Manas Kaushik Week 1

My building

I arrived at my apartment the Saturday before I started in lab and was unsure what to expect. Kasper and I are living together in the Philadelphia area and we moved in the Saturday before our lab started. Saturday we settled into our apartment and on Sunday we explored the area and visited our respective labs to make sure we knew the locations. 

I woke up on that Monday and headed over to the lab safety training session. The safety training session was at a different location than my lab and took about two hours. After it ended at around 11, I headed over to my lab and the nerves were beginning to kick in. I got to the building and took the elevator to floor 11 where my lab was located. Once I got to the lab I met with the Principal Investigator, Mary Mullins, and she made me feel extremely comfortable in the lab. We had a long conversation which helped us get to know each other and she showed me around the lab. I was introduced to my graduate student, Hannah, and she led me to my desk. I unpacked my materials and she started telling me about everyone in the lab. I walked with her to the fish facility and watched her collect the embryos she needed to study. Since it was only the first day and I still had some safety training to complete, there was not much hands on activity I could do. I spent the afternoon learning how to identify the embryos and then helped Hannah with her embryos since she had hundreds to look through. 

My desk area

My second day at the lab started only at 10 which allowed me to sleep in. There was another undergraduate student named Rendy who also started the same day as me. Hannah helped us both set up our lab benches which consisted of all the materials we would need throughout our time at the lab. She also showed us where other materials were located if we ran out or needed to borrow something. Even though it was only my second day in the lab, I was getting a good feel of where everything was located. Once we set up our lab bench, Rendy and I began calibrating all of the pipettes to make sure they were accurate. In the afternoon, Rendy and I attended more safety sessions which were required by the lab. We were done around 5:30  and only had one safety session remaining before we could work with the fish and all the machines.

Wednesday of my first week was when most of my actual lab work began. I received the hand on fish safety training in the morning which allowed me to work directly with the fish. Hannah and Ben (another graduate student) helped Rendy and I learn to inject the embryo. Since we were just learning, there was no goal with the particular injection it was just for practice. We collected the embryos from the fish tanks and began injecting. It is a very intense process which requires lots of patience and attention. After injections, we put the embryos in an incubator to allow them to develop faster. Later that same afternoon, the embryos were developing so we practice counting/sorting the embryos. After about 4 hours it is easy to tell the embryos apart and identify if they are fertilized or unfertilized along with their pattern formation. We had thousands of embryos to identify so this process took almost three hours. 

My lab working space

On Thursday I came in at 10 once again but my day was extremely busy. In the morning I learned to fin clip. Fin clipping is important for collecting DNA from the fish to identify their genotype. To fin clip you create a solution using water and tricaine which temporarily knocks the fish out so it feels no pain. The tail fin is able to regrow so the fish only loses its tail temporarily. Using a net, you grab a fish and place it in the tricaine solution for approximately 20-30 seconds depending on the size of the fish. Once the fish stops moving, you grab it with a spoon and place it on top of a petri dish. Using a fresh razor, you cut a piece of the tail fin and store it for later use. The fish goes back in its tank and continues living normally. Later in the afternoon, Rendy and I continued practicing identifying embryos because it is a hard process that you improve on through lots of practice. The fin clips were placed in a solution over night which will dry them out. 

Learning to identify different embryos

The last day of the first week came fast. Although it was just the beginning, I felt as if it could have still been the first day. Every Friday we have lab meeting where one member of the lab presents on the research they are conducting and takes suggestions on how to improve their studies and results. Today we created a master mix using RAPD digest for our fin clips. Once we put all the fin clips in the RAPD digest, we sealed all the wells that contained the solution shut so that nothing would evaporate during PCR. We put the wells through the PCR machine and received our results. Unfortunately we were not successful using the RAPD digest. We knew this because in four of the wells we only put the master mix and water but no fin clip. In spite of this, it was still fluorescing on the computer meaning something went wrong with the procedure. We spent a full day on this just to find out we were unsuccessful. We did not have enough time to repeat the process this week so we decided we would hold off until next week. My first week was filled with lots of thrills and I look forward to what the next few weeks hold.