zygote2k February 12, 2009 Share February 12, 2009 I remember seeing this in FAMA a number of years ago, but figured it was too advanced for me. Now, I think I can do it. Has anyone ever tried? Link to comment Share on other sites More sharing options...
jason the filter freak February 12, 2009 Share February 12, 2009 clearing and staining what? Link to comment Share on other sites More sharing options...
reefhunter February 12, 2009 Share February 12, 2009 did I miss something? Link to comment Share on other sites More sharing options...
wdytat February 12, 2009 Share February 12, 2009 (edited) did I miss something? See here http://www.wamas.org/forums/index.php?show...amp;hl=clearing Edited February 12, 2009 by wdytat Link to comment Share on other sites More sharing options...
zygote2k February 12, 2009 Author Share February 12, 2009 (edited) clearing the flesh and staining the skeletal and vascular systems. Clearing and Staining for bone and cartilage adapted from Dingerkus and Uhler Procedure: 1. Fix fresh material in 10% formalin 2-3 days or longer. Specimens already in formalin or alcohol, ignore step 1 and go to step 2. 2. Wash in several changes of distilled water for 1-2 days. 3. Place directly into a mixture of 10 mg Alcian Blue 8GN (supplied by Matheson, Coleman and Bell), 80 ml 95% ethyl alcohol and 20 ml glacial acetic acid 10-12 hours. (This solution must be fresh) 4. Transfer to two changes of 95% ethyl alcohol, 1-2 hours per change. 5. Transfer through an alcohol series of 75%, 40%, 15% ethyl alcohol 2-3 hours in each, or until specimen sinks. 6. Transfer to distilled water 2-3 hours, or until specimen sinks 7. Place in an enzyme solution of 30 ml saturated aqueous sodium borate, 70 ml distilled water, and 1 g trypsin enzyme. Change solution every 2-3 days, or sooner if solution turns a bluish color. Continue until bones and cartilage are clearly visible, and the flesh retains no blue color. NOTE: the skin of some gobies remains blue at this stage. 8. If bone is present: Transfer to 0.5% aqueous KOH solution, to which enough Alizarin Red S stain has been added to turn solution a deep purple-red. Leave 24 hours or until bones are distinctly red. If bone is not present go to step 9 9. Transfer through a 0.5% KOH - glycerin series of 3/4 KOH - 1/4 glycerin, 1/2 KOH - 1/2 glycerin, 1/4 KOH - 3/4 glycerin, and finally to pure glycerin. 10. Store specimen in pure glycerin to which a few crystals of thymol have been added to inhibit molds and bacteria. Edited February 12, 2009 by zygote2k Link to comment Share on other sites More sharing options...
lanman February 13, 2009 Share February 13, 2009 Ah... I see - the ultimate cure for ich. bob Link to comment Share on other sites More sharing options...
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