Tracking image correlation: combining single-particle tracking and image correlation

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • A Dupont - , Center for NanoScience (CeNS) Munich, Ludwig Maximilian University of Munich (Author)
  • K Stirnnagel - , Institute of Medical Microbiology and Virology (Author)
  • D Lindemann - , Institute of Medical Microbiology and Virology (Author)
  • D C Lamb - (Author)

Abstract

The interactions and coordination of biomolecules are crucial for most cellular functions. The observation of protein interactions in live cells may provide a better understanding of the underlying mechanisms. After fluorescent labeling of the interacting partners and live-cell microscopy, the colocalization is generally analyzed by quantitative global methods. Recent studies have addressed questions regarding the individual colocalization of moving biomolecules, usually by using single-particle tracking (SPT) and comparing the fluorescent intensities in both color channels. Here, we introduce a new method that combines SPT and correlation methods to obtain a dynamical 3D colocalization analysis along single trajectories of dual-colored particles. After 3D tracking, the colocalization is computed at each particle's position via the local 3D image cross correlation of the two detection channels. For every particle analyzed, the output consists of the 3D trajectory, the time-resolved 3D colocalization information, and the fluorescence intensity in both channels. In addition, the cross-correlation analysis shows the 3D relative movement of the two fluorescent labels with an accuracy of 30 nm. We apply this method to the tracking of viral fusion events in live cells and demonstrate its capacity to obtain the time-resolved colocalization status of single particles in dense and noisy environments.

Details

Original languageEnglish
Pages (from-to)2373-82
Number of pages10
JournalBiophysical Journal
Volume104
Issue number11
Publication statusPublished - 4 Jun 2013
Peer-reviewedYes

External IDs

PubMedCentral PMC3672895
ORCID /0000-0002-0320-4223/work/150885035
Scopus 84878829404

Keywords

Keywords

  • Biological Transport, Cell Survival, Color, Cytosol/metabolism, Fluorescent Dyes/chemistry, HeLa Cells, Humans, Imaging, Three-Dimensional/methods, Microspheres, Reproducibility of Results, Spumavirus/metabolism