Abstract
The manner in which the nervous system regulates animal behaviors in natural environments is a fundamental issue in biology. To address this question, C. elegans has been widely used as a model animal for the analysis of various animal behaviors. Previous behavioral assays have been limited to two-dimensional (2-D) environments, confining the worm motion to a planar substrate that does not reflect three-dimensional (3-D) natural environments such as rotting fruits or soil. Here, we develop a 3-D worm tracker (3DWT) for freely moving C. elegans in 3-D environments, based on a stereoscopic configuration. The 3DWT provides us with a quantitative trajectory, including the position and movement direction of the worm in 3-D. The 3DWT is also capable of recording and visualizing postures of the moving worm in 3-D, which are more complex than those in 2-D. Our 3DWT affords new opportunities for understanding the nervous system function that regulates animal behaviors in natural 3-D environments.
Highlights
The nematode C. elegans has been widely used as a model animal for behavioral neuroscience due to its experimental amenability, fully sequenced genome and simple nervous system
We introduce a 3-D worm tracker (3DWT) for freely moving C. elegans in 3-D environments based on a stereoscopic configuration
Using the 3DWT, we provide a quantitative analysis of C. elegans locomotion in 3-D environments in terms of its trajectory and kinematics
Summary
The nematode C. elegans has been widely used as a model animal for behavioral neuroscience due to its experimental amenability, fully sequenced genome and simple nervous system. A single-worm tracker tracks each worm precisely by following it with motorized stages This method permits high-resolution motion analysis and optical manipulation of neural activity [2,3]. In a multi-worm tracker, in contrast, multiple worms are simultaneously monitored on a single plate or multiple wells and are tracked by image-analysis software with a high throughput [4,5,6,7]. The imaging in these two approaches, has been limited to two dimensions (2-D). Using the 3DWT, we provide a quantitative analysis of C. elegans locomotion in 3-D environments in terms of its trajectory and kinematics
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