ELECTRON MICROSCOPIC STUDIES OF CRYSTALLINE LENSES

描述(调查人员摘要)：这项工作的长期目标 项目是获得对人类晶状体的定性和定量知识 形态学与晶状体功能和功能障碍有关。具体目标 1)裂隙灯生物显微镜(直接照明和反向照明) 用于识别不透明和特定的间断区( 前区和后区、前区和后区 成体核区和胎儿核内区) 皮质(C)、皮质和核混合(CN)和核(N)人 白内障。囊外摘除后，这些晶状体将被 由调查人员开发的一种方法解剖而成 对这些相同的不透明和不连续区域的完整恢复 进行相关的EM分析。然后将使用立体扫描电子显微镜来表征 不透明和间断区的粗略和精细结构 具体参考晶状体缝合解剖。这些信息将是 用于制作晶状体缝合“平面”解剖的3D CAD重建图 在不透明和特定的不连续区域内。2)氦-霓虹灯 研究人员自己设计的激光扫描装置将被用来量化 晶状体缝线解剖对晶状体焦点变异性的负面影响 (即球差)在不同年龄、正常的非白内障中 灵长类(猕猴)镜片。经过激光分析，这些镜片将被 根据研究人员的方法解剖以取回完整的缝合线 来自胎儿和成人晶状体核以及成人晶状体皮质 进行相关的EM分析。然后将使用立体扫描电子显微镜来表征 这些晶状体中缝线的粗略和精细结构 发育(胎儿核)、生长(成人核)和年龄( 成人皮层)。正常灵长类晶状体的这些与年龄相关的区域是 分别相当于和胎儿细胞核的内区， 成人核的前区和后区，以及前区和后区 分离的后部区域。这些信息将被用于生产 晶状体缝合“平面”解剖与晶状体功能关系的三维CAD重建 发展、成长和年龄。3)最后，立体扫描，透射电子显微镜[中压 和常规]和冷冻蚀刻分析将被用来表征 上述晶状体缝合和核解剖的超微结构 隐形眼镜。具体地说，这些研究将定性和量化差距 晶状体缝合和内吞的连接、方阵列膜和内吞 核解剖是发育、生长、年龄和病理的函数。 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获得资格 和量化人类晶状体中的结构变化作为 与浑浊和特定间断区相关的病理学 裂隙灯生物显微镜观察；2)这些研究的结果 将允许对结构变化进行鉴定和量化 正常灵长类晶状体的可比面积随发育而变化， 生长和年龄可以与特定的光学功能相关联 (焦点可变性)。因此，通过外推，这两个集合的结果 的研究可以相互关联来识别人类晶状体的持续变化 预示而不是结果的发育、生长和年龄期间的结构 来自晶状体病理学(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).