Supplement for Purchase of a Cell Sorter for R35GM133797

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PROJECT SUMMARY Cellular heterogeneity is ubiquitous across all domains of life. Genetically identical cells can display heterogeneous metabolic activities, even when grown in identical environments. In bacteria, metabolic heterogeneity can shape the ensemble growth rate and affect antibiotic tolerance. In bacteria, metabolic heterogeneity shapes the ensemble growth rate and affects antibiotic tolerance. However, many questions remain regarding metabolic heterogeneity. For instance, what determines the size of metabolic fluctuation? How is metabolic heterogeneity regulated? Can we control metabolic heterogeneity and therefore eliminate drug-tolerant cells? The lack of fundamental understanding has severely limited the development of effective treatments for multiple diseases in which a small number of transiently tolerant cells often cause disease recurrence. The current MIRA project aims to obtain a systematic understanding of bacteria metabolic heterogeneity by using single-cell metabolic analysis methods. Using an engineered metabolic pathway that produces a unique fluorescent metabolite―betaxanthine, this project aims to determine the origin, dynamics, and propagation of metabolic heterogeneity. Furthermore, this project studies how transcriptional regulations affect metabolic heterogeneity, and explores the influence of metabolic heterogeneity on drug tolerance and strategies to reduce metabolic heterogeneity. This Equipment Supplement application requests fund to purchase a new flow cytometer to replace an old flow cytometer in the PI’s lab that was broken and is not repairable. Flow cytometer is essential to the parental project in studying metabolite heterogeneity. Obtaining a new flow cytometer will substantially accelerate the productivity of this project and enable new capabilities to the PI’s research group to explore new areas in metabolite heterogeneity.