Week 5.

22 July 16

I finished the derivatization process and completed the BMAA detection protocol. Samples were sent to core facilities to be ran. We use a dansyl-chloride derivative with an  integrated internal standard transition using dueterated BMAA. Elution times were used as a confirmation for peaks that correspond to BMAA, with peak BMAA₇₁ being our confirmation ion. We can then find the unknown concentration of BMAA between the ratio of the spike, and sample. We started analyzing  previous results from other samples so I could learn how to use MultiQuant IMQ4.  Multiple problems were observed with the previous standard curves. On Tuesday, I continued analyzing the data. The machine malfunctioned during our sample run, producing skewed peaks, or no peaks at all. This was upsetting because the preparation time is tedious and long. We will rerun the same samples once the machine is fixed. We were able to generate a standard curve with an R value of 0.99985.

The gas pressure nepholometer apparatus we constructed has shown great promise. We were able to generate a gas vesicle collapse pressure curve. We are still manipulating multiple factors to find the best reading. We will look at the differences in pressure collapse curves with samples that are spiked with BMAA, and another that contains a .5 M sucrose solution that would remove the turgor pressure of the gas vesicle.

We have completed small tasks throughout the week. Samples were obtained from Ross Island and Williams Lake. The samples were filtered onto a filter paper to determine the amount of Chlα in the sample. This was followed by nutrient anaylsis prep, Lugols prep, and a glutaraldehyde fixation for future use. Dishes had to be completed. Sheree gave a great presentation to us interns. It was focused on helping us identify our weaknesses and strengths for graduate school, and possible career paths that would best match your personalized skill set and interests.