CONFOCAL SCANNING MICROSCOPY & HI RESOLUTION SEM OF CEREBELLAR CORTEX

共焦扫描显微镜

• 批准号: 6117299
• 负责人: ORLANDO J CASTEJON
• 金额: $ 1.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6117299
• 关键词: animal tissue; bioimaging /biomedical imaging; biomedical resource; microscopy; nervous system

In the last two decades we have reported the conventional and high resolution scanning electron microscope studies of the cerebellar cortex of several vertebrates, including man. These studies have shown mainly cytoarr-hitectonic arrangement, tracing of intracortical circuits and comparative features with correlative techniques, such as transmission electron microscopy, either by thin sections or freeze-etching replicas. Now, we want to examine in detail synaptic morphology and imaging outer surface membrane macromolecules, such as glycoproteins and proteoglycans, using high resolution (SEM) field emission scanning electron microscopy. For sample preparation, I would like to use cryotechniques, gold labeling and chromium coating. Sample preparation techniques will be carried out in the Albrecht Laboratory and at the Integrated Microscopy Resource, using rodent cerebellum. The main objectives of the project will be: 1) To study pre- and post- synaptic ending organization in cryofractured nerve cells. 2) To image the glycocalix in unfi-actured nerve cells and characterize at the outer surface membrane glycoproteins and proteoglycans (hyaluronic acid, chondroting 4- or 6-sulfate.) Also, we will use confocal microscopy. The objectives are: to study the three-dimensional cytoarchitectonic arrangement of granule, Golgi, Purkinje, stellate and basket cells. To trace the intracortical circuits formed by the afferent mossy fibers with granule cell dendrites, climbing fibers and granule cell axons with Purkinje cell dendrites. The scanning confocal images obtained will be compared with previously obtained scanning electron and freeze etching images.

在过去的二十年里，我们报道了常规和高 小脑的分辨率扫描电子显微镜研究 包括人类在内的几种脊椎动物的皮层。这些研究 主要表现为胞质结构排列、皮质内 电路和相关技术的比较功能，如 透射电子显微镜，无论是薄切片或 冷冻蚀刻复制品 现在，我们想详细检查突触 形态学和成像外表面膜大分子，如 糖蛋白和蛋白聚糖，使用高分辨率（SEM）场 发射扫描电子显微镜 对于样品制备，我 想使用冷冻技术，金标签和铬涂层。 样品制备技术将在Albrecht 实验室和综合显微镜资源，使用啮齿动物 小脑 该项目的主要目标是：1）研究 冷冻损伤神经突触前和突触后末梢的组织结构 细胞 2)对未断裂的神经细胞中的糖杯进行成像， 在外表面表征膜糖蛋白， 蛋白聚糖（透明质酸，软骨素4-或6-硫酸盐。）另外我们 将使用共聚焦显微镜。 目的是：研究 颗粒，高尔基体， 浦肯野细胞、星状细胞和篮状细胞。 为了追踪大脑皮质内 苔藓纤维传入与颗粒细胞形成回路 树突、攀缘纤维和颗粒细胞轴突与浦肯野细胞 树突 将获得的扫描共焦图像进行比较 与先前获得的扫描电子和冷冻蚀刻图像。