After ETC clearing my brain. I mounted it in 85% glycerol (v/v).
The brain was rotating in an incubator at 37°C for 2d.
After 1d the brain was almost completely clear. Just some spots inside looked opaque.
After 2d the brain became opaque.
I was expecting the opaque spots from day one were uncleared parts where the glycerol should diffuse,
but it was the other way around. The brain became opaque and just the outside parts look clarified.
Somebody had the same issue, or has an explanation?
I cleared my brain for 5 days at 37°C, following the protocol and with a standard chamber.