I would like to share my current experience on clearing mouse brain. I was also wondering what are your thoughts on these points?
1. After clearing mouse brain (23V, 1.03A, 37C), I can easily look through when holding the sample against daylight. However, when I continue following the protocol with washing the sample in PBST (24hrs x 2), the tissue turns turbid (which resolves after mounting).
2. I usually mount my sample in 80% glycerol to see if the clearing was sufficient and only for imaging in focusclear. The mounted tissue usually becomes pretty translucent. However, my last sample was very clear at the first couple of hours. When I looked at it the next day, it almost seemed like it has never been cleared at all.
I am wondering if anyone has similar experience?
Looking forward to reading your comments.
2. The same could have happened for this point.. I'd try washing the tissue more thorough at RT.
Hope this helps you! Cheers..
For me a 45ºc waterbath for a couple of days does the trick.