One of the reasons undergraduate research is valuable is because things do not always go the way the researcher hopes.
Procedures can take a lot longer to perform than the researcher has planned. Hypotheses cannot always be proven true or not true. And sometimes, the tiny, knocked-out subjects of the experiment stand up and fly away.
All three happened to senior biology major Christina Brunecz of Bear Lake this summer as she designed and conducted her own experiment using Drosophila (fruit flies) to test the anti-inflammatory properties of garlic.
Dr. Lauren Yaich, associate professor of biology, oversaw her research.
“Most of our students plan out their own experiments,” Yaich said. “In Christina's case, I suggested that she could test out either inflammatory or anti-inflammatory foods to see if they affected the incidence of tumors in fruit flies.”
After choosing garlic powder - she used the kind from the grocery store - she determined the concentrations of garlic powder to use in the flies' food.
“It's important to try a wide range of concentrations, since too little will usually not give you any effect at all, and too much could be toxic,” Yaich said. “The students have to play around to find the sweet spot in the middle.”
Brunecz felt she was making progress toward determining that sweet spot by studying her five groups of flies that had each ingested different concentrations of garlic. It appeared that some of the flies' tumors were shrinking, but she ran out of time to finish the study conclusively.
“I wish I could have followed them longer,” she said, but the beginning stages of the experiment took longer than she had expected.
Like so many projects, most of the work involved in Brunecz's experiment came in the set-up, because, unlike garlic powder, one cannot just buy fruit flies with tumors at a store. She had to breed them.
Fruit flies are a favorite of researchers because they are inexpensive and produce a new generation every 10 days. Brunecz began with two fly lines, one of which produces a protein that can activate a tumor-causing gene. By themselves, each fly line looked perfectly normal. However, when mated together, their offspring would grow tumors on their eyes.
To make things even trickier, the experiment required that the female flies be virgins. Here, the flies' fleeting life cycle becomes an advantage. A beaker full of male and female flies mated, and then the females laid their eggs in a food medium (to simulate fruit) in the bottom of the container. Females lay the eggs in the medium.
After about 10 days, larvae hatch from the eggs and make their way up the side of the beaker to become pupa, which are similar to a butterfly's chrysalis.
Brunecz removed the parent flies from the beaker before the pupa hatched. Once the pupa hatched, she had eight hours until the young flies reached sexual maturity - eight hours to collect the hatchlings and separate the males from the females under a microscope.
“What I really came to understand is that timing is everything,” Brunecz said. “You have to know everything about the cycle of a fly's life.”
She managed to successfully mate her virgin flies and produce flies with tumors on their eyes. Then she fed them varying amounts of garlic and studied the results under a microscope.
To study the flies, she placed them on a special plate that piped a steady flow of carbon dioxide past the flies to keep them anesthetized. Examining the flies happens several times during the process -- first, when Brunecz sexed them for the breeding portion. Male flies have visible, if microscopic, genitalia and structures called “sex combs” on their front legs.
Later, Brunecz would examine the offspring for the yellow, bumpy eyes indicative of tumors. Finally, she would check them out to see the effects of the garlic consumption on the tumors.
It was during one of the earlier stages that her carbon dioxide canister ran out of gas. “I was looking at them under the microscope, and they just started to get up and fly away,” she said of her tiny subjects.
Thinking quickly, she rant to another room and put them on ice. Her prompt action saved her from losing all of her subjects, but she did lose some, and it would be something she would have to account for in her results.
Even with its challenges, the experience whet Brunecz's appetite for research. She would like to work on a similar project more in depth, but if it involves fruit flies, she is sure she will check the level of her carbon dioxide canister first.