Internship

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Showcase Summary

SARS-CoV-2, first described in 2019, can cause severe illness, with many suffering from persistent respiratory diseases such as pulmonary fibrosis or cryptogenic organizing pneumonia (COP). One explanation is that virus proteins may persist after infection, continuing to stimulate the immune system and cause inflammation. Bronchoalveolar Lavage (BAL) fluid was collected from patients with chronic pulmonary disease averaging 178 days following severe Covid-19 infection. Preliminary data showed persistent SARS-CoV-2 RNA levels and spike protein in acellular BAL fluid. We hypothesized that persistent RNA and antigens are contained within extracellular vesicles (EVs), providing a protected environment from degradation and allowing persistence. When body tissues are exposed to pathogens, EV production increases, along with the release of proinflammatory markers, leading to further inflammation and disease progression. In our project, we used size Exclusion Chromatography (SEC) to isolate EVs from BAL fluid, which were analyzed for SARS-CoV-2 RNA. These EVs will be further investigated to determine the mechanism by which they stimulate inflammation. Understanding the mechanisms of persistent viral expression and resultant inflammation could lead to treatments for patients with post-Covid pulmonary fibrosis or COP. This model may also apply to other chronic lung diseases post infection.

LHSI Showcase 2025 (1)

Workplace

September Skills

Entry micropipette skills - skills taught in some of my PTLW classes in high school. I use these skills frequently in my internship, as I have to do several assays which require it.
- - - I use these skills in BCA assays, and the preparation of the western blot samples.
Communcation - I have used throughout my time in school, or work, with professors, employers, and leaders. Regularly use this skill in discussing requirements, or protocols at the internship.
- - - Communicating with my mentor when I am unsure of the steps in a protocol. I also have to communicate the results from experiments to other people so they too can understand what has been done, and what the results mean.
Time management - Throughout my time in school I've had to manage work, homework, all while having a busy personal life, and excellent time management played a crucial role. Learning how to balance the internship, classes, and homework requires time management.
- - - I am currently using a task manager extension on canvas that displays all my assignments. I also make weekly to-do lists with everything I need to get done for the week including both school and personal activities.

All helpful entry level skill because they give me entry level knowledge, and basic helpful skill to work from. During the past few weeks, I have already expanded many of those skills, while even developing some new knowledge and skills as well.

Throughout my time working in this site my roles and responsibilities have changed. In the beginning I was a quiet observer, who was watching and picking up different techniques to efficiently do the protocols. At this point in the lab, after working there for nearly 6 months, I am now responsible for all the protocols that have previous been taught such as small levels of Nano site work (counting particles from a sample), Western blots (used to detect proteins), IZON columns (separating particles by size in samples), and BSA assays (detecting trace amount of protein in a sample). While I am responsible for these, I am also responsible for teaching new people in the lab the proper technique, and methods to use to execute them as well. The lab operates on the method "see one, do one, teach one," which is used to reinforce our learning on the protocols. My contributions have impacted the internship team, the results I have gotten from western blots, or the counts gotten from IZON columns are going to be published in a paper when all of the work is finished. Even though some of the protocols are small, and simple such as a BSA, they are needed to be done on every sample, and doing several of these ensures that other lab employees can focus on the bigger questions.

Successes

During my time during this internship, I have felt successful when throughout the semester I have been given more responsibility and independence. In the beginning months it was very much being a shadow who sometimes participated in simple experiments. One of the benchmarks I was being measured on was a BSA assay, which is used to detect the amounts of proteins in unknown samples. Doing these assays requires precise technical skills, the slow development of “lab hands,” and when done correctly can result in a direct linear relationship of protein concentration. Mark Fraser made a comment that when I completed a BSA assay with an accuracy of .99 is when he would be ready for me to move to go further in practice. I was very grateful for the different tips and tricks the people in the lab taught me to optimize things such as my form, and mechanics to try to improve my "lab hands." I was not there when this plate was read, but I can say I jumped up and down when I received this email.

This email was sent a few weeks before my birthday, and I repeatedly made the joke that my birthday cake should say “I got a .99!” For all those curious it did not in the end say that. Following this success I was taught how to do size exclusion chromatography (SEC) which separates particles by size, and western blot protocols which look for specific markers. Just as the BSA, I was supervised at first, and then gradually gained independence. It is now at the point where I am sometimes left for hours of the day unsupervised and trusted to follow the protocol.

I have also had the opportunity to shadow Dr. Twigg and Fraser in the hospital this semester. During these times it has solidified the fact that I do not want to be a doctor and would much rather be in some type of laboratory. It has also allowed me to gain knowledge on different career pathways I was unaware of before that do not require medical school. Currently my major is Cytotechnology, and after hearing different people's opinions I am going to stay on the Cytotechnology degree track since it is a laboratory-based career and use my knowledge on my personal preferences gained from this internship to further guide my choices in education.

Since January we have also had a few new people start in the lab. I have helped train them in protocols such as the IZON columns, BCA assays, some parts of the Western Blotting, and nano site detection. I feel accomplished when they do something well, sometimes even better than I did the first time I ever did the protocols. Such as when Haylee got a .99 R squared value on her BSA assay and it was the first one she had done in months that I had to supervise, or when Karla knew how to use the Nano site after I had explained it, and she had observed a few sample detections. Having this knowledge can help me with career goals because I now know I would like to work in a lab were everyone is aiming for the same goal, where everyone can talk about results and feel supported in their work instead of everything being individualized.

Challenges

After all this time I can do a protein assay, assist in a western, and do a SEC column. However my problem lies within the fact that sometimes I do not understand the purpose of what we are doing. I understand the overall goal or end point the laboratory team is trying to achieve or understand, however sometimes the reasoning or significance of the results is lost on me. Such as, asking me questions on why one antibody used for binding a western membrane is lighting up in a lane but is doing so at the wrong molecular weight. Or when I am asked to justify a concept using the data that is in front of me, like why is this antibody considered a universal marker. The gradual independence, although a good thing, is also a terrifying thing, and is something I have to get accustomed to since that is the end goal. For instance, the time I was left to imagine a western blot for the very first time by myself. For every occurrence before then Mark stood behind me, analyzed the blot, and told me how long to expose it for a reading. The end goal of a western image is to have a near completely white background, no visual border of the membrane, a clear ladder, and black lines in lanes with no smearing. My first time imaging none of them turned out correct, all were grainy, with random black smears, and were quick to be oversaturated (blot turning red) in the ultraviolet light. I luckily could turn to the other person in the lab, Aimee, to come take a look and try and explain what I could possibly be doing wrong. We had a discussion about the blots, and the disadvantages to the marker we were testing for. I unfortunately had to leave before I could have Mark look at them for discussion, but the next time in the lab we will have a discussion about what happened, and how I could do better the next time, as there is always a next time when it comes to western blots. There is always room for improvement, and development of skills when working in a laboratory which is exactly what I am striving to do. I am heavily reliant on the people I work with to help achieve success, I want to be a part of the success, I want everything they do to be a success, and I can always lean on them to help me succeed since that is their goal for me.

A challenge I have faced recently, even if I put it under success is effectively managing to do my own work while teaching someone else a different protocol. The success with Haylee receiving a .99 as an R squared on a BSA assay, I was attempting to prepare all that is required for a Western blot protocol, as well as writing down all the all the in-depth protocols for a common use lab drive. It was difficult to multitask, and keep track of what I was doing effectively, while also taking breaks to help another person with their own work so it turns out correct. Although it was in the end done correctly, it was a different type of challenge than what I have done in the laboratory previously. What had help face these challenges, was experiencing other people from the team having to do the exact thing for me when I was initially training, and what they still sometimes have to do when I have a specific question. Following their example they have set for me, doing things such as asking periodically if they need help when I have a moment, or making it clear they can ask me anything is very helpful when trying to teach someone how to do things correctly the first time.

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