This is a revised SBIR Phase II application. The proposal seeks to develop an automated cell culture and analysis system for single cells and apply this system to study of the growth of human hematopoietic stem cells. The approach is based on the hypothesis that an automated single cell bioreactor can be designed and constructed and that this system will provide a tool to advance the basic scientists' ability to dissect the basic biology of cell division and differentiation and to discover how to expand human hematopoietic stem cells in vitro. The system to be developed is a robotic system that intends to automate the processes of: 1) of cell culture and provide information at the level of individual cells; 2) screening of the effects of biological or chemical moieties on tissue-specific cells; and 3) manipulating the fluid microenvironment of cells based upon real-time identification of changes in cell state (e.g., cell division, cell phenotype, apoptosis, etc.). In this Phase II project, the performance and image processing capability of the existing system will be improved and validated; once these tasks are completed, the system's capabilities will be expanded in terms of throughput and the quality and quantity of information obtained. Applications and testing protocols developed in the Phase II program will be directed at the system's use to understand the cell biology of hematopoietic stem cells although the system ultimately would have a number of biological and drug discovery applications.
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