Re: [Histonet] VonKossa's calcium stain. Counterstain etc
Any haemalum will do, but blue is not the most pleasing contrast to go with the black silver deposits. Pink or light red is better. Here are three pink to red counterstains, all traditional. Check in a textbook or manual for instructions. 1. Neutral red (CI 50040) is good: 0.5% in water; adjust to pH4 with acetic acid; stain for about 2 minutes; the solution keeps for at least 5 years and can be used repeatedly. 2. Safranine O (CI 50240) can be used similarly but needs a longer staining time. 3. Nuclear fast red (CI 60760) is also OK: NFR 0.2G, aluminium sulphate crystals 10G, water 200ml; heat until it boils, cool overnight, decant and filter; stain for 5-10min. The solution is good for about a year; always filter before using. The Biological Stain Commission has standards for certification of all three of these dyes. Certified neutral red and safranine O have been available for many (50+) years. Nuclear fast red was only recently added to the BSC's list. See Frank et al 2007. Certification procedures for nuclear fast red (Kernechtrot), C.I. 60760. Biotechnic Histochemistry 82: 35-39. Certified NFR powder may not yet be available to labs or to vendors of stain solutions. If you buy a ready-made solution of any dye you should choose one that was made from a BSC-certified batch of the powder. To any vendors of dye powders who read this message: email me for more information about the certification criteria for nuclear fast red. Also, check out http://www.biologicalstaincommission.org and click on one of the Vendors tabs or links. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: karine cadoret kcado...@amc.edu.au Date: Thursday, July 16, 2009 22:36 Subject: [Histonet] VonKossa's calcium stain To: histonet@lists.utsouthwestern.edu Hi, When doing a VonKossa stain in order to demonstrate calcium in tissue, does it matter much if I use Mayer's hematoxylin instead of Ehrlich's hematoxylin (which takes 6 months to ripen) ? Also, can I simply use homemade scott's tapwater for blueing instead of using a lithium carbonate solution ? Thank you for your help, Karine Cadoret Fish health laboratory manager National Center for Marine Conservation and Resource Sustainability Newnham, TAS Australia ___ 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] VonKossa's calcium stain
Hi, When doing a VonKossa stain in order to demonstrate calcium in tissue, does it matter much if I use Mayer's hematoxylin instead of Ehrlich's hematoxylin (which takes 6 months to ripen) ? Also, can I simply use homemade scott's tapwater for blueing instead of using a lithium carbonate solution ? Thank you for your help, Karine Cadoret Fish health laboratory manager National Center for Marine Conservation and Resource Sustainability Newnham, TAS Australia ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] VonKossa's calcium stain
Karine, Either Hx will do, though I would not have used a haematoxylin since it will lake with the calcium forming a blue stained deposit. I would expect it to mask the silver of the von-kossa stain. I would recommend 1% neutral red, ethylene green or even a light eosin counterstain. The silver stained calcium deposits should then stand out quite well. 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 karine cadoret Sent: Friday, 17 July 2009 12:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] VonKossa's calcium stain Hi, When doing a VonKossa stain in order to demonstrate calcium in tissue, does it matter much if I use Mayer's hematoxylin instead of Ehrlich's hematoxylin (which takes 6 months to ripen) ? Also, can I simply use homemade scott's tapwater for blueing instead of using a lithium carbonate solution ? Thank you for your help, Karine Cadoret Fish health laboratory manager National Center for Marine Conservation and Resource Sustainability Newnham, TAS Australia ___ 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] VonKossa's calcium stain
What is your tissue of interest? Why not do the Von Kossa stain first and then counterstain with MacNeal's tetrachrome. This way you employ the use of a metachromatic stain for the rest of the tissue instead of just a nuclear staining hematoxylin. Jack On Jul 16, 2009, at 9:34 PM, karine cadoret kcado...@amc.edu.au wrote: Hi, When doing a VonKossa stain in order to demonstrate calcium in tissue, does it matter much if I use Mayer's hematoxylin instead of Ehrlich's hematoxylin (which takes 6 months to ripen) ? Also, can I simply use homemade scott's tapwater for blueing instead of using a lithium carbonate solution ? Thank you for your help, Karine Cadoret Fish health laboratory manager National Center for Marine Conservation and Resource Sustainability Newnham, TAS Australia ___ 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
