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

Jindaporn Yaothak, Jeremy C. Simpson, Linda F. Heffernan, Yuh Show Tsai*, Chung Chih Lin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Purpose: 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. Methods: 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. Results: 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. Conclusions: 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.

Original languageEnglish
JournalJournal of Medical and Biological Engineering
DOIs
StateE-pub ahead of print - 26 Sep 2021

Keywords

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

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