Protein Patterning using Parylene

• 1. Prepare protein solution (usually 20ng/ml of TexasRed-BSA, 50ng/ml of FITC-BSA)

  2. Prepare hydrophobic substrate for parylene membrane, either PDMS on glass, polystyrene, or treated glass. For PDMS on glass, clean both surfaces thoroughly before bringing into contact.

  3. Add parylene membrane into contact with hydrophobic surface using sharp tweezers.

  4. Add protein solution on top of membrane, enough to cover entirely, but not necessary to use more.

  5. Incubate for 15 to 30 minutes. It is ok to incubate longer.

  6. View with fluorescent microscope.

  7. Remove parylene using tweezers, being careful to lift it off rather than drag it.

FOR CO-PATTERNING

• 1. Add second protein on top of the first protein pattern.

  2. Incubate for 15 to 30 minutes, depending on relative strength of the fluorescent signals.

  3. View with fluorescent microscope.

     1. View each fluorescent signal separately and merge images.