Hello everybody,
For my research I have to optimize the growth in liquid cultures of a bacterium for which oxygen availability is very important. Long story short, there will be other factors to test later (I need decent throughput) and space is an issue, so I need/want to keep working volumes low, let's say max 10mL.
I am considering batch fermentation strategies such as the use of baffled Erlenmeyer flasks; however, they do not seem to come in volumes lower than 250mL. Pre-experiments in round 24-well plates were unsuccessful, and I know that the round base plays a big role in limiting gas exchange. I have used plastic cell culture flasks with vented caps such as the ones used for eukaryotic tissue cultures and they work very well, but they are disposable (and expensive) and I am producing a looot of waste. I was wondering if anyone has had the same issue and has used, for instance, square-base plates or similar, and if it worked. Or can you please suggest alternative strategies?
Thanks in advance.
I have had good luck using 10ml culture volumes in 125ml flasks. Even if they are not baffled flasks you will get plenty of aeration (assuming a good speed on your shaker platform) because of the small volume to head space ratio. However you have to shake and can't just let the flasks sit in the incubator.
I know I've used 125 mL baffled shake flasks (Erlenmeyer), which is definitely a good option if you're looking to reuse them. The supplier Chemglass appears to offer baffled flasks down to 50mL. I would just verify it's an appropriate type of glass for your purposes and cleaning methods. If you really like the cell culture flasks, I'm fairly sure those can also be made with glass, which could be autoclaved, but reusing them is apparently a bit contentious depending on your application (Look up "Can tissue culture flasks be reused?" by Alexis Weston on ResearchGate; one of the popular answers contains some helpful resources). However, it should be fine in the case of bacteria. I'm not sure if the vented caps can be autoclave-safe or not, so it would be good to check with the supplier for appropriate cleaning methods.
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