I am just wondering what might have caused this as this is happening many number of times and also in 12 wells and 6 wells plates. I know it might be because of edge effect/evaporation but the medium volume looks very much the same in all wells but i could see a change in color on the side wells. Incubator we use is a new one and everything seems to be fine. How to rectify these problems or else any other alternatives. greatly appreciate your answers.
thanks,
durai
To avoid my plates drying out, once I've seeded the plates I usually put them into a container - a zip-lock lunchbox which we've modified to have a filter in the side, before putting them into the incubator. Maybe that will help?
Stacey
I agree with Stacey, it could be coz of drying of the wells. Try putting only media in those wells, and check if the color change problem persists.! You might just get rid of it!
Make sure that your incubator is humidified and CO2 % is OK! Is your water tray, in the incubator, filled with water??? Also check the CO2!
You should check which material is used for the chamber slides. Oftentimes, cell dye because they cannot attach. If you are using plastic bottom, you may want to try glass botton. Also, you may want to try additional coating materials, like gelatin, collagen and fibronectin. If cells are dying on plates as well after transfection, this could be due to the effect of transgene or toxicity of transfection procedure. Many times, you don't see this after you trypsinize and split cells.
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