This experiment was designed to eliminate as many sources of error as possible but not every part could be entirely controlled.

• Cell Counting: Since a visual method was used to conduct the proliferation assay, the counts may not be 100% accurate. To minimize error, a manual cell counter was used to assist in keeping track of counted cells. As well I was the only person to count cells to decrease the chance of counter bias.

• Accuracy of Well Representation: Each well of the 6-well dish could not be completely counted due to microscopic visibility, and it would be unrealistic in terms of time constraints to visibly image and count 6 wells a day. In order to overcome this source of error, 3 fields of view from each well were imaged in bright field channels, and GFP and RFP channels where necessary. This resulted in 3-9 images per well, or about 40 images per time period. This amounted to approximately 1000 images over the course of experimentation, which is an amply large sample size.

• Plating Efficiency: Although calculations were made for every splitting procedure into a 6-well dish to ensure that the same amount of cells were being deposited into each well, the graphs show that some wells contained a different number of cells at time period one than others did. If the experiment were to be performed again, the calculations would be performed twice. However, the instances of lower plating efficiency that occurred had no significant effect on the outcome of the data.

• In Vitro vs In Vivo: The limitations of this experiment include the fact that this experimentation was performed in vitro and so genetic variations among individuals cannot be analyzed. However this is an early step for the future of KM110r as an oncolytic virus treatment and clinical trials to test KM110r in vivo may not be too far off.