Development of automated microfluidic devices for quantifying nutrient uptake and release from murine adipose tissue with fluorescence imaging

Abstract

This study aims to advance the knowledge of diabetes-associated biological assays such as white adipose tissue secreted glycerol and fatty acid uptake and secretion assays by leveraging the incorporation of microfluidic platforms. The investigation involves several dimensions, spanning from diabetes and obesity to the innovation of modern microfluidic devices. The first chapter initiates a comprehensive exploration of the relationships among diabetes, obesity, and adipose tissue. These relationships lay the groundwork for the ensuing chapters and underscores the pivotal significance of microfluidic tools as unique instruments in addressing complexities of adipose tissue function. In the second chapter, automated microfluidic devices, microfluidic digital analog converters (μDAC), are designed to expedite cell and tissue stimulation at high temporal resolution (5 seconds). This pioneering approach not only streamlines experimentation but also showcases the potential for modulating stimuli, thereby offering deeper insights into dynamic cellular responses and the intricacies of disease (diabetes) mechanisms. Chapter three unveils the application of valve-controlled droplet-based microfluidics, enabling precise sampling from ex vivo adipose tissue. Through multiplexed sensing of glycerol and fatty acid secretion, this technique presents a novel avenue for dissecting adipose tissue dynamics, unveiling implications for metabolic disorders such as diabetes. Shifting focus to nucleic acid driven sensing through electrochemistry, the fourth chapter introduces an innovative automated microfluidic device for biosensor preparation on electrode iii surfaces. This approach streamlines the process of electrode fabrication, potentially elevating the efficiency of nucleic acid-based sensors and paving the way for more precise diagnostic outcomes. Cumulatively, this research underscores the pivotal role of microfluidics in propelling forward bioanalysis, particularly when applied to diabetes research. By combining chapters that span from comprehending diseases to automating assays and multiplexed sensing, this dissertation contributes to the evolving landscape of diabetes research and diagnostic methodologies. It is our hope that the advancements showcased herein will stimulate novel insights, streamlined techniques, and transformative breakthroughs in the future to aid in managing diabetes and its related metabolic intricacies.