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Fig. 3 | BMC Biology

Fig. 3

From: A multi-animal tracker for studying complex behaviors

Fig. 3

The Multi-Animal Tracker is particularly suitable for studying complex behaviors such as chemotaxis. a An image of the experimental chemotaxis plate. Agar plugs soaked with the attractant odorant (red circle), or the control buffer (yellow circle), are placed on the plate’s lid. Neither the attractant, nor the buffer, come in contact with the agar on the plate. Approximately 150 worms are loaded at the starting point (blue circle). The chemoattractant source, the buffer source, and the starting point form an imaginary equilateral triangle with an edge of 4 cm. b A quantitative cumulative dynamics of worm position in the chemotaxis assay over the course of approximately 15 minutes. The lines indicate the number of worms in each of the regions of interest throughout the experiment (color coded). In this experiment, nearly 180 worms were loaded on the assay plate. About two-thirds of them reached the chemoattractant during the first 15 minutes of the assay. c Images were taken at the specific time points (I–IV) throughout the assay (indicated as dashed lines in b) to illustrate chemotaxis progression in the assay plate. Shown are also the trajectories (red) as identified by the tracker software. dg The software suite includes a module to generate Attraction Fields (AF), a visualization designed to provide spatial representation of the chemotaxis process throughout the experiment. Shown here are AFs of two chemotaxis assays in which the isoamyl-alcohol attractant was used in 10−4 (d) and 10−2 (f) dilutions. The experimental field is binned to squares (35 × 25 in this case, but any binning defined by the user is possible). Arrows represent the average direction of the worms and the color code indicates the overall occupancy throughout the course of the experiment (15 minutes, ~1000 frames). e, g Chemotaxis dynamics as detailed in section b. Together, these plots provide a full spatiotemporal representation of the chemotaxis behavior of multiple animals over the course of thousands of captured frames

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