Acquisition of a State-of-the-Art Electron Cryomicroscope
购置最先进的电子冷冻显微镜
基本信息
- 批准号:9977556
- 负责人:
- 金额:$ 80万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:1999
- 资助国家:美国
- 起止时间:1999-10-01 至 2003-09-30
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
9977556Abstract This project involves the acquisition and installation of a 300 kEV field emission gun (FEG) transmission electron microscope (TEM) to determine the structures of cellular machinery. Cellular machines such as the actin cytoskeleton are often large structures difficult to study by methods other than electron cryomicroscopy. For example, it has been possible to dock atomic models for the cytoskeletal components obtained by x-ray crystallography into the molecular maps of the cytoskeletal complexes obtained by electron cryomicroscope. Analysis of filamentous structures has been extended to 10 A resolution in many cases. A state of the art microscope is essential for these studies. The microscope to be obtained and installed during this project will be utilized for the study of the splicesome, voltage-gated ion channels, the actin bundle of the intestinal microvillus, the bacterial flagellar motor and filament, the bacterial gas vesicle, the receptor-kinase signaling complex and the complex of actin with myosin. The aim of these studies are atomic models of the structures. In some instances the structure will be obtained to atomic resolution using electron cryomicroscopy alone. In other cases, atomic models obtained by x-ray crystallography will be docked into molecular maps obtained by cryomicroscopy. The 300 kEV FEG TEM is essential to these projects because the quality of the micrographs limits the resolution of the map in the same way that the quality of the objective lens limits the resolution of the light microscope. No matter how sophisticated, computer processing cannot extract what is not present in the images. The two key features of this microscope in this regard are the higher voltage and the FEG. With these, it is possible to take highly defocused images which have strong contrast at low resolution but which preserve high resolution detail. The strong contrast at low resolution is essential for accurate alignment of images prior to averaging. If the alignment (or correction for distortion) is not accurate, one will average non-equivalent features and resolution will be lost. This microscope will extend the resolution of current maps and therefore, the ability to obtain accurate atomic models whether it be directly or by the docking of atomic models into molecular maps. The cryomicroscope is essential to the training of young scientists. Since the kind of microscopy being used represents a frontier in structural studies of cellular machinery, students and post doctoral fellows must be trained in these techniques. The need for such scientists is increasing rapidly now that the power of cryoelectron microscopy has been demonstrated. There are currently few places equipped for this kind of training.
摘要:本项目采用300 kEV场发射枪(FEG)透射电子显微镜(TEM)采集和安装,用于测定细胞机械的结构。细胞机器,如肌动蛋白细胞骨架,通常是大型结构,很难用电子冷冻显微镜以外的方法来研究。例如,已经有可能将x射线晶体学获得的细胞骨架成分的原子模型与电子低温显微镜获得的细胞骨架复合物的分子图对接。在许多情况下,对丝状结构的分析已扩展到10a分辨率。对这些研究来说,最先进的显微镜是必不可少的。本项目获得并安装的显微镜将用于研究肠微绒毛的剪接体、电压门控离子通道、肌动蛋白束、细菌鞭毛马达和细丝、细菌气囊、受体-激酶信号复合体和肌动蛋白复合体。这些研究的目的是建立这些结构的原子模型。在某些情况下,仅使用电子低温显微镜即可获得原子分辨率的结构。在其他情况下,通过x射线晶体学获得的原子模型将与低温显微镜获得的分子图对接。300 kEV FEG TEM对这些项目至关重要,因为显微图的质量限制了地图的分辨率,就像物镜的质量限制了光学显微镜的分辨率一样。无论多么复杂,计算机处理都无法提取出图像中不存在的东西。在这方面,这台显微镜的两个关键特征是更高的电压和FEG。有了这些,可以在低分辨率下拍摄高度散焦的图像,这些图像具有强烈的对比度,但保留了高分辨率的细节。在低分辨率下的强烈对比度对于在平均之前精确对准图像是必不可少的。如果对准(或对失真的校正)不准确,则将平均非等效特征,并且将失去分辨率。该显微镜将扩展当前地图的分辨率,因此,无论是直接还是通过原子模型与分子图的对接,都能够获得精确的原子模型。低温显微镜对培养年轻科学家是必不可少的。由于所使用的显微镜代表了细胞机制结构研究的前沿,学生和博士后必须接受这些技术的培训。由于低温电子显微镜的威力已经得到证实,对这类科学家的需求正在迅速增加。目前很少有地方配备这种培训。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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David DeRosier其他文献
David DeRosier的其他文献
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收缩机制的物理化学基础
- 批准号:
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$ 80万 - 项目类别:
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X-Ray Crystallographic Studies of the Multi-Enzyme Complexes
多酶复合物的 X 射线晶体学研究
- 批准号:
7513635 - 财政年份:1976
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