Collagen coating of glass bottom dishes and glass bottom plates
Rat tail collagen (type I) solution
It is recommended to purchase collagen solutions, so that sterilization is not needed since filter sterilization of collagen could be problematic.
Dilution the stock solution to 50 ug/ml with 0.02M acetic acid or 0.01M HCl.
- Add solution (50 ug/ml) into a glass bottom dish or glass bottom plate at 5 ug/cm2. If cells will be cultured only on the glass surface, then only the glass surface needs to be covered with collagen solution.
- Incubate for 1 hour at room temperature in a hood.
- Carefully aspirate the collagen solution.
(alternatively, the glass bottom dish/plate can be left in the hood to dry overnight, and the following washing step is not needed)
- Rinse 3 times with PBS or serum-free media.
- Collagen coated glass bottom dish or glass bottom plate can be air dried and 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.