About My Internship

At my internship, Dr. Singh and I research how epigenetic mechanisms such as DNA methylation affect the wound healing mechanisms of people who are afflicted with diabetes, and simple and fast methods by which to treat those afflictions.  One such method is tissue nanotransfection, which utilizes an implanted nanochip and noninvasive electric sparks to transport growth factors to areas of injury to induce cell division and angiogenesis (blood vessel creation). Diabetes is a globally prevalent disease, with over 537 million people having the disease around the world, and a projected 783 million people will be diagnosed with the disease by 2045. My role in the lab involves the completion of procedures such as gel electrophoresis of DNA samples, DNA methylation, PCR amplification, and IHC analyses in order to obtain and provide viable results. These procedures help in gathering data to help in supporting the hypotheses formed by my team to further our research and gain further funding for this groundbreaking technology and innovation. 

Learning and Skills

As an aspiring doctor, I believe that communication and collaboration are valuable skills which I hope to be able to build on and expand in my time at this internship. However, I sometimes feel too dependent on a team, which is also something I want to improve upon in this experience. I would like to build on my ability to think and do things independently, free of any negative thoughts and need for approval from a superior or team member. Building ones confidence in themselves is crucial, and I hope to do so at this internship.

In this internship, I hope to gain the ability to discuss scientific results in depth with others. Additionally, I want to learn the methods for many different scientific procedures, including Western Blot as well as PCR. I also want to be proficient in my use of said data to create presentations and research papers which bear significance in the scientific community. As of recently, I have been able to complete PCR and interpret the results of certain datas including sequences of genes and their significance to the research being done.

I am ecstatic to work with esteemed doctors and researchers on such a groundbreaking project. Seeing how my work and the work of others around me contributes to the grand scheme of this research is heartwarming and invigorating, as it just fuels my passion for helping and contributing to my community as well as the scientific community. I also feel myself becoming a more proficient worker in general, which is a skill I have always aspired to achieve and strengthen. 

Upon assessing my skills and current work value, I have seen some common themes. Something that I have been able to improve upon is my ability to work independently and set a goal for a day and get it done in a timely manner. I never had the opportunity to set my own goals or complete my own tasks without supervision until attending this lab. However, after learning the basics of the lab and general techniques required for the lab, I am now able to undertake several procedures on my own without the instruction or aid of supervisors who also have their own tasks to complete. Additionally, I feel like I am able to communicate in a more efficient and meaningful manner since I have joined the lab. For example, I am now able to explain a procedure, what the end goal of said procedure is, and what the results of the procedure indicate based on previous completions of the procedure. This level of understanding and communication was something I could have only hoped to be able to do before, and this development is something I am very proud of. Despite this progress, I can always be better than I am currently, and there are still certain aspects within these skills that I lack proficiency in. One such area is asking questions. I am sometimes intimidated by the standing of the doctors who work at the lab (to no fault of their own, my respect for them makes me feel like a bother), which can be a hindrance to my ability to communicate as well as to create independence and standing of my own. If I am not able to ask them what to do without them explicitly telling me, then how am I to develop any courage or independence on my own? My advancement in this aspect of my skills has reaffirmed my desire of being a doctor, seeing as how I am able to overcome the challenges involved with communication and am able to effectively convey the details required. This development has helped me overcome one of my larger insecurities about becoming a doctor, that being my inadequate communication skills could be a hindrance on my dream career.

Despite my accomplishments and progress in areas which I previously needed to improve upon, there are still certain skills which I need to work on. One such skill I would like to develop further is to undertake a task without specific instruction on how to, or what task to complete. I feel like in certain aspects, I rely on the doctors instruction more than I should. I would like to be able to set my own goal for a day, and complete it based on what the supervisors and doctors expect without them having to explicitly tell me what to do to satisfy their needs. To do this, I must continue to ask questions and also request that the doctors place me in charge of a more high stakes procedure and to complete it on my own based on my previous experience and skill. I will also continue to seek out opportunities to complete the tasks with no instruction, and to accept failures and setbacks as they come without completely halting my progress. Such progression is necessary for my future career path. Throwing in the towel after making an incorrect diagnosis or not receiving expected results from a test would defeat the whole purpose of being a doctor. The challenge lies in persistence through all difficulty, and I hope to keep developing this skill to further strengthen my security in my future career.

This internship has also had unprecedented relation with the classes I am taking. For example, cell biology has had many topics which I am currently learning in my internship, one of them being the science behind the PCR technique. Although the class teaches the basic principles of its function and the scientific basis behind its mechanisms, the internship takes this knowledge to the next level and allows me to apply it. Any questions the professor could throw at me regarding PCR, I could answer all thanks to the lab. Another example of similarity was in the idea of DNA methylation. In the course, we discussed how DNA methylation can lead to the silencing of certain genes, preventing their expression. DNA methylation is one of the primary subjects being studied in the lab, with methods such as bisulfite sequencing detecting methylated base pairs within an examined sample of DNA. Understanding the mechanisms behind how a methyl group could silence the expression of an entire gene was something I did not fully understand before applying my knowledge from both the class and what I learned in the lab. Sooner or later, I will be able to find connections between classes such as organic chemistry and certain tasks done in the lab, which is part of the beauty of working and applying my knowledge. 

I have found myself developing all of the skills I had hoped to in the beginning of my internship. I collaborate with my fellow interns on not just our lab work, but also on any school work which we have in common. We strengthen each other's weak points in certain school topics, and also help each other find better ways to perform experiments or interpet results. One of my favorite days working in the lab was when all of us were present, and we were shadowing another researcher executing a procedure known as PLA (protein ligation assay). We all took note of every step meticulously, and made sure to watch exactly how it was done in order to perform it ourselves the following days. Seeing how passionate the rest of the interns were, and how much fun it was to do so with them made me realize how much better I have become at working in a group and accomplishing tasks with others. Additionally, my skill in performing other procedures such as PCR and gel electrophoresis has increased since my arrival at the internship. I had ran a gel before in one of my lab classes, however I could not have imagined myself making the solution for a gel and then running it with my own resources. I have become increasingly adept at including the right quantities into the gel mixture in order to create enough of the solution to run however many samples and wells I need. I have also become better at reading the results once the gel run had taken its course. In addition to my increasing skill in gel electrophoresis, I have also gotten much better and faster at performing PCR procedures. When I first started at the lab, it would take me at least a few hours to fully prepare the samples, make sure to label them appropriately, and run them through the thermocycler. However, it has almost become something of a second instinct, where I find new ways to make the procedure and its preparations more efficient and less time consuming. Although the development of these skills may seem to be only lab related, I find myself applying the skills in other facets of my life. For example, making my PCR procedure more efficient has helped me realize that I can find ways to make my gym routine as well as my studying patterns more effective. With all of these skills having been further developed, I feel more and more secure in the career path I have chosen and my desire to become a doctor grows stronger with every day I am in the lab. The work I do there feels like it has meaning, and being able to understand the epigenetic mechanisms behind diseases or ailments like diabetes gives me a deeper repertoire than what I would want as just another doctor. My time at this lab has also taught me that being able to cure someone or treat their disease is a noble cause, but the true pioneering work occurs in the labs which work to eventually eradicate these diseases all together. Such practice has made me see that being a doctor is not the only route to take in order to help advance the fields of medicine and regenerative sciences. Although my main vision is still to become a doctor, my time in this lab has opened my eyes to other career paths I could take while still pursuing my dreams. 

My time in this lab has indisputably been one of the most moving experiences of my life, and every moment there is a pleasure. However, there were some experiences where I thought I couldn't possibly be happier anywhere else. One of my favorite experiences was my first time running a gel. When I turned on the electrodes, the liquid which disperses the charge to the appropriate locations creates these lines of air bubbles which travel in the direction of the electricity flow. The sight was one to behold, and asking my coworkers about the science behind it was invaluable on its own. The learning and admiration I took in this one experience cemented my desire to be in the field of science and medicine. Seeing how physics, chemistry, and biology all worked in tandem on that gel was something I will never forget. 

Throughout my time at the internship, I have always felt like my contributions were miniscule. My tasks onbiously carried weight and were a crucial part of advancing the projects in the lab, seeing everyone elses work made it feel like the work I completed was insignificant. However, our lab recently acquired a sizeable and important grant, and Dr. Singh made sure to personally make me feel recognized and made sure to help me understand that no matter how small the work i completed feels, it is a part of the work that advances our mission. Doing those small procedures, or completing the analyses that felt like a droplet among a sea of others to be completed ended up being worthwhile. 

As my time at the internship progressed, I started making noticeable strides in many aspects of my workplace skills. I was able to make a presentation which Dr. Singh expressed satisfaction with, and assured me of my progress made throughout my time there. I plan to transfer these skills with me in every future endeavor I undertake. 

The Workplace

As a student, it is imperative to develop transferable skills both for a workplace and for outside experiences beyond your career. The person in my lab who I have seen embody the mastery of these skills the most is my supervisor, Dr. Singh. When Dr. Singh walks into a room, his seniority is well respected based on the fact that he made it to the position he is in through hard work and commitment. However, he does not intimidate or look down upon any of the lab members, but rather does the opposite. He uplifts everyone present, answers any questions, and teaches everyone necessary skills as if they were his family. His communication of the tasks he would like completed are assertive, but not aggressive or demanding. His balance of friendliness and professional attitude in the workplace is also something which I sincerely admire, and is something I aspire to have one day. Dr. Singh also treads his own path and plans what he needs to do in order to achieve his goals, which is also an area I want to improve upon myself.  

Working in lab as diverse as mine has been a true privilege. People from all walks of life and of different cultures and ethnicities clock in and out daily not just to make money or get by, but to advance our knowledge of the human body as well as ways to heal it. Just the thought of this collaboration of everybody regardless of their backgrounds to achieve one common goal makes me realize how privileged I am to work in such an environment. Along  with learning how to conduct lab procedures from other researchers in the lab, I have also learned how similar our day to day lives are compared to mine. One of the senior doctors in the lab goes to the gym right after work and then makes himself something to eat, just like I do! We talked about different programs of weight lifting, how it has impacted our work and social lives, and gave each other advice for different challenges we faced. This experience alone strengthened my bond with the doctor, but also helped me realize how at our core, we are all people with similar ideals and goals. I even find out that one of my fellow interns attended sunday school with me when we were children, and I had not even realized it!

When I started at the internship, I expected there to be a certain sense of awkwardness within the interns and the researchers. I did not feel like I could socialize efficiently with my fellow interns or with the respected doctors in the lab. However, overcoming this fear and talking with everyone has made the lab environment extremely enjoyable. I socialize with the other interns on a daily basis, and have even made one of my best friends from this lab. Additionally, I expected the lab to be an excessively serious and somber environment. I felt like such work would require one to be serious and focused at all times. Although taking the work being done seriously is an obvious prerequisite to working anywhere and especially in a lab, I discovered to my surprise that there was a perfect balance between work and social life in the lab. Whenever we see or find something interesting, we call each others attention to the item at hand and discuss how beautiful it is or how it works. This has without a doubt been a highlight of the lab for me. 

A lot of the people who work at the lab with me have faced severe hardships in getting to where they are now. Hearing their stories of the challenges which they faced in coming to America or finding work that aligned with their studies was enlightening in a sense. It resonated with me because it was the same story my parents would tell me about how hard they worked to bring us somewhere with greater opportunity and a chance to actually be someone and make a living and support a family. However, its different hearing it from someone you're not related to. For one, most of my coworkers came from a different country than where I originated. They had their own unique challenges and difficulties in getting to where they were now. Though my family faced its own set of challenges, we also had extended family who were already in America that could help us with many parts of the process, even after the difficult parts of acquiring a visa and lawfully entering the country. My uncles helped find us a home and even paid most of it off until my father was able to find work. They got my parents a car to take them to and from wherever they needed to go. This is a stark contrast from what most of my coworkers faced. Most of them did not have any connections or family here when they arrived, and had to fend for themselves in many of the aspects I described above. Although I've always been grateful for my position in life, hearing these stories has given me a new perspective on life and made me more aware of those around me, both professionally and socially. This newfound awareness has given me this hunger to socialize and learn more about my colleagues, as this one realization came from a few conversations with my coworkers. The things I could learn not just in my profession, but also about the world around me are invaluable. 

Successes and Challenges

One of my greatest successes so far has been inventorying 22 boxes full of anywhere between 40-80 slides of mouse tissue within two weeks in a satisfactory manner. Doing so was one of my greatest accomplishments, even impressing Dr. Singh enough to offer me a job for my junior and senior years. I had never done inventory on a box, and had learned through already inventoried boxes as well as Dr. Singh's instructions what to expect. In addition to ordering the slides in numerical order and listing all the slides present or absent on the boxes, I also had to make an excel sheet containing the slides, their numbers, and their orders. As a result, doing future work with these slides such as staining and PLA will be much easier and more efficient. Additionally, I had created one of my best gels since joining the lab a few weeks prior and got clear and precise results which helped indicate what samples contained mutant DNA vs. the wild type DNA. For almost all researchers, this is a basic skill which is a requirement for any lab. However, seeing how well it turned out was a very reassuring result for me. As I progressed and honed my skills in the basic procedures in the lab, I also found success in other areas in my internship which have translated into applicable skills beyond the lab environment. Dr. Singh asked me to create a presentation about bisulfite sequencing showing my progress and learning about the procedure and how it applies to the work of the entire lab. Dr. Singh was very impressed with the research I demonstrated and had few critiques and things for me to add. This skill is invaluable for any workplace, as being able to present your findings about any subject is important in every field. This success also pertains to one of my goals regarding being able to present my findings from my research. This task and the success with it gave me the opportunity to achieve that goal. 

Although I have seen many successes in my time at the lab, I have also seen my fair share of difficulties. One such difficulty was mastering the gel electrophoresis and PCR. When I first started doing the procedures, they were extremely time consuming and had so much room for small errors. I kept messing up the numbering on a PCR tube, or adding a ladder with too few wells, which eventually became frustrating. However, I have found that repetition and learning from those who have done these procedures frequently and efficiently is the best way to improve my method in these crucial procedures. Once the time comes when I can say that I am proficient in these procedures, not only will I be able to do these procedures in other work environments, I will also be able to transfer the ability to learn from my mistakes and adapt to other challenges whenever they may appear. In addition, my supervisors and the other post docs at the lab were always my helping hand in the lab, giving me valuable advice and always reminding me of those minute intricacies to watch out for and make sure to include within my procedure. These reminders always stick with me, and I have been able to almost completely eliminate any mistakes in those procedures. 

Project Summary

Diabetes Mellitus is an increasingly concerning issue within the field of medicine. Though the disease is manageable, the associated secondary complications become progressively debilitating as the patient’s lives on. One complication is the inability to create functional blood vessels to ischemic tissue. This increases the risk of life-threatening bacterial infections which without treatment could require amputation of the afflicted extremity or even death. At our lab, we explore the molecular basis of vasculopathy in affected ischemic (blood starved) tissue from diabetics as compared to healthy skin. This includes examination of gene expression changes and promoter methylation status of vasculogenic (blood flow related) genes. Our lab has recently shown that decreased blood flow in diabetics is linked to increased vascular gene suppression. Using this technique is key to uncovering a potential long-term treatment for diabetic vasculopathy.  During this internship I learned to examine gene promoter methylation status using bisulfite sequencing. This procedure utilizes treating DNA with chemicals that can differentially react with methylated and unmethylated cytosine residue. The quantification of methylated CpGs (cytosine and guanine pairs) in vascular gene promoter can reveal the amount of silencing in diabetic ischemic tissue. The findings from this project could have significant implications for diabetic patients, particularly those with critical ischemic limb and non-healing diabetic ulcers. Eliminating diabetes is one thing, but reducing the risks of complications that come with it is also invaluably important, which is why the work we do is so crucial. 

Acknowledgement: The laboratory of Dr. Kanhaiya Singh is supported by U.S. Department of Defense grant W81XWH-22-1-0146. We would like to thank Dr. Chandan K Sen, Director, ICRME for his kind support during this internship duration.

Final Project