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==Microarray Fabrication== ===Casting=== # In a dish, mix 45 g silicone elastomer with 4.5 g PDMS curing agent (10:1 silicone:curing agent). # Mix well with plastic knife until bubbles form, approximately 1 min. # Place dish into vacuum chamber and close lid. # Ensure that all valves are in the correct position. (NEED FIGURE) # Turn on pump and wait for bubbles to rise to the surface of the mixture. # Close the small plastic valve on the vacuum chamber and shut off pump. If the pump runs for too long, the rising bubbles will cause the mixture to overflow the dish. # Allow the dish to seat in the sealed vacuum chamber for 20 minutes. Do not worry if there are still bubbles remaining. # Open the small plastic valve on the vacuum chamber. The rapid change in pressure should cause the rest of the bubbles to burst. Open the release valve on the tubing to fully equilibrate. # Pour the mixture over the silicon mold. # Vacuum again, 10 minutes. The pump does not need to be shut off this time as there is no risk of overflow. Do not worry if there are still some bubbles in the material. These bubbles will not affect the wells because the plate is being cast upside down; bubbles are rising to the eventual bottom of the plate. # Bake for 3 hours at 60 deg C. # Use a scalpel to cut around the edge of the dish first, then around the edge of the silicon mold. Peel away and discard this ring. # Use the scalpel to peel the edge of the PDMS disk away from the silicon mold. Carefully work your way around the entire disk. # Carefully peel the PDMS disk away from the silicon mold and place in a new dish. Cut along the edges of the plates. Each disk yields 2 plates. # Transport the excised plates in a covered dish with parafilm around the edge to keep out dust. ===Mounting=== # Use red, 2mm (height) rubber gasket, with 16 squares. Peel off white backing to expose glue. # Place gasket sticky side up on hood shelf, along with PDMS plate. UV for 10-15 minutes. # Attach gasket to PDMS, aligning the edges. Trim any excess PDMS (e.g., due to sloppy cutting after casting). # Clean glass slide and mount PDMS/gasket assembly, gasket side up. The PDMS will stick to the glass. # Cover with scotch tape to keep out dust.
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