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Live tracking of moving samples in confocal microscopy for vertically grown roots
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 displace...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5498147/ https://www.ncbi.nlm.nih.gov/pubmed/28628006 http://dx.doi.org/10.7554/eLife.26792 |
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author | von Wangenheim, Daniel Hauschild, Robert Fendrych, Matyáš Barone, Vanessa Benková, Eva Friml, Jiří |
author_facet | von Wangenheim, Daniel Hauschild, Robert Fendrych, Matyáš Barone, Vanessa Benková, Eva Friml, Jiří |
author_sort | von Wangenheim, Daniel |
collection | PubMed |
description | 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. DOI: http://dx.doi.org/10.7554/eLife.26792.001 |
format | Online Article Text |
id | pubmed-5498147 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-54981472017-07-07 Live tracking of moving samples in confocal microscopy for vertically grown roots von Wangenheim, Daniel Hauschild, Robert Fendrych, Matyáš Barone, Vanessa Benková, Eva Friml, Jiří eLife Developmental Biology and Stem Cells 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. DOI: http://dx.doi.org/10.7554/eLife.26792.001 eLife Sciences Publications, Ltd 2017-06-19 /pmc/articles/PMC5498147/ /pubmed/28628006 http://dx.doi.org/10.7554/eLife.26792 Text en © 2017, von Wangenheim et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Developmental Biology and Stem Cells von Wangenheim, Daniel Hauschild, Robert Fendrych, Matyáš Barone, Vanessa Benková, Eva Friml, Jiří Live tracking of moving samples in confocal microscopy for vertically grown roots |
title | Live tracking of moving samples in confocal microscopy for vertically grown roots |
title_full | Live tracking of moving samples in confocal microscopy for vertically grown roots |
title_fullStr | Live tracking of moving samples in confocal microscopy for vertically grown roots |
title_full_unstemmed | Live tracking of moving samples in confocal microscopy for vertically grown roots |
title_short | Live tracking of moving samples in confocal microscopy for vertically grown roots |
title_sort | live tracking of moving samples in confocal microscopy for vertically grown roots |
topic | Developmental Biology and Stem Cells |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5498147/ https://www.ncbi.nlm.nih.gov/pubmed/28628006 http://dx.doi.org/10.7554/eLife.26792 |
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