My Senior Project began on Tuesday the 7th, I had just flown into New York and was excited to begin my experience at the Department of Chemistry and Life Sciences at the United States Military Academy. Throughout the next three months, I am looking forward to learning all about bio-engineering, as well as how a real-life lab operates. After this first week, I am happy to say that I am well on my way to doing just that.
For the majority of the past four days, my time in the lab has been spent shadowing Dr. Alex Mitropoulos. Dr. Mitropoulos’s work has centered on the development of an “eNOSE”, a device that can utilize the biological pathways that allow living things to detect odorants in order to identify volatile substances. However, before a physical prototype can be constructed, it is important to remember that the biology behind it must be proven effective. To show that it is possible to engineer a cell to respond to an odorant (and for that response to be monitored and interpreted), a workflow had to be developed for the lab. My first day in the facility was spent learning about the process by which this lab’s research is conducted. Firstly, a strand of DNA that codes for an olfactory receptor is received from another USMA lab. A good amount of this DNA is needed, so the original copy is inserted into bacteria which will readily divide and amplify the genetic material. A small beaker ripe with nutrients is the perfect environment for this to happen, it only takes one night for the solution to turn cloudy with bacteria. Although the target DNA is being produced by the bacteria, it is mammalian cells that will be used as sensors in the eNOSE. As such, the DNA must be extracted from the bacteria in a process referred to as “purification” so that it can be transferred into the mammalian cells (this is referred to as “transfection”). A series of solutions purify the DNA by discarding the unneeded cell membranes, proteins, and RNA strands. Transfection is accomplished by a compound known as “Lipofectamine”. This incredible substance forms micelles around the DNA, mimicking a lysosome that eukaryotes will readily accept. This allows the DNA of interest to “Trojan Horse” its way into the target mammalian cells. Finally, I got some experience in the simple (but very important) task of keeping the mammalian cells alive. While bacteria are hardy, the cells which will act as sensors in the eNOSE are quite needy. Every few days, their media must be replaced in order to provide them with enough nutrients to survive. I am very excited to observe the last steps of the workflow in the coming days and weeks. Specifically, I am interested in seeing the different ways that the success of the transfection can be gauged. From what I have heard, performing a Western Blot can reveal if the DNA is being expressed by showing the production of a target protein, but an analysis of the cells’ electrical activity in the presence of an odorant will prove the existence of the desired physiological response. After this successful week, my motivation to keep learning and eventually take up a more active role in the lab can only increase. Thank you for reading the first installment in my blog, feel free to comment below if you have any feedback!
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AuthorIsaac Dabkowski - 2017 BASIS Oro Valley senior Archives |