Incorporation of a Morphologically Controlled Ice Grain Boundary into a Microfluidic Device for Size-Selective Separation of Micro/Nanospheres

A frozen aqueous solution was integrated into a microfluidic device as a size-tunable separation field for the size-selective separation of micro/nanospheres. The width of the ice grain boundaries formed in frozen aqueous solutions could be altered by controlling the operating temperature. A freezing chamber was placed adjacent to the microfluidic channel. A sample-dispersing aqueous sucrose solution was injected into the chamber and frozen, allowing the freeze-concentrated solution (FCS) to run vertically to the microfluidic channel, where the eluting solution flows. The operating temperature can be used to control the physical interaction between the ice wall and micro/nanospheres, enabling size-selective migration. The eluted micro/nanospheres in the microchannel were passed through the eluting solution collected from the outlet. We achieved size-selective separation and collection of microspheres and nanospheres. We separated the exosomes and yeast cells to demonstrate their applicability in bioseparation. The present method is suitable not only for size-selective separation but also for evaluating the biological expression of extracellular vesicles under cryogenic conditions.