So, for my independent project, I starved flies for two days and allowed them to recover for one week. I tested female fertility right before recovery began after starvation, five days after (a medium recovery time), and seven days after (a long recovery time). I did this by counting the number of embryos that were laid by the female flies, and then finding out how viable the embryos were by quantifying the percent of embryos that hatched in two days. From my results, I found that the flies were essentially infertile before they were allowed to recover from starvation. However, female fertility improved when the flies were given enough recovery time. In fact, the fertility rates of the starved flies returned to the rates of the control flies by Day 7. However, the percent hatching data was inconclusive, as my data suggests that starvation may not have as big of an impact on embryo viability as it did on fertility.
I can’t wait to share my experiment with everyone at the symposium!
Hello! I am back to give a final, bittersweet update regarding my independent project as well as my reflections on my time at TRIP. It is amazing how in as little as five weeks, I have acquired more understanding of the techniques and culture of the lab than I ever thought I could obtain before a college experience.
Picking up from my last post, I had just conducted my first female fertility assay, and was eagerly awaiting the results of the percentage of embryos that had hatched, which would dictate the overall embryo viability of each condition. As of now, I have conducted six female fertility assays total, eight vials per assay, and twice ran two assays consecutively. By now, I could probably do these assays in my sleep! Although there were some hardships-- collecting developmental data on a maximum of 32 vials, sorting on ice as not to affect the fertility of the flies, working against time with the two hour incubation period per trial, experiencing two complete failures with two of my trials-- I was able to gather an immense amount of data, and procure some interesting suggestions. I had hypothesized that the use of individual supplements would increase fertility, while combined supplements would reduce fertility, however, was proved slightly wrong. Separately, fenugreek was able to increase both the number of embryos laid and embryos hatched; black cohosh and folic acid decreased fertility. Even in combinations, fenugreek was not enough to counter the negative effects of the other supplements. These findings then lead to more inquiries such as the effects of different concentration levels, how male fertility is impacted, or how these herbal supplements affect flies over generations. Although suggestions at this point, my data demonstrates that the world population must be extremely cautious when it comes to using herbal supplements, especially in combination, and must keep in mind to consume in moderation.
Hello again, thanks for coming back to round two! I have to say that while the last time I wrote a blog I was sure I was winning, this second time around I think the flies have gotten the best of me… for now ;)
Anyways, time to move on to the really fun stuff--my independent project, which is testing the courtship abilities of male fruit flies under exposure to various drugs. Now, I will admit that this wasn’t my first idea; however, after realizing that there were many complications with using CBD as a drug (the cost, the law, and all those other inconvenient things), I decided that I wanted to study and learn about something we hadn’t really discussed during the first few weeks: courting. Male fruit flies have a courtship dance that is identical to every other male’s. This dance involves moves such as “bumping” the female, “singing” a single-winged song, and physically chasing the female. The male continues to do this in order to gain a positive response, or, more simply put, until she agrees to copulate with the him, or, even more simply put, until the two flies have sex (cue candles and “Careless Whisper” by George Michael).
Thus, after completing some preliminary research, I settled on how I would apply courtship into my remaining two weeks at TRIP: fruit fly breeding. I noticed that between the beginning and [possible] success of a male’s courtship dance, there is an X amount of time. I decided to explore what different drugs affect this amount of time and came up with using caffeine and alcohol, used both separately and together. If there is a known substance that can essentially decrease the amount of time it takes for a male to receive a positive sexual response from a female, this could possibly be applied to scientists who commonly use fruit flies in their research. If the time it took to produce fruit flies could be lessened, then the efficiency of fruit fly breeding increases.
I’m mostly still in the planning stages of this project. There’s going to be a lot of steps for this experiment to be completed right, and I haven’t even begun them! In fact, the other day I sorted my first round of flies into four vials for each condition, but the most challenging thing was actually creating what I’m going to call the “courtship chambers”. There has to be a fairly small space, since I essentially want to force the male to attempt to court the female. This means designing my own chambers for my courtship assay, which has been pretty tough. Turns out cutting glass is harder than it looks, so I’ve decided to try and utilize transparent Legos to build my chambers; however, after a few ideas, I think I’ve figured it out, much to my satisfaction. Below is a video that better explains my various attempts (and, of course, failures!) to create courtship chambers… excuse the poor vlogging skills; my career as a YouTuber has yet to take off, and I’ve been just a bit busy with all these fruit flies.
On Thursday, I began my project by making stock solution with the different types of oils. This turned out to be rather complicated, as the solutions had to be very diluted to account for the flies’ small sizes. After making the solutions and mixing them with the fly food, I sorted my flies and made agar plates. I plan on using the agar plates on Tuesday when performing the Larval Memory Assay.
If there is one thing I learned from Thursday, it is how fast time flies when you have a lot to do. I found myself scrambling to finish in time, and had to ask a couple instructors to help (thank you!!). In future sessions, I’m going to try to be faster at the beginning of the session, even if it feels like I have time to spare. Hopefully this way I will be able to complete everything on time.
To prepare for this, I created 4 experimental conditions in terms of what I am injecting in the fly’s food: bitter gourd juice only, sugar only, bitter gourd and sugar, and a control condition. I had to dilute the serving sizes with water since a fly’s serving size is way smaller than a human’s.
“Mom, I need cigarettes”
“You need WHAT? YOU MAY NOT BE SMOKING”
“Mom, it's for a science project”
Back to the drawing board I went.
At this point, It was the day before our first lab day, so I opted for putting nicotine in the fly food. This is where my mom came in. After a few minutes of discussion, she obliged. It turned out that there was a nicotine solution in the lab on Thursday, meaning I didn’t need the cigarettes. This was probably for the better, mostly because I have no clue how to get the nicotine out of one...
Troubleshooting these problems makes me even more excited to see how my experiment turns out. I don’t know if the nicotine will have an effect at all, or if the 10x solution is lethal, or how closely this would simulate humans, but I can’t wait to find that out. As I head into my first data collection day, I can address issues that come up, solve problems I don’t even know exist yet, and continue to refine my project until the final presentation.
You may ask, what about my predictions? I think that my flies will begin to produce very rapidly and that the offspring with have developmental defects such as slow development or altered behaviors. I do not have any data to share yet but I will soon.
My collective experience here at TRIP has been very different than what I expected and very different than the last summer program I did at Temple University. The days are not too long which is a plus, and we are able to go get lunch which is really nice and refreshing. My favorite part is how there is always something hands-on for me to do. I have been learning a lot so far and I hope that continues to happen as the program progresses.
Antioxidants have been shown to be able to protect against free radical damage in cells, and protect them from some types of damage. In my Independent Project I will be looking at which types of antioxidants reduce damage done to the eyes by blue light. In my project I will be exposing adult flies to blue light, but some of the flies will have certain antioxidants in their food. I will have three separate antioxidants, Holy Basil, Selenium, and Vitamin E, and some containers have combinations of two of these antioxidants, and one vial has all three combined. So, while the flies are being exposed to this blue light they will be ingesting antioxidants to hopefully combat the damage. There is one control group that will not be ingesting antioxidants, but will be exposed to the blue light, and there is another control group that will not be eating antioxidants nor will it be exposed to blue light.
Also, I can sometimes be slower than people when doing projects, and I try my best to try and keep up with everybody while not lowering the focus and precision I use when doing the projects. However, one day, I happened to go over the time allotted, and I needed to put the drugs in the fly food, and then put in the flies. Yet, there were lots of people who stayed as long as they could to help me finish up, and with the help of everybody I was able to finish my work that day even though I still went over the time.
Here is my presentation where I explain in more detail the findings of my project:
Moving on, I have loved each and every moment of TRIP. A friend of mine recently asked me how my July went. My response? I told him that every part of July that involved TRIP was fantastic, great, and amazing--everything else was was just alright. Coming into the program, I was extremely apprehensive that I would be placed in an environment where everyone around me would understand things much more than I; ultimately, however, I’ve come to learn so much from this experience. Not only did I gain basic lab skills (micropipetting, sorting flies, making stock solutions, etc.), but I also faced many, many challenges. The best example I can think of is my failed courtship project, which showed me that even though I ended up doing something completely different than I had wanted, I didn’t feel dissatisfied with my work--in fact, I felt even more motivated to do something that worked.
The “failure” didn’t prove to be a roadblock, but rather a detour.
In addition, Dr. Amanda Purdy, Mr. Robert Herbstritt and all of the TAs helped challenge and push me to think outside of the box, to problem solve, and to set my own standards high. If it wasn’t for them, I do not think I would have given a presentation as confidently as I did. Finally, my classmates, from both sessions, were extremely supportive as well! While we may have not been the most talkative group, we all were very ready to help the other out if need be, which was a very nice environment to be exposed to.
Thank you all for taking the time to read my blogs, I hope you’ve enjoyed them!