Huang: Amino Silane Treatment of Glass Slides
This protocol allows the immobilization of bacterial cell on glassware and improves the following imaging. We use amino silane instead of poly-lysine to achieve robust graft.
- 1 ml amino silane (we use (3-Aminopropyl)triethoxysilane from Sigma-Aldrich)
- 100 ml acetone
- 5 Glass microscope slides (or cover slips)
- Staining Jar
- Mix 1 ml silane with 50 ml acetone in the staining jar (2% silane).
- Clean the glass slides or cover slip. To achieve this, you can either
- rinse the glass with ethanol and wipe with Kimwipe, and then blow dry. Or
- ultrasonic cleaning with 10% micro90 detergent for 30 min, rinse with water and then blow dry. Or
- plasma cleaning. Or
- follow the protocol in Pet Brown's lab at Stanford
- Immerse the glass in the jar for 20 min.
- Rinse the glass in a conical tube with 50 ml acetone.
- Blow dry.
- Bake at 120°C for 20 min (optional).
- Silane is very toxic! All the procedure must be done in the fume hood!
- Silane is ultra sensitive to humidity. To avoid hydrolysis and the following crosslink, use inert atmosphere or aliqot into plastic containers. Also, anhydrous acetone is recommended.
- If the staining jar you used is glass, it has been coated with silane! Seperate it from other glassware!