Application of heat-induced antigen retrieval on whole mount immuno-staining for adult mouse cerebellum

The present study examined the application of heat induced antigen retrieval techniques on whole mount immunostaining for the adult mouse cerebellum. After a high-temperature antigen retrieval procedure, cerebella were processed for whole mount immunostaining using a modified protocol developed by Sitolloe and Hawkes (J Histochem Cytochem 50:235-244, 2002). The expression of compartmentation antigens, zebrin II and heat shock protein 25, was analyzed. Characteristic striped patterns of these two antigens were obtained by the present modified protocol. Vibratome sections of whole mount cerebella showed a positive staining in dendrites of particular subsets of Purkinje cells. This optimized protocol circumvents the need to process the cerebella through cycles of chilling and thawing prior to the addition of primary antibody. Thus, heat induced antigen retrieval provides a more rapid and efficient way to perform whole mount immunostaining of adult tissues. This optimized approach allows rapidly and easily for investigation of the spatial distributions of various antigens in adult tissues or organs using whole mount staining.