Poly-lysine coating of glass bottom dishes and glass bottom plates

Materials

Poly-lysine solution

Both poly-L-lysine and poly-D-lysine of MW between 30,000 - 150,000 can be used for coating. Poly-D-lysine is recommended since it is not subjected to protease released from cells.

Prepare 1mg/ml stock solution by dissolving 100mg Poly-D-lysine hydrobromide(Sigma P-0899) in 100 ml water. Filter sterilize the solution then store aliquots at –20℃.

Working solution is obtained by diluting the stock solution with sterile water to around 0.1mg/ml (0.05-0.5mg/ml).

Procedure

  • Add working solution of poly-lysine into a glass bottom dish or glass bottom plate. If cells will be cultured only on the glass surface, then only the glass surface needs to be covered with poly-lysine solution.
  • Incubate for 1 hour at room temperature in a hood.
  • Remove poly-lysine solution. Poly-lysine solution can be re-used for a few times(depending on its concentration).
  • Rinse 3 times with PBS
  • Poly-lysine coated glass bottom dish or glass bottom plate can be kept at 4℃ for about a month. Note that coating on glass surface is less stable than coating on plastic tissue culture dishes or plates. Thus it is recommended to coat the glass bottom dish/plate right before cell culture.

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