CONFOCAL SCANNING MICROSCOPY & HI RESOLUTION SEM OF CEREBELLAR CORTEX

在过去的二十年里，我们报道了常规和高级 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质。这些研究已经 主要表现为细胞胞浆排列，皮质内示踪 电路及其与相关技术的比较特征，例如 透射式电子显微镜，通过薄片或 冷冻蚀刻复制品。现在，我们要详细检查突触 形态和成像外表面膜大分子，如 糖蛋白和蛋白多糖的高分辨率(扫描电子显微镜)研究 发射扫描电子显微镜。在样品制备方面，我 希望使用冷冻技术、金标和镀铬。 样品制备技术将在阿尔布雷希特进行 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 冷冻断裂神经突触前、后的终末组织 细胞。2)对体外培养的神经细胞中的糖杯进行成像 在外表面膜糖蛋白和 蛋白多糖(透明质酸，软骨化的4-或6-硫酸盐。)另外，我们 将使用共聚焦显微镜。目标是：研究 颗粒、高尔基体、 浦肯野细胞、星状细胞和篮状细胞。追踪大脑皮层内 传入苔藓纤维与颗粒细胞形成的环路 浦肯野细胞的树突、攀援纤维和颗粒细胞轴突 树枝状结构。将获得的扫描共焦图像进行比较 具有先前获得的扫描电子和冷冻蚀刻图像。

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.