Developing an automated yeast dissection system for aging research

PROJECT SUMMARY Developing an Automated Yeast Dissection System for Aging ResearchAging is the single greatest risk factor for diseases that are principal causes of mortalityThe 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 diseasesThe 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 cellsHowevertraditional assays of yeast agingincluding microdissection methodshave technical challengesfor instancethe methods are low throughput and the experimental procedures are laboriousAn experiment typically lasts several weeks or monthsand requires overnight storage of the assayed cells at a refrigerator to pause replication throughout the course of experimentThis tedious procedure has substantially hindered progress in the field of aging researchHereinwe propose to develop automated dissection system that enable continuous and automatic dissection of daughter cells without disturbing mother cells as they budThe system allows an automated whole lifespan tracking with high spatiotemporal resolution and large scale quantification of single yeast cellsresulting in significant reduction of labortimeand costIn additionthe 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 mannerThe quantity and types of data acquired by this system are impossible with the traditional assay methodsIn Phase I of this STTR grant we developed a proof of concept prototype automated dissection chip and demonstrated feasibility of the microfluidics based yeast dissection technologyIn Phase II projectwe will complete the development of an automated dissection system through scaling up production of the automated dissection chip and development of a dedicated software to analyze time lapse images generated from the automated dissection chipIn additionwe will optimize application proceduresassess data variability and finalize quality control standards for commercialization of the automated dissection system NARRATIVE Developing an Automated Yeast Dissection System for Aging ResearchIn this proposalwe seek to develop a commercial microfluidics system to study yeast budding and lifespan in a revolutionary high throughput mannerto provide a better understanding on agingThe commercialized system will offer an unparalleled method to significantly accelerate the discovery of novel conserved aging regulators and pathways by providing a fasthigh throughputand accurate analysis at the single cell level in budding yeast