RE: [Histonet] Formic Acid Recipe
Jesus, The formic acid (5%) will decalcify the hydroxyapatite which can be very hard and crunchy to cut but since you ground the section I guess you don't care about that. I would be concerned though that the formic acid might also cause you to lose staining of some cell components that depend on the calcium. The methyl methacrylate can be removed from the section by xylene like deparaffinizing paraffin sections. It works for methyl methacrylate but nothing removes glycol methacrylate. Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jesus Hernandez Sent: Friday, November 04, 2011 3:41 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Formic Acid Recipe Dear all, I was having trouble staining Human embryonic palatal mesenchymal cells with multi-stain solution. My PI told me to use formic acid on the samples so that it could increase the permeability of the methyl methacrylate. I am not sure if this is enough information, but the cells were loaded on hydroxyapatite scaffolds. I have also tried aniline blue and villaneuva stain. Any information on how I can make a solution of formic acid is appreciated along with maybe other stains I could try using. Each sample was grinded to about 50 microns. Thank you. Best Regards, Jesus W. Hernandez Graduate Student Department of Biomedical Engineering University of Texas at San Antonio jesus.w@gmail.com ___ 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] low profile vs. hi profile blades
It depends on the microtome being used and how the blade holder is set up to take high profile or low, many of them (at least the microms I have can take either) on my microm to use low profile I leave the bar on the knife holder so the holder is set to hold shorter low profile blades, I can take that bar out with a couple of little screws on each side which deepens the knife holder space so it can take high profile knives. One of my cryostats knife holder only takes high profile blades so I like to use Sturkey gold high profile there, the other cryostat I have only takes low profile blades. Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nancy Schmitt Sent: Friday, November 04, 2011 7:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] low profile vs. hi profile blades Hi- Please educate me on this. There seems to be a lot of personal opinion involved in regards to which type of blade to use. Thanks for your help Nancy 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] low profile vs. hi profile blades
In my experience even with the harder high profile blades for denser tissues such as bone I still prefer to use permanent tungsten carbide blades I have to send out to get sharpened. The high profile disposable blades seem to vibrate too much for me even the heavier tungsten ones for bone. Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Friday, November 04, 2011 8:17 AM To: Nancy Schmitt; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] low profile vs. hi profile blades In my opinion it doesn't make a lot of difference which type you use, since all you actually use is the 2mm or so that extends beyond the edge of the knife holder. What is crucial is that you use the type of blade your knife holder is designed for. If you try to use high profile blades in a low profile knife holder, or vice versa, you will have major problems. Also, high profile blades are available in a heavier, less flexible form for cutting denser tissues. I don't think the low profile blades offer that option, though I am not certain since I use only the high profile format. ___ 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] Training and Competency Assessment for HE Slide Review
This was what I was going to suggest, I bet they let you participate in HistoQip even if you are from Canada. Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Janice Mahoney Sent: Friday, November 04, 2011 10:10 AM To: joellewea...@hotmail.com; diana.har...@viha.ca; histo net Subject: RE: [Histonet] Training and Competency Assessment for HE Slide Review A great way to do this is by using the results of the Histo QIP from CAP and the NSH. If you participate you get wonderful study and competency materials to use for routine HE's special stains and IHC. Jan Mhaoney Omaha From: joellewea...@hotmail.com To: diana.har...@viha.ca; histonet@lists.utsouthwestern.edu Date: Thu, 3 Nov 2011 17:42:53 + Subject: Re: [Histonet] Training and Competency Assessment for HE Slide Review CC: Any certified histologist has gone through this, but the NSH has good resources for this. Sent from my Verizon Wireless BlackBerry -Original Message- From: Harris Diana diana.har...@viha.ca Date: Thu, 3 Nov 2011 17:34:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Training and Competency Assessment for HE Slide Review Any suggestions for creating a Training and Competency (TC) Assessment for HE slide review? We currently QC all HE slides macroscopically and 15% microscopically. I would like to have all Histotechs trained and competent to QC HE slides. Has anyone gone thru this process? Thanks Diana Harris QC Method Development Technologist Royal Jubilee Hospital Victoria, BC Canada ___ 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] Re: Celestin blue B (was Help)
Corrie Vernick writes: I am currently a histology student at Keiser University. I am doing a project for my routine staining class about Celestine Blue. I've been able to find information on why it was created, the chemical make up, and some of it's uses including the trichrome stain. I am having trouble finding images of slides stained with Celestine Blue. Any additional information about the uses would be helpful as well! Thank you, Corrinne Vernick, Keiser University FL U.S.A. I don't have access to my library this week, but you can get a good bit information by Googling celestin blue B. This dye was often used as a sort of backup or substitute for hematoxylin, particularly in the old outmoded Pearse stain for pituitary cells. R.D. Lillie as I remember didn't think much of the dye, and I don't think this dye is a very good topic for a study such as the one you describe. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: processing program for biopsies
Only 40 minutes in formalin is a problem. Increase the formalin time to several hours and this should suffice for small 2-3mm punch biopsies of skin. For larger skin sections the alcohol, xylene and wax times will need to be tripled at least (as well as fixing the skins for longer (3-4 hours on the machine would be great)). 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 Carol Bryant Sent: Saturday, 5 November 2011 3:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing program for biopsies Importance: High Would you experienced histotechs mind to look at this program and see if it would cause skin biopsies to be poorly processed? I think too much time in alcohols. The pathologists say they look bad. Thanks in advance for your help! Station SolutionTime Temp P/VMix --- -- - - - -- 1Formalin 0:20 OFF P/VSlow 2Formalin 0:20 OFF P/VSlow 370% Alcohol 0:15 OFF P/VSlow 480% Alcohol 0:15 OFF P/VSlow 595% Alcohol 0:15 OFF P/VSlow 695% Alcohol 0:25 OFF P/VSlow 7100% Alcohol 0:25 OFF P/VSlow 8100% Alcohol 0:25 OFF P/VSlow 9Xylene 0:35 OFF P/VSlow 10Xylene 0:35 OFF P/VSlow 11Paraffin 0:15 63C. P/VSlow 12Paraffin 0:15 63C. P/VSlow 13Paraffin 0:15 63C. P/VSlow 14Paraffin 0:15 63C. P/VSlow Carol Bryant, CT (ASCP) Cytology/Histology Manager Lexington Clinic Phone (859) 258-4082 Fax (859) 258-4081 cb...@lexclin.com NOTICE OF CONFIDENTIALITY This message, including any attachments, is intended only for the sole use of the addressee and may contain confidential or privileged information that is protected by the State of Kentucky and/or Federal regulations. If you are not the intended recipient, do not read, copy, retain or disseminate this message or any attachment. If you have received this message in error, please call the sender immediately at (859)258-4000 and delete all copies of this message and any attachment. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. Neither the transmission of this message or any attachment, nor any error in transmission or misdelivery shall constitute waiver of any applicable legal privilege. ___ 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
Re: [Histonet] Re: Celestin blue B (was Help)
With the hematoxylin shortage of a couple of years ago (real, not imagined in about 2007-2008), several companies tried to come up with a synthetic dye substitute. A little background: Celestine blue (CI 51050, also known as Mordant blue 14) is a substitute touted many years ago (late 1960s/early 1970s if I remember, when there was another shortage for different reasons). I've used it years ago, in a stain called MSB for fibrin (look in a Bancroft book). The MSB stain used a double nuclear stain of aluminum hematoxylin (e.g. Mayer or Harris) and celestine blue. We didn't like the celestine blue, because when we mixed the iron mordant salt with the celestine blue dye, it worked right away, but, the next time someone asked for a MSB stain (several months later), the celestine blue was overoxidized, and wouldn't work. So we would have to stop and make the dye up immediately. It was a nuisance. But the double nuclear stain to to try to keep the nuclei a blue color, after going through the next 3 dyes (MSB is sort of a quatrachrome). Eventually, we just started using the Weigert hematoxylin from the trichrome stain in place of the double nuclear stain in the MSB. Our pathologists like the MSB stain better with the Weigert hematoxylin. Current use: Since 2008, three companies that I know of came up with hematoxylin substitutes, in response to that shortage. - Anatech Ltd, which used Mordant blue 3, CI 43820, and called it Tango Blue. So this is NOT celestine blue. http://www.anatechltdusa.com/MSDS_pdf/TangoStainMSDS.pdf - Newcomer Supply, which calls their substitute Newly Blue, but says it is a Celestine blue in one solution, ferric ammonium sulfate in the other solution. I don't know if you mix the two solutions before use, or if you dip the slides in first one solution and then the other. I didn't get around to their booth this year at NSH (sorry Marcia), and have never seen a procedure sheet on this stain. http://www.newcomersupply.com/products/standard-special-stains?page=N#181 - ThermoFisher/Richard Allan, which calls their substitute Phoenix blue.The problem is, they are being secretive (proprietary) about what dye/reagents are in their solutions. I looked at one time, and couldn't find MSDS on their websites And their flyer doesn't mention the dyes name or CI #. However, the photo on the flyer, at least to me, looks like celestine blue. That's not proof that it IS celestine blue. Could be a close look-alike cousin. http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_6906.pdf So look to these companies for more information. Anatech has a newsletter, available on their website, about the hematoxylin shortage (and their Tango blue product, of course). (Not that this will help you with your Celestine blue project, but it will fill you in on the hematoxylin shortage history.) http://www.anatechltdusa.com/Innovators/Inn08Summer.pdf Hope that helps some. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology Beaumont Hospital Royal Oak, MI 48073 (Opinions expressed are my own, and not Beaumont Hospitals'.) -Original Message- From: Bob Richmond Sent: Sunday, November 06, 2011 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Celestin blue B (was Help) Corrie Vernick writes: I am currently a histology student at Keiser University. I am doing a project for my routine staining class about Celestine Blue. I've been able to find information on why it was created, the chemical make up, and some of it's uses including the trichrome stain. I am having trouble finding images of slides stained with Celestine Blue. Any additional information about the uses would be helpful as well! Thank you, Corrinne Vernick, Keiser University FL U.S.A. I don't have access to my library this week, but you can get a good bit information by Googling celestin blue B. This dye was often used as a sort of backup or substitute for hematoxylin, particularly in the old outmoded Pearse stain for pituitary cells. R.D. Lillie as I remember didn't think much of the dye, and I don't think this dye is a very good topic for a study such as the one you describe. Bob Richmond Samurai Pathologist Knoxville TN ___ 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] Microwave processing
Hello, I am a histology technology student and I need information on the melting point of the paraffin used in Microwave Processing… Freida Carson stated in her book something regarding the paraffin temperature set at 84 degrees. The class had a discussion regarding the same and concluded that the tissue morphology would be damage at such high temperature. Can someone please provide me with some input on this issue. Thank you, Jamie G. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] microwave processing
Hi there, On an Instrumentation course at keiser, we had a discussion about the right temperature fro the paraffin when the proceesing is done using microwaves. Can some tell me what should be the ideal temperature with out damaging the processing. The text book mentions 84 degrees, but isn't this to high; perhaps if the time is less, it wont affect it? Thanks, Ismael Lopez ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] microwave processing
I believe that the higher temperature is needed to evaporate off the isopropanol. Just make sure that your tissue is well fixed. 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 Zoe rosa Sent: Monday, 7 November 2011 1:41 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microwave processing Hi there, On an Instrumentation course at keiser, we had a discussion about the right temperature fro the paraffin when the proceesing is done using microwaves. Can some tell me what should be the ideal temperature with out damaging the processing. The text book mentions 84 degrees, but isn't this to high; perhaps if the time is less, it wont affect it? Thanks, Ismael Lopez ___ 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
Re: [Histonet] Re: Celestin blue B (was Help)
Hello I have used Celestin Blue-Haemalum (Mayers) considerably in the past as a substitute for Weigert's and as you say the Celestin Blue stain does not keep and is best made fresh for each use. Our method to simplify this was to make separate double strength solutions of the Celestin blue and the mordant which were mixed 50/50 just prior to use to make a solution of the correct strength. Both of these solutions store well and the preparation time is minimal. regards Tony Tony Reilly B.App.Sc. , M.Sc. Chief Scientist, Anatomical Pathology Pathology Queensland-PA Laboratory _ Clinical and Statewide Services Division| QueenslandHealth Level 1, Building 15,Princess Alexandra Hospital Ipswich Road,WOOLLOONGABBA Qld4102 Ph: 07 3176 2412 Mob: 0402 139411 Fax: 07 3176 2930 Email: tony_rei...@health.qld.gov.au Web: www.health.qld.gov.au/qhcss/ Lee Peggy Wenk lpw...@sbcglobal.net 11/7/2011 8:36 am With the hematoxylin shortage of a couple of years ago (real, not imagined in about 2007-2008), several companies tried to come up with a synthetic dye substitute. A little background: Celestine blue (CI 51050, also known as Mordant blue 14) is a substitute touted many years ago (late 1960s/early 1970s if I remember, when there was another shortage for different reasons). I've used it years ago, in a stain called MSB for fibrin (look in a Bancroft book). The MSB stain used a double nuclear stain of aluminum hematoxylin (e.g. Mayer or Harris) and celestine blue. We didn't like the celestine blue, because when we mixed the iron mordant salt with the celestine blue dye, it worked right away, but, the next time someone asked for a MSB stain (several months later), the celestine blue was overoxidized, and wouldn't work. So we would have to stop and make the dye up immediately. It was a nuisance. But the double nuclear stain to to try to keep the nuclei a blue color, after going through the next 3 dyes (MSB is sort of a quatrachrome). Eventually, we just started using the Weigert hematoxylin from the trichrome stain in place of the double nuclear stain in the MSB. Our pathologists like the MSB stain better with the Weigert hematoxylin. Current use: Since 2008, three companies that I know of came up with hematoxylin substitutes, in response to that shortage. - Anatech Ltd, which used Mordant blue 3, CI 43820, and called it Tango Blue. So this is NOT celestine blue. http://www.anatechltdusa.com/MSDS_pdf/TangoStainMSDS.pdf - Newcomer Supply, which calls their substitute Newly Blue, but says it is a Celestine blue in one solution, ferric ammonium sulfate in the other solution. I don't know if you mix the two solutions before use, or if you dip the slides in first one solution and then the other. I didn't get around to their booth this year at NSH (sorry Marcia), and have never seen a procedure sheet on this stain. http://www.newcomersupply.com/products/standard-special-stains?page=N#181 - ThermoFisher/Richard Allan, which calls their substitute Phoenix blue.The problem is, they are being secretive (proprietary) about what dye/reagents are in their solutions. I looked at one time, and couldn't find MSDS on their websites And their flyer doesn't mention the dyes name or CI #. However, the photo on the flyer, at least to me, looks like celestine blue. That's not proof that it IS celestine blue. Could be a close look-alike cousin. http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_6906.pdf So look to these companies for more information. Anatech has a newsletter, available on their website, about the hematoxylin shortage (and their Tango blue product, of course). (Not that this will help you with your Celestine blue project, but it will fill you in on the hematoxylin shortage history.) http://www.anatechltdusa.com/Innovators/Inn08Summer.pdf Hope that helps some. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology Beaumont Hospital Royal Oak, MI 48073 (Opinions expressed are my own, and not Beaumont Hospitals'.) -Original Message- From: Bob Richmond Sent: Sunday, November 06, 2011 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Celestin blue B (was Help) Corrie Vernick writes: I am currently a histology student at Keiser University. I am doing a project for my routine staining class about Celestine Blue. I've been able to find information on why it was created, the chemical make up, and some of it's uses including the trichrome stain. I am having trouble finding images of slides stained with Celestine Blue. Any additional information about the uses would be helpful as well! Thank you, Corrinne Vernick, Keiser University FL U.S.A. I don't have access to my library this week, but you can get a good bit information by Googling celestin blue B. This dye was often used as a sort of backup or substitute for hematoxylin, particularly in the old outmoded Pearse stain for
