Development of a novel low-cost capillary-based cell and tissue acquisition syste

开发一种新型低成本毛细管细胞和组织采集系统

• 批准号: 7329850
• 负责人: Lili C Kudo
• 金额: $ 12.48万
• 依托单位: NEUROINDX INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-08-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7329850
• 关键词: Advanced Development; Affect; Animal Experimentation; Architecture; Basic Science; Biological; Biological Markers; Biological Preservation; Blood capillaries; Cell Separation; Cell physiology; Cells; Cellular Morphology; Clinical; Collection; Computer Systems Development; Computer software; Condition; DNA; Data; Development; Devices; Disease; Dissection; Electronics; Ensure; Environment; Ethanol; Fluorescence; Freezing; Funding; Gene Expression Profile; Genomics; Goals; Heterogeneity; Human; Institution; Invasive; Label; Laboratories; Lasers; Life; Maintenance; Manuals; Metabolism; Methods; Microdissection; Microscope; Molecular; Molecular Analysis; Performance; Phase; Play; Price; Process; Proteins; Proteome; Proteomics; RNA; Range; Regulation; Research; Role; Sampling; Sorting - Cell Movement; Speed; System; Techniques; Technology; Testing; Time; Tissue Banks; Tissue Fixation; Tissues; Vacuum; Work; abstracting; animal tissue; base; capillary; cell type; concept; cost; drug development; improved; instrument; laser capture microdissection; macromolecule; novel; novel therapeutics; prototype; research study; sample collection; therapeutic target; tissue fixing; tissue preparation; tool; user-friendly

DESCRIPTION (provided by applicant): 1. ABSTRACT Tissue heterogeneity has always limited the information obtainable from genomic or proteomic analysis of biological samples in the study of disease. Tissue microdissection and cell sorting technologies have advanced tremendously over the last decade from simple manual tissue dissection to sophisticated laser capture microdissecting (LCM) instruments and high speed fluorescence assisted cell sorting systems (FACS). In combination with genomics and proteomics technologies it is now possible to generate cell specific transcriptome/proteome data advancing the identification of disease biomarkers and novel therapeutic targets. Currently, LCM and FACS are two main technologies for isolation of specific tissues and cell types. However, it is clear that these technologies are not sufficient to fully support the growing need for cell specific molecular data. This process is hampered by extremely high costs of LCM and FACS machines, high cost of their maintenance and their sophisticated interface. Further, each of them has specific limitations: 1) LCM performs cell and tissue collection using fixed tissues that often affects the quality of biological material such as RNA; 2) FACS requires fluorescent labels and works with dissociated cells. The need for the low-cost, reliable and simple-to-use instrument that would offer capabilities similar to LCM and FACS is evident. Here, we propose a novel capillary-based vacuum-assisted cell and tissue acquisition system (CTAS) that is envisioned as an attachment to existing inverted microscopes and will allow specific cell and cell cluster collection from fresh, fresh frozen and fixed tissues for downstream genomics and proteomics applications. This instrument is estimated to be in the price range of $5,000, therefore it will be affordable for any laboratory. This machine will have interchangeable key components and will be user friendly. Cell specific sorting/capture technology is a prerequisite for precise characterization of the specific cell types for understanding their function, regulation of the metabolism and drug development. Currently two major approaches for acquisition of specific cells are available: fluorescence assisted cell sorting (FACS) and laser-capture microdissection (LCM). These technologies are sophisticated, have high cost of maintenance and the instruments are very expensive. We are developing a low-cost vacuum-assisted capillary-based cell and tissue acquisition system (CTAS). It is a simple, non-invasive (unlike LCM it does not require tissue fixing and drying) technology that can be easily automated and offers wide range of cell- and tissue-specific separation parameters. In addition, it is estimated to be at least 20 times cheaper than any existing devices such as LCM or FACS. This simple, rapid, microcapillary-based technique will be capable of isolating cells and tissues from animal and human material for further downstream molecular analysis without compromising the preservation of macromolecules in the tissue. Low-cost CTAS featuring open system architecture and interchangeable components will play a key role in collection and processing of cell-specific genomics and proteomics data in biomedical experiments.

描述（由申请人提供）：1。摘要在疾病研究中，组织异质性一直限制着从生物样品的基因组或蛋白质组学分析中获得的信息。组织显微切割和细胞分选技术在过去的十年中已经从简单的手动组织切割发展到复杂的激光捕获显微切割（LCM）仪器和高速荧光辅助细胞分选系统（FACS）。与基因组学和蛋白质组学技术相结合，现在可以生成细胞特异性转录组/蛋白质组数据，从而促进疾病生物标志物和新型治疗靶点的鉴定。目前，LCM和FACS是用于分离特定组织和细胞类型的两种主要技术。然而，很明显，这些技术不足以完全支持对细胞特异性分子数据日益增长的需求。这一过程受到LCM和FACS机器的极高成本、它们的高维护成本和它们的复杂接口的阻碍。此外，它们中的每一个都具有特定的局限性：1）LCM使用固定的组织进行细胞和组织收集，这通常会影响生物材料如RNA的质量; 2）FACS需要荧光标记并与解离的细胞一起工作。对低成本、可靠且使用简单的仪器的需求是显而易见的，该仪器将提供类似于LCM和流式细胞仪的功能。在这里，我们提出了一种新的毛细管为基础的真空辅助细胞和组织采集系统（CTAS），设想作为现有的倒置显微镜的附件，并将允许特定的细胞和细胞簇收集新鲜，新鲜冷冻和固定的组织下游基因组学和蛋白质组学应用。该仪器的价格估计在5，000美元左右，因此任何实验室都能负担得起。这台机器将有可互换的关键部件，并将是用户友好的。细胞特异性分选/捕获技术是精确表征特定细胞类型以了解其功能、代谢调节和药物开发的先决条件。目前，用于获取特定细胞的两种主要方法是可用的：荧光辅助细胞分选（FACS）和激光捕获显微切割（LCM）。这些技术复杂，维护成本高，仪器非常昂贵。我们正在开发一种低成本的真空辅助毛细管为基础的细胞和组织采集系统（CTAS）。它是一种简单的非侵入性技术（与LCM不同，它不需要组织固定和干燥），可以轻松自动化，并提供广泛的细胞和组织特异性分离参数。此外，它估计比任何现有设备（如LCM或FACS）便宜至少20倍。这种简单、快速、基于微毛细管的技术将能够从动物和人类材料中分离细胞和组织，用于进一步的下游分子分析，而不会损害组织中大分子的保存。具有开放系统架构和可互换组件的低成本CTAS将在生物医学实验中收集和处理细胞特异性基因组学和蛋白质组学数据方面发挥关键作用。