An automated tracking system to measure the dynamic properties of vesicles in living cells

Tien Chuan Ku, Yun Nih Huang, Chien Chang Huang, De Ming Yang, Lung Sen Kao, Tai Yu Chiu, Chung Fan Hsieh, Pei Yo Wu, Yuh Show Tsai*, Chung Chih Lin

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

28 Scopus citations

Abstract

Recent technological improvements have made it possible to examine the dynamics of individual vesicles at a very high temporal and spatial resolution. Quantification of the dynamic properties of secretory vesicles is labor-intensive and therefore it is crucial to develop software to automate the process of analyzing vesicle dynamics. Dual-threshold and binary image conversion were applied to enhance images and define the areas of objects of interest that were to be tracked. The movements, changes in fluorescence intensity, and changes in the area of each tracked object were measured using a new software system named the Protein Tracking system (PTrack). Simulations revealed that the system accurately recognized tracked objects and measured their dynamic properties. Comparison of the results from tracking real time-lapsed images manually with those automatically obtained using PTrack revealed similar patterns for changes in fluorescence intensity and a high accuracy (<89%). According to tracking results, PTrack can distinguish different vesicular organelles that are similar in shape, based on their unique dynamic properties. In conclusion, the novel tracking system, PTrack, should facilitate automated quantification of the dynamic properties of vesicles that are important when classifying vesicular protein locations.

Original languageEnglish
Pages (from-to)119-134
Number of pages16
JournalMicroscopy Research and Technique
Volume70
Issue number2
DOIs
StatePublished - 02 2007
Externally publishedYes

Keywords

  • Fluorescence microscopy
  • Green fluorescent protein
  • PC12 cells
  • Vesicle trafficking

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