Miller, Jordan S2019-05-172019-05-172018-052018-01-22May 2018Albritton, Jacob L. "Production and Characterization of Uniform and Heterogeneous Cancer Multicellular Aggregates for Longitudinal Studies of Epithelial-To-Mesenchymal Transition." (2018) Master’s Thesis, Rice University. <a href="https://hdl.handle.net/1911/105628">https://hdl.handle.net/1911/105628</a>.https://hdl.handle.net/1911/105628Epithelial-to-mesenchymal transition (EMT) is a proposed mechanism for initial metastatic invasion. Tumors are highly heterogeneous mixtures of tumor cells, and heterogeneous multicellular aggregates (MCAs) have emerged as in vitro surrogates for heterogeneous tumors. We custom-modified a commercial laser cutter to produce microwells by laser ablation of poly(dimethylsiloxane) (PDMS), which could generate MCAs with EMT phenotype. Tumor invasion is a dynamic process, but methods for longitudinal characterization of heterogeneous MCAs are lacking. We propose improvements to quantifying 1) MCA size using 2D maximum Feret diameter measured by automated image analysis; 2) Cell sub-population ratio determined from Imaris-mediated nuclei counting of cells with nuclear localized fluorescent proteins; and 3) Segmentation efficiency based on cell population overlap volume. Finally, we discuss directions for future longitudinal studies of EMT. Improvements to MCA production throughput and longitudinal characterization methods will improve studies of EMT with heterogeneous MCAs.application/pdfengCopyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.multicellular aggregatetumor heterogeneitymicrowell3D image analysisepithelial-to-mesenchymal transitionThesis2019-05-17