking. Let's assume that I take my slides through the xylene/ethanol steps and that after the 70% etoh wash step, they are now in TBST. I could see adding methanol if you were doing the H2O2 blocking as part of the xylene/etoh process and maybe you had the methanol as a substitute for the 70% etoh. My xylene/ETOH looks like this. 3 x 5 min xylene. 2 x 5 min 100% etoh 2 x 5 min 95% etoh 1 x 5 min 70% Then my slides are washed and then epitope retrieved. After epitope retrieval they are again washed and then I do my H2O2 block without methanol. Thanks Patrick. CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IVD P63
I just spoke with my Biocare rep. He said it is untrue at this time, and they are still selling p63. Clare J. Thornton, HTL(ASCP),QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Tuesday, July 10, 2012 12:03 PM To: Cathy Crumpton; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IVD P63 Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 ___ 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] IVD P63
Ventana has gotten the sole patton for that antibody now I beleive, or will have soon. From: Cathy Crumpton To: "histonet@lists.utsouthwestern.edu" Sent: Tuesday, July 10, 2012 11:51 AM Subject: [Histonet] IVD P63 I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 ___ 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] human vimentin in mouse tissue
Greetings! I want to stain for human vimentin in mouse tissues (human fibroblasts). I many years ago, I used a biotinylated V9 clone for just this purpose and it worked great. I tried the biotiylated V9 clone from abcam, but I am not getting any staining at all. Any suggestions? Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IVD P63
I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes? Cathy Crumpton HT(ASCP), Lead Histotechnician Tuality Community Hospital 503-681-1292 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CD40
What are you using for Endogenous blocking? If you are using methanol with H2O2 that may be your problem. Many CD antibodies are very sensitive to the methanol. Try PBS or whatever buffer you are using to dilute the H202. I would also try going straight form the -20 into cold acetone without air drying and fix for10 min. Donna Reynolds HT (ASCP) U. T. M.D. Anderson Cancer Center Chief histology Tech, Department Cancer Biology 713-792-8106 -- Message: 3 Date: Mon, 9 Jul 2012 07:55:16 -0700 From: Bea DeBrosse-Serra Subject: [Histonet] Need help with CD40 on frozen sections To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <493caa64f203e14e8823737b9ee0e25f092ba8f...@exchmb01.isis.local> Content-Type: text/plain; charset="iso-8859-1" Histonetters, I need some help with CD40 on frozen sections (spleen, non-treated). I cut my frozens, let them briefly air dry and keep them at -20?C. When I use them for staining, I air dry them, fix them in cold acetone for 5 minutes and start the staining process. I use the primary AB from BD Pharmingen, # 550285 at various dilution, incubate them overnight at 4?C, use the secondary HRP conjugated antibody, DAB, counterstain and don't get any staining. Does anyone have any pointers, suggestions? Thank you in advance, Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 *** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Beecher tissue array 1 mm. punch set.
Hi Histonet members, I am in trouble. I need 1 mm. tissue array punch set ( Beecher manual instrument ). Either you can sell us one or exchange for a box of ten sets of 0.6mm size punch sets. Please contact me at sayeedud...@sbcglobal.net Thanks in advance for your response. Sayeed. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Subject: Re: [PATHO-L] refreshing perspective on formaldehyde
For the paranoid out there: imagine what a lawyer will do to a hospital in court when Formalin (or other proper fixative) was not allowed to be utilized and the specimen is ruined. The lawsuit settlement will make a fine from OSHA look like buying a candy bar. More than likely it will be a member of or government or high society type who is "wronged" before changes are made. It is hard to believe that a hospital would allow the fixative to be removed from all areas. Yes everyone who is utilizing the fixative needs to be educated in spill clean-up and have spill kits maintained in their area. It is a small price to pay to protect the integrity of the specimen. What if it was your loved ones specimen? I have heard of others leaving the taps open. The remedy was to buy bottles with screw caps with the bonus of the fixative not being as heavy. Lisa White, HT(ASCP) Supervisory HT James H. Quillen VAMC PO Box 4000 Corner of Veterans Way and Lamont PLMS 113 Mountain Home, TN 37684 423-979-3567 423-979-3401 fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] AUTO: Ramona Nelson is out of the office. (returning 07/16/2012)
I am out of the office until 07/16/20= 12. I = will respond to your message when I return. Note: This is an automated response to your message &quo t;Histonet Digest, Vol 104, Issue 10" sent on 7/9/2012 1:00:01 PM. = This is the only notification you will receive whi= le this person is away. _ * ** IMPORTANT MESSAGE FOR RECIPIENTS IN THE U.S.A.: This messa= ge may constitute an advertisement of a BD group's products or services or = a solicitation of interest in them. If this is such a message and you wou= ld like to opt out of receiving future advertisements or solicitations from= this BD group, please forward this e-mail to optoutbygr...@bd.com. * ** T his m= essage (which includes any attachments) is intended only for the designated= recipient(s). It may contain confidential or proprietaryinformationan=d may be subject to the attorney-client privilege or other confidentialit= y protections. If you are not a designated recipient, you may not review= , use, copy or distribute this message. If you received this in error, pl= ease notify the sender by reply e-mail and delete this message. Thank you . ***= Corporate Headquarters Mailing Address: BD (Becton, Dickinson and Compan= y) 1 Becton Drive Franklin Lakes, NJ 07417 U.S.A. *** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
