DEV OF ADVANCED MULTIDIMENSIONAL MICROSCOPY TECHNOLOGIES FOR MESOSCALE IMAGING

University Of California, San Diego, La Jolla CA

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Abstract

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. DEVELOPMENT OF ADVANCED MULTIDIMENSIONAL MICROSCOPY TECHNOLOGIES FOR MESOSCALE IMAGING The correlation of data acquired across scales and technologies has the promise to reveal relationships among subcellular components, the detailed organization of constituent protein building blocks, and the functional role of each in normal and abnormal biological systems. Continued development and deployment of enhancements to the resource imaging instrumentation is critical to improve and further accelerate the capabilities to investigate biomedical research questions, especially those where insight is obtained by establishing structure and function relationships across scales. To this end, are maximizing the impact of our recently deployed 300KV energy filtering TEM/STEM instrument, integrating its advanced production capabilities and commissioning new imaging modes enabled by NCMIR-specified customizations. We are also refining our existing IVEM and ultra-wide-field light microscopy resources to solidify the diversity of techniques that can be performed at this unique facility. Moreover, we continue to develop and integrate new instrumentation technologies to bridge resolution gaps and augment our ability to conduct seamless multiscale, multi-resolution microscopy. For each instrument, we are focusing on TR&D that increases imaging throughput and fidelity and extends the field-of-view achievable at high resolution. Throughout these activities, we balance engineering development with collaborative use and connect efforts to exploit new labeling and specimen development technologies. This includes the development, refinement, and commissioning of new imaging features and functionality of the resource FEI Titan 80-300 C-TWIN STEM/TEM energy filtering IVEM. We are also propelling the use of high-throughput, mesoscale volumetric and wide-field analytical capabilities of energy filtering 300kV and 400kV JEOL IVEMs at NCMIR, implementing and refining procedures to image thick biological specimens in wide-field by integrating most-probable-loss tomography with image mosaicing on the JEOL 3200EF. This, is supporting numerous multiscale studies benefiting from correlated light and electron microscopic image acquisition by refining NCMIR platforms developed for precision ultra-wide-field microscopy. Finally, we are exploring and deploying emerging microscopy technologies that will extend the Center's mesoscale data acquisition capabilities and complement its specimen development and image analysis and database activities.