Lab and Field
Microscope Techniques
Taking General Biology one and General Biology two courses prepared me for the basic understanding of using a microscope. I later took Microbiology which broadened my knowledge of using microscopes, because I had to complete different techniques prior to viewing organisms under the microscope. An example of one of these techniques would be gram staining. We had to do the procedure of a gram stain, which happens to be many different steps, and then view them under our microscopes. If we did not do the gram stain appropriately, we would not be able to see anything under the microscope. At first, it was very intimidating, but once I got it, I understood how to do it forever. Dr. Overholt's Investigation and Research III class also helped me to be proficient using the microscope. We had to view the electrodes under a microscope and perform a patch-clamping technique.
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Making Electrodes
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This was the very first thing we did in Investigation and Research III with Dr. Overholt. It actually took us so many attempts to understand this technique and once we got it, we realized how easy of a task it was. Electrodes are very thin and fragile. They can break at any moment, with the slightest touch. We used a machine, placing a thin piece of glass between the metal pieces within it. We then had to tighten the levers, not tightening them too tight, so the glass could stay in while the electrodes were being made. We would then close the machine and program it to "49", then press enter and wait. My lab partners and I would peek in and watch how the electrode heated up. We saw an orange color every time and when we heard a "pop" we knew they were ready. We could then open up the machine and carefully remove electrodes.The machine basically heats the class into two different electrodes. After having the two sharp pieces of glass, which is what I liked to think of as, we had to heat them up to ensure they were more flat at the ends, using what I liked to call the "heated microscope". This technique was also used to make sure there was a smooth tip to have a whole cell, basically for the patch-clamping technique. They were then ready to use for the patch-clamping technique.
Solution Dilutions
Taking the General Biology one lab, I learned how to create different solutions early in my college experience. I also learned how to do the math behind the solutions. Once again, in my Investigations and Research III course, we made solutions for the extracellular matrix for the cells we used. We used this solution during our patch-clamping technique. I enjoy math so this was my cup of tea.
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Patch-Clamping Technique
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The main goal of this technique is to suction to get a membrane electrically tight seal. In this technique, we took the electrodes, previously created, using Dorsal Root Ganglion cells, or cancer cells. At the start of Investigation of Research III, we were unsure which type of cells would be used. Eventually, we used the cancer cells. Under the microscope, there were so many cells! It was very interesting to me because they looked almost like many bubbles scattered around. We also had to use the extracellular solution we created previously in the semester. The suction was on the opposite end from where we had the extracellular solution coming in. Patch-clamping specifies which membrane potential to go to and injects the opposite amount of current to stay where everything needs to be. This was what we studied with our research and later presented on Scholarship Day with our posters. The hardest thing about patch-clamping for us, was making sure we went down far enough to touch the cells but did not break the electrodes. If the electrodes were broken, there would be glass all throughout our cells. As you could imagine, this was something that happened very often for us.