I'm doing antiproliferative assays with different incubation times. I had the compound (previously diluted in DMSO) together with cell culture medium for 24h, 48h or 72h doing nothing during that time. For the two longer times, the compound crystallizes. How can I get through this problem? I mean, how should the incubation be done for longer time periods.
Thank you in advance!
You can use Eppendorf "chimney" well plates and feel the spaces between the wells with PBS - this should help with evaporation of the media from the wells and temperature variations - I suspect this is the reason for crystallization. Do you see crystals mostly in edge wells or equal amount in all wells?
https://online-shop.eppendorf.com/OC-en/New-Products-91810/Cell-Culture-Consumables-109945/Eppendorf-Cell-Culture-Plates-PF-68136.html
Thank you for the help! I will try that but the wells that have more crystallization are the ones with higher concentration of the compound...
Yes, of course; -) You may want to place your high-concentration wells in the middle of a plate. Many scientists would not use outer wells for any assay >48h, leaving you with 60 wells to use. The outer wells are filled with 300 ul of PBS. Google "edge effect" to see what I'm talking about. Let us know what worked for you. Good luck!
Yes the evaporation is quite possible to be the reason for crystallization in wells, especially in higher conc. As Anton suggested put PBS in all the empty wells on the plate to avoid it...
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