Enter the appropriate extinction coefficients of alamarBlue in the reduced and oxidized form for the wavelengths used in your experiment. The extinction coefficents can be found on page 5 of the technical datasheet (www.serotec.co.uk/minisite) Enter the coefficients with no commas or spaces.

"Experimental parameters" refers to the variable that is being altered for the experiment.

To determine the optimum plating density and incubation time of your chosen cell line the experimental parameters should be several different cell densities, For example 500 cells/ml, 1000 cells/ml and 5000 cells/ml. See (worked example at www.ab-direct.co.uk/microsite).

To determine the cytotoxicity of a chosen agent the experimental parameters should be varying concentrations of the cytotoxic agent. See (method 5 www.ab-direct.co.uk/microsite )

The experimental parameters only need to be entered into the lower wavelength table as they are automatically copied across to all other relevant positions. If the experimental parameter titles are too long they will not be displayed correctly until the results page is printed in landscape.

The absorbance results from the spectrophotometer for each time interval should be entered in a format with the decimal place eg. 0.356

To determine the optimum cell density for the cell line that is being used, a control of alamarBlue in culture media, with no cells must be used. (appendix 1 at www.ab-direct.co.uk/microsite).

To determine the cytotoxicity of a chosen agent a control that contains none of the cytotoxic agent, alamarBlue and the cells must be used.