Julia Hu Entry #2 -- Surgeries and Fake Fires

My second and third weeks at the Fuccillo lab have been quite eventful and equally as enjoyable as the first week. I am gradually wrapping up the molecular cloning portion of my project, which involved the formation of four DNA constructs that will be used to create viruses. Instead of running an abundance of PCR reactions and gels these past two weeks however, I have been doing more DNA sequence analysis, bacterial transformations, and both mini and midi preps (which involve the isolation of plasmid DNA from bacteria) as well. In these past two weeks, I have also learned how to take images using the episcope and accessed the confocal, which is a more advanced microscope than the episcope. The confocal enables the reconstruction of three-dimensional structures from the obtained images by collecting sets of images at different depths of the brain slices. It was fascinating to see the power of the confocal and the beautiful images of the brain slices that it can capture.

Me looking at a brain slice through the episcope

Image of a brain slice under the episcope

Last week, I had the opportunity to witness a graduate student, Ope, perform a surgery on a mouse and inject a Cav Cre virus into the mouse's brain. As he executed the surgery, Ope thoroughly explained each step in the process to me, starting from the anesthetization of the mouse with isofluorane to the injection of the virus. I was surprised to learn that one of the most difficult parts of the surgery is ensuring that the mouse brain is lying on a flat plane before a hole is drilled in the brain for the virus to be injected. In order to ascertain that the brain is lying completely flat, Ope compared the heights of two different brain landmarks, named bregma and lambda, using a measuring device. He then adjusted the clamps holding the mouse's head accordingly to align these two landmarks on the same plane. I was also surprised to learn that the skin can simply be glued back together after the surgery is finished and does not require sutures.

Ope performing the surgery

The mouse undergoing the surgery

Another photograph of the mouse undergoing surgery

I also had the opportunity to witness two perfusions last week. While for a surgery one wants to avoid killing the mouse, a perfusion does involve the death of the mouse. In a perfusion, the mouse is killed either with a lethal injection or by anesthetizing it, making a lateral incision, and then cutting its diaphragm. While the heart is still beating, a solution is injected into the left ventricle of the mouse's heart, and the heart then pumps the solution throughout the mouse's body, draining the body of all its blood. It is crucial that the perfusion is performed quickly since the heart needs to be alive and beating in order for the solution to spread through the mouse's body. The final step is the removal of the mouse's brain, which will be sliced with a vibratome either immediately after the perfusion or on another day.

Outside of the lab, I experienced frantic fake fire incident at my apartment. At 3:30 AM last Friday, someone on the 9th floor of my apartment hit the fire alarm by accident, forcing everyone in the apartment to evacuate. Of course, not knowing it was a false alarm, I freaked out and sprinted down 13 flights of stairs (the highest floor is 14) in flip flops and my pajamas to find out that there was no emergency. Later on, after everyone was finally let back into the apartment, the alarm went off again at 4:30 AM because of a maintenance issue. At this point, I was quite infuriated and fatigued. After storming out of my room again to find out what was going on this time, I very intelligently left my keys in my room and locked myself out. To make this long story short, I ended up staying awake from 3:30 AM until 5:00 AM and having an overall panicked Friday morning.

Despite this experience, I have really enjoyed living in Philadelphia and spending time and working with the people in my lab. Our postdoc, Kyuhyun, came back from Korea last week, and Felicia and I bonded with him very quickly. It is hard to believe that three weeks have already passed!

Photos next to the popular LOVE statue