It is hard to believe that these 10 weeks of summer research have already come to an end! Although the summer has flown by, thinking back to the first few weeks I recognize how much I’ve learned and grown as a scientist and student since the beginning of the summer. Throughout the course of the summer, working in the lab full time, I have really solidified my understanding of my own research and mastered the laboratory techniques that I use regularly. I feel even more comfortable working independently and tackling new projects and procedures. It really has been so valuable to engage in science on a full-time basis because it has allowed me to see the ins and outs of laboratory work and the scientific process.
In terms of my research projects, these past few weeks have had many ups and downs. After collecting interesting data from newborn bees fed double stranded RNA to inhibit the heat shock response of the parasite Nosema, I attempted to replicate the experiment to confirm the data. In the second trial of the experiment however, the results did not match those of the first experiment. Although disappointing, I’ve learned that replicating results is an important part of research, and when data does not repeat itself, you can reevaluate the experimental design and scientific rationale. Often these experimental disappointments can lead to new insights and questions for future research.
For example, I similarly attempted to use newborn bees replicate my experiments using eugenol to inhibit Nosema from transporting amino acids under heat shock conditions. Like the double stranded RNA experiment though, these eugenol treated bees did not show equivalent Nosema spore levels to those of bees in earlier experiments. Despite this frustrating result after three experiments with matching data, instead of giving up on the whole idea, Professor Snow suggested we consider if using newborns may have affected the experimental outcome. Although we did not gather the results we expected, we can still learn new things from the data to apply to future experiments and questions.
My final project, using flow cytometry to characterize Nosema developmental stages, has also both yielded exciting results and presented many challenges. After successfully extracting RNA from our sorted samples a few weeks ago, we attempted to increase the volume of RNA so that it would be detectable using quantitative PCR (qPCR). To accomplish this, we sorted more cells and put higher volumes of the sorted samples through the RNA extraction process. By altering this extraction procedure, we were able to extract appreciable amounts of RNA transcripts of the Nosema β-actin gene, measured by RT-qPCR. When attempting to do qPCR on other genes suspected to be active at different developmental stages, however, we had not extracted enough RNA for detection. We also tried to add a cell cycle dye to our procedure in order to further characterize each distinct life stage, but this didn’t quite work as expected. Although this is also a bit disappointing, procedures often don’t work on the first try, and, for me, trying to iron out the problems is one of the most exciting and rewarding parts of scientific research.
Despite the few minor setbacks with the flow cytometry project, I still had more than enough success and data to present this project at the Summer Research Institute poster session. Of all my projects, I chose to present on this one because of its exciting techniques, broad implications, and successful results. In making my poster, I had to consider how to best convey information to a wide variety of people including students, professors, and non-scientists alike. I learned how to interweave text, diagrams, and pictures of my research in a way that would engage an audience. The poster session itself was such a rewarding and valuable experience. I loved sharing the work I completed and progress I made in lab this summer with my peers, professors, and other attendees. Because of the differences in scientific backgrounds of the poster session audience, I had to be able to explain my research at many different levels. Thinking about how to best accomplish this type of presentation really helped me become a better teacher and scientist, while also solidifying my own understanding of my project.
Overall, this summer of research has been an invaluable and transformative experience. I am surer that science and education are fields that I want to pursue in my future and I feel far more equipped to tackle these paths. By researching alongside the other Barnard biology majors in my lab I have found incredible new peers who inspire me as a woman in the sciences. Looking forward to this upcoming academic year, I am extremely excited to continue my research in the Snow lab. I hope to keep making steps forward in my flow cytometry project, wrap up my other projects, as well as begin a new project! I am so grateful for Barnard’s Summer Research Institute and the Noyce Summer Internship for giving me this incredible opportunity! I feel like I have learned so much in just 10 weeks and feel more confident in my teaching, learning, research, and scientific skills.
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.
This summer I am very excited to be continuing my work in Professor Jon Snow’s lab at Barnard. In the Snow lab we study the cellular stress response pathways of the honey bee and of a parasite that infects honeybees called Nosema ceranae. Nosema belong to a class of organisms known as Microsporidia, which are obligate intracellular eukaryotes which infect various animals from crustaceans to humans to honey bees. Compared to other microbial parasites, the cellular biology of microsporidians has been relatively understudied. Nosema infection is widespread in honeybee colonies across the United States, and is one of an interacting set of stressors which is theorized to contribute to individual death and colony collapse. By characterizing the stress response pathways of both honey bees and their parasite in the Snow lab, we can understand the cellular interactions between these two organisms and find targeted therapies for infected colonies.
In addition to continuing the work on my project from this past academic year, I am also taking up a few additional projects in the lab ter. During the school year I analyzed data from RNA sequencing of Nosema under heat shock conditions to look for novel genes involved in the heat shock response and set up experiments to test them. Because so little is known about Microsporidia, I consulted gene homologs in yeast to determine the potential function of each gene. To continue this work over the summer, I have been conducting experiments treating infected honeybees with a drug called eugenol, which may inhibit the ability of Nosema to harvest nitrogen from its host under heat shock conditions. The hope is that eugenol may be able to serve as an alternative treatment for Nosema infections in honeybees.
Related to this project, I am also conducting experiments to determine if the heat shock response of Nosema can be inhibited using RNA interference (RNAi). This technique involves feeding bees double stranded RNA with the hope that it will inhibit the translation of target genes in the parasite into protein. In addition to reducing infection, this experiment will also help us better understand the role of certain proteins in Nosema heat shock response.
In addition to these ongoing projects, I am also helping Professor Snow with a completely new project to identify and sort the different life stages of Nosema using flow cytometry. Currently in our lab we can only quantify infection through counting mature spores using light microscopy or through quantitative PCR (qPCR). Spore counting neglects to include other life stages of the parasite. By staining samples with different dyes, we will hopefully be able to use the flow cytometer at the medical campus to sort different Nosema life stages.
Finally, I am running trials of potential new drugs to target Nosema infections in bees. These drugs were created in labs in both India and Japan to target human eukaryotic infections, such as malaria, so it is exciting to attempt to apply these drugs to an entirely different animal and parasitic disease.
So far, I have really enjoyed my research in the Snow lab this summer. Having so many projects to work on means that when one experiment is going slow, I always have another to turn to! Although I have spent the past year working in the Snow lab, being here full time, this summer has really allowed me to dive headfirst into my research. Because I only work a few hours a week during the school year, it takes a while to conduct and complete experiments, but during the summer I have been able to get so many things done!
It has also been rewarding to gain increasing familiarity with and learn entirely new laboratory techniques, such as bee collection, bee dissections, spore counting, RNA extraction, cDNA synthesis, quantitative PCR, double-stranded RNA synthesis, DNA purification and many more. Because we use many of these same techniques day-to-day and week-to-week, I have been able to sharpen and perfect my skills at many of them. One of the most fun new skills I’ve learned this summer has been how to collect bees from the hive with a homemade aspirator. By securing a mesh cage inside a plastic tube, we use our lungs to suck up bees to use in our experiments!
Another rewarding part of this summer so far has been spending time with and learning from the other members of the lab. Just like how working full time has allowed me to engage more fully with lab techniques, it has also let me get to know the other members of my lab much better. All of us are undergraduate science majors at Barnard, so we share tips on everything from the best way to dissect bees to must-take biology classes. Being surrounded by fellow women in science has been valuable to my own research and love of biology!
Although all of us in the Snow lab are continuing our work from the school year, and therefore familiar with how the lab works, Professor Snow has implemented a new weekly lab meeting for us to share our progress and findings. Each week one of us does a longer Powerpoint presentation on our research and the remaining members give a brief update on their projects. I think that one of the most important tests of your own knowledge on a subject is your ability to teach it to others, so preparing for my presentations these past few weeks has really forced me to deepen and solidify my own understanding of the biology behind my research projects.
In the upcoming weeks of the summer, I plan to continue to make progress conducting experiments and gathering data for each of my projects. Through this research, as well as through preparing for lab meetings and sharing my project with others, I hope to learn more about the cellular stress responses of honey bees and Nosema and also improve my skills at sharing my findings with others. Additionally, I am excited to begin putting together an informational video about the Snow lab bees and my own research to share with the wider community!