“If you can’t call a professor by his/her first name by the end of your 4 years, you’ve wasted your time at Duke.”
Hey guys!! I’m pretty excited right now because in a few hours my lab is having a Fajita Fiesta on Harriett’s farm!! Anyway, I thought I should make an update about my research. It has been a very intense week in terms of work! I stayed at the lab pretty late most days but once you get into it, you just don’t feel like dropping everything and going home. The project has gotten significantly more interesting with the use of two animal models. The zebrafish is fantastic because it is transparent, breeds fast, and readily available. Nevertheless, it is SO small. We are using a strain of fish known as cmlc2s which have their hearts stained with Green Fluorescent Protein (GFP). They look so cool under the fluorescent microscope!!
We spent a good amount of time extracting the hearts for protein extraction. We have someone from Proteomics helping us with this part so it’s nice to have experts around. Protein work is probably the most tedious of genetic techniques. Although the concept of Western blots is simple (you buy an antibody that recognizes your protein and forms bands if it’s present), it’s actually very complicated and sensitive, especially with tiny proteins. Right now our problem is that we are getting a lot of background and too many non-specific bands, so the next few days will be a troubleshooting period for us. That’s why I’m really happy we have the chick model, as well. The chick cardiomyocytes have the potential to be a good model since we at least know how to culture them. Using a compound called BrdU, we can tell which cells are proliferating (the DNA of a dividing cell uptakes the BrdU and stains it green).
If we can show that there are less PCB-dosed cardiomyocytes than normal cardiomyocytes, it will tell us that the chick might be a good model for hypoplastic left heart as well. Still, it’s important that we don’t get too caught up in this while collecting data. To prevent bias during cell counting, we randomize the wells so that we don’t know if we’re looking at PCB cells or normal cells. I think it’s important that all researchers make sure they take these sorts of measures because it prevents them from affirming their hypotheses for comfort.
Hopefully, we will get some results after modifying the Western protocols for zebrafish protein. It would be great to have since Peggy (Dr. Kirby) is applying for an in-house grant next week and we can show our need for a 2-D gel. The title of this post is actually something my developmental psychology professor, Gary Feng, said.
I’ll post some pictures from the fiesta in the next post! take care
July 4th, 2007 at 10:00 pm
Hey Trisha! I’m glad you like the Kirby lab. I have been there during the past semester and will be next semester as an independent study, and it’s a joy to work there. I work with zebrafish too, but I’m looking at arterial pole formation, as opposed to hypoplastic left heart. They are hard to manipulate because of their size, but you seem to be doing fine. Your pictures look good, too!
July 5th, 2007 at 11:05 am
hey Ashley!! I’ve heard a lot about you! How did you find my blog? Aren’t you in Ecuador or somewhere working with a medical team right now? I saw that letter you sent to the Kirby lab.
I hope that’s a lot of fun…take lots of pictures!!! oooh arterial pole research sounds cool…I feel like the majority of people in the lab study something to do with it. Our project is more of an epidemiological/environmental health project. Yes, working with zebrafish is tough!! They’re just so small. We’re working with chick too so the two models complement each other well…I’ll be doing an independent study too next semester! see you soon