2005 RET Participant Chrysti Carte
Hendersonville High School
Biology and Physical Science
Biomedical Optics Lab
I was involved in a research project in the Optics lab of the Biomedical Engineering Department. I worked in collaboration with a graduate student on a project involving a heat-shock protein known as Hsp70. When cells are subjected to a heat stress, proteins in the cells begin to be denatured, which means they lose their structure (and subsequently their function). Hsp70 is produced by the cell as a repair mechanism, in an attempt to salvage the denatured proteins. In my project, we worked with genetically modified cells. These cells had a luciferase gene (like that in fireflies) added to a part of their DNA responsible for producing Hsp70. In our tests, we subjected these cells to various heat stresses using a water bath. We then added luciferin, a substrate, to these cells. If the cells were stressed enough to produce Hsp70, the luciferase would be expressed and would react to the luciferin, producing a fluorescent glow. Using a special camera that detects luminescence, we would image the cells at various time increments post heat-shock in order to track the production of Hsp70. We then modified the experiment by administering a mild heat shock, known as pre-shocking, a few hours prior to actual heat-shock. This is designed to be mild enough not to damage the cells, but encourages the cells to begin producing Hsp70 in hopes that the actual heat-shock will not be as stressful to the cells. We conducted numerous tests to determine optimum times and temperatures of pre-shock and heat-shock in order to obtain the most Hsp70 production and the least cellular damage. Towards the end of my time in the lab, we also began working with an infrared laser to heat-shock cells, as opposed to using the water bath.
My curriculum is a mosaic based on the reactions of Luminol, a substance used in Forensic Science to detect the presence of blood. When in contact with blood, the Luminol begins to fluoresce. The mosaic, which is designed for a Physical Science class, focuses on the properties of light, fluorescence, and the chemical reactions involved in the practical application of Luminol.