Scanning Microscopy


Apex regions of continuously growing incisors of Wistar rats were quickly dissected, shock-frozen in liquid nitrogen-cooled propane, freeze-dried at -80 °C and infiltrated with Spurr's resin . 400nm thick dry sections were cut with a diamond knife on an ultramicrotome . Relatively flat sections were transferred with an eye lash onto collodium coated aluminum grids. They were flattened with a glass stick and by placing another collodi um coated aluminum grid just on top of the first one, exerting a uniform pressure . After carbon coating the sections were observed using the backscattered and secondary electron signals in a scanning microscope. The predentine was analyzed for calcium and potassium with an energy dispersive x-ray analysis system. The xray spectra revealed in the predentine regions with beginning dentine formation, near the apex, an uneven K-distribution with very low as well as more prominent x-ray peaks. The K peaks were always lower than those of calcium. In areas with advanced dentine formation, prominent K-peaks were always observed. They were normally higher than the Ca-peaks up to a distance of 5- 10 µ.m from the dentine border. Closer to the dentine border the K concentration decreased while the Ca-peak increased. This might indicate that (besides Na) K is used to balance the negative charges of the macromolecules till K is replaced by Ca at the onset of apatite crystal formation.

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