1. Developmental Biology and Stem Cells
  2. Plant Biology
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Live tracking of moving samples in confocal microscopy for vertically grown roots

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Cite as: eLife 2017;6:e26792 doi: 10.7554/eLife.26792

Abstract

Roots navigate through soil integrating environmental signals to orient their growth. The Arabidopsis root is a widely used model for developmental, physiological and cell biological studies. Live imaging greatly aids these efforts, but the horizontal sample position and continuous root tip displacement present significant difficulties. Here, we develop a confocal microscope setup for vertical sample mounting and integrated directional illumination. We present TipTracker - a custom software for automatic tracking of diverse moving objects usable on various microscope setups. Combined, this enables observation of root tips growing along the natural gravity vector over prolonged periods of time, as well as the ability to induce rapid gravity or light stimulation. We also track migrating cells in the developing zebrafish embryo, demonstrating the utility of this system in the acquisition of high resolution data sets of dynamic samples. We provide detailed descriptions of the tools enabling the easy implementation on other microscopes.

Article and author information

Author details

  1. Daniel von Wangenheim

    1. Institute of Science and Technology Austria, Klosterneuburg, Austria
    Competing interests
    The authors declare that no competing interests exist.
  2. Robert Hauschild

    1. Institute of Science and Technology Austria, Klosterneuburg, Austria
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon 0000-0001-9843-3522
  3. Matyáš Fendrych

    1. Institute of Science and Technology Austria, Klosterneuburg, Austria
    Competing interests
    The authors declare that no competing interests exist.
  4. Vanessa Barone

    1. Institute of Science and Technology Austria, Klosterneuburg, Austria
    Competing interests
    The authors declare that no competing interests exist.
  5. Eva Benková

    1. Institute of Science and Technology Austria, Klosterneuburg, Austria
    Competing interests
    The authors declare that no competing interests exist.
  6. Jiří Friml

    1. Institute of Science and Technology Austria, Klosterneuburg, Austria
    For correspondence
    1. jiri.friml@ist.ac.at
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon 0000-0002-8302-7596

Funding

Marie Curie Actions (FP7/2007-2013 n 291734)

  • Daniel von Wangenheim

European Research Council (ERC-2011-StG-20101109-PSDP)

  • Jiří Friml

Austrian Science Fund (M 2128-B21)

  • Matyáš Fendrych

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Christian S. Hardtke, Reviewing Editor, University of Lausanne, Switzerland

Publication history

  1. Received: March 14, 2017
  2. Accepted: June 17, 2017
  3. Accepted Manuscript published: June 19, 2017 (version 1)

Copyright

© 2017, von Wangenheim et al

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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