Youtor
Diaphonization EP09 Results
>> YOUR LINK HERE: ___ http://youtube.com/watch?v=fH_5GvM8UdI
Follow my journey into diaphonization/clearing and staining. These are my first specimens :) • WRITTEN PROTOCOL CAN BE FOUND HERE: https://www.scribd.com/document/35471... • —IN THIS VIDEO:— • EP09: Results • In this episode we finish up the specimens, and take a look at how pretty they’ve become. • —TUTORIAL:— • Materials: Specimens, glycerin, thymol crystals, jars for storage, scissors, tweezers, protection* • *WARNING!: These are toxic chemicals, so be sure to wear protective gear and work in a well ventilated area. • 0) After a few weeks of clearing, my specimens look transparent and are ready to be stored • 1) During the process I’ve noticed some loose skin that floats around and doesn’t look good. Therefore I start by inspecting the specimens and cutting off anything that doesn’t look right (00:18) • 2) Find some storage jars that will fit the specimens, and transfer the specimens here (01:00) • 3) Fill the jar with glycerin (01:25) • 4) Add a pinch of thymol crystals (01:41) - this will work as an anti-fungal agent • 5) Use tweezers to play around with the placement of the specimen in the jar (02:05), and put the lid on once satisfied. Sometimes the specimen can be a little floaty and hard to work with (02:09). This is due to tiny air bubbles stuck inside cavities - don’t worry, they will disappear over time. Just leave the specimen for a few days/weeks to allow the bubbles to clear, and then try to reposition it again. • 6) Done! Now go look at the pretty things • —DISCUSSION:— • I am currently trying to optimize the protocol. Although I am overall happy about the outcome of my first specimens (they do look pretty cool, right?), there are some things that I definitely want to improve: • a) Skinning: If you look closely the mouse has a broken leg and no ears (the tail was broken before skinning), and the chicken has a broken wing. Oops! I also didn’t remove the skin on the chicken’s feet because I was afraid to loose the toes - however the rough skin really took to the Alizarin red, making it look like it has small purple boots on :). I’ve already gotten better at skinning, and my new specimens don’t have these issues. • b) Staining: The staining varies a bit between the species. The mouse has almost no Alcian Blue staining, and may need a longer incubation time than the chicken. I may experiment with incubation time and/or a single animal per jar for staining to see if I get better results. • c) Clearing: These specimens have areas with slightly brown tint, that could have been cleared better. Several factors could make this better: Longer trypsin digestion, higher KOH concentration during clearing, more H2O2, longer incubation times in clearing baths. I will need to see what works best. • d) Storage: The current jars are not 100% optimal as they are too wide and not tall enough for these types of specimens. However this is an easy fix that can always be done at a later point. • If you have any other tips or ideas for improvement, I’d love the feedback :) • —NEXT UP:— • If I manage to do some good optimization, I might do EP10 - Follow up Optimization
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