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Week 1: Becoming Familiar With Testing Methods
I am really excited about this opportunity to perform research through CMOP. I am interning in Dr. Brad Tebo’s lab. My frontline mentor is Dr. Matthew Jones, a post-doctoral researcher studying manganese cycling in the environment.
I began my internship on June 15th and was able to meet several of the other interns in the morning. After becoming oriented to the program and getting my ID badge, I spent several hours becoming familiar with the lab. The afternoon was spent traveling with Vanessa and the other interns to Pacific University where we listened to Jon Waterhouse talk about Indigenous Perspectives and the importance of maintaining strong and respectful relationships with indigenous populations.
On Tuesday, I began my first full day in the lab. My project involves studying certain bacterial species and their ability to oxidize manganese. Before I am able to begin studying this, it is important that I understand and know how to perform various tests that will allow me to measure the types of manganese present in different samples. I spent Tuesday performing and becoming familiar with the LBB (Leucoberbelin Blue) assay. This assay allows me to measure the amount of manganese oxides and manganese (III) present in a sample.
On Wednesday, I created a calibration curve from my LBB spectrophotometer data. This curve will allow me to determine the unknown concentration of manganese oxides or manganese (III) from my samples after collecting data on the absorbance values. I also learned how to use the autoclave as Kati, another researcher in the lab, had media that needed to be autoclaved. In the afternoon, I learned how to perform the formaldoxime assay. This test measures the total amount of dissolved manganese present in the sample of interest. This assay requires several solutions to be made up and mixed and so I made up the first part before leaving for the day.
On Thursday, I continued testing with the formaldoxime assay. I finished making the other solutions and made up several working standards. I then transferred my samples to a 48 well plate before adding the Formal B reagent to them. I used the spectrophotometer to measure the absorbance values of each sample at different wavelengths and created a calibration curve for this assay. The rest of the day was spent learning another testing method—measurement of H2O2 by Amplex red. Matt informed me that this test is more difficult and fiddly than the previous two and I have found this to be true. This test requires the production of two phosphate buffers, which I made up. However, I had a difficult time getting these buffers to the required pH.
On Friday, I continued preparing and performing the Amplex red test. I quickly learned the importance of taking the A-red reagent out of the fridge well in advance since it takes quite awhile to melt and it must be liquid before being added to the working reagent. I created H2O2standards at different concentrations and mixed them with my working reagent in a 96 well plate. After allowing the sample to shake for 20 minutes, I used the spectrophotometer to measure the fluorescence of my samples. The calibration curve was not very good for this trial run as my linear correlation was relatively low. Therefore, I spent the afternoon repeating this test in hopes of getting a better calibration curve. Unfortunately, my second calibration curve was worse than my first which may have been due to allowing the working reagent to sit in the light for too long. I was able to meet most of the others involved in research in the Tebo lab at the weekly lab meeting. Matt, Kati and Christine presented their research areas, allowing the interns to gain a better understanding of what others are working on in the lab. This ended my first week of my internship. Everyone in the lab has been so nice and helpful and I am excited for the upcoming weeks!
