AW: [Histonet] Bone/cartilage/epithelial tissue stain
Rui, in your first post about your problem, you wrote about decalcification of your samples. Are these samples, where you want to demonstrate bone, decalcified? Which decalcifier? Haematoxylin stains only non-decalcified bone, due to the Ca-ions. Strong acids for decalcification (like HNO3 or HCl) may alter the tissue-stainability. They are said to solve some amount of proteoglycans. If they are not decalcified, you can combine a von-Kossa-stain with a trichrome-stain or alcianblue. Calcified bone stains black. An easy stain would be the combined Alcianblue-PAS. Alcianblue demonstrates acid proteoglycans, PAS demonstrates neutral glycoproteins like collagen. But I think, proteoglycans and collagen are in such a tight junction in cardilage, that it will give a mixed dye-appearance. But worth a try. Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Rui TAHARA Gesendet: Freitag, 23. August 2013 09:49 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Bone/cartilage/epithelial tissue stain Hi, Would anyone suggest me what staining is best to color differentiate between cartilage and bone and epithelial tissues in avian embryos? I have been trying Mallory Trichrome for embryos but recently I was suggested that Mallory Trichrome stains cartilage differently in embryos compared to adult samples since Aniline blue stains fiber that may not develop in early embryos. There is some protocol that modified the Mallory Trichrome that could be applied to embryos. However, the resulting colors of all tissues look all purple-ish and difficult to tell the cartilage from the weak blue stain from aniline blue. Currently I am thinking to try out Alcian blue/Hematoxylin and Eosin stain (Ehrlichs hematoxylin). The purpose of the staining is to look at interaction between ossification and epithelial development so I think alcian blue for staining cartilage works but I am wondering if there is any other staining combination with alcian blue exist for visualizing bone and epithelial tissue (e,g. alcian blue/alizarine red with other staining?). Any suggestion would be appreciated! Rui TAHARA PhD Candidate Biology Department McGill University ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Fixing whole testes
I would immediately bisect them longitudinally with a sharp blade, then continue to fix for at least two days ( length of fixation time is irrelevant for anatomical purposes). You should have done much earlier ( sure, you run the risk of loss of anatomical relationships but.I can't see what choice you have, given the size of the tissues. I am always interested in others' views. Yes and, fix on a rocker roller at RT. A faster penetration fixing fluid would have been betteryou would have to test a few, under your conditions. NB: Formalin penetrates a 1cm cube of gelatin 250mm in 24hrs so, penetration through a dense heterogeneous tissue, especially the initital penetration of the T albuginea, will be much slower, allowing for autolytic ( necrotic, really, as the tissue is dead) changes towards the centre of the tissue. Best wishes, Carl Carl Hobbs FIBMS Histology and Imaging Manager Wolfson CARD Guys Campus, London Bridge Kings College London London SE1 1UL 020 7848 6813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Fixing whole testes
My sincere apologies...I left the HTML option on, for the 1st post. I would immediately bisect them longitudinally with a sharp blade, then continue to fix for at least two days ( length of fixation time is irrelevant for anatomical purposes). You should have done much earlier ( sure, you run the risk of loss of anatomical relationships but.I can't see what choice you have, given the size of the tissues. I am always interested in others' views. Yes and, fix on a rocker roller at RT. A faster penetration fixing fluid would have been betteryou would have to test a few, under your conditions. NB: Formalin penetrates a 1cm cube of gelatin 250mm in 24hrs so, penetration through a dense heterogeneous tissue, especially the initital penetration of the T albuginea, will be much slower, allowing for autolytic ( necrotic, really, as the tissue is dead) changes towards the centre of the tissue. Best wishes, Carl Carl Hobbs FIBMS Histology and Imaging Manager Wolfson CARD Guys Campus, London Bridge Kings College London London SE1 1UL 020 7848 6813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Bone/cartilage/epithelial tissue stain
A HE will allow you to identify those three tissue types, in normal tissues, of most organisms ( certainly in avian embryos, imho), IF you know your Histology. There is no stain/demonstration technique that will substitute for this knowledge, imho. Sure, a special stain will help to confirm the tissue type to a high degree. I usually start with staining the section with Alcian blue pH3, then place in Hx ( I use Gill's 2sure, they state you must use Iron Hx...not necessary, for me) for 10 mins. Then place in Van Gieson's solution until what I know is collagen type I is red. I hope this adds to previous excellent posts. Best regards, Carl Carl Hobbs FIBMS Histology and Imaging Manager Wolfson CARD Guys Campus, London Bridge Kings College London London SE1 1UL 020 7848 6813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Wny job opportunity
We have the following need for Histotechnologists. If interested reply to:mkman...@earthlink.netmailto:mkman...@earthlink.net and provide your resume. You will be called promptly to discuss this opportunity. Western NY State- Our large regional reference Lab is seeking 2 HIstotechnologist JOB DESCRIPTION: Collect and prepare/perform tissue specimens received from surgical and non-surgical specimens. Daytime* Full time Monday-Friday*Salary and full benefits. Candidate must have a NY license or ability to get one. OUR NEED IS IMMEDIATE so to expedite consideration reply to: mkman...@earthlink.netmailto:mkman...@earthlink.net and send your resume or call 314-409-8800tel:314-409-8800 This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. - This message was secured by ZixCorp(R). ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
