Developing an automated yeast dissection system for aging research

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Project Summary Developing an Automated Yeast Dissection System for Aging Research Aging is the single greatest risk factor for diseases that are principal causes of mortality. The objectives of aging research are to discover key genes and pathways related to aging that may eventually contribute to retardation of aging and a delay in the onset of age-associated diseases. The budding yeast Saccharomyces cerevisiae has been a powerful model for the study of aging and has enabled significant contributions to our understanding of basic mechanisms of aging in eukaryotic cells. However, traditional assays of yeast aging, including microdissection methods, have technical challenges; for instance, the methods are low-throughput and the experimental procedures are laborious. An experiment typically lasts several weeks or months, and requires overnight storage of the assayed cells at a refrigerator to pause replication throughout the course of experiment. This tedious procedure has substantially hindered progress in the field of aging research. We have developed an innovative and highly effective microfluidic platform for continuous and automatic dissection of daughter cells without disturbing mother cells as they bud. The platform allows an automated whole-lifespan tracking with high spatiotemporal resolution and large-scale quantification of single yeast cells, resulting in significant reduction of labor, time, and cost. In addition, the high-resolution florescence imaging of yeast cells grown in constant and dynamically changing environments offers the ability to examine the dynamics of gene expression and signaling networks in a high-throughput manner. The quantity and types of data acquired by this platform are impossible with the traditional assay methods. Herein, we propose to develop automated dissection platform that enable fast and high-throughput studies of yeast aging. We will validate the reliability and capability of the proposed platform We propose to validate the prototype automated dissection platform for two important assays in yeast aging research: determination of replicative lifespan for mutants in an automated and unattended fashion, and continuous tracking the changes in abundance and cellular localization for specific GFP fusion proteins during the entire aging process. Longer term plan of this proposal is to commercialize a fully automation benchtop system for high-throughput studies of cellular aging.