ImarisTrack is the most powerful commercially available live cell imaging, tracking & analysis software that rises to the challenge of monitoring temporal changes in biological systems (2D and 3D images over time). ImarisTrack is based on a choice of multiple sophisticated manual and automatic tracking algorithms and also includes an intuitive and interactive track editing tool if manual correction is necessary. Together this set of tools offers researchers the confidence and support necessary for analysing extremely large and complex data sets. ImarisTrack also provides translational and rotational drift correction which allows researchers to unravel key individual biological events which take place within a complex and dynamic environment (e.g. track a group of defined cells within a developing embryo).
A major challenge to any tracking software is the fact that segmented objects at different time points have no relationship between the time points. The job of a good tracking software package is (i) to correctly identify the objects that should be all part of the same track, (ii) to accurately join them as part of a track, and (iii) to precisely track the objects position even if on occasion the object may disappear or reappear. Tracking software should also deal with objects that remain stationary and happen to be in the path of another mobile object. This may lead to artefacts and inaccuracies in the tracking process that can be easily spotted and corrected manually.
ImarisTrack allows selection from a variety of manual and automatic tracking methods to join these independent objects into tracks easily. Sophisticated algorithms in ImarisTrack can handle objects that appear or disappear. It can use previous object speed and directionality for predicting future positions as well as weighted intensity information to enable the most accurate tracking possible even for complex, densely populated, dynamic and noisy images. In addition to these automated tracking options, ImarisTrack offers an intuitive (click-drag-drop) and flexible track correction tool to allow users to manually edit individual tracks once the automatic tracking is finished. The track editing window shows an illustration of each object on the screen and its connection (track) to objects at other time points. Changes or selections made in the editing window immediately appear in the image view in 3D space providing real-time interactive editing.
In combination with Imaris MeasurementPro, ImarisTrack provides statistical data that is specific to tracking such as straightness and length of tracks; speed, variability and duration tracks; displacement of movement of the object and many more. Because ImarisTrack links independent objects together in a track, users can look at changes in all previously calculated statistical parameters. You can measure distances in 3D and 4D, areas, volumes, voxel intensities, objects’ tracks as well as identify objects based on intensity, size and their changes over time for each set of objects. Like for any other statistical parameter calculated with Imaris MeasurementPro, tracks can be sorted, filtered, classified, selected and grouped with the exact same interface described for Imaris MeasurementPro, but based on these track specific statistics. Likewise, track statistics can be exported to external programs for analysis.
Additionally, ImarisTrack provides a variety of methods to display the tracked results with the original data. This allows for easy verification of the tracking results and the possibility to present the data in a manner that outside observers can quickly understand.
ImarisTrack functionality requires Imaris. For statistical output, ImarisTrack functionality requires Imaris MeasurementPro.
(1) Track Construction – Algorithms
Integrated into the object “Creation Wizard” of Imaris for both “Spot Objects” and “Surface Objects”, ImarisTrack provides an advanced set of algorithms to manually or automatically group segmented objects into tracks over time. All of the tracking methods can be applied to 2D time series or 3D time series (4D) images.
Inspecting or presenting a tracking result visualized as a line connecting all the positions of an object is a natural starting point, but more innovative and flexible options are needed to make sense of images as the X, Y, Z, and t dimensions increase in size, or as the density of objects in the image increases. ImarisTrack not only provides the right variety of tools to clarify complex positional data, but also can map object size, time index, and other statistical information directly to the tracks.
The main point of tracking objects is almost always to measure how they change over time, ImarisTrack, in combination with Imaris MeasurementPro, provides a variety of measurements to facilitate analysis.
ImarisTrack offers a set of tools to modify tracks once they have been created. This feature allows manual creation of tracks, or editing of automatically generated tracks, or for the correction of drift in parts of the image that are moving that should not be.
- All segmented objects (Spots or Surfaces) initially have no relationship to one another between time points
- Tracking links the objects, thus identifying one object as really being the same object at subsequent time points
- Tracks allow the quantification of movements of objects and changes in size, shape, and intensity
Track Path Display
- Display as a line or a cylinder
- Change the diameter of the cylinder or line
- Turn on or off as needed
- Color code the line or cylinder based on time, a single base color, or
on any calculated statistics (Requires Imaris MeasurementPro)
- Display the line as a “dragon tail”- the connections for the last X
number of time points rather than all time points
Displacement
- An arrow showing the distance and direction between the first and last
point of the track
- Turn on or off as needed
- Color and change diameter as desired
Object Display – As described for Imaris, Spot and Surface objects can be displayed along with the track as a center point representation, the actual segmented object, a scaled down version of the actual segmented object, or can be hidden to show only the original data and the track
Track Specific
- Duration, length, straightness
- Displacement, X, Y, Z, total displacement, squared displacement
- Instantaneous velocity X, Y, Z, velocity, average speed, instantaneous
speed, speed variability
- Quality of fit to autoregressive model- per-axis or mean of all axes
Object Specific – Spots or Surfaces
- Changes in size, shape and intensity over time.
- All statistics as described for objects in the MeasurementPro module
become linked on a per track basis
- A plot of each object's statistics per time point is created when the
object is selected. Selection in the graph changes the visualization
time point.
Export
- Export for one track, a group of tracks or all tracks
- Exports both track specific and object specific statistics for each
track
- Export one, a selection or all calculated parameters
Annotation
In combination with Imaris InPress, once objects of interest have been selected the image can be annotated with any calculated statistical parameter in 3D space. The resulting annotations can be exported as a figure with Imaris InPress.
Tracking algorithms are used for the creation of tracks for surface or spot objects
Brownian motion algorithm
- Finds the most likely path connection in any direction, within a distance constraint.
Autoregressive motion algorithm
- Utilizes direction and speed information from previous motion of the
object to predict future locations to search of an object
- Searchers within a distance constraint around the predicted location
- Excellent of objects that have directed movement
Autoregressive motion expert algorithm
- Utilizes all the parameters of the autoregressive motion algorithm
- Adds the possibility to use object intensity as a criteria for
tracking
- Excellent for when objects of varying intensity cross
Connected components algorithm
- Connects objects that would physically overlap if adjacent time points
where superimposed on each other
- Good only for extremely high time resolution
- Allows for merge and split of objects
Manual tracking
- It allows the user to track spot or surface objects within a time-resolved data set by manually specifying the object location in each time point. This mode can be applied when automatic algorithms need user input and/or modifications.
- Auto-connect to selected spot box option activates manual tracking and advances to next time point when a new object is added or an existing is selected
MaxDistance
- The maximum distance that an object moves between two adjacent time
points
- Utilized in the Brownian motion and autoregressive algorithms as the
distance constraint
MaxGapSize
Fill gaps with detected objects
Uses the Autodepth algorithm for easy selection of tracks or objects in 3D space for editing
Fix misconnected/disconnected tracks in the intuitive Track Editor panel.
- Allows for manual correction of problematic areas and connections within tracks
- Point and click method is easy to use
- Click-drag-drop to create new connections
- Allows for objects that split and merge
Correct object’s translational and rotational drift
- After the objects are tracked you can select an object (or group of objects) that should not be moving during your time series and use it (them) as an anchor/reference point(s).
- Images are shifted on a frame to frame basis so that the selected anchor objects no longer move and the true movement of the other objects can properly be assessed.
Rebuild – The tracking algorithm may be repeated without the need for recreating / re-segmenting the objects
Like surfaces and spots objects, tracks may be filtered as part of the creation wizard based on any of the statistical parameters calculated for those tracks.
Tracks may also be filtered within the filter tab after creation is complete.
Watch this video tutorial and learn how to interact with tracks if tracking needs to be adjusted.
Watch this video to learn how to identify, follow 3D-objects over time, display their paths and analyze their movements.
Watch this video to learn how to identify, follow 2D-objects over time, display their paths and analyze their movements.
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