Diagnostic Microperfusion Platfom for Functional Screening of Thick Preparations

描述(由申请人提供)：用于厚制剂功能筛选的诊断微灌注平台项目的目标是开发一种复杂的、用户友好的高通量诊断仪器，该仪器为厚组织样本提供长期的代谢支持，并允许对培养参数和诊断体系进行无与伦比的控制的功能筛选。 该项目的第一个目标是延长超过500米厚的组织制剂的生存能力，并使至少5天的体外可重复性研究成为可能。扩散受限的间隙质量传输会导致过早腐烂，这种情况在厚样品中尤为严重。我们建议通过强制间隙对流来克服扩散限制，以实现营养物质和气体在间隙中的高浓度。目前，复杂而昂贵的灌流/记录室(哈佛仪器)可以进行长达十个小时的研究。切片正在变薄，其腐烂过程从一项研究到另一项研究都是值得怀疑的，这使得重复实验、验证和解释结果变得困难。为了能够成功完成已开始的实验并增加已进行研究的重现性，我们将大大减缓培养物腐烂，并开发先进的流体体系结构，以便在邻近培养物(S)失败时无菌分离活菌株并保持不变的灌流。 该项目的第二个目标是通过实现对厚组织制剂的无与伦比的访问，克服体外功能诊断方面的技术限制。这将由用户选择是通过功能成像还是使用多电极阵列的电刺激和记录来监控活动。长期的、可重复的研究将为电生理学创造一个新的基准。可能受益于神经活动的长期灌流和监测的样本应用是神经元的再生、发育和可塑性，因为它们涉及到在较长时间内发展的过程。 第三个目标是开发一种经济的制造方法，将该诊断平台转变为具有流体、气体、光学和电气可访问性的廉价、可扩展的高通量设备。这种方法将使我们能够快速且廉价地更改我们的设计，集成不同的构建块，并支持附加功能以满足特定的用户需求。最终产品将是一种简单易用的微型仪器，其模块化结构可以很容易地进行调整，以适应各种应用挑战，包括但不限于对神经和心脏疾病、体外功能药理学、生理学、组织工程学和移植的理解和治疗。 公共卫生相关性：本文提出的新型诊断平台将通过为治疗性研究提供简单、可靠和廉价的试验台来促进公共健康。最终，这一发展将促进医学和科学发现，从而有利于神经和心脏疾病的治疗。

DESCRIPTION (provided by applicant): The objective of the project Diagnostic Microperfusion Platform for Functional Screening of Thick Preparations is to develop a sophisticated, yet user friendly, high-throughput diagnostic instrument that provides long-term metabolic support of thick tissue samples and allows functional screening with unparallel control of culture parameters and diagnostic architecture. The first goal of the project is to extend the viability of greater than 500 ¿m thick tissue preparations and enable at least 5 days long, reproducible studies in vitro. Diffusion limited interstitial mass transport causes premature decay that is particularly exacerbated in thick samples. We propose to overcome diffusion limits by forced interstitial convection to achieve high concentration of nutrients and gas interstitially. At present, complex and expensive perfusion/recording chambers (Harvard Apparatus) allow ten-hour-long studies. Slices are thinning and their decay progress is questionable from study to study making it hard to repeat experiments, validate and interpret results. To enable successful completion of started experiments and increase the reproducibility of performed studies we will substantially mitigate culture decay, and, develop advanced fluidic architecture to aseptically isolate viable cultures and maintain unaltered perfusion if neighboring culture(s) fail. The second goal of the project is to overcome technological limitations in functional diagnostics in vitro by enabling unparallel access to thick tissue preparations. It will be up to users to select whether to monitor activity by functional imaging or electrical stimulation and recording using multi-electrode arrays. Long-term, reproducible studies will create a new benchmark for electrophysiology. Sample applications that may benefit from long-term perfusion and monitoring of neural activity are neuronal regeneration, development and plasticity as they involve processes that develop over extended periods of time. The third objective is to develop an economical fabrication approach that will turn this diagnostic platform into inexpensive, scalable, high-throughput devices with fluidic, gas, optical and electrical accessibility. Such approach will allow us to quickly and inexpensively change our designs, integrate different building blocks, and enable add-on-functionalities to satisfy specific user requirements. The end product will be a simple to use, miniature instrument whose modular architecture can be easily adjusted to suit various application challenges including but not limited to the understanding and treatment of neural and cardiac disorders, functional pharmacology in vitro, physiology, tissue engineering and transplantation. PUBLIC HEALTH RELEVANCE: Novel diagnostic platform proposed herein will advance public health by providing a simple, reliable and inexpensive test-bed for therapeutic studies. Ultimately, this development will facilitate medical and scientific discoveries that will benefit the treatment of neural and cardiac disorders.