Comparison of two 3D tracking paradigms for freely flying insects

In this paper, we discuss and compare state-of-the-art 3D tracking paradigms for flying insects such as Drosophila melanogaster. If two cameras are employed to estimate the trajectories of these identical appearing objects, calculating stereo and temporal correspondences leads to an NP-hard assignment problem. Currently, there are two different types of approaches discussed in the literature: probabilistic approaches and global correspondence selection approaches. Both have advantages and limitations in terms of accuracy and complexity. Here, we present algorithms for both paradigms. The probabilistic approach utilizes the Kalman filter for temporal tracking. The correspondence selection approach calculates the trajectories based on an overall cost function. Limitations of both approaches are addressed by integrating a third camera to verify consistency of the stereo pairings and to reduce the complexity of the global selection. Furthermore, a novel greedy optimization scheme is introduced for the correspondence selection approach. We compare both paradigms based on synthetic data with ground truth availability. Results show that the global selection is more accurate, while the previously proposed tracking-by-matching (probabilistic) approach is causal and feasible for longer tracking periods and very high target densities. We further demonstrate that our extended global selection scheme outperforms current correspondence selection approaches in tracking accuracy and tracking time

FIM Imaging and FIMtrack: Two New Tools Allowing High-throughput and Cost Effective Locomotion Analysis

The analysis of neuronal network function requires a reliable measurement of behavioral traits. Since the behavior of freely moving animals is variable to a certain degree, many animals have to be analyzed, to obtain statistically significant data. This in turn requires a computer assisted automated quantification of locomotion patterns. To obtain high contrast images of almost translucent and small moving objects, a novel imaging technique based on frustrated total internal reflection called FIM was developed. In this setup, animals are only illuminated with infrared light at the very specific position of contact with the underlying crawling surface. This methodology results in very high contrast images. Subsequently, these high contrast images are processed using established contour tracking algorithms. Based on this, we developed the FIMTrack software, which serves to extract a number of features needed to quantitatively describe a large variety of locomotion characteristics. During the development of this software package, we focused our efforts on an open source architecture allowing the easy addition of further modules. The program operates platform independent and is accompanied by an intuitive GUI guiding the user through data analysis. All locomotion parameter values are given in form of csv files allowing further data analyses. In addition, a Results Viewer integrated into the tracking software provides the opportunity to interactively review and adjust the output, as might be needed during stimulus integration. The power of FIM and FIMTrack is demonstrated by studying the locomotion of Drosophila larvae