Microfluidics-Based Single-Cell Chemical Analysis of Cyanobacteria

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Single-cell analysis is an emerging technology that promises to provide new perspectives on biology. Because single-cell analysis observes cells as individuals, it eliminates ensemble averaging and resolves the heterogeneity present within a sample population. This heterogeneity can provide useful information with regard to important cellular processes, especially if a significant number of cells display a marked deviation from that of average-cell behavior. This research project will be to develop single-cell analysis technology on a microfluidic platform to study the phycobilisome (PBS) degradation process of a unicellular cyanobacterium Synechococcus. With the single-molecule detection, very low copy numbers of fluorescent molecules can be directly counted from a single-cell lysate, which is the case for bleached cyanobacteria cells. These tools will be used to understand the biochemical mechanism of PBS degradation at the single-cell level.Broader ImpactsThis project will have a significant impact on many different areas of research. First, the biological insight gained from this research will illuminate a new way of viewing ecological diversity, microbiology, and the interrelationship between them. Second, single-cell technology will be advanced toward the integration of cell culture, isolation and manipulation of cells, and the analysis of intracellular contents (both proteins and nucleic acids). With this integrative approach, it should be possible to obtain accurate biochemical information from individual cells as well as simple phenotypic changes. This strategy can be extended to other types of cells and should provide a new paradigm for studying biological heterogeneity. Third, this project is interdisciplinary in essence, involving chemical analysis with biological outcomes. This combination should induce a synergistic conversation between many fields, including ecology, population biology, and biophysical chemistry, resulting in more productive directions and motivations for future research. Fourth, there will be a strong component of teaching and training both at the undergraduate and graduate level as well as annual multidisciplinary workshops to introduce participants to the potential uses of single-cell technology in biology.