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Discussion in 'Tutorials' started by TacoKel, Jun 11, 2015.

  1. TacoKel

    TacoKel #DyeingTheDead

    Hello anyone who reads my post,

    My name is Kelly and I have been filming a YouTube series entitled "Dyeing The Dead!" (http://www.youtube.com/TacoKellz) I am filming my journey through diaphonization!

    As of today, I have completed all of the steps following up to the Trypsin Digestion. It's on this step that I am stuck on.
    The protocol I am using is entitled "Give New Life to Old Specimens Through Clearing & Staining" by Bob Weck and Pam Miljak.

    The protocol says:
    "The specimen is transfered to a trypsin solution for one to several days or until the body is limp and the skeleton becomes visible. Change the solution if the process takes more than three days or if the solution becomes blue. Larger specimens may take more than a week."

    I prepared 30 mL of a saturated Sodium Borate solution, added 70 ml of distilled water, then added 1.0 g of Trypsin, and mixed it as well as I could.
    My solution became a "cloudy yellow" and developed some small floating pieces. I placed my specimens (mice) into the solution and left over night. When I checked on the solution the next day, it had turned quite blue-ish. My mice had become quite limp but the Alcian blue cartilage stain however was still present on the tissues, tail, and other extremities (feet/hands).
    Since my solution had turned "blue," I prepared a fresh Trypsin Solution which I put my mice in and then have been letting them sit for another day.

    Can anyone PLEASE HELP ME and possibly give me some advice on what the mice should look like and if I am in fact doing this step correctly? PLEASE!!
    This is my first time doing this so I'm not really sure what I'm supposed to be looking for.

    Below I have posted some pictures of what my specimens look like as of the second day in a fresh trypsin bath. As you can see, the tail and tissues still appear to be blue. According to the paper, I was under the impression that the Alcian Blue Cartilage Stain was supposed to be removed from the noncartilageous tissue. Looking at the pictures, I still see a lot of blue. The mice also don't seem transparent just yet.
    [​IMG] [​IMG]

    I'd really appreciate it if anyone could help me out. That would be fantastic!
  2. elonj

    elonj New Member

    I know this is old, but I've watched your YouTube videos and was curious if you got this figured out...

  3. marthahill

    marthahill New Member

    Wow... really interesting. Thanx for that guide