We are treating A549 cells with pulsed electric fields. The media becomes more redish after prolonged treatments (11 hours). When the cells are returned to the incubator overnight, the PH changes back to its 'normal' look (less red). Is it possible the electric fields are changing the PH of the media during our experiments? Or perhaps just having them out of the incubator would cause the media to turn more redish? The media has phenol red in it.
I am not sure if the electric field has effect on PH. But if you consider that PH is actually the availability of the H+ ions its safe to assume that there would be certain effect from the electric fields.
but more importantly there is constant supply of C02 in the incubator that also acts like a buffer and maintain PH, unless you are using a different buffering approach than the carbonate buffering system.
Hello Anthony, I agree with what Zeshaan has stated above, but as you have said that you are using Phenol-Red as your media, may be some electrochemical effects of this media are also causing or giving you the effect/pH changes that you are observing . Also, I do not know what is the polarity of your pulses - unipolar changing between 0 to some maximum will certainly create electrochemical effects. Alternately, try using bipolar pulses if possible and suitable for your experimental trials. Such bipolar pulses, that swing between negative and positive levels would reduce or totally nullify the electrochemical effects including electrolysis.
I hope you shall find my answer useful to you. Good luck!
Yes any media removed from the incubator will shift in pH. If you think that the fields are having an effect the answer is simple. Take out 2 flasks of cells. Pulse one flask and leave the other out of the incubator for the same time (do not apply any fields).
Hope this helps,
Gary
I agree with the previous answers. I want to underline the dependency on the kind of electrical pulses you use. Usually when using electrical pulses in cell cultures you do not want any electrochemical reaction. It would lead to a realease of toxic substances. If your experiement is designed correctly there should be no influence of the electric pulses on the pH. More likely it depends if the cell culture is in or outside the incubator. In addition to the previous suggestion I would suggest to leave one culture outside the incubator without electrostimulation and one with. If there are certain differences between those two cultures you have to check your stimulation regime and the suitabilaty of your electrodes for this experiment.
Edit: What also came into my mind is that the electrical fields could change the cell metabolism too and lead to a different pH. But this is out of my subject area...
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