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

SBIR 主题 439 - 用于下游单细胞多组学分析的先进样品处理平台

• 批准号: 10723179
• 负责人: ADAM BISOGNI
• 金额: $ 39.97万
• 依托单位: INSO BIOSCIENCES INC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-19 至 2023-09-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10723179
• 关键词: Attention; Automation; Biological; Buffers; Cell Count; Cell Line; Cell Separation; Cell Size; Cells; Centrifugation; Clinical; Collaborations; Data; Development; Digestion; Dissociation; Enzymes; Erythrocytes; Expression Library; Filtration; Freezing; Gel; Gene Expression; Genomics; Goals; Hour; Human; Image; Individual; Intervention; Libraries; Liquid substance; Liver; Manuals; Modification; Monitor; Mus; Organ; Pancreas; Pathology; Performance; Phase; Preparation; Procedures; Process; Protocols documentation; Reagent; Recovery; Residual state; Resort; Retrieval; Running; Sampling; Selection Criteria; Small Business Innovation Research Grant; Solid; Source; Technology; Testing; Time; Tissues; Tube; Tumor Tissue; Validation; Variant; base; cell fixing; digital; experimental study; gene expression variation; gene panel; human tissue; image guided; improved; multiple omics; operation; protocol development; transcriptome; transcriptome sequencing; tumor

Single cell purifications are usually needed for most single cell multiomics platforms. Unwanted cells such as dead cells, doublet, or residual red blood cells will greatly reduce the effective data. Sample preparation of solid tissues into viable, non-red blood cells consists of three major steps: tissue dissociation, clump/debris filtration and single cells purification. Improvements been made to individual steps to enhance efficiency and reduce time. Though each step can be individually optimized to high completeness and efficiency, extensive pipetting and centrifugation operations are still required to bridge most, if not all, three steps. Therefore, the multistep, loosely monitored, attention-intensive process normally takes much longer in practice, and the material loss during the whole process can be rather high (from 10^8 to 10^4 -10^6 or 99%- 99.99%). Using Enrich TroVo technology, we believe eliminating intermediate steps while keeping all material in one place is a more effective strategy than improving the efficiency of individual steps. And we found the key of eliminating intermediate steps is to enhance the overall debris tolerance of the cell sorting process and to combine multiple purification goals into one single isolation step. Using image guided digital filter, multiple selection criteria such as viability, singularity, cell size, can be combined into one composite filter and directly applied to a complicated mixture of tissue digests. The purpose of this proposed project is to further specialize such image-based technology into a highthroughput, application ready product. With the newly forged collaboration with Yale pathology, Enrich will be able to validate this platform using multiple clinical samples.

大多数单细胞多组学平台通常需要单细胞纯化。不需要的细胞，如死细胞、双重细胞或残留的红细胞将大大降低有效数据。将固体组织样品制备成有活力的非红细胞包括三个主要步骤：组织解离、团块/碎片过滤和单细胞纯化。