Student Research at Duke

Wow! I can’t believe that I have been at school for just three weeks; it feels like I’ve been here for much longer. I had a very relaxing rest of the summer post-Howard Hughes at home for about a month. A bunch of my relatives came to San Francisco to visit us so we took them to Chinatown and had delicious dim sum and took a tour of Alcatraz, which was possibly one of the most exciting things we did- we got to walk around the jailhouse and learn about the different escape attempts, which I found extremely fascinating. Other than that, I mostly hung out with friends, my parents and sisters, and of course ate a lot of good food. And now I’m back at school, going to class and hanging out with friends…only now all hang out sessions occur in the library, usually at around midnight. But I love being back here; I’m in a triple this year right on the main quad, so everything is really convenient. I started back in lab on the first day of school and am there as an independent study student. I’m currently finishing up the project I worked on from over the summer. I really love going to lab- many of my fellow Howard Hughes-ers and I have come to a general consensus that it would be amazing to just work in a lab for the rest of the three years and attend classes on the side. It is such a relief from my day to go into lab and immerse myself in the project I am working on; it’s a good break from all the lectures and studying. However, just from the first three weeks of classes, I know that I need these classes in order to fully grasp everything that is going on in lab. I am in Genetics and Cell Biology and Organic Chemistry this semester, and I am finding the information to be very interesting- stuff that I like learning about- and extremely useful as well. There are some things about genetics and certain signaling pathways for example that I couldn’t grasp when it was first explained to me in lab or was discussed in lab meetings that I am learning about in lecture. And these are only the first three weeks; by the end, I’ll learn so much more that I can apply to my lab work.

I’m going to get back to studying for the aforementioned classes, which I have put off for long enough. My plans for lab are to retrieve my last set of data from the confocal microscope once it gets fixed and to Metamorph analyze them. Hopefully, this will be done in time for lab meeting on Monday because I am schedule to present. That’s all the lab update there is for now. And I don’t think I’ve said this before, but if anyone is thinking about applying for this program for next summer, I would be happy to share my opinions (which is of course that it was an amazing experience and that you should do it) so please feel free to send me an email.

Today was the poster session, which marked the conclusion of our 8-week program. This will be my last post but I wanted to wrap things up and reflect upon my experience this summer. This was probably one of the best summers I have had! It’s been great to experience Duke in a way separate from the normal hustle and bustle of college academics. I feel like working in the research laboratories gives you a place you belong to or a reason to love Duke even more. I loved that I was engaged in something very intellectually stimulating but also had the opportunity to meet peers with interests similar to my own. I met a lot of really cool people, heard a lot of great seminars on a variety of topics, and got to work with so many knowledgeable people fascinated by science. It is inspiring to be surrounded by all of that. After getting results from the confocal and seeing all the pictures and graphs on my poster, I felt like all the hard work paid off. I am excited to continue my research in the same lab as an independent study in the fall. There is still a lot more to be done- it just makes you realize how short 8-weeks really is when it comes to science research.

I thoroughly enjoyed the poster session we had today. Having people who wanted to hear about my work was thrilling. Also, hearing feedback and being questioned by other PIs with a background in my topic really made me think about certain things that could be improved upon or further experiments that could be done. Below are some pictures from the poster session of my fellow Howard Hughes-ers and myself:

Me, Dean Nijhout, Andrew, and Sarah gearing up for the poster session.

Kristin and I were poster neighbors- we both had a blast!

Andrew and I waiting for the poster session to start.

Cat, my roommate, and I.

Priya and I taking a glance at other posters.

Kristin, Racquel, and I.

Jackie, Kristin, and I ready to present our research!

I have one last bit of lab photos to share.

My desk is on the right.

The lab bench I work at.

Vibratome slicing machine that I use to slice mice brains.

The hood

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Electrophysiology recording setup that our lab uses.

Possibly the sweetest shirts ever. Our whole lab got them. I’m probably going to wear it every day for the rest of the summer.

So fitting for our lab.

As I am sitting here making my poster, I realized that I have not yet blogged about career considerations. Coming to Duke, I was (and still am) pre-med and very much interested in the biological sciences. I had always been pretty certain that I was going to medical school and hoped to somehow find out if I would like pediatric oncology or pediatric neurology/neurosurgery. My first lab research experience was the summer after my junior year through Boston University High School Summer Research Internship Program where I conducted neural stem cell research. It was a great introduction to the life of a research scientist and I realized there were so many different fields to choose from based on your own intellectual curiosity. This Howard Hughes Research Fellowship reminded me again that I do enjoy going into lab to find the answer to my project. Especially now towards the end, I am getting results and compiling data, which makes it all the more exciting. One thing I realized during this program is that research is a slow process full of errors and days of troubleshooting, which can be frustrating. But the excitement when you finally do get data is well worth the effort. This has prompted me to definitely consider an MD/PhD program. I still have time to decide whether an MD/PhD is for me- a decision I know many people in this program are considering- but I know that I will pursue a career in the sciences. For now, I definitely intend to continue science research throughout my undergraduate career. But as for the near future, I must go work on my poster, which I have to admit is a lot of fun to put together. Wow, eight weeks really is a lot shorter than I had expected.

…you dream about nuclear stains. When I closed my eyes last night, I saw blue Hoechst stained nuclei on the back of my eyelids. That may be due to the fact that I basically live in the confocal microscope room now.

For the first few weeks, I had major problems seeing my fluorescence in the confocal microscope. The microscope isn’t supposed to work miracles; you have to be able to see the image through the eyepieces before you take a picture of the image. My problem was that the GFP (green) and tomato (red) fluorescence was dull so I couldn’t really see my cells. The only thing I could see was the Hoechst nuclear stain (blue). I learned that it is quite difficult to image and it takes much longer than it is supposed to when the fluorescence is dull. It feels like you’re looking for something that isn’t there; thus, it was difficult to even find the dorsal lateral, dorsal medial, and ventral parts of the striatum quickly. Another thing I had to check for was bleedthrough in the channels. I had to make sure that the blue channel was not bleeding through to the red and the green and the same for the other channels. I did not want to get false positive signals and analyze them incorrectly.

This is when I truly learned that science research goes quite slowly because of all the unexpected issues and troubleshooting that needs to be done. My mentor and I brainstormed about what sorts of issues could have caused this decreased fluorescence (which used to be quite bright). The main issue was whether it was a problem in the fixation/processing of the tissue or if it was a problem with the biology in the animal. We headed to the epifluorescence microscope to take a look at our samples. One of the guys in the lab upstairs looked at our slides and mentioned that it seemed as if our slices were uneven- thinner at the top and thicker at the bottom. The postdoc I’m working with said that even if the fluorescent signal is dull, the cells should still look healthy; instead, our cells looked unhealthy, prompting him to think that it was a slicing issue. It occurred to us that our vibratome was having issues and was significantly affecting people’s results. As soon as we got it fixed, our fluorescence has been much brighter!

I have gone back to the confocal to image the new slices. However, the Zeiss 410 confocal that I have been using is becoming obsolete because there is a new confocal, the Zeiss 510. The Zeiss 410 confocal scans a little off- the top of my blue channel for my nuclear stain is bright but it dims as it continues scanning to the bottom, so I have a hard time adjusting the brightness and contrast to an appropriate level. Thus, I will be training to use the Zeiss 510 on Monday (the earliest training date available because they are backed up with all the new users). Then, I will need to count my cells and look at colocalization.

I definitely have my plate full for the next two weeks. I know there will be many hours spent in front of the confocal. In addition, I have been learning to clip the toes of my mice and genotype them for my project.

The Zeiss 510 confocal that I will be using shortly.

On Tuesday, our lab took a day off from lab and spent a day at the Eno River. It was so much fun- conclusion: scientists DO know how to have fun! We went “wafting” which means we paddled in our inflatable kayaks down the river and listened to the tour guide give us advice on how to get in tune with nature. It was very relaxing. When we got to the swimming hole, we all climbed across the slippery rocks to swim around in the swimming hole. Then we headed back. The tour guide said that the “winners” of the wafting tour were those who knew how to relax and enjoy nature- those who arrived back at the dock last were the winners. Ying, a grad student in my lab, and I decided to float all the way back, so we got back a lot later than everyone else- well, we also got out at the wrong place and pulled our kayaks onto the trail and then realized that we were not at the correct dock. Nevertheless, we were definitely the “winners”. The upside- they carried our kayaks back to the meeting area; the downside- everyone else had already started eating the food.
We had a potluck-type picnic. The food was DELICIOUS. Everyone brought in some of their favorite dishes: we had grape leaves and tzadziki, tandoori chicken, samosas, shanghai noodles, hummus and pita bread, and strawberry pie. After lunch, we played croquet. I had never played croquet before but it was a lot of fun playing it with my lab. I got fourth….out of four people!

It was a great way to get to know the people in my lab better. Everyone in my lab has funny idiosyncracies that I would not have found out about otherwise- I couldn’t ask for a better bunch of people to hang out with for 8 hours a day!

Ying (left) is a grad student in the lab. Meng (right) is a postdoc who I am working directly with. Meng’s a lot of fun to work with. Now that we’ve gotten to know each other, we know that we both love food, so we often talk about what kind of foods we’re craving.

Viren and Yehong “wafting”. Viren is the other undergrad in the lab; he’s a year older than I am.

On the left is my PI Nicole and across from her is her son Tyler in the dinosaur costume. He was hilarious- he showed up in a dinosaur costume and didn’t want to take it off even though it was really hot outside. Behind them are David, our lab tech, and his son Nathan.

Me, Nicole, and her son getting ready to play croquet.

Viren, Audrey (our new lab tech), David, and his son playing croquet.

So, I’m excited because I finally know how to post pictures on this blog. So, in my lab, I am studying the D1 (direct) and D2 (indirect) pathways of the basal ganglia. I am looking at D1 and D2 fluorescent cells in the striatum using confocal microscopy. I achieve this by perfusing and fixing the mice with 4% PFA and then make brain slices 50 micrometers thick and mount them on slides. I have a few (I will take more) pictures of my lab.

This is me with my brain slices in the 24-well plates in the ice bucket. Behind and to the left of me is the vibratome slicing machine that we use.

This is me with my mouse. I am obviously enamored with the cute animal.

Last week’s seminars on research ethics were very thought-provoking; we were all probably aware of what ethical issues researchers face but talking about them, defining them, and discussing them during Dave McClay’s interactive seminar made us realize that it relates directly with our lives in the lab this summer. We started the week out with a movie that I thought was going to be pointless; however, it is seriously one of my favorite movies- as in I may add it to my favorite movies on facebook perhaps. It is called And the Band Played On (1993) and is based on a nonfiction book written by Randy Shilts. The movie, which I urge everyone to go see, follows the spread of HIV and how the CDC dealt with what was initially known as the “gay disease.” It also discusses how research institutions, government agencies, political motives, and the news media all played roles in the unfolding of events.

The next day, we heard a seminar by Colleen Denny on ethical research in global health. This talk was particularly interesting because first semester of freshman year, I was in the Global Health Focus. This means that I took three classes related to global health- biology of diseases, economics of health policy, and writing 20: public health ethics. Our writing class and the whole FOCUS cluster in general was dedicated to talking about ethical research on human subjects especially in Third World countries. Using research guidelines such as The Belmont Report and the Declaration of Helsinki, Denny discussed risk/benefit, informed consent, etc. Although ethical issues in human research are important, most of us are not doing clinical research and would not need to consider many of the aforementioned ethical issues. Dave McClay came in the next day and talked with us about how ethical considerations, although slightly altered, are still very much applicable at the lab bench. For those of us, myself included, who work with animals, it is important to conduct ethical research on animals. Duke has modules online at the OESO website that everyone doing animal research must complete. It tells you how to minimize the pain the animals experience, how to sacrifice them, and certain proper techniques- making sure to use a secondary method in addition to CO2 chambers. In addition, we discussed how unethical data analysis can be obtained- using photoshop inappropriately or taking pictures of two gels at once and pretending it was one gel.

I interviewed my PI, Dr. Nicole Calakos. Her story was very interesting because she did not grow up knowing she wanted to be where she is now. Throughout junior high and high school, she was always competitive in academics: linguistics, spelling bees, math team. Her high school bio teacher advised her to apply to a research program at a nearby university. She worked with recombinant DNA. This was her first research stint. During her undergraduate years, she did work-study in a molecular biology lab where she made DNA plasmids. After her undergraduate years, she went to Stanford and got her MD/PhD. When asked whether the fact that she would be in school for a long time was a deterrent, she answered that it was not because she enjoyed doing it. What’s interesting is that she was able to balance rigorous academics with other activities. During her undergraduate years, residency, and PhD years, she was a river kayak guide, participated in half iron man marathons, and took part in bicycle racing. When asked why she got involved in neurology, she said that she was interested in the nervous system and got her PhD in neuroscience. She decided to do a rotation in neurosurgery/psychiatry and enjoyed it. She likes that it is very analytical and based in problem solving. Now, she splits her time between research and clinical 90/10 respectively. She sees patients with epilepsy, migraines, Parkinson’s, etc.

It was great to hear my mentor’s story and everyone else’s mentors’ stories. It was interesting to see what they had in common and what was unconventional. They are all inspiring because it lets us know that there isn’t one “right” path to becoming a scientist.