While trying isolation of thermophiles, we came across a difficulty: when we incubated our sample at 50 degree celsius, almost most of the media dried up.
How do we prevent the drying up of media (agar media)?
We place the plates in a plastic hermetic container, and then into the incubator. (I't big enough for allowing good air flow.)
Have you tried sealing plates with parafilm or incubating them inside trays with lid containing water to maintain relative humidity at a high percentages?
Do you buy your culture media or do you prepare it yourself? If its home made, then try pouring as much culture media into the plates as you can (~ 20/25 ml). This combined with a high relative humidity might help to solve your problem.
Thank you very much for all your suggestions. we tried enclosing the plates inside a Styrofoam box and incubating (the styrofoam was to reduce temperature loss during times of power shedding; this was an impromptu idea and i wonder if that was a good idea). We tried sealing the plates with parafilm, but incubating for 3 days at a constant temperature of 50 degree celcius melted the parafilm.
We did not buy the media but it was home made.. I will try increasing the relative humidity as well as the amount of media used. Thank you again.
I have never worked at so high temperatures, but I found that puttinge the plates in a sealable plastic bag containing a wet cloth works pretty well for incubations at 35°C for 3-4 days. It should also work at higher temperatures. I used it for small microcosms (250µL agar /well in 24-wells microtiter plates) and it worked pretty well.
I think you can use water bath for incubation and use parafilm to seal the plates.
You have to increase the air relative humidity around your plates and trying to do what is done in an incubation chamber. Putting the plates in a box (or a sealed bag) is a good solution in order to reduce the air around and regulate RH. You can then put an aqueous solution at a given water activity in the chamber to increase the relative humidity of the chamber atmosphere. I would not use pure water because you'd certainly obtain a lot of condensation into the box but also on the plates.
we will try increasing the RH and incubating the plates inside a closed container. Thank you again for all of your suggestions.
It is better to put the plate upside down in the incubator to reduce the evaporation.
thank you all for your valuable suggestions. We incubated our plates increasing the RH of the chamber and we succeeded in preventing the loss and cracking of media due to over heating and evaporation.
After how long does the agar media crack if kept at room temperature?
We had problem of drying of culture media poured in stainless steel plates.When we increased the RH by placing cottonwool soaked in water in a separate container the drying was found with a few plates.We attributed the difficulty in monitoring the volume of medium to be poured in a SS plate compared with a glass plate.The volume less than the optimal may also lead to drying and cracking.
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