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.
