Clark J M
J Orthop Res. 1985;3(1):17-29. doi: 10.1002/jor.1100030102.
Adult rabbit articular cartilage was prepared for scanning electron microscopy using, in order, glutaraldehyde fixation, enzymatic removal of proteoglycan, dehydration in ethanol, cryofracture in liquid nitrogen, and critical-point drying. Enzymes were effective in fixed material. Fixation, cryofracture, alignment of fracture surfaces with "split lines," and retention of subchondral bone were found to be necessary steps for the preservation of collagen detail. The fibrous framework was found to be similar to that proposed by Benninghoff and favored by more recent phase-contrast microscopic studies. Vertical fibers extending from subchondral bone and a network of tangentially oriented superficial fibrils converge in the transitional zone. No random layer is seen. Pericellular capsules interdigitate with the vertical fibers. When cartilage is prepared in a manner that minimizes tissue damage, scanning electron microscopy provides useful, unique information.
依次用戊二醛固定、酶法去除蛋白聚糖、乙醇脱水、液氮冷冻断裂以及临界点干燥。酶在固定材料中有效。发现固定、冷冻断裂、使断裂面与“分裂线”对齐以及保留软骨下骨是保存胶原细节的必要步骤。发现纤维框架与本宁霍夫提出的并为最近的相差显微镜研究所支持的框架相似。从软骨下骨延伸的垂直纤维和切向排列的浅表原纤维网络在过渡区汇聚。未见随机层。细胞周围囊与垂直纤维相互交错。当以最小化组织损伤的方式制备软骨时,扫描电子显微镜可提供有用的独特信息。