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measuring cell proliferation with SpectraMax M5, M2 and Gemini microplate readers using the CyQUANT Cell Proliferation Assay

le of a template with standards and unknowns is shown in Figure 1. Please note that this is the view of the template setup window that appears when the Ctrl and Shift keys are held down simultaneously to display the values assigned to each well. The assigned names of unknowns (Un01-Un12) do not appear in this view.

Step 4. Place the microplate in the microplate reader. Use the appropriate adapter to correctly position the microplate for top reading.

Step 5. Click the softwares Read button. The instrument will read the plate, then the relative fluorescence units (RFUs) will be displayed in the Plate section of SoftMax Pro.


Analyze the data
Step 1. After the microplate has been read, the data will be analyzed in the Group table that was automatically created when you set up the template.

Step 2. In the Graph section, plot the Mean RFU Value versus Concentration (number of cells/well) from the Standards group. Configuration of the Graph Options and Edit dialog boxes is shown in Figure 2.

Step 3. Choose the appropriate curve fit from the drop-down Curve Fit menu in the Graph sections tool bar. When plotting the standard curve for this application note, we used a log-log curve fit. (See Figure 3.) The quadratic curve fit may also be suitable.

Step 4. Determine the number of cells per well for experimental cell samples (unknowns). The SoftMax Pro CyQUANT assay protocol calculates these values from the cell-based standard curve and lists them in the Concentration column of the Unknowns group section.


RESULTS FOR
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