For the past month, I have been analyzing red blood cells (RBCs) to try and better understand their membrane shape and how it is affected by mutations. There is research at The Scripps Research Institute exploring whether mutations in myosin filaments within the membrane skeleton have an affect on the RBC's shape and if they cause anemias. The red blood cell's biconcave disk shape and deformability rely on the membrane skeleton, a viscoelastic network of short actin filaments cross-linked by flexible spectrin tetramers. In this structure, nonmuscle myosin (NMIIA) motors exert force on diverse F-actin networks to control cell shape. Based on past research, it has been found that there is a general role for NMIIA contractility in promoting membrane stiffness and maintaining RBC biconcave disk cell shape.
In order to better understand how different types of mutations affect the red blood cell's shape and membrane, I measured multiple microscopic images from both mutated and non mutated patients. Collecting data would involve a long process of analyzing the RBC images on a computer program, recording cell data from multiple images per patient, and ultimately organizing the data of over thousands of cells. With the data I found on the RBCs, I compared the mutated and non mutated patients cells by area and aspect ratio (major axis/minor axis.) I created separate graphs for both the area and aspect ratio. I organized the data on the graph to compare each of the different types of mutated cells to the median of the non mutated cells. What I found when comparing the RBC data in this way was that there was a more drastic change in the median areas, but less so for the aspect ratios. Depending on the type of mutation, some measurements were higher in value than the non mutated patient, while others were lower in value. The data I collected on mutated red blood cells will be important for the future of the lab when they continue researching anemia more and how mutations in myosin filaments affect the overall shape of the cell membrane. The graphs I made reveal the significant changes in area and aspect ratio when comparing the different mutated cells to the non mutated cells. To complete this project, the majority of my work was analyzing the images of red blood cells from blood smear disks. The process I went through in the program was eliminating the unusable distorted cells that were misshaped when smeared on the disk, then collecting the usable RBC measurements, and organizing the data to create the box-plot graphs. To put in perspective how many cells I needed to measure, there were around 50-110 cells per image, about 5 images per patient, and over 12 patients to analyze. Most of month was dedicated to eliminating the unusable data from the RBCs and after all the data was collected and entered into box-plot graphs, analysis was finally able to begin. My final product includes multiple box-plot graphs I am unable to show you for confidential reasons. Two graphs are for area and two graphs are for aspect ratio. The first area and aspect ratio graphs include cells from 1 non mutated patient and 10 mutated patients. The second round of area and aspect ratio graphs organize the mutated cells into their specific domain (area of the myosin filament that is mutated) and includes data for 3 non mutated patients.
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Being at The Scripps Research Institute has made me think a lot about my future and pursuing a career in the science field. At the beginning of the month, when I first started interning in the lab, I wanted to gain experience in a lab and learn more about the field of Molecular Medicine. The impact this lab has left on me is giving me more of an interest in science and experimenting. I’ve been able to see the significance of scientific research, outside of school, and it’s been fascinating. I’m interested in Biology and other sciences and in college I’m planning to study these subjects. I mainly think about pursuing Nursing as a career. I’d like to work in an environment with more energy and movement. The lab environment is a very calm environment and from what I’ve observed the majority of your work is independent. I want a career where I can move around and talk with people. However, I would try and pursue a career in a lab if at any point the future, I began being interested in a very specific field of science. Individual labs have very specific focuses, I was studying the Red Blood Cell membrane for my internship.
The majority of the people I have spoken with or worked with at TSRI have their PhD or are pursuing their PhD. They’re all very focused and passionate about what they’re researching, which makes them enjoy their jobs. I’m not sure for myself if there is an area of science I am passionate about, mainly because I haven’t had the opportunity to research different branches of biology. The work they do is very independent, but involves collaboration from others when considering results and new theories in their projects. I’m looking forward to future science classes in college and possibly future internships in more labs. From this internship I’ve gained the ability to collect data and analyze, which has given me insight into what experimenting in the Biology field is like. It can be very time-consuming and difficult to know when any change in data is significant, but after a month of working in a lab I can say that I have found significant data on the red blood cell membrane. I’m grateful for this opportunity and I’m excited for what is to come in the future. |