Suction Technique
1. Clean PDMS surface with ethanol and water ~ use a tweezer to hold PDMS and dip it in a beaker of ethanol (200 proof) and then in a beaker of distilled water.
2. Dry PDMS with nitrogen air.
3. Dilute 1 mg/ml fibronectin 1:10 with PBS (1X).
4. Coat the surface of a microscope cover slide with the fibronectin solution (dipping slide in solution or spreading solution over slide).
5. Place the PDMS mold on the fibronectin coating of glass slide.
6. Put a drop of boiling 1% aqueous agarose solution along one of the open edges of mold.
7. Apply vacuum suction on the opposite edge ~ fill the channels of PDMS feature with agarose by pulling the solution through the channels with a glass pipet connected to moderate vacuum.
8. Allow agarose solution in the channels to dry for 12 hours.
9. Detach agarose from the PDMS mold.
Stamping Technique
10. Clean PDMS surface with ethanol and water ~ use a tweezer to hold PDMS and dip it in a beaker of ethanol (200 proof) and then in a beaker of distilled water.
11. Dry PDMS with nitrogen air.
12. Dilute 1 mg/ml fibronectin 1:10 with PBS (1X).
13. Coat the features of the PDMS surface with droplets of fibronectin solution (» 600 ml) using a micropipet.
14. Let PDMS sit for 15 minutes to dry.
15. Remove the excess fibronectin solution on PDMS with vacuum suction.
16. Clean PDMS surface with PBS and water. Use a tweezer to hold PDMS and dip it in a beaker of PBS (1X) and then in a beaker of distilled water.
17. Dry PDMS with nitrogen air.
18. Use the feature surface of PDMS and stamp it on a microscope cover slide (placed in a 6-well plate).
19. Cover the cover slide with cell culture (» 2 ml in each well of a 6-well plate).