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!