Andi, Your PI has requested the Goldner's trichrome stain because it provides a stark contrast between the green (light-green SF yellowish) stained mineralized component of bone and the red (acid fuchsin-ponceau) stained newly formed unmineralized "bone-like" dense collagen fibers (osteoid) that will eventually mature and mineralize to form bone. Now if you subject mineralized bone to an acid solution, the process of demineralization begins and acts to remove the mineral (calcium and phosphorus) content. The acid will also have an effect on the soft tissue components, but the effect varies depending upon the acid concentration and certain properties of tissues. Nevertheless, the most notable effect of acid decalcification will be seen as a change in the density of bone and demonstrated by stain uptake and penetration. If you have to demineralize your bone in order to cut sections because you do not have the capabilities to cut undemineralized bone, then you need a stain that provides the best contrast for this now demineralized state of bone. Once the bone has been decalcified, you essentially have now demineralized bone (less dense) and still unmineralized "bone-like" collagen fibers, all of which will now be contrasted differently with the Goldner's stain. While an H&E can be used to distinguish bone that was once mineralized (old bone) from unmineralized "bone-like" collagen fibers (osteoid or newly forming bone), there is not enough of a stark contrast and especially if you are wanting to use some form of image analysis software. With that said, the best stain then to use in this situation is a Masson's trichrome where the bone stains blue (aniline blue) and the osteoid stains red (biebrich-scarlet). With regards to the demineralization method, the EDTA will take too long to arrive at the same result that a simple 5% formic acid will provide. Also, your sections will be cut at a standard 5 microns. Hope this information helps you to get what you need and understand a little more of why you need what you need! :) Best Regards, Jack
From: algra...@email.arizona.edu To: histonet@lists.utsouthwestern.edu Date: Thu, 8 Mar 2012 08:00:58 -0800 Subject: [Histonet] Goldner's Trichrome for Osteoid Hopefully a hard tissue guru is out there and can help with this. A PI here is asking about doing a Goldner's stain on mouse femur. I'm not set up to cut sections of undecalcified bone so my questions are: 1. can this stain be done successfully on decalcified bone? 2. if so, what type of decal should be used - EDTA? 3. I have googled the stain but didn't see what the preferred thickness was for this stain. Need this info fairly soon! The bones are on the way. Thanks!!! Andi G. _______________________________________________ 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
