PT - JOURNAL ARTICLE AU - Daniel von Wangenheim AU - Robert Hauschild AU - Matyáš Fendrych AU - Vanessa Barone AU - Jiří Friml TI - Live tracking of Moving samples in confocal microscopy for vertically grown roots tips AID - 10.1101/116285 DP - 2017 Jan 01 TA - bioRxiv PG - 116285 4099 - http://biorxiv.org/content/early/2017/03/14/116285.short 4100 - http://biorxiv.org/content/early/2017/03/14/116285.full AB - 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.