Since I have returned to school after the conclusion of my winter break, I have been super excited to return to my research. Since I have returned, I have been conducting in a few new techniques and utilizing several different cell lines through experimentation. Dr. Krucher explained to me her hopes for her current work with 3-D’s. At the end of last semester, she informed me that another method of killing off cancerous cells in 3D cultures was needed and that she was looking into one that involved some drug. Currently, we are working with special plasmids of different cell lines and adding the drug called Dox (Doxycycline) to slowly diminish those cells. The plasmids are injected into the cell lines and must have a signal bind to the promoter region to initiate a response. I believe there is a fluorescence (Dharmafect) involved that will glow a green shade with the presence of protein or Rb in the breast cell lines. With PNUTS KD initiated in the cells lines again, there should be less of a green presence in the cells because they are undergoing apoptosis, or at least that is the hopes of this experiment. I am still new to this concept especially due to the fact that this technique is not set in stone with Dr. Krucher in regards to its functionality.
Currently, I am working with hanging-drop plates, which is probably one of the more tedious and challenging techniques I have been taught so far. Hanging-drop plates are difficult because they contain spheres of clumped cells that can easily drop through the wells and onto the bottom part of the plate. Eventually the spheres must be poked through each well, collected in a tube, run a protein assay to see how much protein I have in total, then run a western blot to see if Rb is present in my sample. However, I have retrieved some exciting results for Dr. Krucher through my most recent western blot. In essence, I feel I am finally contributing worthy results for my faculty advisor, even though she always does tell me I am doing a wonderful job and to not get discouraged when something doesn’t seem correct. Recently, Dr. Krucher has asked me to help out with her experiments as well while she is in lecture, which elates me and shows she is comfortable with me in the lab and the utilization of all materials on my own.
As I continue in my research, I have come to find that I enjoy it more and more each day. There is always something new to learn in research in general, and being that I am an avid learner, I am always willing to soak in new information in an exciting manner. Additionally, even though I am still learning new techniques here and there, I have becoming increasingly better in my performance in general which essentially makes me feel a bit more confident and have a more enjoyable experience. My coworker and I have even become closer and connect very well with each other. Even though there have been some challenges or set backs here and there such as the 3D’s not allowing enough cell death by PNUTS KD, this experience has been a positive, pleasing, and intellectual one so far.