When creating a new analysis setting for your images, you can select the following segmentation methods:

• Segment Region Classes Independently: This method allows you to define several classes and subclasses. With this method, you can define the segmentation algorithm for each class independently.
• ZOI (Zones of Influence): This method constructs a zone of influence (ZOIs) and a ring around each primary object. The primary objects are generated by segmenting the selected image channel with the selected class segmenter. The ring is defined by its width and distance from the primary object. The distance from the ZOI border from the ring can be specified. The ZOI area also incorporates the primary object and ring area.

1. On the Analysis tab, in the Image Analysis tool, click and select New from the dropdown.
2. In the Settings field, enter a name for your image analysis setting and click Save.
3. A new *.czias file is created and saved in the ...\ZEN\Documents\Image Analysis Settings folder.
4. You have created a new image analysis setting. As default the method Segment region classes independently is used.
3. For Method, click .
4. The Segmentation Method Selection dialog opens.
4. From the Method drop down menu, select a method and click OK.
5. You have created a new image analysis setting using the method of your choice.
5. In the Image Analysis tool, click Edit Image Analysis Setting.

1. The Image Analysis Wizard opens with the Classes step and includes already a predefined set of classes, depending on the selected method. Follow the steps in the wizard to define your image analysis. Each method comes with a predefined set of steps which allows you to make all necessary settings for image analysis.

• Classes: Allows you to add classes and subclasses.
• Frame: Allows you to define a measurement frame. Only the area of the frame will be analyzed.
• Region Filter: Allows you to define simple or complex conditions to filter the detected objects according to their parameters.
• Automatic Segmentation: Allows you to set the parameters for the automatic segmentation.
• Interactive Segmentation: Allows you to modify the results of the automatic segmentation or draw/delete objects. Note: this step only generates relevant results in Start Interactive Analysis run.
• Features: Allows you to select measurement features from an extensive list and to define measurement features for classes and subclasses independently.
• Statistics: Allows you to define custom statistical features for your regions or objects.
• Results Preview: Shows a preview of your measurement results for the current view port.

For more information, see the following examples:

• Measuring Fluorescence Intensity in a Multichannel Image
• Counting Number of Fluorescence Signals per Nuclei
• Measuring Mean Fluorescence Intensity on a Ring around the Primary Object
• Counting the Number of Objects in a Ring around the Nucleus