I coat my cell culture flasks with a mixture of poly-L ornithine and fibronectin overnight then aspirate the coating solution and dry it open in a laminar flow cabinet under UV light .
Is the drying step always necessary or it is a need only for short term storage in the refrigerator? Can I use my flasks for cell culturing instantly after aspiration of the coating solution without drying? Is the drying step necessary for the proper assembly of proteins and polymers on the coated surface?
P.S As different protocols for different coating procedures are available in the internet I will be happy if someone could provide an explanation for the role of drying step. For example my protocol for fibronectin coating involve the drying step but in the others one can find information that drying should be abandoned when coating with fibronectin?
Hi Tomasz,
In one of the our project I used collagen coated dishes. You must completely dry them before use. I tried to use them freshly coated but cells don't like it. Also I used sterile collagen solution and just dry dishes in the hood under air flow for 15 min as UV light may be harmful for your protein. Also coated dishes I don't keep in a fridge, they were fine after keeping at RT.
I have coated my dishes with poly-D-lysine and then fibronectin before and did not dry any of them. My cells behave differently on theses dishes than on dishes coated with poly-D-lysine only. So I assume the fibronectin coating worked.
I was looking at how different extracellular matrix molecules could affect cell behaviors and drying protein will most likely disrupt its structure. In fact in our protocol if you coat with protein it is essential that it never get dried out (even between washes). But if you just need to coat so that your cells stick it may not be a concern.
So in my opinion it is not necessary to dry our your coating. And if you need your coating molecule to be biologically functional you should not dry it.
In my case (I coat with Poly-L-lysine) the drying step is necessary because Poly-L-Lysine has been described as a source of toxicity for cultured cells, so we need to wash with miliQ water before seeding cells; here the drying step is just to avoid deletereous effects of water. However, if your substrate is non toxic I think you can try it without drying step...
Does anyone know approximately how long the drying process takes? I assume that an overnight drying step is used for convenience...
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