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.
Field
L H
LH
Department of Zoology, University of Washington, Seattle 98105.
D000818:Animals; D001181:Arthropods; D004210:Dissection / Q000295:instrumentation / Q000379:methods*; D051959:ELAV Proteins; D004864:Equipment and Supplies; D005615:Freezing; D006023:Glycoproteins / Q000032:analysis; D006135:Gryllidae; D007150:Immunohistochemistry; D019476:Insect Proteins; D008465:Mechanoreceptors / Q000378:metabolism; D009420:Nervous System / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000032:analysis; D012679:Sense Organs / Q000378:metabolism; D013194:Staining and Labeling
D051959:ELAV Proteins; D006023:Glycoproteins; D019476:Insect Proteins; D012261:Ribonucleoproteins; C085190:glionexin protein, insect