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

用于活体脑细胞采集的低成本显微切割系统 (CTAS-Live)

• 批准号: 8690150
• 负责人: Stanislav Karsten
• 金额: $ 50.68万
• 依托单位: NEUROINDX INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-18 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8690150
• 关键词: Adult; Affect; Area; Autopsy; Basic Science; Blood capillaries; Boxing; Brain region; Budgets; Calibration; Cell Culture Techniques; Cell Survival; Cell model; Cells; Coculture Techniques; Collecting Cell; Collection; Communities; Complex; Computer software; Computers; DNA; Development; Ensure; Ethanol; Evaluation; Feedback; Freezing; Genomics; Growth; Housing; Human; Image; In Vitro; Individual; Label; Lasers; Legal patent; Life; Marketing; Methods; Microdissection; Microscope; Modeling; Modification; Neurosciences; Organism; Performance; Phase; Price; Procedures; Process; Proteins; Proteomics; Protocols documentation; Questionnaires; RNA; Recruitment Activity; Research; Research Personnel; Resolution; Role; Sampling; Side; Site; Small Business Innovation Research Grant; Sorting - Cell Movement; Specimen; Sucrose; Suggestion; System; Technology; Testing; Time; Tissue Sample; Tissues; Training; Translational Research; Vacuum; Work; base; brain cell; brain tissue; capillary; cell culture collection; cell type; commercialization; cost; cost efficient; digital; instrument; macromolecule; manufacturing process; minimally invasive; nerve stem cell; nervous system disorder; novel; performance tests; progenitor; prototype; public health relevance; relating to nervous system; research study; satisfaction; sensor; sound; tissue culture; touchscreen; user-friendly

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.

描述（由申请人提供）：为了推进当今的神经科学的基本和转化研究，必须进行细胞/区域特定研究。当必须得出细胞特定信息时，脑组织的复杂性会带来重大挑战。目前尚无商业可用的低成本仪器，可以在细胞分辨率下进行复杂的组织显微解剖，更重要的是收集活细胞 来自天然脑组织和细胞培养皿。此外，现有的激光辅助显微解剖仪器是复杂的，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开发了一种基于使用一次性毛细管单元（DCU-LIVE）和短而仔细调节的真空脉冲的活细胞和组织采集的方法。原理实验的证明表明，该方法对于天然和预处理的脑组织的显微解剖非常有效。用CTA的LIVE收集的细胞适合重新培养。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了CTA-Live原型的发展，该原型具有来自组织和培养物的细胞分辨率显微解剖功能，而不会影响收集的样品的可行性。该SBIR项目的II阶段将开发为两个商业模型：一种基本模型：具有半自动功能和微型分解和活细胞收集的能力，以及通过计算机控制校准过程，目标识别和通过触摸屏界面的触摸屏界面来通过计算机控制的校准过程，可以通过计算机控制的校准过程从多个样品中进行快速细胞和组织习惯进行全自动版本的ctas-live版本。拟议工具的一个重要优势是它们的低价允许有效的市场入口和商业化。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美元之间，至少显示了至少四倍和两倍的价格优势，而基于现有的激光系统。 CTA-LIVE的基础模型和自动化模型均应通过各种组织样品和细胞培养物进行彻底测试，这些细胞和细胞培养物的重点是可获得细胞的生存能力以及大分子的基因质量（例如RNA和蛋白质）。将测试表散荧光的模块，以收集从组织中收集荧光标记的细胞。将优化人类验尸组织的显微减退，RNA和蛋白质的质量将用于基因组学和蛋白质组学应用。因此，除了对CTA-LIVE原型的初步测试外，还将开发几种应用说明，为潜在客户提供“样本到归因”协议。开发和“内部”测试的CTA-LIVE仪器将发送到Beta测试站点，以确保最终用户满意度并获得有价值的反馈。这些研究的结果将是在该项目的第三阶段中成功商业化。