The microfluidic layers were fabricated from polydimethylsiloxane (PDMS; GE RTV615) by standard soft lithography techniques. The molds were fabricated by coating photoresist SU-8 25/50 with desired thicknesses on a silicon wafer and photolithographic patterning (fig. S1). Premixed PDMS prepolymer and curing agent (ratio, 5:1) were poured onto the mold for the valve layer and cured. Premixed PDMS prepolymer and curing agent (ratio, 20;1) were spin coated on the mold for the channel layer at 1500 rpm for 1 min and incubated at 65°C for 20 min. Then, the valve layer was released from the mold and placed on the channel layer with alignment. The assembly was baked at 65°C overnight for efficient bonding. The assembly was further bonded with alignment to the microwells layer cured from premixed PDMS prepolymer and curing agent (ratio, 20:1) by baking at 65°C overnight. The obtained device was further bonded to a glass slide to prevent deformation when applying a pressure to the valves.

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