ELECTRON MICROSCOPIC STUDIES OF CRYSTALLINE LENSES

描述（研究者摘要）：长期目标 本项目的目的是获得有关人类透镜的定性和定量知识 因为它与透镜功能和功能障碍有关。 具体目标 1）裂隙灯生物显微镜检查（直接和逆向照明）将 用于识别不透明和不连续的特定区域（ 前、后分离区 成人核的区域和胎儿核的内部区域）， 皮质（C）、皮质和细胞核混合（CN）和细胞核（N）人类 白内障 在囊外手术取出后，这些晶状体将 通过研究人员开发的方法进行解剖， 这些不透明和不连续区域的完整恢复 进行相关电磁分析 然后使用立体SEM表征 不透明区和不连续区的粗结构和细结构 具体参考透镜缝合解剖。该信息将 用于生成透镜缝线“平面”解剖结构的3D CAD重建 在不透明和不连续的特定区域内。 2)氦氖 将使用研究者自己设计的激光扫描设备来量化 透镜缝解剖对透镜焦点变异性负面影响 (i.e.球面像差）， 灵长类动物（猕猴）晶状体。 经过激光分析，这些镜片将 根据研究者的方法进行解剖，以取出完整的缝线 来自胎儿和成人透镜核以及来自成人透镜皮质 进行相关电磁分析 然后使用立体SEM表征 这些晶状体中缝线的大体和精细结构作为功能 发育（胎儿核），生长（成人核）和年龄（ 成人皮质）。 正常灵长类动物透镜的这些与年龄相关的区域是 分别相当于，和胎儿核的内部区域， 成人核的前区和后区，以及前区和后区， 分离的后区。 这些信息将用于生产 透镜缝线“平面”解剖结构的三维CAD重建作为透镜的功能 发展、成长和年龄。3)最后，立体SEM、TEM [中压 和常规]和冷冻蚀刻分析将用于表征 上述透镜缝合和核解剖的超微结构 眼镜. 具体而言，这些研究将定性和量化差距 透镜缝和核的连接、方形阵列膜和内吞作用 核解剖学作为发育、生长、年龄和病理学的函数。 这些研究的重要性源于以下事实：1） 这些研究的结果将首次允许资格 以及作为以下函数的人透镜中的结构变化的量化 与不透明和特定不连续区域相关的病理学 通过裂隙灯生物显微镜观察; 2）这些研究的结果 将允许定性和定量的结构变化， 正常灵长类动物透镜的可比面积作为发育的函数， 与特定光学功能相关的生长和年龄 （焦点变化）。 因此，通过外推，这两组的结果 的研究可以相互关联，以确定人类透镜的持续变化 结构在发展，成长和年龄，预示，而不是结果 透镜病理学（老花眼和白内障）。

DESCRIPTION (Investigator's Abstract): The long term objective of this project is to gain a qualitative and quantitative knowledge of human lens morphology as it relates to lens function and dysfunction. Specific aims are; 1) Slit lamp biomicroscopy (direct and retro-illumination) will be used to identify opacities and specific zones of discontinuity (the anterior and posterior zones of disjunction, the anterior and posterior zones of the adult nucleus and the inner zones of the fetal nucleus) in cortical (C), mixed cortical and nuclear (CN) and nuclear (N) human cataracts. After extracapsular surgical removal, these lenses will be dissected by a method that the investigators have developed that permits that intact retrieval of these same opacities and zones of discontinuity for correlative EM analysis. Stereo SEM will then be used to characterize the gross and fine structure of the opacities and zones of discontinuity with specific reference to lens sutural anatomy.This information will be used to produce 3D CAD reconstructions of lens suture "plane" anatomy within opacities and specific zones of discontinuity. 2) A helium-neon laser scan device of the investigator's own design will be used to quantify the negative influence of lens sutural anatomy on lens focal variability (i.e. spherical aberration) in variably aged, normal non-cataractous primate (macaque) lenses. After laser analysis, these lenses will be dissected according to the investigator's method to retrieve intact sutures from the fetal and adult lens nucleus as well as from the adult lens cortex for correlative EM analysis. Stereo SEM will then be used to characterize the gross and fine structure of the sutures in these lenses as a function of development (the fetal nucleus), growth (the adult nucleus) and age (the adult cortex). These age-related zones of normal primate lens are the equivalent of respectively, and the inner zones of the fetal nucleus, the anterior and posterior zones of the adult nucleus, and the anterior and posterior zones of disjunction. This information will be used to produce 3D CAD reconstructions of lens suture "plane" anatomy as a function of lens development, growth and age. 3) Finally, stereo SEM, TEM [medium voltage and conventional] and freeze-etch analysis will be used to characterize the ultrastructure of lens sutural and nuclear anatomy in the above described lenses. Specifically, these studies will qualify and quantify the gap junctions, square array membrane and endocytosis of lens sutural and nuclear anatomy as a function of development, growth, age and pathology. The importance of these studies is derived from the following facts:1) the results of these studies will permit for the first time the qualification and quantification of structural changes in the human lens as a function of pathology that correlate with opacities and specific zones of discontinuity as observed by slit lamp biomicroscopy; 2) the results of these studies will permit the qualification and quantification of structural changes in the comparable areas of normal primate lens as a function of development, growth and age that can be correlated with a specific optical function (focal variability). Thus, by extrapolation, the results of these two sets of studies can be correlated to identify ongoing changes in human lens structure during development, growth and age that herald rather than result from lens pathology (presbyopia and cataract).