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

路易斯安那州立大学获得了一项奖项，以开发一种仪器，用于从组织中的单个细胞中定位取样基因组物质。提出的空间分辨率单细胞基因组学仪器将能够在单细胞水平上以高分辨率研究细胞过程。该仪器将有助于单细胞的局部取样和确定异质群体中细胞间的遗传差异。它将增强研究干细胞发育的基因组学和研究大脑中的遗传嵌合体的能力，这对理解神经系统疾病及其治疗具有重要意义。该项目将涉及学生和教师，以及本科生、研究生和博士后，这些项目将推动路易斯安那州和墨西哥湾沿岸地区的STEM教育和职业发展。该项目的成果将通过一系列社会媒体平台进行传播，旨在向公众广泛展示该方法的能力。单个细胞之间的遗传差异导致其生物功能的差异；然而，从大量细胞中提取DNA和RNA模糊了单个细胞的重要信息。高性能的下一代测序可以在分离的细胞上进行，但只有在它们被提取出来之后，才会失去它们在生物体中的位置信息。该项目旨在创造一种设备，可以从单个细胞中提取物质，同时保留其在组织中的位置。该仪器使用原子力显微镜对组织成像，并使用高精度尖端增强激光烧蚀选择材料进行定量和测序。材料在空间选择区域的单个细胞中进行高精度采样。项目目标是建立激光消融系统，实现自动成像和样本采集，并对该仪器进行乳腺癌组织样本的演示。

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.