When you create a region, it appears in the Region List. To
display the list, click the Region List icon in the tool bar or choose Tools > Show Region List. The Region List displays a list of all regions and stat markers (see page 7-30) in the selected Analysis Method, their type, the plots axes, and the region’s color.
• To display all the columns in the Region List window, click Resize.
• To delete a region from the plot, select the region in the Region List and click
Delete. Click Yes in the confirmation box. The region will be deleted from all associated plots. The region name in the Analysis Method will appear as ???.
Global and Local Region Adjustments
You can make adjustments to regions for specific samples within a data set. For example, you may wish to tweak a region for the data in one or more samples to compensate for a shift in the data.
1 To change a region for an individual sample, press and hold the Ctrl key while manipulating the region (elliptic, rectangle, or histogram).
To change the region for multiple samples, highlight the desired wells, then hold the Ctrl key while manipulating the region. The change will be applied to all the
highlighted samples.
The region will turn from red to black, indicating that it is a local region—it applies only to the sample data displayed in the plots.
NOTE: If you manipulate a global region without holding down the Ctrl key, the
change will automatically apply globally (to the data for all samples in the run).Once a local region is created, if you attempt to manipulate any region again (local or global) without holding down the Ctrl key, the following message appears:
• If you wish to keep all local regions local and apply the change to all of the global regions, click Keep Local.
• If you wish to apply the change to all samples in the run, click Make Global. If you want to make an adjustment to a local region to manipulate it further, hold down the Ctrl key to keep the change local and not affect the global regions.
After making a region(s) local, you can still make adjustments to global regions without holding down the Ctrl key. When the dialog box appears, click Keep Local. The adjustment will apply to all global regions. The local regions will remain local and unchanged.
Gates
A gate can be as simple as a single region, or a complex combination of regions and multiple parameters. Gates allow you to further characterize and isolate populations based on the regions. A maximum of 32 gates can be created for one Analysis Method.
Defining a Gate
The gating process has been simplified by dragging and dropping regions. Any region can be dragged from one plot to any other plot. This process allows you to create gates defined with the “AND” operator. You can also create gates using the OR and NOT operators. Refer to “Gate List” on page 7-28.
1 Click to select the region (handles appear), then click in the center of a region and drag it to another plot.
This “applies” the data within the boundaries of the region to the second plot. For example, if you created a region on live cells in plot 1 and drag the live-cell region to plot 2, only the live cells will be displayed in plot 2. A New Gate Name dialog box
prompts you to rename the newly created gate. You can enter a more meaningful name or use the default.
NOTE: You can also define a gate by typing the regions and the operators AND, OR, and NOT. Refer to “Gate List” on page 7-28.If the Analysis Method has gates already defined, you can also select a gate from the
Plot gate menu in the plot tool bar.
When you apply a gate, the following changes occur:
• The events displayed in Plot 2 will automatically change to reflect only those defined by the region that was dragged in. To view the ungated data again, click the Plot gate icon to the right of the plot and select <ungated>.
• The color of the newly gated data will reflect the color of the gate defined in the Gate List window. The default color for the first gate is red.
• The plot heading will reflect the gate applied (for example, Plot P01, gated on P02.R1).
• The newly created gate will appear in the Analysis Methods pane under the Method, in the Gate List (see "Gate List" below), and the Plot gate pop-up menu to the right of the plot.
NOTE: Changes made to the position or size of any region(s) used to define a gate
will automatically be reflected in all plots gated by that region.Select the region, then drag it from one plot to another to apply a gate.
Gate List
When you create a gate, it appears in the Gate List. To display the list, click the Gate List icon in the tool bar or choose Tools > Show Gate List. The Gate List displays a list of all gates in the selected Analysis Method, their definition, and color.
• To display all the columns in the window, click Resize.
• To delete a gate from the list, select the gate and click Delete. Click Yes in the confirmation box.
• To change the gate color, double-click the color in the Color column. The
ColorPickerDialog appears. See “Changing the Color of Gated Data” on page 7-29 for information on using this dialog box to change the color.
• You can turn a region into a gate by entering a name for the gate in the Name field and typing the region name (for example, R1) into Definition field.
• You can also define a gate using the operators AND, OR, and NOT. Create regions to identify the subpopulations of interest. Open the gate list and type an appropriate name for the gate. Type a definition, for example:
• R1 AND R2 means the event must be in both the R1 and R2 regions to be included in the gate.
• R1 OR R2 means the event must be in either the R1 or R2 region to be included in the gate.
• R1 AND (NOT R2) can be used if R1 and R2 overlap and you want to include events in R1 but not in R2.
The gate can now be applied to any plot by clicking the Plot gate icon to the plot tool bar and selecting the gate from the list.
• The ColorGate box allows you to view a backgate. Start by creating a gate in one plot. Then, drag that gate to the plot interest. This applies the gate to the second plot and shows only the gated events. Select Tools > Show Gate List and check the
color. View the second plot without a gate (select <ungated> from the plot tool bar) to see the color gated events in relationship to all events.