Low-cost microdissection system for live brain cell collection (CTAS-Live)

描述(由申请人提供)：a为了推进当今神经科学的基础和翻译研究，必须进行针对细胞/区域的研究。脑组织的复杂性对细胞特异性信息的获取提出了巨大的挑战。目前还没有商业上可用的低成本仪器，可以在细胞分辨率下进行复杂的组织显微解剖，更重要的是，可以收集活细胞 从天然脑组织和细胞培养皿中提取。此外，现有的激光辅助显微解剖仪器是复杂的，并受到特定的样品预处理方案的限制，一般禁止将其用于天然脑组织。在第一阶段，NeuroInDx开发了一种基于一次性毛细管单元(DCU-Live)和短而仔细调节的真空脉冲的活细胞和组织采集方法。原理实验证明，该方法对原始脑组织和经过预处理的脑组织进行显微解剖是非常有效的。用CTAS-Live收集的细胞适合再培养。采集的样品可为下游应用提供高质量的RNA、DNA和蛋白质。该项目的第一阶段导致了CTAS-Live原型的开发，该原型具有从组织和培养中进行细胞分辨率显微切割的能力，而不影响收集的样本的生存能力。该SBIR项目的第二阶段将把CTAS-Live开发成两种商业模式：一种是具有半自动功能和能力的基础模型，用于显微解剖和活细胞采集；另一种是全自动化版本的CTAS-Live，能够通过计算机控制的校准过程、目标识别和通过触摸屏界面控制的连续采集从多个标本中快速采集细胞和组织。拟议工具的一个重要优势是其价格低廉，从而可以有效地进入市场和实现商业化。基础型号的价格低于2.5万美元，高端自动版本的价格将在6.5万美元左右，分别显示出至少四倍于现有激光系统的价格优势和两倍的价格优势。CTAS-Live的基础模型和自动化模型都将通过各种组织样本和细胞培养进行彻底的性能测试，重点是获得的细胞的活性和大分子(例如RNA和蛋白质)的高质量。表观荧光模块将用于从组织中收集荧光标记的细胞。人类死后组织的显微解剖将得到优化，RNA和蛋白质的质量将为基因组学和蛋白质组学应用进行评估。因此，除了CTAS-Live原型的初始测试之外，还将开发几个应用笔记，为潜在客户提供“样本到结果”协议。开发和“内部”测试的CTAS-Live仪器将被送往测试版测试现场，以确保最终用户满意并获得宝贵的反馈。这些研究的结果将是CTAS-Live在该项目第三阶段成功商业化。

DESCRIPTION (provided by applicant): A In order to advance today's basic and translational research in neuroscience, it is imperative that cell/region specific studies can be conducted. Complexity of brain tissues poses a significant challenge when cell specific information has to be derived. Currently there is no commercially available low cost instrument that would permit complex tissue microdissection at cellular resolution and more importantly collection of live cells from both native brain tissues and cell culture dishes. Furthermore, existing laser assisted microdissection instruments are sophisticated and limited by the specific sample pretreatment protocol generally prohibiting its use on native brain tissues. During Phase I, NeuroInDx developed a method for live cell and tissue acquisition that is based on the use of disposable capillary units (DCU-Live) and short, carefully regulated vacuum impulses. Proof of principle experiments demonstrated that the approach is extremely efficient for the microdissection of native and pretreated brain tissues. The cells collected with CTAS-Live are suited for reculturing. Collected samples yield high quality RNA, DNA, and protein for downstream applications. Phase I of this project resulted in the development of a CTAS-Live prototype that features cellular resolution microdissection capabilities from both tissues and cultures without affecting the viability of collected samples. Phase II of this SBIR project will develop CTAS-Live into two commercial models: a base model featuring semiautomatic functions and capability for both microdissection and live cell collection, and a fully automated version of CTAS-Live capable of fast cell and tissue acquisition from multiple specimen via computer controlled calibration process, target recognition and continuous collection controlled via touch screen interface. An important advantage of the proposed instruments is their low price that permits efficient market entrance and commercialization. The base model is priced under $25,000 and the high end automatic version would be priced in the range of $65,000, demonstrating at least four and two times price advantage, respectively versus existing laser based systems. Both base and automated models of CTAS-Live will be thoroughly tested for its performance with a variety of tissue samples and cell cultures focusing on the viability of the acquired cells and hig quality of the macromolecules (e.g. RNA and protein). The epifluorescent module will be tested for the collection of fluorescently labeled cells from tissues. Microdissection of human postmortem tissues will be optimized and the quality of RNA and protein will be evaluated for genomics and proteomics applications. Therefore, aside from initial testing of CTAS-Live prototypes, several Application Notes will be developed offering "sample-to-result" protocols for the potential customers. Developed and "in-house" tested CTAS-Live instruments will be sent to the beta testing sites to ensure end-user satisfaction and obtain valuable feedback. The result of these studies will be successful commercialization of CTAS-Live in Phase III of this project.