Principal Investigator/Program Director: Haussler, David Project Summary The focus of our CEGS Center for Live Cell Genomics is to build new methodology and capacity for large-scale, long-term, inexpensive, modular, customizable, shared, Internet-of-Things- controlled, reproducible live cell culture and tissue-based experimental genomics disease models. Tissue models include traditional cell culture as well as organoid and primary tissue explants obtained from surgery or biopsy. A particular focus is the integration of organoid factories that support tissue growth and maintenance with external and on-chip electro- optofluidic analytical modules to become part of an ecosystem that is modeled after open- source software. This system will use commodity sensors, cameras, and computers linked in platforms that are flexibly designed using simple, widely available techniques potentially in order to stimulate rapid innovation in experimental platforms for tissue culture. This novel technology will allow us to address major scientific issues in neurodevelopment and pediatric cancer. These include questions about what genes contribute to human brain development, or what specific molecular pathways are disrupted in individual pediatric cancer cases. Over the first year of our project, it has become clear that in order to advance our research agenda on both the scientific and technological sides we require an established, multi-modal nanoparticle analysis method. Specifically, we are facing two challenges. We need to be able to assess the nanoscale extracellular vesicles (EVs), specifically exosomes, that are being produced in our organoid factories as a measure of organoid cell types, metabolism, and health. As these measurements need to be carried out frequently over long periods of time, a fast and relatively easy-to-use technique is required. Secondly, in order to develop optofluidic devices that can carry out measurements on individual EVs and exosomes and their molecular content, we need to be able to establish ground truth references against which the performance of these devices can be compared. To this end, we are requesting supplemental funding to acquire a Nanoparticle Tracking Analyzer (NTA) instrument that will provide these capabilities throughout our project. An NTA is a unique instrument that provides multi-modal analysis on populations of biological and inorganic nanoparticles. Pertinent capabilities include buffer calibration, measurement of EV concentration and size distribution, surface charge measurements, assessment of labeling efficiencies, and more. As such, an NTA is uniquely suited for our requirements and will dramatically accelerate the progress of our research as well as expand its outcomes. 1