Analytical Imaging

项目概述（见说明）：成像在许多AECC成员的研究项目中起着至关重要的作用。分析成像设施（AIF）为整个AECC社区提供常规和复杂的成像技术。显微镜技术范围从传统的明场，到外显荧光，再到激光扫描共聚焦，再到高分辨率透射和扫描电子显微镜，包括大分子的低温电镜，在这个真正全面的共享资源中。这种广泛的成像模式允许研究疾病的动物模型。表达或标记荧光报告分子的细胞可以在整个动物、整个器官、单个细胞或细胞区室中成像。活细胞可以实时成像以监测生理过程。光激活荧光分子可以通过光操纵进行动态体内研究。共聚焦显微镜和外显荧光与反褶积配对，可以对固定材料进行3D成像或对活细胞进行4D成像。传统的透射电镜超微结构分析通过电子断层扫描扩展到三维。最近安装的场发射扫描电子显微镜（SEM）扩展了该设施的SEM能力，包括自动大面积绘图，3D图像重建，x射线微分析，相关荧光和SEM成像以及冷冻水合样品成像。AIF工作人员协助用户进行实验设计、数据收集、定量图像分析和演示。新用户被教授为其特定研究目标所需的成像技术，而有经验的用户可以定制任何成像站，利用该设施的大量光学和配件库存。该设施为电子显微镜样品制备提供定制的全方位服务，以确保质量控制并提供最广泛的技术。AIF工作人员具有通过化学固定，树脂包埋，超薄切片，免疫金标记和阴性染色等方法制备样品的专业知识。该设备为电子显微镜提供全方位的低温技术，包括快速冷冻，金属镜或高压冷冻。冷冻样品可以进行冷冻替代、冷冻断裂、旋转阴影或冷冻切片，然后在环境或低温下进行TEM或SEM成像。AIF为分子细胞遗传学和组织病理学共享资源的显微镜要求提供支持。自上次监测小组审查以来，已增加了重要的新设备、人员和服务，大大提高了非洲影像中心的成像能力。

PROJECT SUMMARY (See instructions): Imaging plays a vital role in the research programs of many AECC members. The Analytical Imaging Facility (AIF) makes routine and complex imaging technologies available to the entire AECC community. Microscope technology ranges from traditional brightfield, to epi-fluorescence, to laser scanning confocal, to high resolution transmission and scanning electron microscopy including cryoEM of macromolecules in this truly comprehensive shared resource. This wide range of imaging modalities allows for the study of animal models of disease. Cells expressing or labeled with fluorescent reporter molecules may be imaged in the whole animal, whole organ, single cell or cell compartments. Live cells may be imaged in real time to monitor physiological processes. Photo-activatable fluorescence molecules may be manipulated by light for dynamic in vivo studies. Confocal microscopes and epi-fluorescence paired with deconvolution enables 3D imaging of fixed material or 4D imaging of live cells. Traditional ultrastructure analysis by transmission electron microscopy is expanded to 3D by electron tomography. The recent installation of a field emission scanning electron microscope (SEM) has expanded the facility's SEM capabilities to include automated large area mapping, 3D image reconstruction, x-ray microanalysis, correlative fluorescence and SEM imaging and imaging frozen hydrated samples. AIF staff assist users in experimental design, data collection, quantitative image analysis and presentation. New users are taught imaging techniques required for their specific research objective, while experienced users may customize any imaging station, utilizing the facility's large inventory of optics and accessories. The facility offers customized full service sample preparation for electron microscopy, to ensure quality control and to offer the widest range of techniques. The AIF staff has the expertise to prepare samples by methods that include chemical fixation, embedding in resin, ultrathin sectioning, immunogold labeling and negative staining. The facility offers a full range of low temperature techniques for electron microscopy including quick freezing by plunge, metal mirror or high pressure freezing. Frozen samples can undergo freeze-substitution, freeze fracture, rotary shadow or cryosectioning and are then imaged in the TEM or SEM at ambient or low temperature. The AIF provides support for the microscopy requirements of the Molecular Cytogenetics and Histopathology Shared Resources. Since the last CCSG review, important new additions of equipment, personnel and services have been made, which have substantially enhanced the imaging capabilities at AECC.