Immunostaining Helobdella (leech) embryos

The purpose of this staining protocol is to detect the localization of specific antigens in the Helobdella (leech) embryo. Immunostaining protocols must be optimized for each antibody and embryonic stage. Here, as a starting point, we present a general-purpose immunostaining protocol. Immunostaining of the leech embryo can be performed on the intact embryo or on dissected parts. For post-gastrulation embryos (stages 9 and 10), if the area of interest is in the germinal plate, immunostaining can be performed on the dissected germinal plate. It can also be performed on the ventral nerve cords dissected from late-stage embryos or juveniles.