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

样本制备

• 批准号: 8361906
• 负责人: GINA E SOSINSKY
• 金额: $ 16.04万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361906
• 关键词: Area; Biological Preservation; Cells; Complex; Detection; Electron energy loss spectroscopy; Freeze Substitution; Freezing; Funding; Grant; Image; Image Analysis; Label; Maps; Metals; Methods; Microscopy; Molecular; Multiprotein Complexes; National Center for Research Resources; Pathway interactions; Preparation; Principal Investigator; Proteins; Quantum Dots; Research; Research Infrastructure; Resources; Source; Specimen; Staining method; Stains; Technology; Tissues; United States National Institutes of Health; cost; pressure

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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 利用多标记和多尺度成像绘制蛋白质、细胞复合物、途径和组织超分子组织的标本制备、保存和染色方法。 我们在这一领域的努力集中在继续开发技术，使蛋白质和大分子复合物的成像使用多个标签和多尺度显微镜。 这些方法包括：（1）扩大我们用于多尺度成像的荧光化合物库;（2）将高压冷冻/冷冻取代与分子标记方法相结合，以进一步增强超微结构的保存;（3）采用二氨基联苯胺的金属螯合衍生物，以实现使用电子能量损失光谱（EELS）和其他检测方法的同时多蛋白质EM标记;（4）探索了利用量子点、纳米荧光粉和荧光蛋白进行阴极发光成像的可行性。