Tim, I do not have the paper, but a few years ago I worked the kinks out of that stain. Your acetic acid is too long. Try for the amount of time that you usually do for a one step. A few dips would be a good start. Then rinse in dH2O, not wash. Also, I do not remember using the Weigert's and Verhoeff's solution. No need. Just the Bouins mordant, VVG and did a trichrome as a counter stain. Also, do not differentiate as much with the Ferric. Under differentiation will compensate for the other reagents. How long do you usually leave the slides in one step when doing a regular one? Those would be good starting points.
Toysha N. Mayer, MBA, HT (ASCP) Instructor Program in Histotechnology School of Health Professions MD Anderson Cancer Center (713) 563-3481 [email protected] ------------------------------ Message: 10 Date: Thu, 5 Jan 2012 09:45:25 -0800 From: "Morken, Timothy" <[email protected]> Subject: [Histonet] Elastichrome paper, and troubleshooting To: Histonet <[email protected]> Message-ID: <8d7c2d242dbd45498006b21122072bf89f5ee...@mcinfrwem003.ucsfmedicalcenter.org> Content-Type: text/plain; charset=us-ascii Does anyone have a pdf (or could fax me) the original paper for "Elastichrome" stain: Richardson, L. "Combination Elastic Trichrome Stain," Laboratory Medicine, 6.1, 1975 We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past. I do have another paper in the same vein, "A combination verhoeff's elastic and masson's trichrom stain for routine histology," O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible. Our problem is that the elastic stain washes out during or after the trichrome. Procedure: Bouins, 56C one hour Wiegerts hematolylin, 3min Verhoffs elastic stain, 15 min 2% ferric chloride differentiation (so far so-good) Gomori's trichrome Blue, 15 min 0.2% glacial acetic acid, 1 min Wash dH20, dehydrate, clear, mount Results: trichrome looks great, no elastin stain. I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions. Thanks for any help. Tim Morken Supervisor, Histology, IPOX UC San Francisco Medical Center Box 1656 1600 Divisidero St, B217 San Francisco, CA 94115 USA 415.514.6042 (office) 415.885.7409 Fax [email protected]<mailto:[email protected]> ************************************* _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
