What you've done for us--Our next steps
Anna and Gene are in front of the newly obtained Microplate reader (blue and white, right behind the diskette that Anna's holding). This was purchased with your donations and now makes our next set of experiments possible. Anna has already tested the equipment and her calibration curves are in Gene's hands.
The Microplate reader is capable of reading optical signals ('ordinary' light), fluorescence (as in many lightbulbs) and luminescense (as from fireflies). The technology available to measure these variations of light signals are used today to measure many diverse biological processes. Richard is eager to explain these light phenomena in the next lab note, so I'll limit my remarks here to what this equipment enables us to do: a) we can now measure reactive oxygen species (ROS) by a fluorescent technique; uncoupling 2 (UCP2) by an optical technique; and ATP by a luminescent technique; and we can do all of the above on our own instrument in a timely fashion. The ROS concentrations, we believe should parallel the UCP2 values, as described briefly in lab note 16 of this Experiment.com and in Lab Notes 8 and 9 in our first Experiment. ROS should be high in cancer cells but not in controls and we've shown this in just 2 cancer lines and one control line so far.
Our laboratory neighbors and friends have been extremely generous with their assistance, but it's much better to have our own equipment going forward. We can now perform our 'grand experiment' in which we will repeat all measurements on all 7 cancer cell lines, and 3 control cell lines, not limited to just three cell types. We can now measure ROS and can compare to UCP2 measured using a dual antibody technique (superior to our previous method). And we will also be performing metabolomics on all the cell lines to measure metabolites of glycolysis vs. respiration.
Thank you yet again as we are now on our way to finishing this most important phase of cancer cell experiments that we've been working on for years.