Poster Presentations from the 2023 Microscale Innovation in Life Science Symposium
Contour Analysis to Quantify Tumor Cell Invasion in Organ - On - Chip Models
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Open to download resource. Microphysiological systems (MPS), including organ-on-chip (OOC) models, are in vitro systems designed to replicate properties of a specific organ or tissue and have recently gained attention as potential tools for drug discovery. OOC models of cancer can enable drug discovery and mechanistic investigations of tumor metastasis by modeling local tissue invasion. The OOC developed by Emulate, Inc. utilizes a two-channel system with endothelial cells in the bottom channel and epithelial cells in the top channel and incorporates physical forces such as fluid flow and peristalsis-like deformations. We have adapted this system to model CRC by including Green Fluorescent Protein (GFP)-expressing patient-derived colon cancer (CRC) organoids in the top channel and human intestinal microvascular endothelial cells (HIMEC) in the bottom channel. After six days, invasion from the epithelial compartment into the endothelial compartment can be visualized by fluorescence confocal microscopy (using the Operetta CLS High Content Analysis Platform) and the number of invaded GFP+ tumor cells can be quantified via image analysis software (Harmony Software). Traditional image processing and manual analysis can be time-consuming, subject to technical variation and can only be performed on high-quality images. A particularly challenging task is to accurately and precisely identify the boundary created by the endothelial layer, so that tumor cells invading into the endothelial compartment can be reliably identified and quantified. We propose an application of topographical analysis as a robust method of data analysis that can account for technical artifacts such as variation in the tilt in the microscope stage while also capturing fine details in the contours of the endothelial tissue, leading to a more accurate measurement of invasion. To evaluate its performance, we applied our analytical approach to the CRC OOC model and revealed persistent texture in the tissue surface that can be utilized to capture actively invading objects missed by alternative methods. This workflow was developed into an assay-agnostic web tool requiring only the coordinates of the epithelial and endothelial cells, thereby allowing scientists to quickly and accurately quantify data and interpret results using interactive data visualization. Requiring solely positional information allows this workflow to be applied to data collected with other microscopy and image analysis tools and other OOC models investigating tissue invasion. This tool, the Chip Invasion and Contour Analysis (ChICA), provides spatial information within OOC models and enables higher throughput analysis of invasion, making these chips more viable models for drug discovery and cancer research applications.
Elevating & Automating Cell and Extracellular Vesicle Purification with Digital Magnetic Sorting
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Open to download resource. Ferrologix Digital Magnetic Sorting (DMS) technology, also known as ratcheting cytometry, is an innovative platform that allows for the quantitative sorting of magnetically labeled cells based on their surface-level expression and the number of magnetic beads attached to each cell. The process involves transporting magnetically tagged cells across a substrate containing ferromagnetic microstructures, which are subjected to a cyclic magnetic field. Transport within sterile cartridges and across microstructures with a gradient pitch enables the sorting of cells based on the level of bound magnetic content. This technology combines traditional magnetic sorting with flow cytometry-like capabilities with the scalability of magnetic sorting, allowing cells to be fractionated rapidly based on one or several surface markers. Ferrologix is commercializing this technology into multiple products at the benchtop/process development scale and full manufacturing (whole leukopak) scale. Relevant applications in cell therapy discovery and manufacturing include rapid & high throughput sorting of rare immune cell types, simultaneous multi-marker sorting, cell fractionation based on antigen density level, and extracellular vesical purification.