Manipulating Variables of the 6-Well Microtiter Dish Biofilm Assay Protocol: The Effect of Temperature on the Amount of Variability within the Data
Caroline Barnett
Honors Project: Manipulating Variables of the 6-Well Microtiter Dish Biofilm Assay Protocol: The Effect of Temperature on the Amount of Variability within the Data
Honors Course: Honors Interdisciplinary Undergraduate Laboratory Research
Instructor: Suzanne Ziegenhorn and Gary Mines
My name is Caroline Barnett and and I’m pursuing an Associate in Applied Science in Medical Laboratory Technology. I will graduate in the Spring of 2025 and transfer to a university the following Fall to complete my Bachelor of Science in Medical Laboratory Science. Afterwards, I hope to explore specialties in my field and pursue a graduate degree. I can say with certainty that the work I’ve done in the Honors Program is why I’m studying to become a Medical Laboratory Scientist. One of the first Honors classes I took when I started at Oakton was Microbiology. Getting to do the detective work of finding out what bacteria I’m testing for the “Unknowns Project” and figuring out what bacteria was causing my patient’s illness for my Honors Case Study is what ignited my passion for science and the medical field. One of the most pivotal opportunities given to me by the Honors Program was taking the Interdisciplinary Undergraduate Laboratory Research class. The research I worked on and the skills I learned in this class are ultimately what led to my decision to join the Medical Laboratory Technology Program. Being in the Honors Program has given me the privilege to learn from some of the best professors I’ve gotten to work with. These professors continue to be an inspiration as I further my education. I look forward to starting my career as a Medical Laboratory Scientist and exploring what the field has to offer.
In the Interdisciplinary Undergraduate Laboratory research class, our focus was on studying biofilm, a slimy substance produced by certain types of microorganisms as a defense mechanism to protect themselves. One type of experiment we can do to study biofilm is a 6-well Microtiter Dish Biofilm Assay. In my experiment we incubate the bacteria so it can grow and form biofilm in each of the wells, and then after incubation we rinse the wells, only leaving the biofilm behind. From here, we use a dye called crystal violet to stain the biofilm, which will allow us to get absorbance value readings from a spectrophotometer so we can see how much biofilm was formed. However, an issue arose when our data sets started lacking consistency and we were seeing a lot of variability between each of our data points. From here, we decided to focus our research on the biofilm assay protocol, so we could take a deeper dive into what could be causing this variability. We each took on different variables that could be contributing to the variability in our data sets, mine being the temperature that the bacteria is being incubated at, and also the biofilm-forming bacteria we are experimenting on. However, my research took a turn when I started getting constant contamination in my control wells and had to figure out what was causing the contamination.
