[Histonet] Direct immunofluorescence question
Barb, Have you tried a diamond pencil? Available from a variety of sources, these pencils can be used to etch a circle or other shape around the tissue to be viewed. I believe they use lesser-quality industrial diamonds to make the tips on the pencils (i.e. not the diamond ring quality). Coming across the etched line under the fluorescent microscope produces brightness that helps the pathologist find the tissue. We circle our DIF's on the back of the slide - it won't wash off and won't interfere with reagents this way, but is still visible in a darkened room. Hope this helps, Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Rio-Hortega Silver Carbonate Stain Polak's variant?
Hi All,
Question - Has anyone heard of a Rio-Hortega Silver Carbonate stain (Polak's
variant) for mitochondria staining? Any suggestions?
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
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wk email mailto:pru...@ihctech.net pru...@ihctech.net
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RE: [Histonet] Fibronectin AB for human tissues
I use AbCam 32419 (rabbit monoclonal, clone F1) on the Bond at a dilution of 1:300 after EDTA retrieval; very clean, crisp results Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Igor Deyneko Sent: Tuesday, October 05, 2010 1:07 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Fibronectin AB for human tissues Dear Histonetters! I've been looking for a good Fibronecting antibody to stain some human tumors for connective tissues. I've tried 6 from Abcam, all either not working or giving high background. if anyone knows of a good antibody, i would really appreciate it. Thank you. Igor Deyneko Infinity Pharmaceuticals Molecular Pathology Cambridge, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 83, Issue 4
Barb, Eric, all- We also use a diamond pen/pencil-and they do contain an industrial grade diamond, so they are quite reasonably priced; I looked one up on the VWR website (just as a reference, not necessarily an endorsement! :o) and they run about $20-30 each. They last a very long time, too-We¹ve had ours for several years. We use them for our direct IFs, FISH, anything dark field. Stephanie Stephanie Rodriguez, HTL(ASCP), QIHC Lead Molecular Technologist-FISH IHC Technologist III Phenopath Laboratories Seattle, WA (206) 374-9000 On 10/5/10 10:15 AM, histonet-requ...@lists.utsouthwestern.edu histonet-requ...@lists.utsouthwestern.edu wrote: Message: 11 Date: Tue, 5 Oct 2010 10:53:36 -0400 From: Gagnon, Eric gagn...@kgh.kari.net Subject: [Histonet] Direct immunofluorescence question To: histonet@lists.utsouthwestern.edu Message-ID: f93bd6329fc3ae4c8db116b985fbc31327c3a...@kghmail.kgh.on.ca Content-Type: text/plain; charset=iso-8859-1 Barb, Have you tried a diamond pencil? Available from a variety of sources, these pencils can be used to etch a circle or other shape around the tissue to be viewed. I believe they use lesser-quality industrial diamonds to make the tips on the pencils (i.e. not the diamond ring quality). Coming across the etched line under the fluorescent microscope produces brightness that helps the pathologist find the tissue. We circle our DIF's on the back of the slide - it won't wash off and won't interfere with reagents this way, but is still visible in a darkened room. Hope this helps, Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada _ Message: 1 Date: Mon, 4 Oct 2010 12:49:44 -0500 From: ba...@gundluth.org Subject: [Histonet] Direct immunofluoresent question To: histonet@lists.utsouthwestern.edu Message-ID: of9419a09e.870a453a-on862577b2.00614e6b-862577b2.0061e...@gundluth.org Content-Type: text/plain; charset=US-ASCII Hi all - The pathologist that reads our direct IF slides likes to have the sections of tissue circled on the slide so that they are easier to find. We currently use black marker on the back of the slide, but find that many times it smears and are looking for a better solution. What kind of slides do others use, and if you circle your sections what marker/pen/etcher do you use? Any thoughts are greatly appreciated! Barb Moe Gundersen Lutheran Medical Center La Crosse WI This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Rabbit Brain Tissue Processing Schedule
Hello everyone, While I am well versed in protocols to process bone from a variety of species, my experience in processing soft tissue is less extensive. Currently, I do have some soft tissue protocols which I inherited from my predecessor, and with which I have generally had success, but I am know that there exists more tissue specific protocols than I am currently using. More specifically, I would like to find a protocol to fix, infiltrate and embed rabbit brains in paraffin. Would anyone have a protocol that they would not mind sharing? Thanks again to all of the folks on histonet who share their vast knowledge with those of us who are still learning. Best regards to all, ~Sean Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-9807 (fax) smcbr...@andrew.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Rio Hortega (Polaks)
Patsy, Here is THE citation and abstract for this publication in J Histochem Cytochem (found it by Googling staining method). You can download the pdf at no charge. The photographs are excellent. Journal of Histochemistry and Cytochemistry Volume 52 (2): 211-216, 2004 Silver Carbonate Staining Reveals Mitochondrial Heterogeneity José M. LópezCepero Summary Silver staining methods, when selective, yield a high-contrast and high-resolution image in optical microscopy. A classical method for silver impregnation of mitochondria has been applied to murine tissues and reveals a marked heterogeneity among mitochondria in single cells. This heterogeneity can be detected in the optical microscope but is even more evident at the ultrastructural level. The differences in staining intensity may reflect different stages in the mitochondrial life cycle. The progressive accumulation of uranylargyrophilic material may be a marker of mitochondrial aging. This highly selective staining procedure may be of use in studies of mitochondrial changes under pathological conditions and during apoptosis. (J Histochem Cytochem 52:211216, 2004) Key Words: mitochondria silver carbonate mitochondrial life cycle mitochondrial heterogeneity mitochondrial fusion Gayle M. Callis HTL/HT/MT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Osteoclast
We need to find a way to stain mainly Osteoclast. Any suggestions? Thank you very much. Lin Lin S. Bustamante, B.Sc.; HT(ASCP) Research Associate Histology Lab Supervisor Veterinary Integrative Bioscience Texas AM University College Station, TX 77843-4458 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Osteoclast
You can do TRAP ( Tartrate-resistant Acidic Phosphatase) stain for osteoclasts. On 10/5/10 4:58 PM, Lin Bustamante lbustama...@cvm.tamu.edu wrote: We need to find a way to stain mainly Osteoclast. Any suggestions? Thank you very much. Lin Lin S. Bustamante, B.Sc.; HT(ASCP) Research Associate Histology Lab Supervisor Veterinary Integrative Bioscience Texas AM University College Station, TX 77843-4458 ___ 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] embedding beads/tissue marking dyes
Hi We may be encountering some issues with our tissue marking dyes 'clogging' up our processors. Ive heard of a system where a coloured 'bead' is embedded right beside the tissue, and subsequently cut onto the slide. This does not mark the margins obviously, but is used as a method to track like specimens that are grossed, embedded and subsequently cut in a row. (red, orange, green, blue.) I explored the archives for embedding beads, but only found reference to a glass bead that is placed in the wax inside the block, but not subsequently cut. PS: we are still troubleshooting our dyes (dilutions, brand, etc.), but if anyone has a brand of dye that they like and have no issues with, I'd be thrilled to get the information as well! Thanks in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology, Tech III Diagnostic Scientific Centre Calgary Laboratory Services Phone: 403-770-3588 Pager: 403-212-8223 X07630 P Please consider the environment before printing this email. This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: embedding beads/tissue marking dyes
When you place dye on any tissue, make sure you pipette some bouins on to the tissue... Apparently the bouins sets the stain... afterwards dab the tissue with paper towel to sop up the excess dye/bouins. Some dyes are better than others, but try the bouins first Call me or email me directly, I can help you with details :) Maria Katleba MS HT(ASCP) Pathology Dept. Mgr Queen of the Valley Medical Center 1000 Trancas Street Napa CA 94558 (707) 252-4411 x3689 direct (707) 226-4385 pager (707) 294-9229 cell- anytime -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jacqueline Farnsworth Sent: Tuesday, October 05, 2010 2:14 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] embedding beads/tissue marking dyes Hi We may be encountering some issues with our tissue marking dyes 'clogging' up our processors. Ive heard of a system where a coloured 'bead' is embedded right beside the tissue, and subsequently cut onto the slide. This does not mark the margins obviously, but is used as a method to track like specimens that are grossed, embedded and subsequently cut in a row. (red, orange, green, blue.) I explored the archives for embedding beads, but only found reference to a glass bead that is placed in the wax inside the block, but not subsequently cut. PS: we are still troubleshooting our dyes (dilutions, brand, etc.), but if anyone has a brand of dye that they like and have no issues with, I'd be thrilled to get the information as well! Thanks in advance, Jacquie Jacqueline Farnsworth Anatomic Pathology, Tech III Diagnostic Scientific Centre Calgary Laboratory Services Phone: 403-770-3588 Pager: 403-212-8223 X07630 P Please consider the environment before printing this email. This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice from St. Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] DIF Transport Media
Vinnie, I hope you are well. The following might be of use: Specimens for immunofluorescence are usually submitted to the laboratory in cell culture fluid (eg Hanks or RPMI) or on saline soaked gauze. For transport to other institutions, Michel's Transport Medium has often been used: Michel's Buffer 1M potassium citrate, pH 7.02.5 ml 0.1M magnesium sulphate 5.0 ml 0.1M N ethylmalemide5.0 ml Distilled H2O 87.5 ml * Mix well and store at 2 8oC. Exp. 1 year Michel's Transport Medium Michel's Buffer 100ml Ammonium sulphate 55gm Adjust pH to 7.0 7.4 with 1M KOH. Store at 2 8oC. Exp. 1 year Unfortunately Michel's Transport Medium has erroneously been called Michel's Fixative. None of the components of Michel's Transport Medium is a fixative. Ammonium sulphate precipitates antigen-antibody complexes in diseased skin and renal tissues. N ethylmalemide modifies free sulphydryl groups of cysteine residues in proteins (Fischer 2006). Michel's Transport Medium has been shown to be deleterious to morphology. Ultrastructurally, complete destruction of plasma membranes and intracytoplasmic organelles occurs after 48 hours storage. On the other hand, antigenicity is well preserved even after many days storage (Fischer 2006). Fischer (2006) Intern J Surg Pathol 14(1):108. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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 Della Speranza, Vinnie Sent: Wednesday, 6 October 2010 6:10 AM To: 'kristen arvidson'; histonet Subject: RE: [Histonet] DIF Transport Media Zeus was a company that used to market michel's under their own label. I don't believe zeus still exists. Michel's is the name associated with the author of the original paper. I don't have the reference. This solution does not require refrigeration. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, South Carolina 29425 Tel: (843) 792-6353 Fax: (843) 792-8974 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Monday, October 04, 2010 2:30 PM To: histonet Subject: [Histonet] DIF Transport Media Is Michel Medium the same as Zeus? Do these need to be refrigerated? Thanks. ___ 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 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
