Susaki, E. A. et al. Nat. Commun. 11, 1982 (2020).

Rendering large tissue samples such as organs or whole organisms transparent yet suitable for antibody or dye staining remains challenging despite the wealth of tissue-clearing protocols. In particular, full tissue penetration of staining reagents merits further research. Susaki et al. have developed an improved clearing pipeline based on the established CUBIC technique. Their modified CUBIC-HistoVision approach optimizes ionic strength, temperature and chemical composition of the tissue-clearing cocktails, as well as antibody staining conditions and other steps in the pipeline. Furthermore, the researchers tested several nuclear stains as well as 44 antibodies relevant to the neuroscience community for their performance in CUBIC-HistoVision. The researchers primarily showcase the pipeline for immunostaining or nuclear labeling of mouse brains, but also demonstrate it on an intact infant marmoset specimen.