Duke University | Howard Hughes Undergraduate Program

I spent 7 weeks of my precious summer vacation for the Howard Hughes Precollege. Most mornings, I would wake up at 5:45am so I could get ready. Daily, I would spend about 2 hours on the rocky public bus commute. On days that I woke up 10 or 15 minutes late, I would have to wait what seemed like forever for the next bus to come. Due to the days that I ended up being late, I learned to sprint in flip flops down Research drive. Even through all these troublesome events, I wish Howard Hughes Precollege Program never ended. I wish the program did not stop at 7 weeks, but went on and on and on. This may sound really weird, but yes, I just wish I was locked up forever at this time period.

Oh how I just wish…..

I remember my first day of the Howard Hughes Precollege Program. I tried to hide the balls of sweat on my head after running around Duke looking for the Biology Building. I thought the meeting was going to be at the Bryan Center (please don’t ask me where I got this from, I don’t even know) and just getting there took me ages! When I found out my error at 8:20am, just 10 minutes before I would punish myself in shame for being late to the very first meeting, I rushed around trying to get to the actual location. In desperation, I was on the verge of pressing one of those ‘emergency’ buttoms to ask for directions! Good thing I found out where I was….
First week was lab orientation. Oh how lost I was. Protein, PCR, enzyme, lysozyme, pipette, cells, plasmid, ect ect ect! I went home and watched an hour worth of youtube videos just to review on my freshman biology. For the 5 days of orientation, I came into the lab lost to the new materials we covered but everyone that I asked questions was really helpful and patient with me. Thinking back now, if I was Lavendar or Connie, who were the ones that helped me the most, I would just ignore me.

I was also introduced to the Civil and Environmental Engineering Department this week too. When I first found out that I will be in the Engineering deparment, I was absolutely horrified. ENGINEERING?? I didn’t know a thing about building. I think Dr.Gunsch knew this and so when I had my first meeting with her, she sat me down and told me the importance of a woman role model. Oh Dr.Gunsch, she is such an amazing woman. In this category goes Christina, Lindsey, and Kauro aswell. Without their help, I would never have been able to experience and explore the great opportunity to its maximum.

EVERYBODY: NANOSILVER IS COOL!

I think the best part of the HH program was the people I got to meet. The 14 other HH members were all extraordinary students and all who became a stellar example in my life and helped me to squeeze out of my comfort zone at my high school. I truly am going to miss their presence, especially at lunch time. Noon was when we all got together, ate, talked about our research, and about our day. I was able to build my friendshi p with many of the girls (and boys). Do you guys remember our first week of lunch? I have to say, it was kindof awkward at times. When it got silent, we would look at each other and I would do the ‘awkward turtle’…. Memories… Today, I found myself wondering where we would all go for lunch as a group. Blue swings? Twenties? Blue Express? This lingering question went about until I was reminded that HH was over…

I truly hate the world ‘over’. Howard Hughes Precollege program of 2009 is NOT over and it will never be. In my mind and heart, HH will be living forever ( this is so cliché, but who cares, I’m a special person :p)! Everybody, we need to keep in contact! This is what technology was made for!

Before I end this blog, …. nooo I can’t believe it’s coming to this!!!.... I would just like to thank the Howard Hughes Precollege Program advisor and mentors: Dr. Wahl, Kriti, Tanya, and Chris. Without your dedication and help, the Howard Hughes Precollege Program of 2009 would never have worked out the way it did. To the 14 amazing HH students, thank you so much. You guys are all amazing and I hope to hear the best from you all. Here, I would also like to thank Brook Teffera, my lab partner. In the beginning, I have to admit, you were not my best friend (cough cough)… but now Brook, you have proved yourself to be a true scientist who can wear his labcoat unnecessarily in pride. Last but not least, thank you so much Dr. Gunsch, Christina, Kauro, and Lindsey. You all proved to me that Civil and Environmental Engineering is the way for future females to go!!!

Howard Hughes Precollege was the greatest experience that I have ever had. It was worth everything I gave up for the summer. If you are wondering if you should apply for the Howard Hughes Precollege program for the future, you’ve made your 1st mistake in science (well at least in this part of science.. you get my point)… DON’T WONDER!!! Just do it!

I will continue to research at Duke University, working still for Dr.Gunsch in the Civil and Environmental Engineering Department. Like Shalini, I am pretty sure that I will pursue a career as a scientist, working hard for a PhD. I don’t think I would have come to this point of my life so early without HH.

My HH bag smells like laundry detergent. It is soft and you can sense the cleanness of the bag. It has slightly shrinked… oh well. The surface of the bag has changed and it will continue to do so as I use it. It might get another HUGE stain or form a hole somewhere. Regardless, the bag is the bag… it represents and will never be forgotten!

Thank you so much Howard Hughes Precollege Program, I will miss you.

Leighanne

Me next to my poster, "The effects of nanosivler particles on Escherichia coli K12 and Bacillus Subtilis." 

Howard Hughes Precollege Program members of 2009

Brook Teffera, my lab partner, and me. 

(left to right) Christina Arnaout, me, Dr. Gunsch

The Joys Of My Research
It’s almost done! From the first day in my lab, I knew what my research project was and how to conduct it. I’ve leachated all my plates and this project will be completely done with next week. I’m just waiting until next Thursday so I can plate my last set of R2Aagar 10 day. Oh the joys of this project. This research project has really taught me how to be resilient and patient as a person and most importantly, love and enjoy ‘research’.

The Woes Of My Research
Well… I don’t really have any major woes other than the fact that this Howard Hughes Precollege Program is slowly ending, meaning this research project will be over soon too. I very much enjoy coming in the morning and checking on my E.coli, making trips to the autoclave, and plating agar plates. If I did have a tiny little complain about this project, it would be that none of my nanoproducts samples leachated nanosilver! So all my plates that I did with nanosilver products, such as the teddy bear or lotion, I didn’t get any zone of inhibition. Are these companies false advertizing? LIARS!

Kirby-Bauer plating method on teddy bear skin on R2A agar with E.coli K12. The white streaks are the E.coli bacteria and as you can see, there is absolutely no zone of inhibition. 

E.coli 1mg/L AgNO3 on LB agar. You can see

a slight zone of inhibition.

Zone of inhibition can be measured by a caliber. It can measure the inhibition more accurately than a ruler.

Couple of weeks ago, I went to a brand new Asian store in Brier Creek. As I was looking around for some yummy Korean goods (which I didn’t get), my eyes set on agar! Mango agar to make it sound a little better!

It’s agar though, isn't that cool! Agar, which my lab uses all the time, is used to solidify liquids into a jelly substance, such as LB broth. If agar is added, LB broth miraculously turns into LB agar! When I first started to use agar, I did a little research on agar and found out that agar is actually seaweed extract. Agar is often used in Asian dishes and when I shared this information with my lab, I promised them that if I ever found agar, I would buy it and make it for all of us. So I did buy it! And I made it in mango flavor! The only ingredients needed were either cold water or milk and the agar powder. I made mine with both milk and water, just to see if there would be a significant difference in taste and I’m so glad I did because the milk mango agar was so much better! It had more flavor and just a better *ting* to it. Before my lab got a taste of it (which they still haven’t), some girls in the HH program got a taste. Then at lunch, some more kids got a serving. The texture of the agar was basically just like jello. I don’t see the reason to purchase agar, which is relatively pricier than the american brand. But at least I know now… experience is an amazing gift!

Above: Nancy Wang from the HH program had her serving of mango agar.

Below: Guy tasting the agar.

The first thing I do and must do when I enter the building is to change my shoes. In my lab, it is mandatory to wear closed toe foot wear! It’s a safety procedure. If not, Dwina, our lab manager, warned me that she will make us wear these ridiculous boots!

The awesome reject boots you have to wear if you do not have closed toe shoes.

Once my beautiful tennis shoes are on, I go ahead and inoculate my E.coli (which is in the incubator) under the biological hood. Guess what you guys, I no longer despise the scent of the incubator! It still does smell terrible, but it doesn’t cause me to go into a seizure-like-mode due to the excruciatingly painful scent. Now, I voluntarily sniff the incubator and make comments such as, “Oh Brook, today the incubator smells a little salty” or “the incubator is awfully sweet scented today.”

After this, what I do everyday varies. On days I need to plate my leachates, I usually wait until after lunch. This is so that I am sure that my E.coli reaches the lag phase (when the colony multiplies greatly). I then go under the biological hood and spend all afternoon swabbing E.coli, dipping filter plates, and parafilming. The next day, I examine the plates I made and check if there are any zones of inhibition. If I do not plate, I make sure our lab has a good stock of LB broth, LB agar, and R2A agar. If we are short, I go ahead make these. Also, I make sure my lab is spotlessly clean (no dirty dishes, awkwardly placed lab items, or dust). Shadowing Christina, Lindsey, Kauro, or Jenny (a graduate student in the same department) is something I do regularly and in the process I get to learn what they do, their projects in detail, and most importantly, they teach me techniques, methods, and procedures that I can learn for inspiration or so I can possibly help them.

Swirly agar remain. This is hardened agar stuck to the bottom of a flask that I need to get out before I can wash the dishes.

I’m really fortunate to be in the lab I am now. I have a caring PI (Dr.Gunsch), amazing students that help me (Christina, Lindsey, Kauro, and Jenny), and a funny lab partner I’m getting use to working with (Brook). Howard Hughes Precollege Program has past its half-way mark and the 15 of us are getting prepared for our posters. I don’t want to leave…My typical day in the Gunsch lab is so much more interesting that a day at school. Do you not agree? Where else would you be able to work with bacteria, smell a stinky incubator, get locked in a walking freezer, pipette constantly, watch graduate students work on the coolest projects ever, and conduct studies to help our human race live more environmentally friendly on our Earth?
 

 This week I got to work on my Kirby-Baucer plates. Before the advent of miniaturization and computerization of microbial techniques, one of the most common methods used to rapidly determine bacterial sensitivity was resistance to specific antimicrobial drugs was to use small paper disks, each saturated with a specific concentration of these different drugs. In my experiment, these ‘drugs’ were my leachates from nanosilver products and the bacteria was E.coli K12. These Kirby-Baucer plates can be interpreted by its size of the zone of inhibition. The zones are measured in millimeters and the size describes an organism as being susceptible, sensitive, or resistant to a drug.

Right:Kirby-Baucer Plate                            

MY EXCITING EXPERIMENT!

STEP 1: Prepare leachates
I had a total of 21 samples. 1mg or 1ml was extracted from each sample and was inserted into a bottle of 100ml of nanopure water which underwent around 24 hours of mixing on the stirring table.

STEP 2: Prepare the E.coli.
The E.coli needs to be at its log phase, which is when it multiplies exponentially, doubly in number every few minutes. From doing O.D measurements and making an exponential graph with excel last week, I learned that the E.coli is in its log phase about 2 hours after being inoculated. The ‘bug’ needs to be used before it reaches stationary phase, which is when the bacteria’s booming growth stops and the number of bacteria stabilizes.

  Left: This is about what the exponential growth of E.Coli K12 looks like. 

STEP 3: Prepare agar plates /Streak with E.coli/Dip filter paper disk into leachate solution and place it into plate
In this experiment, I used LB agar which I plated previously. So I had 21 samples which to ensure the data, I made triplicates. This is a total of 93 plates! Working under the biological hood, I streaked all the agar plates with E.coli, which was inoculated 2 hours before, and then placed filter paper disks into the center, three disks for each sample.

STEP 4: Para film and store in incubator

So in total, I used:
93 plates
93 swabs
93 sprays of ethanol to clean the forceps which I use to place the filter paper disks
93 filter paper disks
93 strips of parafilm
MANY kimwipes to dry my forceps

Brook, my lab partner, did the same procedures, just with Bacillus, a different bacteria. So that’s even more materials used! While I was doing all this, I just noted to myself how much resources were used due to scientific experiments and research…. All the plates I used cannot even be reused due to sterility issues…Maybe someone should invent an efficient way to conduct experiments!!!

RESULTS
So after 24 hours, I looked at my 93 plates. No big changes could be found from the previous day, other than that the E.Coli had grown into thick colonies. E.coli was resistant to all the ‘drugs’ except for 1mg/L of Silver Nitrate. Christina said that maybe these nanosilver products are scams, but we need to give the plates more time if we want to make a more accurate assumption. So both Brook and my plates were placed into the walk in freezer. It will be in there for a week before we check up on them… until then… :D

 PHOTOGALLERY 

Christina, Brook, and I examining our plates. 

Kirby-Baucer Plates

Brook, Christina, and I in the cold walk-in-freezer!