Probing the Ultimate Limits of Biology: Developing Microparticle Platforms for High-Throughput Single-Cell Assays

Download or read book Probing the Ultimate Limits of Biology: Developing Microparticle Platforms for High-Throughput Single-Cell Assays written by Joseph Michael de Rutte and published by . This book was released on 2020 with total page 122 pages. Available in PDF, EPUB and Kindle. Book excerpt: Techniques to analyze and sort single cells based on complex phenotypes such as secreted products have the potential to transform our understanding of cellular biology as well as accelerate the development of next generation cell and antibody therapies. Microfluidic techniques have emerged over the last two decades that enable such assays by creating small compartments that can isolate individual cells. Despite these technical advances, adoption of this technology has been slow due to the complexity of the approaches and the requirement of specialty instruments. To address this issue, we developed a microparticle based approach to create uniform sub-nanoliter water in oil compartments using only standard lab equipment. We refer to these droplets formed with microparticles, dropicles. In this dissertation I will provide an overview of various strategies used to create microscale compartments, specifically to perform single-cell secretion assays. I will then discuss in detail an approach to fabricate and use 3D structured microparticles to perform single-cell secretion assays massively in parallel with standard lab equipment. By making this microparticle approach compatible with existing high-throughput flow cytometers we are able to analyze and sort of over 100,000 single cells based on their secreted products. Lastly to create a truly democratized platform there is a need to be able to manufacture these particles in a scalable manner. In these last chapters I will discuss approaches to fabricate microparticles with different morphologies in high-throughput.