When I treated my cells with acrolein (less then 1mM) (in a 12 well cell culture cluster) the other cells that were in a tissue culture flask 25 (growing normally without any treatment) were damaged by the aldehyde , because of it's volatility. The test were performed by using 2,4-Dinitrophenylhidrazine and the aldehyde presence in the tissue flask were observed by the formation of a preciptated (by the reaction of aldehydes with 2,4-dinitrophenylhidrazine). How can I treat my cells with in the same incubator as the cells non-treated?
I would propose to use an icubator only for this experimente/these cells to avoid effects on other cells not treated with aldehydes!
It would be perfect, but we don't have this free space to put a new incubator. Thanks for your suggestion Andreas Eisenreich
Lucas,
If your cells grow in suspension, the solution is quite simple. Just put them into sealable tubes and seal the tubes.
However, assuming that your cells are substrate adherent, here are two possibilities for doing this type of treatment in the same incubator as other cells.
1. Use 25 cm² tissue culture flasks for your treated cells and seal the top of the flasks. This should contain the acrolein fumes. The pH of the culture medium will need to be monitored visually and adjustments made as needed.
2. Make or purchase a small sealable acrylic chamber outfitted with an air tight gasket. Ideally this chamber should be large enough to hold several or many multi well plates. Put the multiwell plates into this chamber, seal it and then put the chamber inside of your incubator.
For both of these scenarios, you’ll need to flush your flasks or the auxiliary chamber with 5% CO2 in air before they go into the incubator to maintain the proper media pH. Note that 100% CO2 can’t be used for this, it must be 5% CO2 in air.
Good luck,
Robert Walter
Robert J Walter Thank you for your help. I'll try to do this, I think it will work!
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