The soft microfluidic probe consists of two layers, a molded microchannel layer and a capping layer, bonded by corona treatment. The fabrication process began with formation of a mold in the geometry of the microfluidic channels by photolithography and deep reactive ion etching (STS Pegasus ICP-DRIE; SPTS Technologies, Newport, UK) on the surface of a silicon wafer. A glass slide (length, 75 mm; width, 50 mm; thickness, 1 mm; Corning Microscope Slides, Corning, NY, USA) treated with a Pt inhibitor, a solution of 5% [3-(2-aminoethylamino)propyl]trimethoxysilane (Sigma-Aldrich, St. Louis, MO, USA), and 95% methanol by weight, facilitated the release of a layer of PDMS (elastomer/curing agent ratio, 10:1; Sylgard 184, Dow Corning) cast between the glass slide and the patterned silicon wafer. A layer of Scotch tape served as a spacer at the boundary of the slide to define the thickness of the PDMS. A thin layer (~60 nm) of polymethyl methacrylate (PMMA A2; MicroChem, MA, USA) spin-cast on the silicon mold served to prevent adhesion between the silicon mold and the PDMS. Curing in an oven at 70°C for 45 min and then carefully releasing the glass slide with the patterned PDMS layer from the silicon mold completed the fabrication of the thin microfluidic channel layer (thickness, ~100 μm). A capping layer of PDMS (thickness, ~100 μm) was prepared by spin-casting (elastomer/curing agent ratio, 10:1; Sylgard 184, Dow Corning) on a PC substrate and baking at 70°C for 45 min. Bonding this cover layer to the microfluidic channel after activating the surfaces with a laboratory corona treater (Electro-Technic Products) yielded a microfluidic probe. The PC sheet was then peeled away, and the probes were released from the glass slide with a razor blade.

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