Three-Dimensional Visualization of the Aorta and Elastic Cartilage after Removal of Extracellular Ground Substance with a Modified NaOH Maceration Method

Department of Anatomy, Oita Medical University, Japan.
Journal of Electron Microscopy (Impact Factor: 1.63). 11/1993; 42(5):328-33.
Source: PubMed


A modified NaOH maceration method for removing extracellular ground substance was applied to scanning electron microscopic (SEM) studies of the mammalian aorta and elastic cartilage. Fixed tissues were freeze-fractured in liquid nitrogen and then immersed in 2N NaOH solution for 4-6 hr at room temperature. Since this method selectively digested proteoglycans, cellular and fibrous elements were observable clearly under the SEM. The elastic laminae of smooth muscle cells of the aortic tunica media were concealed by fine collagen fibrils, while chondrocytes in the elastic cartilage were encapsulated by a dense network of fine collagen fibrils and branching elastin fibers.

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    • "They were deparaffinized with xylene, hydrated , and fixed again in a mixture of 2.5% glutaraldehyde and 2% paraformaldehyde (Karnovsky's fixative). After specimens were immersed in 2N NaOH at 37 C for 3 hr to digest proteoglycans [4], they were washed thoroughly in distilled water and placed in cacodylate-buffered 1% osmium tetroxide, 1% tanic acid solution and 1% osmium tetroxide for 1h each. They were then dehydrated through a graded series of ethanol, dried by the tbutanol drying method, coated with gold and observed using a scanning electron microscope (Hitachi S-800, Japan). "
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