Diagnostic Microperfusion Platfom for Functional Screening of Thick Preparations

描述（由申请人提供）：用于厚制备物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代谢支持，并允许通过对培养参数和诊断架构的非平行控制进行功能筛选。 该项目的第一个目标是延长大于500 μ m厚的组织制备物的生存能力，并实现至少5天的体外可重复研究。扩散限制的间隙质量传输导致过早衰减，这在厚样品中特别加剧。我们建议通过强制间隙对流来克服扩散限制，以实现高浓度的营养物和气体。目前，复杂和昂贵的灌注/记录室（哈佛仪器）允许长达10小时的研究。切片变薄，它们的衰变过程从一项研究到另一项研究都是值得怀疑的，这使得很难重复实验，验证和解释结果。为了能够成功完成启动的实验并提高所进行研究的重现性，我们将大大减轻培养物衰减，并开发先进的流体结构，以无菌分离活的培养物，并在相邻培养物失败时保持灌注不变。 该项目的第二个目标是通过实现对厚组织制剂的非平行获取来克服体外功能诊断的技术限制。这将取决于用户选择是否通过功能成像或电刺激和使用多电极阵列记录来监测活动。长期的、可重复的研究将为电生理学创造一个新的基准。可能受益于长期灌注和神经活动监测的示例应用是神经元再生、发育和可塑性，因为它们涉及长时间发展的过程。 第三个目标是开发一种经济的制造方法，将这个诊断平台变成廉价的，可扩展的，高通量的设备，具有流体，气体，光学和电气的可访问性。这种方法将使我们能够快速，廉价地改变我们的设计，集成不同的构建块，并启用附加功能，以满足特定的用户需求。最终产品将是一种使用简单的微型仪器，其模块化结构可以轻松调整，以适应各种应用挑战，包括但不限于神经和心脏疾病的理解和治疗，体外功能药理学，生理学，组织工程和移植。 公共卫生相关性：本文提出的新型诊断平台将通过为治疗研究提供简单、可靠和廉价的测试平台来促进公共健康。最终，这一发展将促进医学和科学发现，这将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.