Arsenault A L, Ottensmeyer F P, Heath I B
Department of Anatomy, University of British Columbia, Vancouver, Canada.
J Ultrastruct Mol Struct Res. 1988 Jan;98(1):32-47. doi: 10.1016/s0889-1605(88)80932-7.
Newborn mice epiphyseal growth plates were preserved by slam freezing/freeze substitution and examined by conventional electron microscopy, stereopsis, high voltage electron microscopy, and electron spectroscopic imaging (ESI). To illustrate the improved ultrastructure of this cryogenic procedure, conventional, aqueously fixed growth plates were included showing collapsed hypertrophic chondrocytes surrounded by a depleted and condensed extracellular matrix. In contrast, the cryogenically prepared epiphyses contain chondrocytes and extracellular matrix vesicles both in direct contact with proteoglycan filaments retained in an expanded state. ESI is an electron microscopic technique which enables the direct localization of atomic elements superimposed over fine structural details. This technique was used to examine the colocalization of calcium and phosphorus within matrix vesicles and within their associated extracellular environments. Matrix vesicles appeared in three distinct diameter ranges. The integrity of the matrix vesicles was examined at various stages of mineralization and also within the mineralized zone of provisional calcification.
新生小鼠的骨骺生长板通过速冻/冷冻置换法保存，并采用传统电子显微镜、立体视觉、高压电子显微镜和电子光谱成像（ESI）进行检查。为了说明这种低温程序改善后的超微结构，还纳入了常规水固定的生长板，显示出肥大软骨细胞塌陷，周围是耗尽和浓缩的细胞外基质。相比之下，经低温处理的骨骺含有软骨细胞和细胞外基质小泡，两者都与保持膨胀状态的蛋白聚糖丝直接接触。ESI是一种电子显微镜技术，能够直接定位叠加在精细结构细节上的原子元素。该技术用于检查基质小泡及其相关细胞外环境中钙和磷的共定位。基质小泡呈现出三个不同的直径范围。在矿化的各个阶段以及临时钙化的矿化区内检查了基质小泡的完整性。
