Cleaning coverslips before seeding cells

[@moinulhoque2160]

It's a nice video. Could you please make another video of how to do the coating?

[@Livecellimagingfacility]

Hi Moinul After cleaning and sterilizing the coverslips, simply put them on a piece of parafilm in a humid chamber (i.e. a box with paper soaked in water on the edges), add the solution you want to coat with, put a second coverslip on top so that you use the solution to coat 2 coverslips at the same time and you prevent evaporation on the edges. Close the humid chamber during incubation time (which is often at room temperature but it depends on what you are coating). When the coating is done, carefully remove the top coverslip, flip it so that the coated side is upwards, remove the solution (often can be stored and reused) from the bottom coverslip. Rinse all coverslips with PBS. They are ready to be used. Make sure you keep track of which side was coated. Good luck!

[@janggeumseo592]

@@Livecellimagingfacility Hello, I am trying to coat 13mm round coverslips with collagen. But I am very new to this area. I tried to add 10-20 microliter collagen solution on top of coverslips but it was very hard to spread them uniformly with pipette tip. Dipping them in solution is not useful because both sides will be coated. In the method you explained; second coverslip is put on top of first coverslip and they are located in the solution so outer surfaces will be coated whilst inner surfaces will remain uncoated. This is my understanding. could you please help me with that? I also don't have humid chamber and never used it before. Thank you!

[@Livecellimagingfacility]

sorry for not answering earlier. Somehow I did not get notified of your comment. Yes you are totally correct in your understanding. We sandwich the coating solution between 2 coverslips so only 1 side of the coverslip gets coated. To make a humid chamber, simply take a Petri dish, put a piece of parafilm completely flat at the bottom, place some absorbant paper at the periphery of the dish and wet them completely but not enough for the water to run out. There you have a cheap and simple humid chamber! :)

[@shaogonelee4481]

very helpful

[@Livecellimagingfacility]

Thanks! :)