From course catalog: BIO497: Provides guided research in biology, molecular biology, microbiology, cell or human physiology, genetics, biochemistry, or biotechnology.
For this course, I continued my STAR and Fall 2022 research in the Petrie lab and applied to have my time recognized for research credit by the BIO department. This counted towards a laboratory elective when I was in the major. Below is my research contract that was sent to my academic advisor when applying:
Student Name- McKayla Procopio
Student ID#- 14474074
Professor- Dr. Ryan Petrie
Term- Winter Quarter 2022/23
For credit (# of crs): 3 credits
Research Contract I will be participating in research in Dr. Ryan Petrie’s laboratory. Below is a description of my research project and the methods that I will perform in the Department of Biology.
The general focus of my research in the Winter quarter is identifying differences in morphology and migration between wild-type mouse embryonic fibroblasts (MEFs) and AIM2-Knockout MEFs. We know that AIM2 (absent in melanoma 2) is a protein that works in conjunction with Tropomyosin 1.6 to form a part of the actomyosin machinery that creates lobopodial protrusions in human fibroblasts. By knocking out AIM2, we expect to see changes in the structure of MEFs as well as the way they migrate. In the Fall quarter, we captured live image movies of both the wild-type and AIM2-Knockout cells to directly compare the two types of cells migrating over a course of both 8 hours and 90 minutes.
During the Winter quarter, I will continue to culture the wild-type and AIM2-Knockout MEFs to capture more live image movies to compare the morphology of these cell types using phase- contrast microscopy. I will analyze their migration by completing velocity assays to identify whether a lack of AIM2 protein affects the speed of cells. To get the velocity of cells from the movies, I will use Fiji to do cell tracking. Additionally, I will be completing immunofluorescence assays to identify where proteins such as tropomyosin and vinculin are. With tropomyosin, we expect there to be localization differences between the wild-type and AIM2-knockout cells since there is a connection between AIM2 and tropomyosin. Vinculin is an integrin that forms linkages between the cell and its environment. I will be looking to identify any differences in localization between the wild-type and AIM2-knockout cells to determine whether vinculin and AIM2 work together to create focal adhesions. I will be imaging the immunofluorescence cells with the Zeiss Axioobserver in the Cell Imaging Center and use Fiji to analyze my images. The immunofluorescence assays will be completed both in 2D and 3D to identify potential differences in polarity of these proteins across different dimensions.
Over the course of these studies, I will participate in 11 laboratory meetings where I will have an opportunity to present my findings and discuss results with other members of the research team. These results will be maintained in my laboratory notebook as an official record to justify my involvement in manuscripts, presentations, and written reports on my work. I anticipate spending an average of 12 hours in the laboratory each week for 3 credits. As Dr. Petrie and I have discussed, my studies take first priority so my laboratory time may decrease during mid-term and final examinations times. I will inform the laboratory staff of these times in advance.
For this course, I continued my STAR and Fall 2022 research in the Petrie lab and applied to have my time recognized for research credit by the BIO department. This counted towards a laboratory elective when I was in the major. Below is my research contract that was sent to my academic advisor when applying:
Student Name- McKayla Procopio
Student ID#- 14474074
Professor- Dr. Ryan Petrie
Term- Winter Quarter 2022/23
For credit (# of crs): 3 credits
Research Contract I will be participating in research in Dr. Ryan Petrie’s laboratory. Below is a description of my research project and the methods that I will perform in the Department of Biology.
The general focus of my research in the Winter quarter is identifying differences in morphology and migration between wild-type mouse embryonic fibroblasts (MEFs) and AIM2-Knockout MEFs. We know that AIM2 (absent in melanoma 2) is a protein that works in conjunction with Tropomyosin 1.6 to form a part of the actomyosin machinery that creates lobopodial protrusions in human fibroblasts. By knocking out AIM2, we expect to see changes in the structure of MEFs as well as the way they migrate. In the Fall quarter, we captured live image movies of both the wild-type and AIM2-Knockout cells to directly compare the two types of cells migrating over a course of both 8 hours and 90 minutes.
During the Winter quarter, I will continue to culture the wild-type and AIM2-Knockout MEFs to capture more live image movies to compare the morphology of these cell types using phase- contrast microscopy. I will analyze their migration by completing velocity assays to identify whether a lack of AIM2 protein affects the speed of cells. To get the velocity of cells from the movies, I will use Fiji to do cell tracking. Additionally, I will be completing immunofluorescence assays to identify where proteins such as tropomyosin and vinculin are. With tropomyosin, we expect there to be localization differences between the wild-type and AIM2-knockout cells since there is a connection between AIM2 and tropomyosin. Vinculin is an integrin that forms linkages between the cell and its environment. I will be looking to identify any differences in localization between the wild-type and AIM2-knockout cells to determine whether vinculin and AIM2 work together to create focal adhesions. I will be imaging the immunofluorescence cells with the Zeiss Axioobserver in the Cell Imaging Center and use Fiji to analyze my images. The immunofluorescence assays will be completed both in 2D and 3D to identify potential differences in polarity of these proteins across different dimensions.
Over the course of these studies, I will participate in 11 laboratory meetings where I will have an opportunity to present my findings and discuss results with other members of the research team. These results will be maintained in my laboratory notebook as an official record to justify my involvement in manuscripts, presentations, and written reports on my work. I anticipate spending an average of 12 hours in the laboratory each week for 3 credits. As Dr. Petrie and I have discussed, my studies take first priority so my laboratory time may decrease during mid-term and final examinations times. I will inform the laboratory staff of these times in advance.