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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 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。 我们在这一领域的努力集中在开发能够使用多标记和多尺度显微镜同时成像蛋白质和大分子复合体的技术。这些方法包括(1)新的二氨基联苯胺金属螯合衍生物，可用于同时使用电子能量损失谱(EELS)进行多蛋白质EM标记；(2)提高用于多尺度成像的荧光化合物库的灵敏度；以及(3)改进树脂包埋样品中探针荧光、抗原性和细胞超微结构的保存，以促进新的成像应用。此外，我们正在努力扩大荧光纳米颗粒(例如量子点)作为多尺度、多模式成像探测器的用途。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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 are focused on developing technologies to enable simultaneous imaging of proteins and macromolecular complexes using multiple labels and multiscale microscopies. These approaches include (1) new metal chelating derivatives of diaminobenzidine that can be used for simultaneous multi-protein EM labeling using electron energy loss spectroscopy (EELS); (2) enhancing the sensitivity of our repertoire of fluorescent compounds used for multiscale imaging; and (3) improved preservation of probe fluorescence, antigenicity, and cellular ultrastructure in resin-embedded samples to facilitate new imaging applications. In addition, we are working to extend the usefulness of fluorescent nanoparticles (e.g., quantum dots) as probes for multiscale, multimodal imaging.