A cryotomic method of hemisecting insect appendages for neuro-immunohistology

A technique is described for longitudinally hemisecting small and often flexible appendages of arthropods, by embedding the material in frozen 30% sucrose which is wax-like in texture. The cleanly cut tissue is then amenable to immunohistochemical techniques since the problem of impenetrability of cuticle to antibodies is by-passed. The apparatus utilizes a simple temperature-regulated aluminium stage cooled by dry ice. Fixed, surcrose-infiltrated appendages are block-mounted on the stage, and cut with a razor blade during visual guidance using a dissecting microscope. A combination of hemisections and thick sections also can be prepared. This method has been used to demonstrate the expression of glionexin, a glial cell-associated glycoprotein, and ELAV, a neuronal cytoplasmic protein, in mechanoreceptor organs and sensilla of the cricket Acheta domesticus.