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。死细胞、双联体或残留红细胞等不需要的细胞将大大降低有效数据。将固体组织样品制备成活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。 对各个步骤进行了改进，以提高效率并减少时间。尽管每个步骤都可以单独优化以达到高完整性和高效率，但仍然需要大量的移液和离心操作来桥接大多数（如果不是全部）三个步骤。因此，多步骤、松散监控、注意力密集的过程在实践中通常需要更长的时间，并且整个过程中的材料损失可能相当高（从 10^8 到 10^4 -10^6 或 99%- 99.99%）。我们相信，使用 Enrich TroVo 技术，消除中间步骤，同时将所有材料保留在一个地方，是比提高各个步骤的效率更有效的策略。我们发现消除中间步骤的关键是增强细胞分选过程的整体碎片耐受性，并将多个纯化目标结合到一个分离步骤中。使用图像引导数字滤波器，可以将多种选择标准（例如活力、奇异性、细胞大小）组合成一个复合滤波器，并直接应用于复杂的混合物 组织消化物。该项目的目的是进一步将这种基于图像的技术专业化为高吞吐量、应用就绪的产品。通过与耶鲁大学病理学新建立的合作，Enrich 将能够使用多个临床样本验证该平台。