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3. Together with the vial, a Petri dish (the container for molding), the master, and the posts for forming reservoirs are placed in a desiccator. The surfaces of the items are exposed to the vapor of silanization agent for 70 min with the pressure pumped down to 20 psi. The silane coating facilitates the separation of PDMS replica from the master. 4. Measure the desired amount of RTV 615. The weight ratio of A and B parts is 10:1. 5-cm-thick PDMS replica in a 4-in. Petri dish. 5. Mix the solutions thoroughly using a glass rod for 3 min to ensure complete and even cure.

After cooling to room temperature, bring out the holder. A schematic of the equipment build in our laboratory is shown in Fig. 3. 42 Tani et al. Fig. 2. Schematic diagram for fabrication of Si microwell array chip. Fig. 3. Schematic of the equipment for thermal oxidation of Si chip. 2. Photolithography of the Si Chip 1. All procedures must be carried out in a yellow-lighted room to prevent exposure of photoresist-coated chips. 2. The Si chip with oxidized layer is washed by soaking in RCA solution at 80°C for 20 min, and then rinsed with water.

A series of samples (antigen) is then brought from a second multichannel chip as the lines of antibody and sample make a grid pattern. The specific binding between immobilized antibodies and antigen molecules takes places at each intersection of the two lines in a combinatorial fashion. The overall mosaic pattern of the chip has been expanded to a three-dimensional (3D) format suitable for stable immobilization of tester strains. For this purpose, the assay format presented here is exploiting the 3D microfluidic network, which consists of the assembly of two multichannel and one perforated microwell chips.

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