Natural Products Synthesis (3)

Research in science has not always been easy. For four weeks, we have been trying to identify our hypothesized product in our new reaction pathway, but all the effort seems to be futile. It turns out that our one step reaction has yield more than 10 byproducts, which is unexpected. That also causes great challenge in separating and finding the molecules we want among endless byproducts. Given the scale of the reaction, numerous products are yielded in the order of milligrams, and the identification of molecules is a real challenge.

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When Something Finally Worked

It happened! Our trial finally worked! After a few weeks straight of straight troubleshooting, we have finally managed to pull off a successful trial! Over the past few weeks we have dealt with problems with cell count. Before plating, we have to use a hemocytometer to count cells so that we can control how many cells we put into each well. When we first started conducting the trials on our own, our counts were so low, making it nearly impossible to start trials. We started to become more careful when culturing flasks and becoming more selective with which flasks to use. We also encountered issues with the fixation process throughout our trials rendering all of our cells dead on our slides. This was a problem that left us at a loss because we would see cells on our slides before we started fixation, but not afterwards. We figured out that when we were aspirating the washes, we were leaving our cells in too dry of an environment, so we began to increase our speed when fixing our cells. We also decreased the amount of washes so that we were not disturbing our cells too much. After our fixation and mounting process, we looked at our cells underneath a microscope and for our previous trials, there were very little neurites on the cells. This was concerning because we stimulated our cells with the proper neural growth factors that should cause more neurite extensions to grow. We ended up increasing the concentration of neural growth factor to help stimulate the growth. After all these changes made to our procedure, we have finally seen a successful trial! The cells look beautiful, healthy, and happy underneath he microscope with plenty of neurite growth. It has been very exciting to see the fruits of our labor finally come into fruition. I look forward to using these experimental techniques on other cell lines.