I am trying to do metabolic study of some cancer cell lines and want to measure the amount of CO2 released during the experiments. Does anyone have the reliable and affordable way of measuring the exact amount of CO2 released from the cells?
Dear Yagya:
Carbon dioxide (CO-2) is an end-product of cellular respiration. CO-2 evolution was originally assessed employing radio-labeled glucose, fatty acids or proteins with carbon isotopes at specific positions. This was followed with the employment of gas chromatography–mass spectrometry (GC-MS), which facilitated the detailed analysis of metabolic flux of carbon through the glycolytic, TCA, pentose phosphate and other relevant pathways. These methods are quantitative and sensitive but are labor intensive. Recently analyzers have been used to elucidate CO-2 release.
As an aside, the following is a method for estimation of CO-2 in microbial ecosystems.
Martin E. Brummell and Steven D. Siciliano, Measurement of Carbon Dioxide, Methane, Nitrous oxide, and Water Potential in Soil Ecosystems. In Martin G. Klotz and Lisa Y. Stein, editors: Methods in Enzymology, Vol. 496, Burlington: Academic Press, 2011, pp. 115-137.
New technologies in trace gas detection are revolutionizing our ability to study soil microbiological ecosystems. Field-deployable infrared-spectroscopy detectors capable of rapidly measuring multiple analyte gases simultaneously allow estimates of soil:atmosphere gas exchange and below-ground gas concentrations, and production dynamics across divergent ecosystems, creating opportunities to study interactions between microorganisms, soils, atmospheres, and global cycling, as well as interactions between different gases. The greenhouse gases CO2, CH4, and N2O can be measured in the field and compared to each other to uncover links between the biochemical pathways responsible for the production and consumption of these gases. We have developed techniques using a nondestructive, Fourier-transform infrared detector under remote field conditions in three campaigns in the Fourier-transform infrared detector under remote field conditions in three campaigns in the Canadian High Arctic to measure highly variable gas processes in soils.
Hi Yagia
There are some sensor devices ( http://us.mt.com/us/en/home/products/Process-Analytics/DO-CO2-ozone-sensor/dissolved-carbon-dioxide.html) but they are expensive and difficult to use.
A suggestion I could make is to indirectly evaluate the CO2 in the culture medium by pH measurments. As you know, CO2 affects the pH of medium and usually the cell culture media are supplemented with phenol red, a molecule which changes color according to pH. As the cells produce more CO2, the pH of the medium decreases (becomes more acidic). On the other hand, sodium bicarbonate, NaHCO3 (used as a buffer), dissociates into sodium and bicarbonate ions. By increasing the bicarbonate ions, the pH increases. The concentration of the sodium bicarbonate in the medium must be matched with the level of CO2 in the atmosphere above the medium (i.e. for media containing 1.5 to 2.2 g/l sodium bicarbonate, use 5% CO2).
If you have these parameters under control, then the approximate pH level can be monitored by observing the color of the phenol red supplemented media. If you need a more precise quantification, digital color measurements (using an ELISA reader for example), may help.
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