Hello again! It's hard to believe it but it’s almost Week 8! My introductory fly experiment presentation went well, and I finished early, so no taking away of the clicker. Now, I’m two weeks into my independent research project and the lab is starting to get less scary. I feel like I’ve gotten to know everyone better now, and being in the lab is so much fun! I’ve become much more efficient in making my fly food, and thanks to Dr. Leystra’s concentration wizardry I only needed to make two drug stocks for each of my conditions.
Speaking of my experimental conditions, I’ve chosen my independent research project! I decided to test the effects of lead poisoning on fruit flies gut microbiome. I did some additional research and found that vitamin C and iron-rich diets were sometimes used to remove lead from the human body, so I used both as drugs. My working hypothesis is that lead will decrease microbiome diversity, and that iron and vitamin C will restore it. If you’ve done the math, you might have noticed that would mean I have 8 vials to make and test every week. And you would be correct. Do I regret my decision? Depends on how Week 8 goes, but so far no. I’ve managed to create an assembly line system to make all my water solutions for my fly food, which cuts down on that step by almost half. Last week, however, I ran into the issue of having no functional pens to label my vials with. After sending over 10 pens to their demise though, I finally found one that worked! I also found out there is a drawer full of brand new pens in the prep room, only a couple of feet from where I was attempting to label my vials. Fun! I also decided to lower the number of flies in my vials from 60 to 30, which makes my sorting time as low as a four vial experiment.
All that time saving is necessary because for my experiment as I am doing the microbiome assay, which is complicated, to say the least. It requires five males from each of my vials to be put in separate microcentrifuge tubes. After that, you need to drown them in ethanol for one minute to kill external bacteria that will interfere with the results. The problem with microcentrifuge vials is that there is no way to knock the flies out once they wake back up. I found that out the hard way when one of my CONTROL vial flies escaped as I pipetted in the ethanol. I guess you could say I have a bit of a love-hate relationship with the flies. Even in death, they manage to be annoying, as two of the CONTROL vial flies managed to fall out of the tube when I flipped it over to air dry. Luckily, it was smooth sailing from this point on, as all you need to do is crush them up into MRS broth and then dilute into a 1/50 mixture. Pipette it into the MRS plate, and then boom, done! If I did that step correctly, I now have data! If I didn’t, then yes, I do regret having 8 experimental vials.