Hang Lu, Ph.D. (Georgia Institute of Technology)
Melissa Kemp, Ph.D. (Georgia Institute of Technology)
Zhexing Wen, Ph.D. (Emory School of Medicine)
Johnna Temenoff, Ph.D. (Georgia Institute of Technology)
Wilbur Lam, Ph.D. (Georgia Institute of Technology)
Microfluidic tools for studying development in embryos and brain organoids
Development in multicellular organisms is a complex process requiring multiple intracellular and extracellular signaling events. High content screening tools enable the cellular and subcellular assessment of developmental changes which in turn have led to a variety of genetic, pharmacological and therapeutic screens involving multicellular organisms. Developing high-content screening tools for both multicellular systems require high resolution imaging of protein and gene expression changes, relatively large number of samples to better characterize inter and intra-populational differences, and multiplexed readouts using the same sample to obtain layered information about developmental changes. Microfluidics can address these challenges by enabling high magnification imaging, parallelization, rapid reagent delivery and exchange and lower reagent consumption. Hence, this thesis seeks to address high content screening challenges that affect the study of development in active areas of research in my lab using microfluidics: C. elegans embryogenesis and cellular development of brain organoids. I will demonstrate this through three specific aims that involve developing and improving microfluidic-based technology for large-scale imaging and characterizing the sample of interest by incorporating assays for probing structural features and functionality. As a result, I will develop a microfluidic-based assay for conducting high resolution measurements of gene expression changes during C. elegans embryogenesis using single molecule fluorescence in situ hybridization (aim 1). Next, I will develop and optimize culture conditions for microfluidic-based culture of forebrain organoids and coculture of microglia and cerebral organoids (aim 2). Finally, I will develop assays for multiparametric and in situ assessment of immune cell and neuronal cell interactions in cerebral organoids (aim 3). Combining high throughput microfluidic technology with high content imaging tools will improve the characterization of factors affecting development in these two biological systems.