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一个大的CCD探测器马赛克，以提供一种改进的胶片替代品，用于生物电子显微镜数据的直接数字记录。这样的系统将立即适用于和有益于大分子的冷冻电子显微镜（cryo-EM）和大分子复合物、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cryo-EM），该系统将能够常规收集高达100万张大分子复合物的图像，促进和加速蛋白质复合物的高分辨率结构的确定，而不会难以生长纯化材料的大晶体（否则蛋白质晶体学的情况）。同样的新检测器技术也将允许收集宽视场图像，用于跨越多个空间尺度的中等分辨率3D显微镜，以确定组织中细胞的大功能域中超分子复合物的精确组织。 在最近成功的透镜耦合CCD相机技术的基础上，我们的目标是建立一个新的探测器，包含9个CCD探测器的马赛克，每个探测器都有自己的透镜，可以覆盖15 cm x l5 cm闪烁体屏幕的重叠部分。该马赛克探测器的功能分辨率为5600 x 5600像素。我们最初将设计和测试一个单一的“模块”，这个探测器与一个背面照明2K × 2K像素（13.5克）的CCD传感器和一个原型透镜。随后将设计和开发一个2 x 2的探测器和透镜镶嵌体。这个中等复杂的镶嵌相机将使我们能够在第三年制作9个CCD镶嵌之前解决所有设计和开发问题。在这三年中，一项重要的工作将是绘制出所有探测器的失真图，并开发软件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.