This summer has been flying by and it is hard to believe I only have 4 weeks left working in the Snow Lab! These past few weeks have been full of exciting findings. After a few roadblocks to my experiments, three of my projects have picked up steam. It’s been so rewarding to see the experiments that I’ve set up and conducted yielding interesting data. I’ve really learned this summer that scientific research is a process with many ups and downs--experiments can fail over and over again, but then a small tweak can produce exciting or even unexpected results.
When I first tried to treat bees infected with the parasite Nosema ceranae with eugenol--a drug we suspected would inhibit the parasite from stealing amino acids from its host--there was no difference in spore counts between the control and treated bees. Because this was one of my first experiment of the summer, I was definitely disappointed with these results. Professor Snow pointed out, however, that just because experimental results don’t fit with the hypothesis, doesn’t mean they are not useful. He reminded me that often the most interesting scientific discoveries arise when experiments don’t go as planned, and when the findings aren’t as expected. Keeping this in mind, we decided to set up a second experiment in which some bees treated with eugenol were simultaneously heat shocked, because we knew that the protein that is targeted by eugenol is upregulated during heat shock. The results of this experiment showed that there were fewer spores in bees treated with eugenol and heat shocked that there were in either bees just heat shocked or just treated with eugenol! I’ve now been working on repeating this experiment to verify these results, which has been going well so far.
Another aspect of this summer of research which had been particularly enjoyable has been working more closely with the bees. Although I’ve worked in the Snow lab for almost year now, most of the academic year is too cold for the bees, so that most of our work is on samples collected in warmer weather. Professor Snow has taught me so much this summer about beekeeping, including how to introduce a new queen to a hive, how to perform a hive inspection, how to differentiate different types of brood cells, and so much more. In learning from him about the bees, I’ve been collecting ideas and strategies for conveying information to others that I plan to use in my informational video. A few weeks ago I got to travel with Professor Snow across town to check on a hive we donated to Harlem Grown, an urban farm serving the Harlem community. When we got there, Professor Snow checked the hive, found that it was doing very well (with many brood cells of new bees) and reassured the farm workers that everything was in order. Because there were so many bees about to be born, we took a frame back to Barnard with us both to lower the volume of bees at Harlem Grown and to use the newborn bees for our experiments. This was particularly exciting because we got to see the newborns emerge straight from their cells and handle them immediately because they do not fly or sting for a few days!
Newborn bees are preferable for longer experiments because they live much longer that the older forager bees that we normally collect from the landing board of our hives. The newborns we retrieved from the Harlem Grown hive allowed me to finally set up my project of feeding double stranded RNA to the bees to attempt to knock down the expression of Nosema heat shock genes. This experiment requires a few days of infection and then over a week of double stranded RNA feeding, and we hadn’t been able to keep the usual foragers alive long enough for an experiment of this length. In past years, Professor Snow has not had much luck raising newborns because they could not feed themselves in the first few days. This year, however, he learned a new method for raising newborns from a colleague at a conference. She suggested that we follow her protocol, in which you put a few foragers in with the newborns to care for them in the first days of their life. This approach, along with our new larger newborn rearing cages, worked fantastically and we were able to keep the newborns alive long enough to complete the study! This experiment led to some very unexpected but interesting results, so in these last few weeks of the summer I am repeating it with a new group of newborns we just collected.
For my third project of the summer, I’ve finally gotten started with flow cytometry, travelling up to the Columbia Medical Campus three times now with Professor Snow to analyze and sort Nosema life stages. It has been so exciting to work on this new project because it is so unlike any other procedure we typically do in lab. After we analyzed the samples on our first visit to the flow cytometer and correlated them to the spore count and Nosema DNA levels in each initial sample, we attempted to use the sorting function of the flow cytometer on our second visit. If we can sort Nosema into distinct stages and extract RNA from the sorted samples, we can them see the differences in gene expression of each life stage! The difficulty is to successfully extract RNA from the sorted samples because RNA is so unstable and the process of sorting is rough on the cells. So, to attempt to accomplish this, Professor Snow suggested we use an entirely different RNA extraction technique than usual so that we could sort the cells directly into the chemical used for extraction. Because this was a new protocol and we were unsure even if the RNA survived the sorting process, Professor Snow and I were extremely excited when our qPCR showed that I had managed to extract a small amount of RNA from the sorted samples. I am looking forward to attempting to sort and extract additional RNA from Nosema samples in the upcoming weeks. Because the life stages of Nosema are so poorly understood, I feel like this project could really make a big impact and provide novel insights into my other projects, as well as the projects of other members of the lab and outside scientists working on Microsporidia!
Besides learning about what it means to conduct research and be a scientist this summer, I have also learned about how to share scientific information with others. In our newly implemented lab meetings every week, each lab member shares a little bit about what they have been working on in the past week while one of us does a longer powerpoint presentation on their project, its background, and the data they have collected. A few weeks ago it was my turn to present at lab meeting and, of all my experiments, I decided to explain my flow cytometry project because it is so different from my fellow lab member’s projects and they had told me that they were eager to learn about the technology and my results. In preparing and presenting, I was really forced to take a big picture look at what I am researching and think about how best to explain in to my peers. After the lab meeting, I found that I had a much better grasp on my topic and felt more confident engaging with it and explaining it to others outside the lab. I’ve realized that the ability to teach others about something you have learned is the best test of your mastery of a subject and can even force you to further solidify your understanding! As I prepare my poster for Barnard’s Summer Research Institute poster session at the end of the summer, I hope to take what I’ve learned about presenting scientific information to others and use it to craft an engaging and informative poster! In the remaining weeks of research in the Snow lab, I am looking forward to running a few more experiments to verify results and wrap up loose ends, as well as preparing for continuing this research during the school year in the Snow lab.