Protocol Citation: Chuyu Chen, Ciarra Smith 2022. Coating coverslips for cell culture. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvornd9v4o/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 15, 2022
Last Modified: May 31, 2024
Protocol Integer ID: 70088
Keywords: ASAPCRN
Abstract
Coated cover-slips provide a nourishing adherent surface for cell culture. This protocol provides step by step instruction on how to coat coverslips for cell culture.
Poly L-Lysine/ Poly D-Lysine coating
Poly L-Lysine/ Poly D-Lysine coating
Precoat the coverslips with poly-L-lysine (P5899) or poly-D-lysine (P7280) at 100 μg/ml.
This is incubated for 30 minutes or more at room temperature. The coverslips are washed with sterile water and allowed to dry.
They can be stored at room temperature until ready to use (up to 1 month).
Day before culturing: Laminin coating
Day before culturing: Laminin coating
Coverslips are then coated with 30 - 50 μl of laminin (50 μg/ml) which has been warmed. This is prepared the day before culturing of cells.
Incubate overnight in a humidified 37°C, 5% CO2 incubator or at 37 °C for 30 minutes.
Remove excess laminin and rinse with PBS twice. Alternatively, wash with DME (50 μL) twice.
Add the cells to the coverslips/slides and culture as needed.