Duke University | Howard Hughes Undergraduate Program

So I already introduced my project in my first blog because I was lucky enough to get started on it from the first week of this program.  Hence in this blog I will give you a brief overview of how my project is progressing (well actually this week I don't know if I can use the word progressing).  The first week I was so excited to be doing experiments for my project, learning new techniques, and just learning everything about being a scientist.  This week the lab is totally different from my first 2 weeks.  In short, it is slow and frustrating.  The experiments we did the first 2 weeks produced mixed results.  Some matched the results of the paper that I am following, but some did not.  So in order to get the same results that the other study obtained, I have realized that I need to use exactly the same materials that the UNC group used.  Some of the substitutes we tried (for whatever reason from convenience to more cost-effective) haven't given us the published results.  So my mentor and I have basically spent all week finding the exact materials needed, ordering them, and waiting for them to arrive.  Today one important order was supposed to arrive which would allow me to start another cell culture, but there was bad weather in Memphis and so now it is coming tomorrow.  Waiting for the materials to arrive makes the day go by so much slower.  In addition, it is frustrating because at this point I am still trying to repeat an experiment that someone else did, and just repeating that experiment and getting the results they got is taking forever!  I want to start doing something new where I don't know what the results are going to be, but I know that getting the foundation correctly laid is necessary.  Without being able to repeat the experiments done by the UNC group, our results would not be as credible.

Another frustrating part about following someone else's experiment is that you don't really know what they did! I mean they wrote what they did in their materials and methods section, but it's not enough to follow.  For example, the paper I am following says that blood was drawn into PPACK, which is basically an anti-coagulant...but there are no tubes with PPACK already in them.  So how did they make the PPACK from the powder it comes in?  How much PPACK did they use?  Was the PPACK just sitting in the tube that the blood was drawn into?  As I try to follow what the paper says, I am often confused about what exactly they did.  Now I understand why in our science classes they tell us to write our procedure very detailed so that anyone could easily follow it.  Recently we emailed the person who wrote the paper a bunch of questions about the procedure.  Hopefully she responds soon so I can get back to doing stuff! Waiting for materials and email responses is not very exciting...

Oh, but yesterday I did learn to use a hemocytometer, which was pretty exciting.  I thought it would be some complicated machine, but it turned out to be a small device about the size of a post-it note that you simply put under a microscope and count cells under.

So to happy news - this past weekend was amazing.  Some of us from Howard Hughes went to the beach for the weekend! It was perfect weather and and I had a lot of fun playing frisbee, playing volleyball, and just swimming in the ocean!  We had seafood later at a nice restaurant and some of us stayed over at a friend's house :)  Also, recently I had a really cool EMT call where I helped deliver a baby!  Amazingness!

So hopefully next time I blog, I will have lots of experiments to talk about because all of my materials will have come in by then!

 Expectations are dangerous things. If you expect too much from someone, something, or some place then chances are that failure to meet those expectations will lead to sadness. But at the same time, it is fun to expect...to look forward to something...and then when it finally comes, enjoy it to the fullest. If you aren't expecting something, then it is hard to be prepared for it.

I am not really sure what I expect from my summer research. It is sort of a blend of things. I want to be able to experience working in a research lab for 8 hours a day 5 days a week to see if I would like to pursue a career in research. At the same time, I want to be able to understand and perform common experiments such as cell culturing, which I can use in any lab. I also want to attain a certain degree of independence in this lab so that if I do continue throughout my Duke career in this lab I will be able to do my work independently. I want to see what it is like to think like a researcher...come up with questions, find ways to explain unexpected data, and come up with clever ways to solve problems. Basically at the end of the summer I want to be able to say that I know what it is like to do research in a lab so that when I am deciding on a career path, I can weigh the options better. Coming up with results at the end of my project besides inconclusive would be nice also.

Besides my experience in the lab, I expect several other things during my summer. I hope to learn to cook a little...I've made chicken and vegetable pasta and omeletes...and today I am gonna attempt to make Indian food (uh oh!). I also want to catch up with Grey's Anatomy (blah...I am so behind!). And of course spend time with my friends without having to worry about homework!

So now a little update on how my project is coming along. One of the antibodies we got is not working as well as we want it to. Hence along with flow cytometry, we are going to use another method to show the presence of aggregates. We are basically creating our own cell line. We took some blood, isolated the white blood cells, and are now growing them on cover slips and glass slides. After they grow, we are going to add red blood cells and look for aggregates. I never knew growing cells was so hard...they have to be fed special media, cleaned, and placed in perfectly sterile environments. Another cool thing we did today was we stained some whole blood to see the different types of white blood cells...I could tell some of the monocytes apart from the others which was pretty cool. We did this because later we will have to verify that the blood cells in the aggregates are actually monocytes. I think soon I am going to stain my own blood for fun...I wonder if my cells will look much different from the cells of the sickle cell patient we stained today?

As I am doing more research I feel like I am straying from my original project. After each data set I receive, I find something interesting about the data that I wonder about. Then I want to explore that mystery...so by now I have so many more questions about my work in general than when I started my project...weird considering that you would expect to have more answers and less questions after 2 weeks. But in this case, I feel like what I thought I knew before I am now beginning to question. I am even questioning papers that were written in the past. But its good exercise for the brain to be always thinking, always pondering, always wondering.

 Hi Everyone!

My name is Shilpa Agrawal and I am a rising sophomore at Duke University. As part of the Howard Hughes Program this summer, I will be working in the lab of Doctor Marilyn Telen. The people in the lab study sickle cell disease. I am really excited because I have never worked in a wet lab before!

So in the first few days I was introduced to my project. Over the summer I had read several articles and one of them (from UNC) showed how sickle red blood cells not only stick to one another but also to monocytes and platelets to create aggregates. These aggregates contribute to the clumping of cells, which eventually can block blood vessels and impair blood flow. In addition, I was told that epinephrine often increases blood vessel blockage as sickle cell individuals have more of a change of blood vessel blockage when they are under stress. So for my project, I am looking at whether and how epinephrine affects these aggregates of sickle red blood cells, monoctyes, and platelets.

My mentor Mardee. who is absolutely amazing, explained to me some of the procedures I would need to use for my project. I learned many things from the basics of how to pipet properly to flow cytometry. One thing really cool about my project was that we started from the very beginning. I had to order the reagents that I would need! I had never thought that a lab wouldn’t have the necessary supplies. On my first day in the lab I saw so many chemicals and supplies that I never even considered having to one day purchase even more. But with the help of Mardee, we ordered some reagents we would need…why, by the way, was so much more difficult than I thought it would be! One antibody has like 20 variations and you have to go through each variation and pick the perfect one! Right now I am on the part where I have to test each antibody separately to make sure it works, which makes me realize that this one project is going to take a very long time! But that’s okay because I love it so far…especially not knowing what is going to happen next and getting to do so many experiments with fresh human blood (okay I sound like a vampire now).

The people in my lab are so friendly and nice. It is a relatively small lab but that makes it nice and cozy. My mentor Mardee answers all my questions so thoroughly that I know in my project exactly why I am doing each step and that feeling is really great because then you aren’t just following a protocol. She is also very patient when I do things like get blood on the Pipetman (which hopefully will start happening less so I don’t waste all of the lab’s ethanol). Susan is also super cool (expect for the fact that she’s from UNC and wears her UNC clothing sometime). Not only does everyone help me around the lab but they also are helping me adjust to living in an apartment for the first time and having to cook! Sometimes Mardee prints out recipes that are not that difficult to follow so that I actually have something for dinner.

Here are some pictures from my lab:

Susan! Who agreed not to wear her UNC attire for this picture :)

Mardee!

Me! Doing flow cytometry with sickle cell blood in the pipet.

Our fridge full of blood and antibodies and other things!