Duke University | Howard Hughes Undergraduate Program

 In a lab with 16 people, each doing many separate projects, discussing what I am working on doesn't do the lab justice. With that disclaimer, I will briefly explain my personal project for this summer.

I am doing in vitro studies in an attempt to characterize one aspect of the Numb protein in mice neural stem cells. Specifically, I create and maintain cell cultures with stem cells harvested from transgenic mice and stain them at various stages (arresting their growth) in order to visualize the presence and localization of various marker proteins. One of these markers, and the one we are most interested in, stains Numb. Using microscopy, we hope to visualize the localization of Numb in proliferating (especially dividing) cells and various types of differentiating cells.

So, why would we want to study that?

Numb has been thoroughly characterized in Drosophila and is known to localize into only one daughter cell of an asymmetrically dividing cell. This observation is correlated to cell fate, as Numb is partitioned into the cell that will differentiate into a mature neural cell. The Numb- cell on the other hand, remains a proliferative cell with stem cell-like properties. Now, keep in mind that all of this occurs in the adult brain--does it interest you to figure out how stem cells self-regenerate in an adult human brain? That is why studies are now turning to mice, the mammalian model organism. Unfortunately, Numb localization experiments in mice has produce variable and inconsistent results. To date, no one has successfully visualized Numb localization in mammalian postnatal neural stem cells. That is, we are unsure as to the function of Numb in mammals, which may or may not be similar to that in Drosophila.

Currently, my experiments have been malfunctioning in certain ways, because the assays are not definitive or perfected. Thus, I am trying out two different Numb markers and altering the protocol in order to achieve optimal staining and visualization results. Don't hold your breath for any conclusive results. It could be a while.

 So, I don't think I've mentioned this, but all of the projects in my lab are dependent on the use of microscopes. Ironically, microscopes have never had an affinity for me and to reciprocate, I've always avoided them. Unfortunately, all of my data will come from the microscope. Great. It's as if life decided to spite me. I recall visions of horror when I sit in front of a light microscope in some high school lab, my glasses occasionally clanking against the eyepiece, squinting a lot, turning the adjusters in frustration, and all in vain because I couldn't see anything. So there, I've used light microscopes and dissecting microscopes in high school. Of course, I had heard of the great and mystical electron microscopes but that was pretty much it. Sometimes I wonder if I'm not meant to do science: I've also discovered a dislike for NMR, the heavenly tool of chemists.

Now, most of you readers are probably expecting some sort of epiphany in the following lines, something like "But now, in my current lab, I learned to use the microscope and love it/am so good at it!" or something like that. Uh uh. Nothing of the sort. Sorry to disappoint, but I'm still a failure at using microscopes.

Instead, I just wanted to share some of my recent microscope experiences. Half the time at my first lab meeting was spent talking about microscopes; with my keen and intelligent mind (this is a joke, I hope you can discern), I understood two things. One, broken glass is bad, and two, we have really expensive microscopes. A few days later, I encountered our room of microscopes. I don't even think most labs have two entire dark rooms outfitted with microscopes. Anyhow, during this initial encounter, I was introduced to my new unwilling idol: the confocal microscope. I don't even know what that is. I still don't know how it works. Anyhow, my first piece of information: "This thing is worth more than your house." And that was it. I didn't touch it that day.

A week later, I had a rendez-vous with the confocal microscope. My postdoc was kind enough to take me into the dark lair of this lofty beast. I wasn't allowed to touch it, but I did get a chance to look into the eyepieces. I saw colored dots! Yay! Perhaps, I am doomed to be unimpressed with seeing microscopic specimens, even those that express different fluorescent markers.

I have yet to face the creature a third time, but rest assured, I will overcome my reservations and stare it down. Literally.

 When I think about what I hope to get out of this summer research experience, I realize that I have pretty modest expectations. From previous research projects, I know that research is repetitive, slow, and frustrating. So, honestly I am first just looking to learn lab techniques and get the hang of conducting my own experiments. Once I accomplish that, I expect to repeat the same tasks pretty much throughout the summer.

Of course, I am hoping to have my own sort of mini-project soon. Currently, I have five neural stem cell cultures to take of (I LOVE it, so fun!), and I am also performing immunocytochemistry (ICC) on these cells to stain them for various proteins. We are particularly interested in the Numb, which is a cytoplasmic fate determinant protein. If my own experiments work out, I would be able to visualize Numb localization in the descendents of neural progenitor cells. I am not exactly sure how these results fit into the whole scheme of things (need to do more reading!), but I know that currently in mammals, Numb localization has not been characterized yet. My main goals research-wise are to be able to answer the following questions when you ask me: What am I doing? Why am I doing it? And why it is interesting?

If I manage to break out of my shell in lab, I would be able to learn many things about the path of science and medicine. The members of my lab are snapshots of all stages in such careers: a rising senior applying to medical school, a graduate student, a pre-pharmacy student, MD/Phd students, an MD/Phd fellow (can you say b-u-s-y), postdocs, and even an MD/Phd instructor. They are all ridiculously busy, weaving in and out of the lab constantly, making them rather elusive when you want to talk to them (I just thought of the Heisenberg uncertainty principle…). But, they are all approachable and often have interesting stories to tell.

 So I had lunch with Jingjun Li, the very knowledgeable and (thankfully) patient postdoc who is teaching me in my lab. I got to interview her, and here is what happened:

Q: How did you get interested in science?
A: Family with science background: mother was a scientist and father was an engineer.

Q: What training/education have you had?
A: Masters degree in Biomedical Research and Cell Biology (in China); Phd in neurobiology (UNC-CH)

Q: Have you experienced teaching and did you enjoy it?
A: At college in China, half a semester was spent teaching high school students, and at UNC-CH, she was a T.A. She did not really enjoy teaching high school students, though being a T.A. was more manageable.

Q: Have you always worked in academia? Any interest in industry?
A: Yes, she has always worked in academia and does not thing her personality suits industry work. She is patient and not ambitious enough. Also, she thinks she is not outgoing enough to deal with the bureaucracy.

Q: What do you like about science/research?
A: She likes to think independently and enjoys reading literature. She also likes to design experiments herself and to prove her own ideas.

Q: What do you dislike about research?
A: The scramble for and lack of funding, and how doing research is based on funding/ She also thinks competition between research groups are a waste of resources and manpower because people don't collaborate and try to hide developments from each other.

Q: What do you think about the dynamics of the lab?
A: It's a bit too busy and people don't have the time to talk/ Also, the labspace is too crowded and lab members come and go too often.

 During this summer's Howard Hughes Research Program, I am working in the Kuo Lab, under the Cell Biology department. One of the most difficult but exciting aspects of this lab is the lack of scientifc literature detailing what my lab is trying to do. Apparently, no group to date has established an immortal cell culture of these neural stem cells; that is, instead of a traditional lab with nicely written protocols that are sure to work, we have to run experiments under different conditions to try and simulate in vivo conditions. It's pretty tough.

Despite reading many articles and even browsing through a neurogenesis textbook, I still fail to grasp details of my lab. Each mouse cross is injected with a new cocktail of genes and each cross is slightly modified from before. One thing I am clear about: I will be creating and maintaining stem cell cultures using primary cells extracted by other lab members, most likely won't have to decapitate mice personally, and will we working on incredible cutting-edge research. Sweet.

The first two days on the job consisted of sitting around and finding things to read. I was getting pretty restless and really wanted to start moving my hands. Today, I learned to make the growth media necessary for my future culture experiments--I still have a long way to go, even with basic lab techniques. Fortunately, the postdoc I am working with is patient with my mistakes. Another exciting but tedious realization: in a biology lab, the biology dictates your schedule. It will often be necessary for me to go into lab on weekends because cell cultures need to be monitored each day. Proliferating cells wait for no one. I am actually looking forward to going in on weekends because I hope that once I can independently conduct experiments, I can help out lab members who have other obligations on weekends (e.g. family and kids, events, etc.). Hopefully I'll get skilled enough someday.

In the meantime, here are some photos of my lab:

a) 

b) 

Fig.1(labeling for kicks and giggles) a) A general view of the lab from my modest space, b) me writing down some protocols in my lab notebook.

a)     

b) 

Fig. 2 a) The cell culture hood I work in, resting state with UV lamps on, b) the same hood on with an experiment running. Note: According to a protocol I read, it has actually been shown that UV light is ineffective in maintaining a sterile environment and is only damaging to eyes. As a result, this function has been removed in newer hood models. Just fyi--I found it to be interesting.

Until next time,

Lisa