The first obstacle was the actual generation of Lm biofilms. With extensive literature review and assistance we were able to combine multiple aspects from different articles to develop a protocol that produced biofilms. Subsequently the problem of quantifying biofilm formation plagued my project. I was using Crystal-Violet and colorimetric assays in an attempt to quantify biofilms but, unfortunately, this reagent imparted too many errors in our results due to the undifferentiation between viable and non-viable cells. I therefore proposed using a flurometric test which would further confirm the viability and activity of the biofilm. Using his previous experiences with fluorometric techniques, Dr. Rolf suggested using the resazurin reagent. To summarize the protocol, starter cells were incubated overnight and then diluted into microtiter plates where they were allowed to adhere to the well wall at specific times depending on the sample, the subsequently inoculated microtiter plates were subjected to a series of washings with PBS buffer to remove free living cells and unattached cells, followed by air drying and staining with resazurin, finally measurements were taken in 30 minute increments for a total of two hours at the corresponding wavelengths for quantify resazurin activity, this was performed at 48 hours, 72 hours, 96 hours, various culture times