Diagnostic Microperfusion Platfom for Functional Screening of Thick Preparations

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.