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Validation
A synthetic video of five cells with changing shape was used in the evaluation of tracking software. Each cell was tracked using respective tracking software, and their coordinates in pixels were obtained.
The following steps were taken to analyse tracking data:
- Convert synthetic trajectory coordinate points into pixels.
- Calculate the displacement in each frame for both synthetic and tracking data
- Calculate the distance travelled in each frame for both synthetic and tracking data
- Obtain the errors in coordinates, dislacement and distance travelled by the cell
The errors associated with manual tracking and Volocity software are shown below. All errors are given in terms of pixels.
Manual Tracking
Error in X Coordinate
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Error in Y Coordinate
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Mean = 1.7354, Standard Deviation = 1.7187
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Mean = 1.2524, Standard Deviation = 1.0568
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Error in X Displacement
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Error in Y Displacement
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Error in Distance
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Mean = -0.0022, Standard Deviation = 1.0002
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Mean = -0.0018, Standard Deviation = 0.6795
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Mean = -0.1506, Standard Deviation = 0.9196
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Volocity
Error in X Coordinate
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Error in Y Coordinate
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Mean = -1.7149, Standard Deviation = 0.1376
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Mean = -0.4907, Standard Deviation = 0.1614
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Error in X Displacement
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Error in Y Displacement
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Error in Distance
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Mean = 8.2732e-005, Standard Deviation = 0.1111
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Mean = 6.1416e-004, Standard Deviation = 0.0712
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Mean = 1.4262e-004, Standard Deviation = 0.0947
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Evaluation
Volocity
Pros
- Lower error in terms of tracking centroid than other tracking software.
- Less time consuming.
Cons
- Failure to track cells continuously over a long number of frames > 70
- Baised to cells which run, rather than tumble
Manual Tracking
Pros
- Able to discern cells visually
- Able to track over larger number of frames
Cons
- Time consuming
- Higher error in terms of tracking centroid as compared to tracking software.
Conclusion
- Manual tracking will be used to track B.Subtilis.
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