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Live tracking of Moving samples in confocal microscopy for vertically grown roots tips

Daniel von Wangenheim, Robert Hauschild, Matyáš Fendrych, Vanessa Barone, Jiří Friml
doi: https://doi.org/10.1101/116285
Daniel von Wangenheim
Institute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, Austria
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Robert Hauschild
Institute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, Austria
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Matyáš Fendrych
Institute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, Austria
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Vanessa Barone
Institute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, Austria
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Jiří Friml
Institute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, Austria
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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.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted March 14, 2017.
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Live tracking of Moving samples in confocal microscopy for vertically grown roots tips
Daniel von Wangenheim, Robert Hauschild, Matyáš Fendrych, Vanessa Barone, Jiří Friml
bioRxiv 116285; doi: https://doi.org/10.1101/116285
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Live tracking of Moving samples in confocal microscopy for vertically grown roots tips
Daniel von Wangenheim, Robert Hauschild, Matyáš Fendrych, Vanessa Barone, Jiří Friml
bioRxiv 116285; doi: https://doi.org/10.1101/116285

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