Authors used this solution to clear mouse brain, mouse embryos and whole body of D. melanogaster.[17] Main drawback of this solution is bleaching of GFP signal and insufficient clearing of highly myelinated tissue of adult animals.[2] Therefore, many other reagents were tested with aim to find GFP compatible and more sufficient clearing. As results tetrahydrofuran (THF) and dibenzyl ether (DBE) were chosen as the best regents for clearing.[18] Based on those findings the 3DISCO protocol was published in 2012.[1]