Microfluidic platform for generating dynamic profile of cell stimuli for non-adherent cells.

A T cell clone is able to distinguish Ags in the form of peptide–MHC complexes with high specificity and sensitivity; however, how subtle differences in peptide–MHC structures translate to distinct T cell effector functions remains unknown. We hypothesized that mitochondrial positioning and associated calcium responses play an important role in T cell Ag recognition. We engineered a microfluidic system to precisely manipulate and synchronize a large number of cell–cell pairing events, which provided simultaneous real-time signaling imaging and organelle tracking with temporal precision. In addition, we developed image-derived metrics to quantify calcium response and mitochondria movement. Using myelin proteolipid altered peptide ligands and a hybridoma T cell line derived from a mouse model of experimental autoimmune encephalomyelitis, we observed that Ag potency modulates calcium response at the single-cell level. We further developed a partial least squares regression model, which highlighted mitochondrial positioning as a strong predictor of calcium response. The model revealed T cell mitochondria sharply alter direction within minutes following exposure to agonist peptide Ag, changing from accumulation at the immunological synapse to retrograde movement toward the distal end of the T cell body. By quantifying mitochondria movement as a highly dynamic process with rapidly changing phases, our result reconciles conflicting prior reports of mitochondria positioning during T cell Ag recognition. We envision applying this pipeline of methodology to study cell interactions between other immune cell types to reveal important signaling phenomenon that is inaccessible because of data-limited experimental design.

Low-cost microfluidic devices are desirable for many chemical processes; however, access to robust, inert, and appropriately structured materials for the inner channel wall is severely limited. Here, the shear force within confined microchannels was tuned through control of reactant solution fluid-flow and shown to dramatically impact nano- through microstructure growth. Combined use of experimental results and simulations allowed controlled growth of 3D networked Zn(OH)F nanostructures with uniform pore distributions and large fluid contact areas on inner microchannel walls. These attributes facilitated subsequent preparation of uniformly distributed Pd and PdPt networks with high structural and chemical stability using a facile, in situ conversion method. The advantageous properties of the microchannel based catalytic system were demonstrated using microwave-assisted continuous-flow coupling as a representative reaction. High conversion rates and good recyclability were obtained. Controlling materials nanostructure via fluid-flow-enhanced growth affords a general strategy to optimize the structure of an inner microchannel wall for desired attributes. The approach provides a promising pathway toward versatile, high-performance, and low-cost microfluidic devices for continuous-flow chemical processes.