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转化研究，必须进行细胞/区域特异性研究。当必须导出细胞特异性信息时，脑组织的复杂性构成了重大挑战。目前，没有商业上可获得的低成本仪器，其将允许以细胞分辨率进行复杂的组织显微切割，并且更重要的是收集活细胞 从原生脑组织和细胞培养皿中提取。此外，现有的激光辅助显微切割仪器是复杂的，并且受到特定样品预处理方案的限制，通常禁止其在天然脑组织上使用。在第一阶段，NeuroInDx开发了一种活细胞和组织采集方法，该方法基于使用一次性毛细管单元（DCU-Live）和短的、仔细调节的真空脉冲。原理实验证明，该方法是非常有效的原生和预处理的脑组织的显微切割。用CTAS-Live收集的细胞适合再培养。收集的样品产生高质量的RNA、DNA和蛋白质用于下游应用。该项目的第一阶段导致CTAS-Live原型的开发，该原型具有来自组织和培养物的细胞分辨率显微切割能力，而不影响所收集样品的活力。该SBIR项目的第二阶段将把CTAS-Live开发成两个商业模型：一个基础模型，具有显微切割和活细胞收集的半自动功能和能力，以及一个全自动版本的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收集，从多个标本中快速获取细胞和组织。所提议的工具的一个重要优点是价格低廉，可以有效地进入市场并实现商业化。基本型号的价格低于25，000美元，高端自动版本的价格将在65，000美元范围内，与现有的激光系统相比，分别显示出至少四倍和两倍的价格优势。CTAS-Live的基础和自动化模型都将通过各种组织样本和细胞培养物对其性能进行全面测试，重点是获得的细胞的活力和高质量的大分子（例如RNA和蛋白质）。将测试落射荧光模块，以从组织中收集荧光标记的细胞。将优化人类死后组织的显微切割，并评估RNA和蛋白质的质量，以用于基因组学和蛋白质组学应用。因此，除了CTAS-Live原型的初步测试外，还将开发几个应用说明，为潜在客户提供“样品到结果”的协议。开发和“内部”测试的CTAS-Live仪器将被发送到beta测试站点，以确保最终用户满意并获得有价值的反馈。这些研究的结果将是CTAS-Live在该项目第三阶段的成功商业化。

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.