A polyclonal antibody raised against the hatching enzyme ofCiona intestinalis(D'Aniello et al., 1997) was used on larvae of different ages in whole mount immunofluorescence experiments in order to localize the cells secreting the enzyme. After staining with FITC-conjugated second antibody, the larvae were observed by confocal microscopy. Larvae just before hatching (9–10 hours after fertilization) showed the presence of the enzyme in the peripheral cells of the adhesive papillae. The newly hatched larvae showed fluorescence also in the epidermal cells of the tip of the tail. Higher magnification confocal images of the papillae revealed bright fluorescence both in peripheral cells of the papillae and in the cavity between the tunic and the apex of the papillae (hyaline cap).