SPECIMEN PREP & STAINING TECH FOR MAPPING PROTEINS, CELL COMPLEXES, & PATHWAYS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 标本制备、保存和染色技术，用于使用多标记和多尺度成像绘制蛋白质、细胞复合体、途径和组织的超分子组织图。 我们在这一领域的工作重点是继续开发技术，以便能够使用多标记和多尺度显微镜对蛋白质和大分子复合体进行成像。这些方法包括：(1)扩大我们用于多尺度成像的荧光化合物的库；(2)将高压冷冻/冷冻取代与分子标记方法结合起来，以进一步加强超微结构的保存；(3)采用二氨基联胺的金属螯合衍生物，使其能够使用电子能量损失谱(EELS)及其他检测方法同时标记多蛋白质EM；(4)以及探索使用量子点、纳米荧光粉和荧光蛋白质进行阴极发光(CL)成像的可行性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. SPECIMEN PREPARATION, PRESERVATION AND STAINING TECHNOLOGIES FOR MAPPING PROTEINS, CELLULAR COMPLEXES, PATHWAYS AND THE SUPRAMOLECULAR ORGANIZATION OF TISSUES USING MULTIPLE LABELS AND MULTISCALE IMAGING. Our efforts in this area focused on continuing to develop technologies to enable imaging of proteins and macromolecular complexes using multiple labels and multiscale microscopies. These approaches include (1) expanding our repertoire of fluorescent compounds used for multiscale imaging; (2) combining high-pressure freezing/freeze-substitution with molecular labeling approaches to further enhance preservation of ultrastructure; (3) adapting metal chelating derivatives of diaminobenzidine to enable simultaneous multi-protein EM labeling using electron energy loss spectroscopy (EELS), and other detection methods; (4) and exploring the feasibility of cathodoluminescence (CL) imaging using quantum dots, nanophosphors and fluorescent proteins.