Observation of Medaka Larval Gonads by Immunohistochemistry and Confocal Laser Microscopy

The combination of immunohistochemistry and confocal laser microscopy enables the observation of cellular structures and protein localization within cells using whole-mount tissues. However, such high-resolution imaging requires several steps, such as proper dissection before fixation and antibody staining, and the appropriate positioning of tissues on a glass slide for observation. Here, we describe the method developed by our laboratory for the immunohistochemistry of medaka embryonic and larval gonads, focusing on the dissection and mounting of tissues for confocal laser microscopy. Positioning the gonad just beneath the coverslips is essential to obtain high-resolution images at a level where cellular components of germ cells, such as germ plasm and nuclear structures, can be clearly observed using an oil immersion objective lens.