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Department: Microbiology and Immunology More Information A Day in the Life of an Undergraduate in the Bloom LabI had been intimidated to join a research lab freshman year. I was not sure if I had enough curiosity to stimulate a research project, and I did not want my nights and weekends to be spent inside the lab. However, I knew that working in a lab would be the only way I could ever decide if research was something I wanted to pursue after graduation. Now that I have learned and worked in the lab for six months, I am curious to know exactly how a biochemical pathway works, I know that the PI will help me to develop a project, and I have never had to come into the lab on a weekend. So a few myths have been busted.When I got over my qualms about research during sophomore year, I used the Undergrad Biology website to search for professors who studied genetics. I spoke with three professors, and I decided that despite the walk out to the Vet Center, I would gain the most experience at Dr. Bloom’s lab. My lab is small; the summer I spent here was in the company of my PI, a second professor, a lab technician, and the Lite Rock radio station. Despite the same soundtrack every day, there were many advantages to my lab situation. I had almost an entire lab bench and a laptop to myself, and my professor himself was always available for questions about protocols and discussions of results. On the other hand, working with cancer cells can mean a lot of down time, so I had to look for ways to occupy myself. I began my own project in the summer following one semester of training. I work with two Burkitt’s lymphoma cell lines. One cell line is a derivative of the other, and both cell lines overexpress Bcl-2, which is an important anti-apoptotic protein. Apoptosis is a form of controlled cell death, so it is a useful pathway for anti-cancer drugs to target. When cells overexpress the Bcl-2 protein however, they can show more resistance towards apoptotic cell death. Our two related cell lines seem to differ both in metabolism and sensitivity to anti-cancer drugs. We are interested in revealing a critical component or pathway that facilitates apoptotic cell death and that is enabling the more sensitive cell line’s bypass of Bcl-2-mediated protection. Our approach to this question is to treat the cell lines with anti-cancer drugs and then measure and compare cell proliferation and cell death. To measure inhibition of cell proliferation I use a Coulter Z1 Counter, which measures the cell density of a sample. To measure cell death, cells are stained with dye that is DNA-specific. Using fluorescence microscopy, viable cells can be discriminated from apoptotic cells, which exhibit fragmentation of the nucleus. One hundred cells are classified as viable or apoptotic, and a replicate count is performed. Measuring the two endpoints has been useful because we have seen that cell proliferation is a much more sensitive endpoint than apoptosis is. Even if the resistant cell line does not show a large amount of cell death, cell proliferation is usually completely inhibited at a certain drug concentration. Not only is this evidence that the drug did properly enter all cells, but this has also been a preliminary indication that the difference between the two cell lines lies in different apoptotic signaling mechanisms rather than in mechanisms for cell proliferation. When I joined the lab, I was in the middle of taking Genetics and I would not be taking Biochemistry until the following semester. The more I delved into the research material, the more relevant genetics became, but I do not think it is necessary to take Genetics before joining Dr. Bloom’s lab. I am looking forward to taking Biochemistry because I am sure that I will learn in depth some of the things that I have read in review articles and taught myself during this summer. Many of the papers I read were about actual clinical treatments, even though we work with in vitro models in the lab. Therefore, I believe that an interest in cancer treatment is the most important factor for joining this lab. |