Hey, I am using 96 well plates to grow Bacteroides samples and tracking their OD using an automated plate reader. I have to take readings for a long period of time (48 or 72h) and so am experiencing evaporation loss from the wells uptown 50% by the end of the growth period. Although I am using BreatheEasy sealing membranes to seal the plates, I still end up with evaporation.
Has anyone experienced this after using a breathable membrane? Any help and suggestions would be much appreciated.
Thanks!
Hello Anubhav Basu
Evaporation of culture media is a common phenomenon, particularly when dealing with experiments over longer incubation periods. This occurs due to differences in the moisture content within the 96 well plate and incubator. Hence, the relative humidity needs to be maintained to prevent this evaporation. I suggest that you place a small stainless steel tray containing autoclaved water that will maintain the humidity within the incubator. It will definitely prevent evaporation from your wells. Wish you good luck!
I also have evaporation issues when working with a 96 well plate in an anaerobic chamber. A few things I've noticed helps:
1. No well is left empty, and the outer-most wells are filled with blank sterile media (these tend to evaporate slower than liquid culture wells, in my experience).
2. Make sure the anaerobic chamber (or incubator) humidity level is on the upper end (60-70% humidity, in my experience within the anaerobic chamber). To raise the humidity level, you can leave a bottle of sterile autoclaved water (or a tray like a previous commenter suggested!) in the chamber.
3. If it's possible to use a 48 well or larger, that will also minimize evaporation loss.
Hope that helps!
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