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ZEISS Microscopy Knowledge Base

Performing manual tracking on existing segments»

This guide explains how to perform object tracking using existing segments (previously detected).

The segments belonging to the track must be manually selected before applying the pipeline to generate the track.

Opening the working dataset

  1. Select Open... from the file menu.
    File menu showing Open... option highlighted with CTRL+O shortcut
  2. Select the dataset from the file browser.
    The dataset is a multi dimensional, discrete, representation of your real sample volume. It can be structured as a Z series of planes (Optical sectioning) of multiple channels (dyes) in a temporal sequence of time points (located in several spatial positions).​
    Usually, the dataset shows a single experimental situation (a complete experiment can be composed by several datasets). The datasets are available as graphic files saved in plenty of file formats (standard formats as well as proprietary formats)​.
    Note: The dataset is visualized according to the current rendering setting parameters. Refer to the arivis Pro Help for more details.
    Red fluorescence microscopy image with punctate and curved bright structures on black background

Detecting the objects

  1. Run the appropriate pipeline to detect the segments over time.
  2. If required, apply the pre-processing steps followed by the Segmentation tools.
    Sidebar titled Segmentation listing Simple threshold, Smart, Import, Special
    Any segmentation operator can be used to generate the segments. Blob Finder, Watershed, Machine learning tools are available and applicable accordingly to the dataset features.
  3. The segments can also be generated importing labeled images.
    Import pane showing Labeled Image Segmenter option and caption Imports label / binary images as segments
  4. The segments must be detected over time. Therefore, all the time points should show the found objects.
    Black background with scattered small multicolored blobs

Selecting objects manually

  1. Set the active time point as time #1.
    The time points can be manually selected in the following ways:
    Using the scroll bar in the Navigator panel.
    Navigator panel: red microscopy preview with bright red spots, vertical scrollbar and time slider with play controls.
    Using the time tools on the main icon bar (must be set in the preferences).
    Row of five small square icons showing arrows and corner overlays on light grey background.
    Text labels 'Go to Previous Time Point' with 'CTRL+B' and 'Go to Next Time Point' with 'CTRL+F'.
    Using short keys. By default, the time changes are assigned to the CTRL + B / Ctrl + F keys (the keys assignement can be changed in the preferences).
    Note: The short key method is the suggested one.
  2. Set the active cursor mode as following (mandatory):
    Cursor mode icons with white arrow, magnifier, move cross, and highlighted black pointer icon with red curved arrow.
  3. Switch back to the white arrow if you need to interact with the volume.
    Interface showing 'Volume:' checkbox and 'Maximum Intensity' dropdown menu.
    Note: The volume visualization can be disabled if necessary (only in 4D view mode).
  4. Click on the segment you want to identify to be tracked.
  5. The segment will be highlighted either on both the image and on the object table.
    Black background with several colored 3D blobs and a yellow circle highlighting a red blob
  6. Go to the next time point (Ctrl + F).
  7. Click on the same segment to identify it.
  8. Keep the Shift key pressed while clicking on the object.
  9. The segment will be highlighted either on the image and in the object table.
    Close-up on colored 3D blobs on black background with yellow circle around a red blob
  10. Repeat the selection until the segment path is completed.
    Note: From the second time point up to the end of the segment selection (over time), the Shift key MUST be kept pressed while clicking on the object.
  11. On the object table, right-click on one of the selected segments (grayed) to identify it.
    Software object table showing segment list with 'Segment #114 (Blob Finder)' highlighted by a red box
  12. From the pop up menu, select Add Tags...
    Context menu over segment list showing 'Value: "Name"' and 'Add Segment Filter' with 'Segment #114 (Blob Finder)' selected
    Tags menu showing an 'Add Tags...' option highlighted
  13. Type in the new TAG name (e.g. Track#1). Press Add Tag.
    Add Tags dialog showing Tags field with Track#1 and Add Tags and Cancel buttons
    Note: Don‘t click anywhere else. This will deselect all the segments.
  14. The new TAG is shown inside the object table. It includes all the selected segments (please check).
    Panel with checked Track#1 tag and list of segments: #114, #271, #427, #638, #798, #958 (Blob Finder)

Multiple segments can be grouped in different tags following the described procedure.

Tracking pipeline execution

The tracking pipeline must be created. Here below an example:

Input ROI panel: ROI 'Current image set', Tag 'Track#1', Brownian (Centroid), Max 2 µm, Store Objects tracking checked

Input ROI

  1. ROI: Sets the processing and analysis target space.
  2. Current Image Set: The complete dataset (XYZ and time) is processed.
  3. Channel: Not important for this task. Select All Channels.

Import Document Objects

  1. Tag Filter: Sets the TAG(s) previously created (Track#1 and so on) from the manual object's selection procedure.
  2. Tracking: Set the parameters to track the selected segments according to the object's behavior.
  3. Store Objects: Set the data to be transferred to the objects table.

The Tag Filter can import multiple TAGS, each one will generate a single track.

Viewing the results

The track is computed and the data will be transfered to the object table.

The needed features can be selected by using the Feature Columns tool.

Dark green irregular blob on black background with a white-outlined green arrow marker near center pointing up-left.
Objects window screenshot showing highlighted Track #001 (Tracking #1) and a table of segments with speed and acceleration.

Editing tracks

  1. To modify the tracks, open the Track Editor.
    Three small toolbar icons, rightmost icon (monitor with blue dot) outlined in red
    Track Editor window showing Active: Segment #114 (Blob Finder), left track list with Track #001 (Tracking #1): 1-6 and timeline
  2. The tracks are listed on the left panel. The tracked objects belonging to the specific track are shown on the right panel.
  3. Click on the track.
    Track Editor window with Track #001 (Tracking #1): 1-6 highlighted by red outline and timeline with green spots
  4. Select the time point and the segment to be edited. The related frame is shown in the viewing area. The track and the segment are highlighted.
    Timeline closeup showing a blue track row with five green spots and a red arrow pointing upward
  5. Right-click on the selected time point.
  6. A pop up menu is displayed.
    Right-click context menu listing Select branch, Remove spot CANC, Split before spot, Split after spot
    The track can be splitted and merged with another one. The segments in the track path can also be deleted.
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