Author(s): Kazuhiko Sawada and Emi Ochiai
This paper introduces a protocol for whole-mount immunostaining adapted for the adult mouse cerebellum. This technique was originally described by Sillitoe and Hawkes (J His-tochem Cytochem 50:235-244, 2002) for screening the parasagittal arrangements of cere-bellar compartmentation antigens, i.e., zebrin II and heat shock protein (HSP25). We fur-ther optimized this technique to apply to heat-induced antigen retrieval, which provides a more rapid and efficient way to perform whole-mount immunostaining of adult tissues (Sawada and Sun, Curr Neurobiol 1:21-24, 2010). This approach allows for the visualiza-tion of spatial organizations of cerebellar examinations of the normal topography, particu-lar components of olivocerebellar tracts, and patterning defects caused by mutations without 3D reconstruction of immunostained serial sections.