[Histonet] Toluidine Blue Dilution
Hey Netters,
Does anyone have a procedure for diluting Toluidine Blue from powder or 1%
aqueaous with 10% NBF. I have always eyeballed the measurement as I was
taught. However, as a new sup, I need to make sure we have an SOP in our manual.
Thanks All,
Natacha Mitchell, BS, HTL (ASCP)
Histology Lab Supervisor
Ben Taub General Hospital
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[Histonet] Toluidine blue stain fo plant tissue
Could anyone give me the concentration (? 0.1-0.05%) and the exact technique/protocol to use. I am staining shoot apical meristems not leaf or root. Regards Peter Noyce PhD student. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] toluidine blue
Hi All- Is anyone using Toluidine Blue on air dried touch prep slides? Specifically for endobronchial ultra sound specimens (ebus). I am wondering if you can tell me what you are using for a toluidine blue recipe. Thank you for your help- Nancy Schmitt MLT, HT(ASCP) Pathology Support Services Manager United Clinical Laboratories Dubuque, IA 52001 Ph. 563-690-4142 NOTICE: This email may contain legally privileged information. The information is for the use of only the intended recipient(s) even if addressed incorrectly. If you are not the intended recipient, please notify the sender that you have received it in error and then delete it along with any attachments. Thank you. NOTICE: This email may contain legally privileged information. The information is for the use of only the intended recipient(s) even if addressed incorrectly. If you are not the intended recipient, please notify the sender that you have received it in error and then delete it along with any attachments. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine blue stain for MMA
Hi, is there anyone have an experience with MMA toluidine staining? Im using T7200, T9100, Osteo-bed resin in lab now. Thanks, Kai Research Histotechnologist ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Toluidine blue stain for MMA
Yes. Probably hundreds of Histonetters stain plastic sections. Let us all hope they don't all bombard the Histonet listserver with replies to your question. Instructions for staining plastic sections with toluidine blue are in every library that contains books with paper pages, and also (albeit with less authority) in great abundance on the Web. Try typing SEMITHIN STAIN into Google. I just did, and an excellent web site came up on top of the heap. John Kiernan = = = On 27/07/15, Kai Hong via Histonet histonet@lists.utsouthwestern.edu wrote: Hi, is there anyone have an experience with MMA toluidine staining? Im using T7200, T9100, Osteo-bed resin in lab now. Thanks, Kai Research Histotechnologist ___ 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] 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] 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
[Histonet] Toluidine blue background
We would like to use Toluidine Blue for staining frozen sections of kidney biopsy. We finally are able to get the stain dark enough, but we can't get rid of the background caused by the OCT.We've washed with water, alcohols, and PBS with no success. Is anyone out there using Toluidine Blue for frozen staining? If so, what is your recipe and how do you get rid of the background? Thanks Beth Ann O'Neil, MT(ASCP)SC, HTL, QIHC one...@wvuhealthcare.commailto:one...@wvuhealthcare.com Histology Supervisor, Technical Specialist Lab: 304 - 293 - 6014 Office: 304 - 293 - 7629 - 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. 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
[Histonet] Toluidine Blue stain for Mohs
Hi All, I am in need of a Toluidine Blue stain for Moh's Lab, would anyone be willing to share with me? It is for a friend. In advance, thank you very much. Debbie Siena 800.442.3573 ext. 229 | www.statlab.comhttp://www.statlab.com/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine blue
Henk Al weer even geleden: weet je al iets over de toepassing van CD4 op varkensweefsel formaline gefixeerd en in paraffine ingebed weefsel? Heb nog een vraag: zijn jullie in het bezit van een cryostaat waar je nog met ouderwetse vaste messen kan snijden? Ik ben op zoek naar de mogelijkheid om m.b.v. een D-mes bothoudend/verkalkte weefsels te snijden. We hebben hier een microm, maar die is zoals die hier nu staat alleen toepasbaar voor disposable mesjes. Een aanpassing om vaste messen te gaan gebruiken kost een ruime 1000 euro en als het niet zou werken is dat zonde-geld. Alvast bedankt Joost TNO.NLhttp://www.tno.nl/ Joost Bruijntjes T +31 88 866 17 38 F +31 30 694 49 86 E joost.bruijnt...@tno.triskelion.nlmailto:joost.bruijnt...@tno.triskelion.nl Disclaimerhttp://www.tno.nl/tno/email/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Toluidine blue
Anyone know Dutch? From: histonet-boun...@lists.utsouthwestern.edu on behalf of Bruijntjes, J.P. (Joost) Sent: Fri 12/9/2011 4:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Toluidine blue Henk Al weer even geleden: weet je al iets over de toepassing van CD4 op varkensweefsel formaline gefixeerd en in paraffine ingebed weefsel? Heb nog een vraag: zijn jullie in het bezit van een cryostaat waar je nog met ouderwetse vaste messen kan snijden? Ik ben op zoek naar de mogelijkheid om m.b.v. een D-mes bothoudend/verkalkte weefsels te snijden. We hebben hier een microm, maar die is zoals die hier nu staat alleen toepasbaar voor disposable mesjes. Een aanpassing om vaste messen te gaan gebruiken kost een ruime 1000 euro en als het niet zou werken is dat zonde-geld. Alvast bedankt Joost TNO.NLhttp://www.tno.nl/ Joost Bruijntjes T +31 88 866 17 38 F +31 30 694 49 86 E joost.bruijnt...@tno.triskelion.nlmailto:joost.bruijnt...@tno.triskelion.nl Disclaimerhttp://www.tno.nl/tno/email/ ___ 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
Yes, I met him in a bar in old Soho.. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: 09 December 2011 14:48 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Toluidine blue Anyone know Dutch? From: histonet-boun...@lists.utsouthwestern.edu on behalf of Bruijntjes, J.P. (Joost) Sent: Fri 12/9/2011 4:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Toluidine blue Henk Al weer even geleden: weet je al iets over de toepassing van CD4 op varkensweefsel formaline gefixeerd en in paraffine ingebed weefsel? Heb nog een vraag: zijn jullie in het bezit van een cryostaat waar je nog met ouderwetse vaste messen kan snijden? Ik ben op zoek naar de mogelijkheid om m.b.v. een D-mes bothoudend/verkalkte weefsels te snijden. We hebben hier een microm, maar die is zoals die hier nu staat alleen toepasbaar voor disposable mesjes. Een aanpassing om vaste messen te gaan gebruiken kost een ruime 1000 euro en als het niet zou werken is dat zonde-geld. Alvast bedankt Joost TNO.NLhttp://www.tno.nl/ Joost Bruijntjes T +31 88 866 17 38 F +31 30 694 49 86 E joost.bruijnt...@tno.triskelion.nlmailto:joost.bruijnt...@tno.triskelion.nl Disclaimerhttp://www.tno.nl/tno/email/ ___ 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] Toluidine blue
I put it in a translator program and it's something like this: long time ago: you know anything about the application of cd4 on varkensweefsel formalin fixed and in paraffin embedded fabric? have another question: are you in the possession of a cryostat where you still with old-fashioned fixed knives, cutting up? I am trying to the possibility of using a D-Knife bothoudend/i fabrics to cut off. We have here a microm, but which is as in here and now only applicable for consumer blades. Babylon 9 Bobbie -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Friday, December 09, 2011 9:48 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Toluidine blue Anyone know Dutch? From: histonet-boun...@lists.utsouthwestern.edu on behalf of Bruijntjes, J.P. (Joost) Sent: Fri 12/9/2011 4:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Toluidine blue Henk Al weer even geleden: weet je al iets over de toepassing van CD4 op varkensweefsel formaline gefixeerd en in paraffine ingebed weefsel? Heb nog een vraag: zijn jullie in het bezit van een cryostaat waar je nog met ouderwetse vaste messen kan snijden? Ik ben op zoek naar de mogelijkheid om m.b.v. een D-mes bothoudend/verkalkte weefsels te snijden. We hebben hier een microm, maar die is zoals die hier nu staat alleen toepasbaar voor disposable mesjes. Een aanpassing om vaste messen te gaan gebruiken kost een ruime 1000 euro en als het niet zou werken is dat zonde-geld. Alvast bedankt Joost TNO.NLhttp://www.tno.nl/ Joost Bruijntjes T +31 88 866 17 38 F +31 30 694 49 86 E joost.bruijnt...@tno.triskelion.nlmailto:joost.bruijntjes@tno.triskelio n.nl Disclaimerhttp://www.tno.nl/tno/email/ ___ 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] Toluidine Blue
Henk - Mast cells, on tissue and/or cytospins, are pH dependent - 2.0-2.5. Tissues are usually already fixed in formalin; cytospins can be fixed in 95% alcohol or formalin. I am sending our directions separately. The stain should less than 5 minutes totally. Cheryl ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine blue
Hi All, Have someone a good workong protocol for staining mastcells in cytospin slides with Toluidine Blue. Mention also the pH and fixation. Thanks, Henk van de Kant Utrecht University, Faculty of Science, Division of Pharmacology, David de Wied Building, Universiteitsweg 99, 3584 CG, Utrecht The Netherlands ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine blue in decaled bone
We may need to do toluidine blue staining to assess cartilage changes on FFPE EDTA decalcified rat joints. We don't have experience w/ T blue and I have read the EDTA will extract proteoglycans. Help? Thanks, Brett Brett M. Connolly, Ph.D. Molecular Imaging Team Leader Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_conno...@merck.com T- 215-652-2501 F- 215-993-6803 Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine blue staining of CFU-F
Hi all, I'm looking for a protocol to counterstain mouse CFU-F grown in tissue culture**. The protocols I've found just say they counterstained with this dye, don't say what it's dissolved in, sometimes list the percentage of it, and never list the time they stained for. The only full protocol I found was for staining of mast cells in sections, but that's dissolved in 70% ethanol and very acidified. I'm not sure that's what I want. Has anyone done this before? Thanks, Adam ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Toluidine Blue and staining archaeological residues.
Hello Paula, Thanks for the reply. We do use IKI regualarly for starch staining, however once the slide is stained it is difficult to counterstain to detect animal residues which might be present. The residues from grindstones have lost a lot of their microsopic diagnostic properties and we are hponing that staining the residues will help differentiate between intensive specialised seed grinders versus multi-purpose expedient grinders. I am finding that the Phloroglucinol stain is light sensitive and therefore not permanent which allows for some counterstaining but still the technique is a work in progress. Any more thoughts most welcome. Regards Birgitta Stephenson Research Microscopy Lab, University of Queensland On Tue, 20 Jul 2010 06:51:09 -0700 (PDT), Paula Pierce cont...@excaliburpathology.com said: Starch (plants) + Iodine = Black solution. From: Birgitta Stephenson bstep...@fastmail.fm To: histonet@lists.utsouthwestern.edu Sent: Mon, July 19, 2010 8:19:27 PM Subject: [Histonet] Toluidine Blue and staining archaeological residues. Hello Histonetters, I am working with archaeological residues lifted from Holocene grindstones. I was trying to find a stain that could differentiate between plant and animal tissue in one hit. I have been using buffered Toluidine Blue solutions however given that these are ancient residues which are lifted using 20ul of water, the subtle colour differences between blues, purples, and violets have not been useful. I was looking at the possibility of counterstaining the Toluidine Blue stained residues with say Phloroglucinol or IKI which would highlight the plant component of the residue and then see if what was left looked like collagen. Has anyone tried this type of counterstaining? OR does anyone know of a stain that would colour differently for plant and animals understanding that this is not a tissue section but microscopic residues in solution? Thanks Birgitta Stephenson The Research Microscopy Laboratory, University of Queensland. -- Birgitta Stephenson bstep...@fastmail.fm -- http://www.fastmail.fm - Access your email from home and the web ___ 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 -- Birgitta Stephenson bstep...@fastmail.fm -- http://www.fastmail.fm - Access your email from home and the web ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Toluidine Blue and staining archaeological residues.
Starch (plants) + Iodine = Black solution. From: Birgitta Stephenson bstep...@fastmail.fm To: histonet@lists.utsouthwestern.edu Sent: Mon, July 19, 2010 8:19:27 PM Subject: [Histonet] Toluidine Blue and staining archaeological residues. Hello Histonetters, I am working with archaeological residues lifted from Holocene grindstones. I was trying to find a stain that could differentiate between plant and animal tissue in one hit. I have been using buffered Toluidine Blue solutions however given that these are ancient residues which are lifted using 20ul of water, the subtle colour differences between blues, purples, and violets have not been useful. I was looking at the possibility of counterstaining the Toluidine Blue stained residues with say Phloroglucinol or IKI which would highlight the plant component of the residue and then see if what was left looked like collagen. Has anyone tried this type of counterstaining? OR does anyone know of a stain that would colour differently for plant and animals understanding that this is not a tissue section but microscopic residues in solution? Thanks Birgitta Stephenson The Research Microscopy Laboratory, University of Queensland. -- Birgitta Stephenson bstep...@fastmail.fm -- http://www.fastmail.fm - Access your email from home and the web ___ 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] Toluidine Blue and staining archaeological residues.
Hello Histonetters, I am working with archaeological residues lifted from Holocene grindstones. I was trying to find a stain that could differentiate between plant and animal tissue in one hit. I have been using buffered Toluidine Blue solutions however given that these are ancient residues which are lifted using 20ul of water, the subtle colour differences between blues, purples, and violets have not been useful. I was looking at the possibility of counterstaining the Toluidine Blue stained residues with say Phloroglucinol or IKI which would highlight the plant component of the residue and then see if what was left looked like collagen. Has anyone tried this type of counterstaining? OR does anyone know of a stain that would colour differently for plant and animals understanding that this is not a tissue section but microscopic residues in solution? Thanks Birgitta Stephenson The Research Microscopy Laboratory, University of Queensland. -- Birgitta Stephenson bstep...@fastmail.fm -- http://www.fastmail.fm - Access your email from home and the web ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Toluidine blue
Just be sure you get dye certified by the Biological Stain Commission. The bottle must have a Certification label or it is not certified to perform as expected. Sigma is reliable. Geoff zodia...@comcast.net wrote: Hello to all, I was wondering if anyone knows where to purchase toluidine blue (for mast cells) other than polyscientific. Thank you in advance for your replies, Jenny ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine blue
Hello to all, I was wondering if anyone knows where to purchase toluidine blue (for mast cells) other than polyscientific. Thank you in advance for your replies, Jenny ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Toluidine blue
Newcomer Claire From: histonet-boun...@lists.utsouthwestern.edu on behalf of zodia...@comcast.net Sent: Wed 4/21/2010 5:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Toluidine blue Hello to all, I was wondering if anyone knows where to purchase toluidine blue (for mast cells) other than polyscientific. Thank you in advance for your replies, Jenny ___ 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] Toluidine Blue
I am currently writing a procedure for the use of Toluidine Blue staining for both FNA's and frozen sections at the request of one of our pathologists. Can anyone guide me in the use of this stain in lieu of Diff Quick or HE for rapid processing (such as references I could use, etc) and does anyone have a procedure I can cite for permanent mounting/storage of these slides? Thanks in advance, Edie Lehman MT(ASCP) Anatomical Pathology Supervisor ed...@fmchealth.org Fairfield Medical Center People you know. Care you trust. Visit us at http://www.fmchealth.org or our online store at http://fairfield.thehospitalstore.com 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. 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
[Histonet] Toluidine blue staining
Hello. Our lab has had highly variable results using toluidine blue to stain 1.5 micron thick cross sections of plastic embedded nerve tissue from both mouse and human nerve biopsy. It seems like the stain is darker in the biopsy tissue than the mouse but both sometimes stain equally poorly. We have tried changing the protocol recipe to make it darker by adding more toluidine blue powder but that has not worked either. We have used 2 grams toluidine blue, and 2 grams of sodium borax with 100 ml of distilled water. We have tried changing the sodium borax. Sodium tetraborate-decahydrate and a sodium tertaborate 99.9% metals basis from sigma Aldrich have both had the same results. Does anyone have any idea how to make it stain darker? I can't really emphasize enough how light the blue is. I did read one protocol that involved adjusting pH, is that a possible factor? Thanks Christopher J. Edwards Research Associate, Sahenk Lab Center for Gene Therapy Wexner Building 2, Room 3224 Lab Phone: 355-5224 - 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
Re: [Histonet] Toluidine blue staining
To get consistent staining you must standardize your procedure. I make 1% aqueous Tol. Blue and 2% aqueous borax (sodium tetraborate) in separate containers. Each day I mix equal parts of each solution so I always use fresh stain. My final conc. of stain is 0.5%, doubling to 1% makes no difference. Stain at consistent temperature in a water bath (60-65 degrees C) for a consistent time. Not dark enough? Stain longer and/or at a higher temp. When I stopped using a drop (how big?) of stain on a hotplate (how hot?) until I though it was stained (how long?) I got much better results. Also, are the biopsy and the mouse tissue fixed and processed identically? Geoff Edwards, Chris wrote: Hello. Our lab has had highly variable results using toluidine blue to stain 1.5 micron thick cross sections of plastic embedded nerve tissue from both mouse and human nerve biopsy. It seems like the stain is darker in the biopsy tissue than the mouse but both sometimes stain equally poorly. We have tried changing the protocol recipe to make it darker by adding more toluidine blue powder but that has not worked either. We have used 2 grams toluidine blue, and 2 grams of sodium borax with 100 ml of distilled water. We have tried changing the sodium borax. Sodium tetraborate-decahydrate and a sodium tertaborate 99.9% metals basis from sigma Aldrich have both had the same results. Does anyone have any idea how to make it stain darker? I can't really emphasize enough how light the blue is. I did read one protocol that involved adjusting pH, is that a possible factor? Thanks Christopher J. Edwards Research Associate, Sahenk Lab Center for Gene Therapy Wexner Building 2, Room 3224 Lab Phone: 355-5224 - 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 -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine Blue stain
Jen - We do T=blue staining almost daily. If you will contact me directly I will send you our directions. It is a very inexpensive stain, and can be reused. You will need to pH it often. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
