I have started using the Thermo Fisher Heracell 240i tri-gas incubator for culturing the cells in hypoxia (5% Co2 and 5% O2). I have connected the incubator to nitrogen gas cylinders to achieve hypoxic condition. To my surprise, the incubator is using a lot of nitrogen gas to maintain 5% O2 inside the chamber. It is using almost one full cylinder of nitrogen (roughly 6500 liters of gas) per day. When I contacted Thermo on this issue, they said that tri-gas incubators use a lot more N2 than CO2 because the displacement/suppression of O2 requires far more N2 than the addition of only 5% CO2. Also, the frequency of door opening significantly alters the use of nitrogen gas. During my experiments, I opened the door hardly twice a day. Does anyone have the same experience with the Heracell tri-gas incubator? I would really appreciate your feedback on this. Thank you for your time.
6500 liters per day?? i will inquire here in our group. generally we switch off the gas supply in the beginning of the day before opening the doors , and switch on the gas supply at the end of the day or end of the work. switch on the gas supply when you are sure that you'll not open it again in the day. Hope it will help you.
I would get away from tri-gas incubators to maintain low O2 levels. Just because the little meter on the incubator reads 5% or 3% O2, this is meaningless to what your cells experience. An incubator reading 5% O2 containing 10 cell culture dishes with different media levels and cell types will all have very different pO2 levels available for the cells. Have a look at this excellent article to see the difference between atmosphere O2, media O2 and cell-available O2 levels.
Article Limitations of Oxygen Delivery to Cells in Culture: An Under...
Plus, with constant door openings and all of the gas exchanges throughout the day, all of the publications that report X% O2 culture are pretty much meaningless in terms of what the cells actually experienced during their culture life.
Ditch the dumb gas-controlled incubators and try culturing in the Petaka G3 in a regular incubator, no supplemental CO2 required. The Petaka is the only cell culture technology that automatically and precisely regulates the O2 level in the culture media and available to the cells. It is also the only technology that allows you to measure the pO2 in your media in real time, outside of the incubator so you have actual O2 levels throughout your culture and during your experimental steps.
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