Whole-mount immunostaining protocol for screening of spatial organizations of cerebellar compartmentation

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 Histochem Cytochem 50:235–244, 2002) for screening the parasagittal arrangements of cerebellar compartmentation antigens, i.e., zebrin II and heat shock protein (HSP25). We further 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 visualization of spatial organizations of cerebellar examinations of the normal topography, particular components of olivocerebellar tracts, and patterning defects caused by mutations without 3D reconstruction of immunostained serial sections.