Scanning Microscopy


The internal three-dimensional organization of secretory and maturation stage ameloblasts was examined using field emission scanning electron microscopy. Particular attention was given to the structure of the Golgi apparatus, the distribution of smooth membrane tubulo-vesicular elements and their relationship with endosomal/lysosomal components. Rat incisors were fixed by intracardiac perfusion with glutaraldehyde, decalcified and divided into segments. The tissues were cryoprotected with glycerol and freeze-fractured. They were then macerated in osmium, and after conductive staining with osmium/tannic acid, the samples were critical-point dried and sputtered with gold. High-resolution scanning electron microscopy showed that ameloblasts contain a well-developed Golgi apparatus, even in the maturation stage where cells are generally believed not to be actively involved in protein secretion. Smooth tubulo-vesicular structures formed a complex network which extended throughout the cell. In secretory stage ameloblasts, this network reached into Tomes' process and consisted of small and large tubules, and distended portions. The smaller tubules radiated from a central core of organelles towards the plasma membrane. Numerous lysosomal/endosomal elements were observed in the Golgi region, and in some cases smooth tubular portions were seen at the surface of multivesicular bodies. These data show that high-resolution scanning electron microscopy can be applied to correlate three-dimensional structural detail with the secretory and resorptive functions of ameloblasts.

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