Live tracking of moving samples in confocal microscopy for vertically grown roots

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

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

    Institute of Science and Technology Austria, Klosterneuburg, Austria
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9843-3522
  3. Matyáš Fendrych

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

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

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

    Institute of Science and Technology Austria, Klosterneuburg, Austria
    For correspondence
    jiri.friml@ist.ac.at
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 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, University of Lausanne, Switzerland

Version history

  1. Received: March 14, 2017
  2. Accepted: June 17, 2017
  3. Accepted Manuscript published: June 19, 2017 (version 1)
  4. Version of Record published: July 5, 2017 (version 2)
  5. Version of Record updated: May 2, 2018 (version 3)

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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  1. Daniel von Wangenheim
  2. Robert Hauschild
  3. Matyáš Fendrych
  4. Vanessa Barone
  5. Eva Benková
  6. Jiří Friml
(2017)
Live tracking of moving samples in confocal microscopy for vertically grown roots
eLife 6:e26792.
https://doi.org/10.7554/eLife.26792

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https://doi.org/10.7554/eLife.26792

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