Hi everybody
I'm facing a serious problem when growing Arabidopsis Thaliana in a square petri dishes vertically.
in few days, a water appears in the bottom of the dish, like in the photo and I have to remove it, and there is a risk to contaminate the culture with fungi and destroy the experiment.
any solutions??
thanks in advance
Hi,
Even if you leave the plates open until they solidify you will get some condensation, especially if you stratify you seeds at 4 degrees. Avoid parafilm and Saran Wrap when sealing your plates and use a breathable tape instead. We use band-aid rolls. If your plates still are collecting water, put them vertically on a pile of paper towels and the water will come out and go into the towels, no need to open the plates or remove the seal. The latter will however only work if you use porous breathable material for sealing the plates.
After you pour the media in the plates (in a sterile laminar flow hood), leave them open (with the lid off) for 20-30 minutes. That will reduce the water content of the media and should reduce or eliminate the water that accumulates in your vertical plate.
Also, if you wrap/seal the plate currently using parafilm, you could switch to something like micropore tape that allows gas exchange from the plate and the outside atmosphere (while still blocking contamination). Use of micropore tape typically reduces condensation in our tissue culture containers, so it would likely help with your problem too.
after having poured the medium into the plates, you must let them solidify half-open next to the flame in order to eliminate all the vapors (when cooling, these vapors form droplets of water). Also you have to close the plates with parafilm and invert them (cover lower face) during incunation
Roger Thilmony Khadidja Side Larbi thank you so much, I really appreciate your answers
Hi,
Even if you leave the plates open until they solidify you will get some condensation, especially if you stratify you seeds at 4 degrees. Avoid parafilm and Saran Wrap when sealing your plates and use a breathable tape instead. We use band-aid rolls. If your plates still are collecting water, put them vertically on a pile of paper towels and the water will come out and go into the towels, no need to open the plates or remove the seal. The latter will however only work if you use porous breathable material for sealing the plates.
Svetlana Boycheva Woltering in fact, although I usually leave the plates opened after pouring the medium for a while I get the water as you said... so thank you so much, I will also take your suggestion into account...
Yup good suggestions. After pouring media in the plates (in a sterile laminar flow hood), leave them open (with the lid off) for 20-30 minutes and then wrap with breathable tape. After practicing this, had never met this situation.
You may use air drier to evaporate the water droplets, no need to open the seal from plates.
Shweta Jha thanks, can you please provide me with a link to an air drier and how to use it?
I think a hair dryer might heat the seedlings too. If you are using a room with standardised conditions or a phytotron you could change the humidity settings to 35-40% instead of 50%. One possible solution that I forgot about in my previous answer is increasing the agar by about 10 - 15% (for ex. 8.5 g/l instead of 8).
Svetlana Boycheva Woltering I also think that the hair dryer would contaminate the medium....
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