When you run a dose response experiment, you can use Cytobank’s Dose Response Analysis functionality (available on hosted models of Cytobank) to generate a dose response curve and determine the EC50, as well as specify a number of statistics associated with the dose response data.
Getting Started with Dose Response Analysis
- Enter the Working Illustration, and proceed with annotation as usual, defining Populations and tagging samples with relevant information such as Stimulation and Timepoints.
- Enable the Dosages figure dimension. Drag the Dosages orange figure dimension box to the second position (this is where it will need to be for the Dose Response View, step 5 below). Click the Setup button to tag your FCS files with dosage information.
- In the Add Doses box, enter a comma-separated list of dosages and click the Add Doses button. When the empty dose boxes appear, assign your FCS files to their appropriate doses. When finished, return to the Illustration.
- Use the Choose button in the orange Dosages figure dimension box to select your desired dosages for use in analysis. Note: Do not choose an 'unstimulated' or '0' dose, as your plots will not render. (True dose response calculations don't use '0' values.)
- Click the Dose Response View link in the blue Working Illustration box on the upper left. Note that the Dose Response page looks very similar to the Working Illustration page; you'll know you are on the Dose Response page because the "Getting Started" box will say "Dose Response" in the header and the first blue box on
the left will be "Working Dose Response.
The first thing you will have to do in order to see dose response curves is to type the numerical dose values into the text boxes in the blue Doses Dimension box on the left, and specify the units in the pull-down menu at the top of that box. Make sure not to enter any letters or special characters into this box – just numbers. Click “Save changes” when done.
- Next, configure your Plot Controls. Specify a statistic (eg, Medians, Percentile, Means), set a Percentile value if you chose this Statistic, select a gate if you chose Percent in Gate as your Statistic, specify a channel from which to pull the data, select a plot size, select a compensation matrix, and choose a dose sort order.
- Click to update the Illustration. You will now see your data points plotted and connected by a dose response curve if the data fit the equation.