[Histonet] Toluidine blue
Hello all, I was taught to do Toluidine Blue O without a control. Is there actual one and what would it be? I'm staining a bone core. Don's ask why, it's research and what a researcher wants... Plus they have a protocol they are following for this cartilaganous defect. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Toluidine blue
We use a canine mast cell tumor as positive control - veterinary lab naturally. Probably looking for mast cells in the core. Tresa -Original Message- From: Bernice Frederick [mailto:b-freder...@northwestern.edu] Sent: Tuesday, June 02, 2015 12:02 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Toluidine blue Hello all, I was taught to do Toluidine Blue O without a control. Is there actual one and what would it be? I'm staining a bone core. Don's ask why, it's research and what a researcher wants... Plus they have a protocol they are following for this cartilaganous defect. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.edu ___ 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] Toluidine blue
Hi Bernice, Your researcher probably has no idea. I can demonstrate cartilage, mast cells, bacteria and mucins depending on the tol blue technique I use. Ie not all tol blue techniques are the same. Ask him what he is after. Without control I would not recommend his work for publication if asked to review it. Lot of luck, Tony. From: Bernice Frederick [b-freder...@northwestern.edu] Sent: Wednesday, 3 June 2015 4:01 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Toluidine blue Hello all, I was taught to do Toluidine Blue O without a control. Is there actual one and what would it be? I'm staining a bone core. Don's ask why, it's research and what a researcher wants... Plus they have a protocol they are following for this cartilaganous defect. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.edu ___ 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 Sydney Children's Hospitals Network. This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network 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] PAS stain
Hi Mica, Most haematoxylins will do eg Harris or Mayer's. I would recommend a light counterstain (it is a counterstain, not the main stain). Differentiate if too strong. Also periodic acid treatment increases nuclear affinity for Hx so shorten the counterstain time. Regards, Tony From: Mica [faith14...@aol.com] Sent: Tuesday, 2 June 2015 1:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAS stain Just a question...what kind of hematoxylin do you use as a counter stain for your PAS stain and do you use a differentiator? Sent from my iPad ___ 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 Sydney Children's Hospitals Network. This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network 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] plants in the lab
Hi Michelle, Why would patients be in a histo lab anyway? From: Michelle Lamphere [michelle.lamph...@childrens.com] Sent: Sunday, 31 May 2015 10:36 PM To: 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] plants in the lab Our hospital Safety and Infection Control departments have policies in place prohibiting any potted plants from being in the hospital, anywhere. We can have them if they are only in water, but the soil presents an infection control issue for patients because of potential mildew, mold, spores, etc. Michelle Lamphere Senior Tech, Histology Anatomic Pathology O: 214.456.2318 | Fax: 214.456.0779 E: michelle.lamph...@childrens.com 1935 Medical District Drive | B1.06 | Dallas, Texas 75235 Message: 2 Date: Fri, 29 May 2015 14:23:00 -0400 From: Blazek, Linda lbla...@digestivespecialists.com To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in the lab Message-ID: 5a2bd13465e061429d6455c8d6b40e391742126...@ibmb7exchange.digestivespecialists.com Content-Type: text/plain; charset=us-ascii Happy Friday all! Does anyone have documentation of the benefit of having plants in the lab? I know this was discusses quite a while ago but I can't find references for it. Any help would be appreciated. Thanks, Linda Please consider the environment before printing this e-mail This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Children's Medical Center Dallas immediately via e-mail at priv...@childrens.com. Children's Medical Center Dallas and its affiliates hereby claim all applicable privileges related to this information. ___ 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 Sydney Children's Hospitals Network. This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network 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] toluidine blue for cartilage with controls and mast cell staining
You wrote: We use a canine mast cell tumor as positive control - veterinary lab naturally. Probably looking for mast cells in the core. Tresa -Original Message- From: Bernice Frederick [mailto:b-frederick at northwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ] Sent: Tuesday, June 02, 2015 12:02 PM To: Histonet at lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Subject: [Histonet] Toluidine blue Hello all, I was taught to do Toluidine Blue O without a control. Is there actual one and what would it be? I'm staining a bone core. Don's ask why, it's research and what a researcher wants... Plus they have a protocol they are following for this cartilaganous defect. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 * Bernice and Tresa, Having done a bone research study like this in the past, controls should be and were carefully done. You need to know normal cartilage from treated or defect in cartilage. The researcher certainly should have set their experiment up accordingly but may have controls in place now???You did not say if this is articular cartilage from exterior joint surfaces where they took the core or deeper in the bone at the growth plate? These two cartilages will stain differently with T blue. Normally and when studying articular cartilage defects, it is wise to also do a safranin O/fast green stain along with the T Blue. Controls are extremely important and need to be carefully set up. Hopefully, you have a contralateral bone normal core from the same animal OR a core sample from an untreated, naive control animal.It was never stated what the experimental animal model is being used? I have done a study like this in the past. When core is decalcified with an acid or EDTA, then the control needs to be decalcified exactly the same way and at the same time as experimental cores with defect. If you are decalcifying with EDTA, then you should have a normal core that is not decalcified. This is difficult with mouse but possible with larger animals. The reason is to see if the proteoglycans in the articular or even the growth plate cartilage will be extracted by EDTA, and not appreciably by buffered formic acid. Articular cartilage where proteoglycans have been removed by a decalcifying agent will have different tinctorial quality (lighter) than cartilage never exposed to a decalcifying agent. EDTA is used by biochemists to extract proteoglycans for biochemical studies, and will the same thing in a cartilage section. Hence, there will be less staining seen with the toluidine blue or the Safranin O/fast green stain after EDTA. Hence you should run two controls, 1)a decalcified cartilage control and 2) an undecalcified control. How you decalcify will be important in order to retain proteoglycans in the cartilage. I strongly suggest using buffered formic acid, available commercially. You will find recipes for buffered formic acid in text books that contain sodium formate or sodium citrate. Look for these ingredients in product MSDS before you buy the formic acid decalcifying solutions. If there is any question about EDTA versus buffered formic acid and other acid decalcifiers i.e HCl, Nitric acid, etc. for cartilage studies, I will be happy to send publications concerning this topic privately. The Toluidine blue that we do for cartilage is designed to show cartilage staining and not mast cells. It could be the mast cells might be seen along with the cartilage staining but that is not the point. The toluidine blue stain I do for mast cells is entirely different from the toluidine blue cartilage staining protocol. I will be happy to send you a toluidine blue stain procedure for cartilage and also the SafO/Fast green protocol. I have a superb T blue mast cell stain from Churukian which allows mast cells to stand out without any blue background in surrounding tissues that is often seen with other T blue staining protocols. Hope this helps. Gayle M. Callis HTL/HT/MT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
