It seems that using the FocusClear(r) reagent is paramount to obtain the best transparency. However, this mounting medium is expensive, and seems to be back-ordered for a while due to large demand. Also, the formulation is proprietary, which makes further optimization harder.
While trying different mounting reagents for the ETC treated samples, I realized that glycerol does not give great results, even after very long incubations. ScaleA2 (4M Urea) works great on ETC treated brains, full transparency is achieved in just hours, but I'm afraid that it could disrupt the molecular integrity of the sample.
Could we try to design a home-made mounting reagent that would give similar results?
It seems that the best way to achieve index matching is to use a solution with ions presents at a high molarity. FocusClear(r) seems to use diatrizoic acid and meglumine to achieve that. Does anyone had any luck in making a good substitute?