3 research outputs found
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Multiwell Culture Devices with Perfusion and Oxygen Control
A microfluidic structure is formed with one or more microfluidic channels for receiving fluid and passing the fluid through a channel in communication with a collection chamber integrated with that channel through a transition stage that allows the collection chamber to gather targets within the fluid, such as cells, including circulating tumor cells in blood. The transition stage may be formed for an asymmetrical configuration with an obstacle and chamfered face configuration. A gas permeable membrane provides perfusion control for directing gas to the collection chamber. Porous elements provide bubble venting from the fluid flow
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Resealable, optically accessible, PDMS-free fluidic platform for ex vivo interrogation of pancreatic islets
We report the design and fabrication of a robust fluidic platform built out of inert plastic materials and micromachined features that promote optimized convective fluid transport. The platform is tested for perfusion interrogation of rodent and human pancreatic islets, dynamic secretion of hormones, concomitant live-cell imaging, and optogenetic stimulation of genetically engineered islets. A coupled quantitative fluid dynamics computational model of glucose stimulated insulin secretion and fluid dynamics was first utilized to design device geometries that are optimal for complete perfusion of three-dimensional islets, effective collection of secreted insulin, and minimization of system volumes and associated delays. Fluidic devices were then fabricated through rapid prototyping techniques, such as micromilling and laser engraving, as two interlocking parts from materials that are non-absorbent and inert. Finally, the assembly was tested for performance using both rodent and human islets with multiple assays conducted in parallel, such as dynamic perfusion, staining and optogenetics on standard microscopes, as well as for integration with commercial perfusion machines. The optimized design of convective fluid flows, use of bio-inert and non-absorbent materials, reversible assembly, manual access for loading and unloading of islets, and straightforward integration with commercial imaging and fluid handling systems proved to be critical for perfusion assay, and particularly suited for time-resolved optogenetics studies