Student Research at Duke

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!

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.