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Development of a 2D Automated Tracking System to Characterize Golgi-Derived Membrane Tubule Fission and Fusion Dynamics



Intracellular organelles play an essential role in regulating the biochemistry of various processes and events in eukaryotic cells. Understanding and quantifying their morphological dynamics should allow for their deeper analysis. However, the development of organelle and membrane tracking algorithms is challenging, especially in the case of studying the regulation of Golgi-derived membrane tubules, which show varied sizes and shapes, continuous fission and fusion events, and different directions of movement. Here we have sought to establish a tracking system for tubular subcellular structures, to characterize morphological changes in membrane fission and fusion events.


The development of a tracking algorithm consists of two methods of particle linking: the k-nearest neighbor method applied to successive images with a high temporal resolution, and a modification method named fixed-lag interval smoothing Kalman filtering, for situations of temporary particle disappearance and forward and backward movement.


The system shows excellent efficiency for tracking. One key advantage of this system is that it not only provides track mapping of tubules for fast and convenient interpretation, but also displays tubule labeling and the type of fission and fusion, with corresponding changes of events displayed in the track mapping view. Moreover, tracking measurements of event type and growth rate in length, motility, velocity, and diffusion features are calculated and saved offline use for further analysis.


This prototype tracking system can be applied to different organelles undergoing fission and fusion events, e.g. Golgi-derived tubules and mitochondria, to help biologists quantify and better understand the mechanisms underlying changes in membrane dynamics.

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C. elegans :

Caenorhabditis elegans






Fission event


Fusion event


Forward direction


Backward direction


Brefeldin A


Yellow fluorescent protein


Reference direction


k-Nearest neighbor


Fixed-lag interval smoothing


Left-to-right direction


Right-to-left direction


Fixed interval smoothing


Orientation value


Total event of fission and fusion


Total length of tubules


Mean total length of the tubules


Total distance traveling


Net distance traveling


Maximum distance traveling with a specific path distance (from frame to frame) and the distance values in pixels


Total trajectory time


Confinement ratio


Mean total distance speed


Mean net distance speed


Linearity of forwarding progress


Mean square displacement


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This work is partially supported by the Ministry of Science and Technology, Taiwan, R.O.C under Grant no. MOST 107-2221-E-033-023-MY2. LFH was supported by a postgraduate fellowship from the Irish Research Council (IRC).

Author information




JCS and LFH collected the image data. Conceptualization and study design was performed by JY, YST, and CCL, methodology by JY and YST, implementation by JY, and validation by all authors. All authors interpreted the results data. JY, JCS, and YST wrote the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yuh-Show Tsai or Chung-Chih Lin.

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Yaothak, J., Simpson, J.C., Heffernan, L.F. et al. Development of a 2D Automated Tracking System to Characterize Golgi-Derived Membrane Tubule Fission and Fusion Dynamics. J. Med. Biol. Eng. (2021).

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  • Tracking system
  • Fission
  • Fusion
  • Fixed-lag interval smoothing
  • Kalman filter
  • Track mapping