Internship

About the Internship

My internship site is focusing on type 1 diabetes research, which is a condition that prevents the body from producing insulin, and Insulin is a hormone that helps regulate blood sugar levels. The focus is on proteins that regulate immunes responses called cytokines and testing for their functions to look for any valuable observations related to their reactivity with beta cells, which are cells in the pancreas which produce insulin. 

The goal of my internship focuses on understanding how certain cells in the pancreas become damaged or die, which is important for the development of Type 1 diabetes. This research is very important because it could lead to new ways to prevent or even cure Type 1 diabetes, improving the lives of millions. Fun fact: I get to look at cells under a microscope and watch how they react when they’re stressed—it's amazing to see how they respond in real time!

Plasmid DNA Transfection Protocol   ("Plasmid DNA Transfection Protocol.” YouTube, YouTube, www.youtube.com/watch?v=cPA2OQv8qA8. Accessed 22 Feb. 2025.)

Video above shows how DNA plasmid transfection is done in INS-1 cells using Lipofectamine 2000, a transfection reagent. My experiment follows a similar process to study biosensor activity under cytokine treatment. 

Project Summary 

• Type 1 diabetes (T1D) is a complex disease influenced by both genetic and environmental factors, leading to autoimmune destruction of insulin producing beta cells. The leading, yet somewhat controversial, hypothesis for this environmental trigger is viral infection. To investigate how cytokines can impact the beta cells directly, we treated INS-1E cells (a rat beta cell line) with IFN-α, a cytokine released during viral infections, to see if it affects cellular reactive oxygen species (ROS) production. 

• To measure ROS, we used fluorescent probes and biosensors coupled to confocal microscopy. I used the probe dihydroethidium (DHE), which lights up in the nucleus when ROS is present. We also used biosensors to track ROS in different cellular compartments: the cytoplasm (AAV8-GRX1-roGFP2), endoplasmic reticulum (MYC-ER-roGFP), and mitochondria (TOMM20-GRX1-roGFP2). These biosensors were introduced using the reagent Lipofectamine 2000. All experiments compared IFN-α-treated cells to corresponding control cells.  

Workplace

My strengths in foundational laboratory techniques, such as pipetting, electrophoresis, and data analysis, comes from hands-on experience in my genetics and cell biology coursework, where I effectively conducted experiments and analyzed data. These skills will allow me to quickly integrate into my internship team and contribute meaningfully to our research projects. To grow, I plan to seek mentorship to learn advanced techniques while improving my communication skills in team settings. This is important for my future career as a physician, where teamwork and technical skills are essential in patient care. By developing these skills, I’ll be better prepared for collaborative work in healthcare. Ultimately, I want to make a positive impact in the medical field.

During my internship, I went from observing and taking notes to independently running experiments and getting results. I’ve been a collaborative team member, troubleshooting on my own but asking for help when needed. My work helped compare biosensor activity under the microscope, contributing to the team’s research. Even small tasks, like setting up experiments, played a role in achieving our larger goals.

Successes and Challenges

In the fall, one of my proudest moments was when I independently split a cell culture flask and followed the experimental protocol to view the results under the microscope. I was able to carry out the task completely on my own, which gave me confidence in my technical skills and understanding of the process.

Another success occurred when I was able to accurately identify and analyze ROS presence in the cells under study. It was a challenging experiment, but I found that by thoroughly reviewing the protocol and asking for feedback from my supervisor, I could perform the experiment successfully.

In the spring, one of my biggest successes during my internship was independently performing a full transfection experiment after initially just observing. I successfully plated INS-1 cells, transfected them with biosensors using Lipofectamine 2000, and later analyzed their activity under the microscope. Seeing clear, measurable results confirmed that I had followed the protocol correctly. This experience taught me the importance of attention to detail, troubleshooting, and confidence in my lab skills. In my future career in medicine, I will apply this independence and problem-solving mindset to diagnosing and treating patients, ensuring I take initiative while knowing when to seek guidance to provide the best possible care.

Balancing multiple responsibilities—school assignments, exams, my internship, and another job—has been a significant challenge. Managing all these commitments at once sometimes feels overwhelming. Moving forward, I plan to create a more structured schedule to better manage my time and reduce stress.

Another challenge has been learning how to deal with unexpected experimental results. Sometimes, things don’t go as planned, and I have to troubleshoot issues like equipment malfunction or inconsistent data. To overcome this, I’ve been actively seeking more hands-on advice from lab mentors and looking for additional resources to strengthen my troubleshooting skills.

Additionally, in the spring, another challenge I faced was troubleshooting transfection issues despite making adjustments. I repeatedly reviewed my experimental procedure and consulted lab members to confirm I was following the steps correctly, yet I still observed no activity during imaging. With my supervisor’s help, I discovered that my cells had died in the flask before transfection, which explained the lack of biosensor activity. This experience taught me the importance of checking cell viability before experiments and considering all possible factors when troubleshooting. With guidance from my team, I improved my cell culture techniques, ensuring more reliable results in future experiments.