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SELECTBIO Conferences Genomic Applications of qPCR, dPCR & NGS


Single Cell Systems - A Step Toward Functional Single Cell Biology

Jay West, Senior Director of R&D, Fluidigm Corporation

The study of single cells has evolved over the past several years to include expression and genomic studies of an increasing number of single cells. Several studies have demonstrated the wide variation and vast heterogeneity within cell populations of similar phenotype. While the characterization of these populations will likely set the foundation for our understanding of genomic- and expression-based diversity, it will not be able to link the functional differences of a single cell to its underlying genomic structure and activity. Currently, it is not possible to perturb single cells in a controlled environment, monitor and measure the response due to perturbation, and link these response measurements to downstream genomic and transcriptomic analysis. The inability to perform these studies is primarily due to lack of a robust experimental platform, which can integrate and miniaturize many of these single cell functional experimental study steps. In order to address this challenge, we developed a device based on the use of fluidic logic which enables the integration of several key aspects of many biological experiments at nano-scale reaction level. This polymer-based Integrated Fluidic Circuit (IFC) has been used to select and sequester specific single cells based on a phenotypic trait for downstream experimentation. Experiments with sequestered cells that have been performed include on-chip culture, exposure to a variety of stimulants, and post-exposure image-based response analysis, followed by preparation of the mRNA transcriptome for massively parallel sequencing analysis. The flexible system embodies experimental design and execution that enables routine functional studies of single cells.

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