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.
