Live tracking of moving samples in confocal microscopy for vertically grown roots
- Institute of Science and Technology Austria, Austria
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
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
- Christian S. Hardtke, University of Lausanne, Switzerland
Version history
- Received: March 14, 2017
- Accepted: June 17, 2017
- Accepted Manuscript published: June 19, 2017 (version 1)
- Version of Record published: July 5, 2017 (version 2)
- 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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Live tracking of moving samples in confocal microscopy for vertically grown roots
eLife 6:e26792.
https://doi.org/10.7554/eLife.26792
