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

描述(申请人提供)：1.在疾病研究中，组织的异质性一直限制了从生物样本的基因组或蛋白质组分析中获得的信息。在过去的十年中，组织显微解剖和细胞分选技术有了巨大的进步，从简单的手动组织解剖到复杂的激光捕获显微解剖(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，现在可以产生细胞特异性转录组/蛋白质组数据，促进疾病生物标记物和新的治疗靶点的识别。目前，LCM和FACS是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定分子数据日益增长的需求。这一过程受到LCM和FACS机极高的成本、高昂的维护成本和复杂的接口的阻碍。此外，它们都有特定的局限性：1)LCM使用固定的组织进行细胞和组织收集，这通常会影响RNA等生物材料的质量；2)FACS需要荧光标记，并与分离的细胞一起工作。显然，需要一种低成本、可靠和简单易用的仪器，提供类似于LCM和FACS的功能。在这里，我们提出了一种新型的基于毛细管的真空辅助细胞和组织采集系统(CTAS)，该系统被设想为现有倒置显微镜的附件，并将允许从新鲜、新鲜的冷冻和固定组织中收集特定的细胞和细胞簇，用于下游基因组学和蛋白质组学应用。这种仪器的价格估计在5000美元左右，因此对任何实验室来说都是负担得起的。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、新陈代谢调节和药物开发的先决条件。目前，获取特定细胞的主要方法有两种：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，仪器也非常昂贵。我们正在开发一种低成本的真空辅助毛细管细胞和组织采集系统(CTAS)。这是一种简单、非侵入性的技术(与LCM不同，它不需要组织固定和干燥)，可以很容易地实现自动化，并提供广泛的细胞和组织特定分离参数。此外，据估计，它比任何现有的设备，如LCM或FACS，至少便宜20倍。这项简单、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，而不会影响组织中大分子的保存。低成本的CTAS具有开放的系统架构和可互换的组件，将在生物医学实验中收集和处理特定细胞基因组和蛋白质组数据方面发挥关键作用。

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.