Student Research at Duke

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.

This past week, I have learned that it is important to know what chemicals you are dealing with- their toxicity, the precautions suggested, and the method of disposal. Dr. Calakos emphasized that one of the first things to do when working with a new substance is to look up its MSDS on the Duke OESO website. Two of the substances I’m working with are 4% Paraformaldehyde and avertin. By reading about them, I learned that 4% PFA causes respiratory irritation if inhaled so heating must be performed under the chemical hood. Avertin, an anesthetic used for IP injections in mice, is good for about two weeks. When it starts to deteriorate, the pH becomes less than 5 and it is no longer effective as an anesthetic.

Knowing the facts about the substances I’m working with makes me feel more responsible for my own project and better able to discuss with others about what the necessary precautions should be when working with a certain substance. Thus, I would like to share my mentor’s advice with others. Below is the link for the OESO website; you can look up the chemicals you are working with, too!

http://www.safety.duke.edu/

I am working in the Bryan Research Building under Dr. Nicole Calakos’s guidance. You can read about the work the lab does here: http://www.neuro.duke.edu/ctn/faculty/calakos/

Today I am vibratome slicing a whole brain (we had been doing half brains) because I need to confocal image 3 different areas of the striatum, which you can’t see if you only use the half brain. A confocal microscope is pretty difficult to use when you use it for the first time. The Zeiss 410 microscope that I use is located in the French Science building. Everyone who uses it is trained because it is a delicate instrument. The confocal gives you clearer images than you would get on an epifluorescence microscope because it gives you an average of multiple layers of tissue or cells so you can see the spatial arrangement (3-D look). I look at brain slices that are 50 micrometers thick and have three fluorescent colors (red, blue, and green). The great thing about the confocal is that you can see all three at once on your slide and if you focus the image correctly (which takes me awhile because I am still getting used to it) the results are pretty neat. Here’s a picture of the confocal I use: