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Calibration 1: OD600 Reference point

• Purpose of this calibration: To transform absorbance data to a OD₆₀₀ measurement, calculate a plate-reader specific (Tecan Infinite 200 Pro) conversion factor for OD₆₀₀ from Abs₆₀₀ calculated for Ludox CL-X on a mass spectrophotometer.
• Beer-Lambert’s law of absorbance dictates that optical path length plays a fundamental role in determining absorbance:

• This is necessary because cuvettes (used in photometer) have a fixed path optical path length when using light scattering to measure absorbance as opposed to the varying path lengths of wells in a 96-well plate which change as the volume of sample added in them changes.
• Results: Cell density readings can thus be converted to OD₆₀₀ by multiplying correction factor value, 4.138.

Table1.

Table shows absorbance measurements (at 600 nm) for LUDOX CL-X and dd-H2O using a plate reader. The corrected Abs₆₀₀ is the difference between the LUDOX CL-X reading and dd H2O reading. Reference OD₆₀₀ is a measurement by a spectrophotometer (provided on iGEM excel sheet). OD₆₀₀/Abs₆₀₀ is the correction factor to convert Abs₆₀₀ to OD₆₀₀, calculated by dividing Reference OD₆₀₀ by Abs₆₀₀.