Summer Research with the College of Pharmacy: Week 3-A Very Happy Birthday and Some pH-un!

This week came to prove that a phenomenal research team and a great work environment really do make an opportunity like this worthwhile. Aside from observing and performing lab experiments, there's a lot of thinking and collaborating. Every person on the research team is examining something different, but at the end of the day, everybody's individual parts are brought together to answer a broader question. Additionally, while research may require a lot of patience and thinking, there is definitely a "fun" dimension involved.

In the process of blogging about my team's specific project details, it's necessary to be wary of what I post. This is because if too much information is publically posted about a particular experiment or our overall research goal, someone else may come to identify what our research project is about, and perhaps, publish it as their own. To avoid this, I must do two things: 1) Avoid detailing the particulars of what my research team and I are investigating before official publication 2) Avoid posting media of our specific procedure on a given specimen. I wish to share more with you, but unfortunately, until publication, I am limited to what EXACTLY I can say. Just know that alongside my lab protocols and techniques I perform, there's a lot more going on that I can't reveal right now. :)

One of the members of my lovely research team had her birthday this week-Andrea! Just because we are also working intensely to meet deadlines and submit papers doesn't mean we can't sit back and have a little bit of fun! To celebrate her birthday, one of the other members of the team, Cori, bought a delicious cake and brought it in. Every person on the research team, including Dr. McConnell, came in to celebrate and sing a version of "Happy Birthday" in their respective languages. After we celebrated, the inevitable discussion of what needs to be done in the lab and problems some of us are encountering in meeting particular deadlines ensued. Even though I have officially been a member of the lab for less than 3 weeks, I already feel like I've been here for years. It's definitely an enchanting feeling.

The Birthday Cake!

Happy Birthday!

The Research Team! (missing a few)

Furthermore, I continued to make more TBST solution in the lab. TBST is probably one of the most commonly used solutions in our research lab because we are dealing with a high volume of Western blots. I also learned how to make a dilute solution of an antimicrobial to place in our cell incubator. It requires 5 mL of a blue substrate called AquaClean and 1000 mL (or 1 L) of distilled water. This may sound easy, but there has to be a lot of precision to how solutions are mixed and what type of "distilled water" is used. There's double distilled water, distilled water, and tap water in the lab. Some solutions require double distilled water (commonly labeled as ddH₂O) while others require only distilled water (commonly abbreviated as DW). The pipetting procedure also is quite fastidious--one must spend a certain amount of time re-filling the pipette and emptying it out before all the substrate is removed. The solution must also be mixed thorougly in order to reach proper homogeneity.

AquaClean Substrate

Dilution

Pan

Cell Incubator

With all of this TBST solution-making, it's not surprising that I also ran out of the 10x TBS, the substrate used to make the TBST solution. This was a rather lengthy and tedious process, especially since it requires the use of a pH meter and careful pipetting. First, I added 24.2 g of Tris base and 80 g of NaCl to a plastic beaker. Then, I stirred the contents after adding 800 mL of ddH₂O . Because TBS is a buffer, I also had to ensure that the pH of was adjusted to a certain level--7.6 In order to do this, I had to standardize the pH electrode with 3 different pH level solutions (4, 7, and 10). I adjusted the pH of my solution by adding HCl to lower the pH and by adding NaOH to raise the pH until I reached 7.6 This took a while because one drop could really make a big difference in what the pH meter read, and I did not want to go over or under the level needed. Last, I added the remaining ddH₂O to the solution to make 1 L total.

Tris Base

Sodium Chloride

pH Standards

Standardizing the pH meter

Adding HCl and NaOH to adjust the pH reading

Final pH-7.6

The Final Product-10x TBS!

This week was also very unique because I experienced my first team meeting! All of the research members and our PI, Dr. McConnell, assembled in a conference room to discuss the progress of our individual projects/experiments. Since there are many deadlines to meet, the tone of the conversation was tense and hasty. Dr. McConnell expressed the necessity to identify specific substrates in one of the members of my research team's experiment. This is extremely important because in order for a research paper to be published, it needs to answer a question with very specific results. If not, then the paper may not be of value to even publish. A general answer or a general piece of evidence is not sufficient enough for publication. Furthermore, we talked about what other experiments needed to be performed while also being mindful of the fact that these experiments must be replicated several times in order to yield a significant conclusion. Although my research team seemed a bit stressed out with what seemed to be a conversation of never-ending experiments and things to do, I promised to help out as much as I could. Dr. McConnell even reminded me to help out with several of the projects so that these looming deadlines could be met.