SBIR PHASE I TOPIC #439 - ADVANCED SAMPLE PROCESSING PLATFORMS FOR DOWNSTREAM SINGLE-CELL MULTI-OMIC ANALYSIS

本提案的长期目标是开发一个快速，免提的平台，集成组织解离，克隆表征和分离，以支持单细胞多组学领域的发展。在提案的第一阶段，enrich将通过开发用于原位组织解离、多标准单细胞分离和无损单细胞检索的基本试剂、方案和消耗品，来改进现有的基于图像、抗碎片的克隆分离机。组织消化、单细胞捕获、清洗和回收的效率将由现有的实时成像功能严密监控。从人肝脏和胰腺肿瘤中分离出足够的、高活力的单细胞，通过10xGenomics平台制备每个样本的scRNA和scDNA测序文库，提交Illumina测序。验证将通过一系列指标来实现，包括总操作时间、人工干预水平、单细胞产量和质量以及sc序列可映射性。如果成功，下一阶段将进行多部位肿瘤类型研究。

The long term goal of this proposal is to develop a fast, hands-free platform that integrates tissue dissociation, clone characterization and isolation to support the growing field of single celll multiomics. During the phase I of the proposal, enrich will adapt the exiting image based, debris-resistant clone isolation machine by developing essential reagents, protocols and consumables for in-place tissue dissociation, multicriteria single cell isolation and lossless single cell retrieval. Efficiency of tissue digestion, single cell capture, washing and retrieval will be tightly monitored by existing real-time imaging functionality. Adequate and high viability single cells from human liver and pancreas tumors will be isolated and scRNA and scDNA sequencing libraries of each sample will be prepared by 10xGenomics platform and submitted for Illumina sequencing. The validation will be achieved by an array of metrics including total operation time, level of manual intevention, single cell yield and quality and sc sequence mappability. if successful, a multi-site, tumor type study will be performed in the next phase.