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Tracking cells with Analysis Pipeline

The segment tracker operator used for connecting moving objects in time, that is finding relationship of objects during a time course. This allows for sophisticated measurements of a group created between a “parent” and “child” objects.
This operator requires two groups of objects, references and subjects, to find relationships between the groups.

Step 1

Prepare Image and Identify Objects



Use Segment generation operators (Intensity Threshold, Blob Finder, and Membrane Segmenter) to identify the objects or cells of interest.

Step 2

Filter Threshold

Add a Segment Filter to exclude over segmented and unwanted objects, or incorrectly detected objects.



Step 3

Setup Segment Tracker Operator



Now add the ‘Segment Tracker” operator, and set the input from the filtered segments, and then select the motion type. Most typical biological samples and direction should select “linear regression” and objects with random motion should select “Brownian motion” option.  Be sure to set the max distance as the maximum distance that an object will move between adjacent time points.  
There is now the choice to use the center of “Intensity” object mass, or the bounding box, or object centroid to use as the center point of the tracked object.

Be sure to rename the operator and color the tracks appropriately




Be sure to rename the operator and color the tracks appropriately. After creating the tracks, new features will be available, e.g. the set of objects over the time course which make up the objects.





Now look in the objects table,  then select the tracked segments, and use the “Drill Down” mode, in conjunction with the Chart, to see features which change over time, such as mean intensity of the selected tracks. Pick and graph as many of the tracks to see information about the moving objects, such as Neutrophils recruited into the epidermis of the zebrafish in this example.




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