IDBR: TYPE A Nanoscale Laser Ablation Capture for Single Cell Genomics

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An award is made to Louisiana State University to develop an instrument for localized sampling of genomic material from single cells in tissue. The proposed spatially resolved single cell genomics instrument will enable studies of cellular processes on a single cell level with high resolution. The instrument will facilitate local sampling of single cells and determination of genetic differences from cell-to-cell within a heterogeneous population. It will enhance the ability to study the genomics of stem cell development and the study of genetic mosaicism in the brain, which has important implications in understanding neurological diseases and their treatment. The project will involve students and teachers as well as undergraduate students, graduate students, and postdocs in several initiatives that will advance STEM education and career development in Louisiana and the Gulf Coast region. The results of the project will be disseminated using an array of social media platforms that are aimed at demonstrating the capabilities of the approach broadly to the general public.Genetic differences between individual cells cause differences in their biological function; however, extracting DNA and RNA from large numbers of cells obscures important information about individual cells. High performance next-generation sequencing can be performed on isolated cells, but only after they have been extracted, which loses the information about their location in the organism. This project is aimed at creating a device that can extract material from single cells while preserving its location in the tissue. The instrument uses an atomic force microscope to image the tissue and high-precision tip-enhanced laser ablation to select material for quantification and sequencing. The material is sampled with high precision from individual cells in spatially selected regions. The project goals are construction of the laser ablation system, automated imaging and sample collection, and demonstration of the instrument with breast cancer tissue samples.