Observation of oral organelle and digestive organelle in Paramecium caudatum using Laser Scanning Confocal Microscopy

摘要

Using laser scanning confocal microscopy, the morphological structures of oral organelles and digestive organelle in Paramecium caudatum were re-observed. In the preparation of samples, 5% KMnO4 was used. In addition, continuous tomoscan and 3-D reconstruction techniques were adopted. The result clearly revealed some structures of oral organelles such as vestibulum, buccal cavity, endoral membrane, quadrulus, dorsal and ventral peniculi, etc. Their positions and morphology basically corresponded with results in previous studies, 3-D conformation of oral structure is also provided. Food vacuoles and their forming process were also clearly observed. The diameter of the cytopharynx expands when it ingests and the food vacuole is formed. This suggests that the membrane of the vacuole may extend into the membrane of the cytopharynx just when the diameter of the cytopharynx expands. Specimens of P. caudatum were collected from a stream in the Wuchang region of Heilongjiang Province. The samples were prepared as follows: Cells were suspended in 0.01% TritonX-100 solution for about 1 min, fixed by 5% KMnO4 solution for 4 min at room temperature, and washed in PBS (pH7.4). They were than exposed to 0.5% TritonX-100 solution for 25 min at room temperature; then incubated overnight in the primary antibody at 4℃ and washed in PBS. Avoiding sunlight, an overnight incubation in the secondary antibody at 4℃ follows, washed by PBS. Mounting the samples onto slides with glycerol[Acta Zoologica Sinica 51(4): 718–722, 2005].