Working at the Mann Lab - full time for 10 weeks - this summer has been exciting, lots of work, thought-provoking, and challenging. I had the opportunity both to carry out a complete project of my own and to work with others in the lab on their long-term projects. Working on my project was both very exciting and very challenging because, for the first time in the lab, I had to take full responsibility. The project started off slowly, as I became familiar with the dissection technique, antibody staining, and using the confocal microscope. I also had a great time creating my poster for SRI. Creating the poster made me more aware of what I knew and did not know about the project. It also allowed to me to gain valuable insight into how to present results and how to provide background information. My SRI poster was the first scientific poster I made, so I learned about making figures and the different ways posters are created in science.
The projects I have worked on independently, such as the genetic cross projects, have also taught me a lot. By monitoring my own flies, I learned about the fly life cycle and genetics. For each cross, I had to determine the possible genotypes of progeny we might get, which reinforced my understanding of genetics. I have always enjoyed genetics, in school; this summer, I got a chance to see a type of genetics research in a lab setting. I enjoyed the challenge of understanding advanced genetics; I am looking forward to taking genetics at Barnard, this upcoming year.
Working at the Mann Lab, I had a chance to speak with and observe many biology graduate students and post-doctoral students. Working in academia, in a STEM field, is by no means easy. Many of the researchers, who I work with are highly determined. However, I think the most successful researchers are the ones, who have a flexible attitude, in addition to perseverance. Frequently, there are unpredictable variables in the research setting. Equipment may break, or genetic crosses may take longer than anticipated, as mine did. People with flexible attitudes seem better able to deal with these ups and downs. Another characteristic that the successful researchers have is curiosity. Curiosity is what drew me to science; science allows and encourages you to ask questions.
This summer, I have worked more independently than I ever have before. In labs at school, the instructions were always very clear, and there was not much to figure out on my own. I could always speak with classmates or consult my book or protocol. But this summer, I could not always find resources easily, and my mentor encouraged me to think about the crosses and other projects on my own. At times, I felt very challenged by the large amount of independent work, but as a result, I now feel more confident tackling scientific questions and experiments on my own. I also learned that I enjoy collaboration. In my last two reports, I discussed collaboration, and its importance to STEM education and research. Collaboration is key for STEM discoveries. However, on the day to day basis, many of the researchers primarily work independently and are in charge of making all or most of the decisions for their work. Working independently so frequently, this summer, also made me aware of the satisfaction in working as part of a team towards a common goal. However, both are important aspects of science.
My experiences, this summer, have reinforced my love of science and the research process! I look forward to continuing to do scientific research throughout my time at Barnard.
In my last report, I discussed my antibody staining project. As of last week, I have imaged all my fly ventral nerve cord samples using the confocal microscope. I used a software program to analyze those images, and discover the results! I spent most of last week making my poster for the Summer Research Institute (SRI) presentation.
Another project I worked on in the lab is setting up and monitoring genetic crosses. The ultimate goal of this experiment is to create flies with a genetic modification. These flies will then be used in behavioral experiments. My graduate student mentor, Clare, studies walking behavior in flies using the FlyWalker: an apparatus designed to allow the study (the walking behavior) to put the flies in a small chamber from which they cannot escape. The FlyWalker is accompanied by a software program that allows us to track the steps and other gait parameters of the fly’s walking. I spent much of my time in the lab during the spring semester going over the tracked videos to ensure the program had tracked the walking accurately.
Scientists have established that three neurotransmitters play a key role in walking: serotonin, dopamine, and octopamine. Researchers in the Mann Lab have already done experiments tracking the walking behavior in flies with one or two of these systems inhibited. The aim of the genetic crosses - I am setting up - is to inhibit all three neurotransmitter systems. When only one or two neurotransmitters are inhibited, there is a possibility that other neurotransmitters have redundant roles or may compensate for the loss. When these flies are ready, they will be walked using the FlyWalker and the data will be analyzed. The way the neurotransmitters are inhibited is by using the Shibire tool: a heat-sensitive tool that stops the formation of synaptic vesicles which carry the neurotransmitters. When the flies are heat shocked, formation of these vesicles stops. However, at non-heat shock temperatures, the flies behave like control wild type flies. Monitoring the crosses takes a large part of my day at the lab. Every morning, I check on the flies to see if there are new progeny, and to make sure they are not overcrowded. Additionally, most crosses require that the females are virgin females because female flies can store sperm. By selecting virgin females, we know that we are selecting female flies who will mate with males to create the exact type of progeny fly we want. Fly crosses normally take 10 days to produce progeny. The progeny are either used for experiments or used for another cross. I enjoy working with the crosses and being exposed to many different types of flies. It’s also always fun to find the progeny I’ve been waiting for!
In my last report, I mentioned how important collaboration is in science. One very interesting collaboration is between my mentor, Clare, and Columbia engineers. This summer, I have had several opportunities to accompany Clare, when she goes to meet with Tanya, who is a Columbia engineer and is great at developing ways to design assays for the FlyWalker, which Clare and other Mann Lab members want to test. Clare wants to design a chamber so she can study the behavior of flies crossing gaps. Tanya and Clare worked together to design an optimal chamber. While I was there, Tanya and Clare laser printed several chambers. It was very interesting to see how Tanya and Clare spoke about designing different aspects of the chamber. Tanya shared the engineering perspective, while Clare contributed the biology perspective. I am enjoying the opportunity to see this type of collaboration that highlights STEM fields being so much more interdisciplinary than they often seem at first.
Since my antibody staining project has reached its end, my goal for the next two weeks is to focus on my crosses. I have also been assisting other members in the lab with some of their projects. One project I am working on uses overlap PCR. I have really enjoyed seeing other researchers’ work because I am still exploring different areas of interest in biology. My other goal is to prepare for my presentation on July 29th at the SRI poster session. I am excited to present and see the work others in the program have completed.
This summer, I am primarily working on two projects. One is an antibody staining project and the other is setting up genetic crosses. For now, I am focused on antibody staining, and I look forward to writing about the genetic crosses for my next report. While the genetic crosses are part of my graduate student research mentors’ work, I am in charge of the antibody staining project. I have worked with my research mentor at the Mann Lab to set up the steps of this project from start to finish. I will create my poster for the Summer Research Institute (SRI) poster presentation on this project. The goal of the antibody staining is to see if neuromodulatory cells in the fly release multiple neurotransmitters. To do this, I must first dissect the flies. The flies are put in a small glass dissecting tray, in liquid, under the microscope and forceps are used to carefully remove the fly’s legs, in order to expose the flies’ ventral nerve cords (VNC): the fly analog of our spinal cord. Dissections can sometimes be challenging as the flies are very small; it is hard to keep them still because I dissect them in liquid.
The flies are then preserved in a paraformaldehyde solution and placed in an antibody solution. The antibody solution contains the antibodies against the two neurotransmitters I want to test. The next step of the process - getting to use the confocal microscope to image the VNCs and brains on the slides - is the most exciting step because after imaging and image analysis I will find out the results of my project. The confocal microscope takes pictures of the sample from top to bottom in small increments. After the imaging for each antibody is complete, the pictures can be overlapped using a computer program to create a full view. Imaging using the confocal microscope can often take many days because it takes 40-50 minutes to image just one VNC or brain.
So far I have practiced each step of the process, including dissecting, putting the flies in the paraformaldehyde solution, and making antibody solutions. I am also learning how to use the confocal microscope. I have begun the procedure for my own samples. My goal in the following weeks is to complete the process of imaging my samples, analyzing my results, and working on a poster to present at the Summer Research Institute (SRI) poster session on July 29thHowever, I have set aside time to allow for the possibility that the procedure does not go as planned, as can often be the case in science.
Having spent a little over a month working here full time (I started working in the lab in January part time), I am learning more about scientific research, and what is means to pursue a Ph.D. in a science field. Patience is key, as experiments can sometimes take longer than one would initially believe. The majority of the people who work in the Mann lab are graduate students or postdoctoral researchers, so I have the opportunity to hear about their accomplishments and the challenges in their research. Even though flies have a short life cycle, there are still challenges with time. Complex genetic crosses with flies can still take weeks or even more than a month to set up, delaying research.
Collaboration is key to both higher education in STEM and research. Researchers in the lab will borrow flies from one another or bounce ideas off one another when a project is not going as well as they would like.
Another important collaborative tool in the Mann Lab is the weekly lab meeting. Each week for two hours, one member of the lab presents their research to all the lab members. Dr. Richard Mann, the PI (Principal Investigator), asks questions and gives his suggestions about further direction for the research. In addition, other members of the lab offer advice. Considering the lab is quite large (more than 20 people), lab meeting is one of the only times everyone is together. The lab includes members from both departments: Biology and Neuroscience, and not everyone works on the same project at the same time. I look forward to lab meetings because I enjoy learning what other people in the lab are doing. Hopefully by the time I write my next report, I will be well on my way to finding out the results of my antibody staining project!