Dissociated neurons of C. elegans embryos develop in tissue culture and exhibit many adult phenotypic traits (Christensen et al., 2002). This allows ready access for electrophysiological recordings. However, cultured neurons may differ significantly from normal adult neurons. An alternate approach, organotypic cell culture, has been used to study the mammalian brain and can provide many of the benefits of tissue culture from fully differentiated adult neurons. We have developed a method of short-term culture of adult C. elegans neurons. Worms were transferred to M9 buffer and placed on an orbital shake-table for 1 hour to loosen adhering bacteria. Worms were then washed with sterile M9 buffer (using a 30-micron nylon mesh as a filter). Then the worms were moved to a cooled plate covered with culture medium. Worms were cut twice with a #11 scalpel blade: once caudally to relieve internal pressure and then just behind the terminal bulb of the pharynx to allow exposure of the nerve ring. Individual heads were then transferred to a sterile chamber containing culture medium. During cutting, the worms were covered with L-15 medium supplemented with antibiotics and sufficient sucrose to bring the osmolality to 340 mOsm. For incubation, worm heads were immersed in a solution identical to that used by Christensen et al. for culture of dissociated embryonic tissue: L-15 medium supplemented with fetal calf serum and antibiotics (340 mOsm). The somata of GFP-labeled AWA chemosensory neurons were often exposed. Exposed neurons maintained dendrites and axons for up to 5 days. Dendrites (responsible for chemosensory transduction) of AWA neurons with exposed somata often retained a normal appearance and contact with the amphid sensilla. This contrasts with our previous physiological preparation, in which recordings were done on worm heads in physiological saline shortly after dissection. In these neurons, the dendrites often appear broken or beaded within hours after cutting. Contraction of the body wall muscles and cuticle continued during the culture period. This culture system may offer a new and possibly more efficient approach to C. elegans electrophysiology.