The WPI-2016 iGEM team developed its unique method of measurement through technological constraints. We lacked proper access to a flow cytometer at our university. However, measuring fluorescence was still required to analyze our experiments. We devised a new way to measure fluorescence of a sample using microscopy and image analysis. By using the program ImageJ, the images taken of fluorescence microscopy were analyzed for brightness on a pixel basis. Through taking images of the same sample of cells at the same exposure times, data was collected and averaged to derive fluorescence data. This data was then checked through a collaboration with the Boston University Weblab, using their flow cytometer. When compared, the flow cytometry data from BU confirmed the legitimacy of the microscopy image analysis method. This technique was also used for the validation of another BioBrick part that focused on arsenic absorption, shown below. It is through this ingenuity that we believe that our team qualifies for the measurement prize.

Quantifying Fluorescence via Microscopy

ImageJ Analysis:

• “Analyze” → “Set Measurement” → select “Area”, “Integrated density”, “Mean gray value” → “OK”
• For each image, “Analyze” → “Measure” (Results should include “Area”, “Mean”, “IntDen”, and “RawIntDen”)
• Save “Measure” results

The Excel analysis was performed as follows:

Arsenic Absorption Experiment