Aivia Software

Phase Cell Tracking

The Phase Cell Tracking recipe in Aivia detects cells and tracks their motility over time in time-lapse phase contrast microscopy images.

This recipe applies a machine-learning-based algorithm for cell segmentation.

The recipe measures the motion attributes of the detected cells for comprehensive characterization of cell migrations.

Parameters and Presets

Parameters

Recipe parameters for Phase Cell Tracking and their descriptions are summarized in the table below.

Preset Group

Parameter Name

Min Value

Max Value

Description

Preset Group

Parameter Name

Min Value

Max Value

Description

Detection

Background Flattening Factor

0

100

Adjusts the sensitivity of the background flattening operation; a lower value will preserve larger objects and more background variations

Contrast Threshold

0

255

Adjusts the detection sensitivity on the machine-learning-enhanced image; a lower value will detect bigger and more objects

Fill Holes Size

0

50,000 px2 or µm2

Adjusts the maximum size threshold for filling in gaps inside a detected object; a lower value will preserve more holes in the detection

Smoothing Factor

0

100

Adjusts the amount of smoothing applied to the outlines of the detected objects; a lower value will preserve more of the objects' morphological features

Tracking

Object Size

0

1,000,000 px2 or µm2

Specifies the range of objects to be included in the analysis results based on the areas of the detected objects

Max Search Range

0

1,000 px or µm

Specifies the maximum distance for track-point matchmaking between successive time frames; a higher value will expand the search distance for fast-moving cells

Min Track Length

0

1,000

Specifies the minimum number of time frames before a detected object is considered a valid track; a lower value will generate more, and often shorter, tracks

Separation Factor

(Cell Partition only)

1

100

Adjusts the sensitivity of the object separation operation; a lower value will preserve larger objects with multiple intensity peaks

Track Lineage Option

Not applicable

Specifies whether to track cell divisions for quantifying lineages

 

Presets

There are two preset groups in the recipe: Detection and Tracking. Each group has three pre-configured parameter groupings to help you get started on the analysis. The default preset values are as follows:

 

Detection

Parameter Name

Low

Medium

High

Background Flattening Factor

50

60

63

Contrast Threshold

185

120

70

Fill Holes Size

50 px2 or µm2

500 px2 or µm2

1,000 px2 or µm2

Smoothing Factor

1

2

4



 

Tracking

Parameter Name

Low

Medium

High

Object Size

1 - 250 px2 or µm2

50 - 1,000 px2 or µm2

150 - 6,000 px2 or µm2

Max Search Range

20 px or µm

20 px or µm

20 px or µm

Min Track Length

8

8

8

Separation Factor

50

70

85



 

Tutorial

Before beginning the tutorial, please download the Phase Cell Tracking Demo image. For information on how to select presets or modify parameter values, please refer to the tutorial on how to use the Recipe Console.

  1. Unzip the demo file and load the demo image, “PhCellTrackDemo.tif,” into Aivia.

  2. In the Recipe Console, click on the Recipe selection dropdown menu and select the Phase Cell Tracking recipe.

  3. Select the Medium presets for both the Detection and Tracking preset groups.

  4. Click the Start button or press the F4 key on your keyboard to begin applying the recipe to the image.

The detected object outlines and tracks will be overlaid on the image.

 

Results

 

Phase Cell Tracking tutorial results

 

Measurements

The Phase Cell Tracking recipe generates track measurements for each detected cell. You can add additional measurements to the analysis results with the Measurement Tool in Aivia and view measurement definitions on the Measurement Definitions page. The measurements generated by the recipe are given in the table below.

Morphology

Position

Morphology

Position

  • Direct Line Length

  • Total Time

  • First Frame

  • Last Frame

  • Direct Line Velocity

  • Acceleration Magnitude

  • Velocity Magnitude