A LARGE MOSAIC CCD CAMERA FOR CRYO EM

描述（由申请人提供）： 拟议项目的目标是设计和建造大型 CCD 探测器，为生物电子显微镜数据的直接数字记录提供改进的胶片替代品。这样的系统将立即适用并有益于大分子的冷冻电子显微镜 (cryo-EM) 以及大分子复合物、细胞器和细胞子系统的 3D 电子断层扫描。对于分子显微镜（使用冷冻电镜），该系统将能够常规收集多达 100 万张大型大分子复合物图像，促进和加速蛋白质复合物高分辨率结构的确定，而无需生长纯化材料的大晶体（与蛋白质晶体学的情况不同）。同样的新探测器技术还将允许跨越多个空间尺度的中间分辨率 3D 显微镜收集宽视场图像，从而能够确定组织中细胞大功能域中超分子复合物的精确组织。 基于最近成功的镜头耦合 CCD 相机技术，我们的目标是构建一种包含 9 个 CCD 探测器镶嵌的新型探测器，每个探测器都有自己的镜头，可以覆盖 15 cm x l5 cm 闪烁体屏幕的重叠部分。该马赛克检测器的功能分辨率为 5600 x 5600 像素。我们首先将设计并测试该探测器的单个“模块”，其中包含一个背照式 2K x 2K 像素（13.5 gm）CCD 传感器和一个原型镜头。随后将设计和开发 2 x 2 探测器和透镜镶嵌体。这款中等复杂的马赛克相机将使我们能够在第三年制作 9 CCD 马赛克之前解决所有设计和开发问题。三年期间的一项重大工作将是绘制出所有探测器失真并开发软件以将探测器子图像无缝拼接在一起。

DESCRIPTION (provided by applicant): The goal of the proposed project is to design and build a large mosaic of CCD detectors to provide an improved film substitute for direct digital recording of data for biological electron rnicroscopy. Such a system will be immediately applicable and beneficial to cryo-electron microscopy (cryo-EM) of macromolecules and 3D electron tomography of macromolecular complexes, organelles, and cellular subsystems. For molecular microscopy (with cryo-EM), this system will enable the routine collection of up to l million images of large macromolecular complexes, facilitating and accelerating the determination of high-resolution structures of protein complexes, without the difficulty of growing large crystals of purified materials (as is otherwise the case with protein crystallography). The same new detector technology will also allow the collection of wide-field images for intermediate resolution 3D microscopy spanning multiple spatial scales, to enable determinations of the precise organization of supramolecular complexes throughout large functional domains of cells in tissues. Building on the recent success of lens-coupled CCD camera technology, our goal is to build a new detector containing a mosaic of 9 CCD detectors, each with its own lens that can cover an overlapping portion of a scintillator screen of 15 cm x l5 cm. This mosaic detector will have a functional resolution of 5600 x 5600 pixels. We will initially design and test a single "module" of this detector with one back-side illuminated 2K x 2K pixel (13.5 gm) CCD sensor and one prototype lens. This will be followed by the design and development of a 2 x 2 mosaic of detectors and lenses. This moderately complex mosaic camera will allow us to work out all design and development issues before making the 9 CCD mosaic in the 3rd year. A significant effort during all 3 years will be to map out all detector distortions and to develop software to seamlessly stitch together detector subimages.