Background
The Cytobank platform provides many different types of gates for defining populations. You can access the different gate types (shapes) in the Gating Editor. These gating tools are standard configurations such as rectangle, ellipse and polygon. In addition, the Cytobank platform also provides tools such as quadrant, split and range gates. The quadrant gate also comes with the hinged option to move each quadrant borderline. You can also move the intersection of the quadrants (the center point). Depending upon the target population you can choose which tool to use to create the gates. While some options such as rectangle, polygon or quadrant are available to gating on dot plots, the split and range options are only available when gating on histograms.
Navigating to the gating tools within the Gating Editor
From within an experiment, click on Gates>Gating Editor in the experiment navigation bar.
The gating tools are located on the top of the Gating Editor page.
Check this article to learn more about the Gating Editor interface.
Gating tools on dot and contour plots
To use the gating tools on dot or contour plots, first select the markers of interest on the axes. Click on the Settings icon to access all plot settings and select the plot type.
Then, select the gating tool with the desired shape by clicking on one of the icons on the top of the Gating editor bar:
- To create a rectangular gate, click on the plot where you would like the gate to begin, drag to the desired size and release the mouse click when finished. Alternatively, click where the rectangular gate would start and click again where it would end.
(Dot plot showing the CD56+ expression on CD3 axes in a CD45+ population. The rectangular gate was used to identify the NKT Cells populations.)
- To create an ellipse gate, click on the plot where you would like the center of the ellipse to be, drag to the desired size and release the mouse click when finished. Alternatively, click where the ellipse gate would be centered and click again where it would end.
(Dot plot showing the CD56+ expression on CD3 axes in a CD45+ population. The ellipse gate was used to identify the NKT Cells populations.)
- To create a polygon gate, click on the plot where you would like the polygon to start, continue clicking to determine the shape of the gate and double-click at the initial point to complete the polygon.
(Dot plot showing the CD56+ expression on CD3 axes in a CD45+ population. The polygon gate was used to identify the NKT Cells populations.)
- To create a quadrant gate, click on the plot where you would like the center of the quadrant to be.
(Quadrant plot showing the CD56+ expression on CD3 axes in a CD45+ population. The quadrant gate was used to identify the NKT Cells populations.)
After implementation of the gating tools a pop-up window appears to include the name of the new gate and population generated. A quadrant gate will generate four populations named:
- Gate name LL = Lower Left quadrant
- Gate name LR = Lower Right quadrant
- Gate name UL = Upper Left quadrant
- Gate name UR = Upper Right quadrant
Once the gate has been created you can modify the size by clicking on one of the orange connections and adjusting it. Click inside the gate and drag and drop it to adjust the location of the gate in the plot.
When using a quadrant gate, moving one of the orange connections will hinge the borderline of the quadrant to adjust the angle. The center dot can also be moved. The movement of each quadrant is limited to the plot axis where it was originally drawn.
(Hinged Quadrant plot showing the different monocyte sub-population generated on monocytes. Each axis can be moved independently to encompass 4 different sub-populations of monocytes.)
(Animation showing steps to generate hinged shaped over a quadrant gating. Each axis can be moved independently to encompass 4 different sub-populations of monocytes.)
Gating tools on histogram
To use the gating tools on histograms, first change the Y axis to counts or use the Settings icon to select the histogram Plot type. The Cytobank platform offers you two gating tools to work with histograms: split and range gates:
- To create a split gate, click on the plot where you would like to split the histogram in two.
(Histogram showing the CD3 expression in a CD45+ population. The split gate was used to identify the CD3+ and CD3- populations.)
- To create a range gate, click on the histogram where you would like the gate to begin, drag to the desired size and release the mouse click when finished. Alternatively, click where the range gate would start and click again where it would end.
(Histogram showing the CD3 expression in a CD45+ population. The range gate was used to identify a CD3+ population.)
After implementation of the gating tools a pop-up window appears to include the name of the new gate and population generated. A split gate will generate two populations named:
- Gate name positive = Right part of the histogram
- Gate name negative = Left part of the histogram
Once the gate has been created you can adjust the size by clicking on one of the orange connections and adjusting it.
*For Research Use Only. Not for use in diagnostic procedures.