Huang: Amino Silane Treatment of Glass Slides

From OpenWetWare
Jump to: navigation, search

Overview

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.

Materials

  • 1 ml amino silane (we use (3-Aminopropyl)triethoxysilane from Sigma-Aldrich)
  • 100 ml acetone
  • 5 Glass microscope slides (or cover slips)
  • Staining Jar

Procedure

  1. Mix 1 ml silane with 50 ml acetone in the staining jar (2% silane).
  2. Clean the glass slides or cover slip. To achieve this, you can either
    1. rinse the glass with ethanol and wipe with Kimwipe, and then blow dry. Or
    2. ultrasonic cleaning with 10% micro90 detergent for 30 min, rinse with water and then blow dry. Or
    3. plasma cleaning. Or
    4. follow the protocol in Pet Brown's lab at Stanford
  3. Immerse the glass in the jar for 20 min.
  4. Rinse the glass in a conical tube with 50 ml acetone.
  5. Blow dry.
  6. Bake at 120°C for 20 min (optional).

Notes

  1. Silane is very toxic! All the procedure must be done in the fume hood!
  2. 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.
  3. If the staining jar you used is glass, it has been coated with silane! Seperate it from other glassware!

Contact