Making Sense of Scents - Week 1 at Duke

I thought I was done with the visa stuff the moment I had sent in the forms to Jason, the human resource manager, but first thing I had to do Monday was to meet with Visa Services. This building was pretty far from my apartment so my dad and I took the bus. After that we went off to the International House (IHouse) for a "scholar orientation". It started pouring while we were walking there so we waited for the rain to die down under a tree. When we got there, I had to fill out some forms. Lisa came to talk to us about how to get around Duke and the Durham area as well as student activities. She gave me a whole stack of information which was really helpful. The first time she talked to me, she called me "my dear Jocelyn" which actually made me feel uncomfortable at first but I got used to it because she kept on referring to and talking to "her dear Jocelyn". It started pouring again and the thunder kicked in too. My dad and I didn't know how to get to the Duke Eye Center from here, but Lisa was so kind in offering to drive us there. She was like a "grandma-you-would-want-to-meet-in-the- woods" as PJC so refers to, although she wasn't too old. She was just really nice.

I called Hiro (my PI) from the Duke Eye Center and he said he would walk over in a bit. Next thing, we're walking to the lab just having a conversation about the summer. We walk into the building and I finally meet Jason! I still have yet to give him a present! Anyway, we have a lab meeting and one of the grad students presented a paper that she read. I realized after a minutes into the meeting that the whole lab was Asian. Not one single non-Asian. I thought that was pretty funny.

After the meeting, I was introduced to the grad student I would be working with: Neha. She showed me around the lab and I was introduced to the other lab members. I talked to Natalie, who is the only undergrad, about her research. I also talked to Mingshan about her topic of research. Then Neha brought me and another high school student to get our Duke cards. Neha also described her project to me.

For the remaining hours of my first day, I counted glomeruli in the olfactory bulb from pictures that Neha had taken with the microscope. Pretty eventful first day. Then the remainder of the week I read some review papers on the olfactory system. Our project investigates the difference in number of glomeruli in the olfactory bulb of wild type mice and knock out mice. The wild type (WT) mice have two proteins: RTP1 and RTP2. The knock out (KO) mice have neither of the proteins. Neha did 6 mice crosses and we genotyped the mice with the gel results we got. She told me that RTP1 and RTP2 genes are linked so if one mice had RTP1 the chance of it having RTP2 would be very likely.

Neha already had the DNA sequences ready for the gel so we made the mix and the gel. Each mice would have 4 columns in the gel: 1 WT, 1 M (mutant/knockout), 2 WT and 2 M. The numbers are the protein RTP1 and RTP2. She told me that they had designed the primers for the sequences so if the band is visible on the gel that means the sequence annealed to the primer. If in one mice, there are bands for 1 WT and 2 WT, that means the mice is homozygous WT, having both RTP1 and RTP2. If bands are all visible for 1 WT , 1 M, 2 WT and 2 M, then the mice would be heterozygous.

Then another experiment she did was put mouse 1 in a paper bucket as the control. She put mouse 2 in a paper bucket, then another paper bucket with a smell (acetophenone). We had four slides of the cross section of their olfactory bulb after the experiment. We did RNA in situ hybridization. For slides 2 and 7, she had a cFos probe that would bind to cFos that has DIG. For slides 3 and 8, she had a Gs probe that would bind to Gs which is present in all the cells. Slides 2 and 3 were from mouse 1 and slides 7 and 8 were from mouse 2. She also added a primary antibody which would bind to DIG. Then a secondary antibody that binds to the primary antibody, which has properties that allow for fluorescence. If the probe has cFos to bind to then there would be a lot of staining (purple in our case).

Apart from these 2 major experiments, I also got to section paraffin blocks of tissues for cross sections to put on slides. I got to use this really cool machine. I thought it was awesome when I first saw Neha use it but now that I have sectioned 4 blocks onto 20 slides of at least 10 cross sections each, it has become pretty tedious and actually somehow painful because the machine is at -20 degrees.

The lab members Mingshan, Ting, Neha, Jianghai, Jessica, Natalie and the other high school students are all very approachable. Yesterday when I was reading review articles, Jessica, the lab technician, asked me if I had plans for the weekend and I had absolutely none. That's why I'm writing this right now! But she gave me a bunch of recommendations to go to Chapel Hill, 9th street and other places in Durham. So far first week has been going well! Hope to do even more hands on work though!