RT Journal Article
SR Electronic
T1 Live tracking of Moving samples in confocal microscopy for vertically grown roots tips
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 116285
DO 10.1101/116285
A1 Daniel von Wangenheim
A1 Robert Hauschild
A1 Matyáš Fendrych
A1 Vanessa Barone
A1 Jiří Friml
YR 2017
UL http://biorxiv.org/content/early/2017/03/14/116285.abstract
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.