This protocol is an excerpt from Sun and Chen,&#160;A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila,&#160;J. Vis. Exp. (2017).
1. Video-assisted Movement Tracking
<ol>
	<li>Fill a 6-cm-diameter Petri dish with transparent silicon elastomer to make the tracking arena. Leave a 3 mm space between the silicon and the dish lid to allow the flies to walk freely but not fly.</li>
	<li>Anesthetize four flies from either the sham or treated group each time and place them in the arena. Leave the flies at 22 &#176;C for 1 h.</li>
	<li>Position a Charge-Coupled Device (CCD) camera above the arenas and record for 5 min.</li>
	<li>Analyze the recorded movement trajectories using Ctrax software (freely available from Caltech). Export the tracked data in a programming language (e.g., Matlab)-compatible format and analyze the data based on distance traveled per frame. Calculate the mean walking distance for each fly and combine it with all other recorded flies/group to obtain a mean cumulative distance travelled by the population of files in the same group.</li>
</ol>

<table><tbody><tr><td>60 mm Petri Dishes</td><td>Fisher Scientific</td><td>FB0875713A</td><td>Used as a tracking arenas</td></tr><tr><td>Flow Regulator</td><td>Genesee Scientific</td><td>59-122WC</td><td></td></tr><tr><td>Fine Brush</td><td>Genesee Scientific</td><td>59-204</td><td></td></tr><tr><td>Flypad</td><td>Genesee Scientific</td><td>59-114</td><td></td></tr><tr><td>Sylgard Silicone Elastomer</td><td>Dow Corning</td><td>4019862</td><td></td></tr><tr><td>CCD Camera</td><td>Microsoft</td><td>HD-5000</td><td></td></tr><tr><td>Ctrax Walking Fly Tracker</td><td>Caltech</td><td>Ctrax 0.2.11</td><td></td></tr><tr><td>MATLAB Image Processing Toolbox</td><td>MATLAB</td><td>R2015b</td><td></td></tr></tbody></table>

tracking freely walking flies: a method to assess locomotion in drosophila

Tracking Freely Walking Flies: A Method to Assess Locomotion in Drosophila

Source: Sun, M. and Chen, L. L. <a href="https://www.jove.com/video/55602/" target="_blank">A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila</a>. J. Vis. Exp. (2017).
This video describes how to track freely walking flies to assess their general locomotion ability. The featured protocol demonstrates the assay&#39;s setup and data analysis using video tracking software.

Source: Sun, M. and Chen, L. L. A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila. J. Vis. Exp. (2017).
This video describes how to ...

- Use a covered arena-- for example, a Petri dish filled with clear silicone-- to ensure flies can walk freely, but are unable to fly inside the arena when it's covered. Place the dish on a white illuminated surface and position a camera directly above. With the lid open, transfer anesthetized flies into the dish, cover it, and allow the subjects to recover. Flies will move freely around the arena during the trial. To assess locomotion, track the position of each fly over time and calculate parameters that characterize locomotion, such as distance traveled by each fly. In this example protocol, we will see a demonstration of the assay setup and analysis using video tracking software. - To track fly movements in a Petri dish, first, fill a 6 centimeter dish with transparent silicone elastomer to make the tracking arena. Leave a 3 millimeter gap between the silicone and the lid, so that flies can walk freely in the gap, but not take flight. Beneath the tracking arena, have a white background. Next, anesthetize four flies from the same treatment group and place them in the arena. Let the flies acclimate for an hour to the arena, which should be fully lit and at about 22 degrees Celsius, not hotter than room temperature. After an hour, use a charge-coupled device camera positioned above the arena to record the fly's activity for 5 minutes. After the 5-minute recording, anesthetize the flies in the arena and return them to a new home vial. Then, dispose of the arena. Later, analyze the recordings using the freely available Ctrax software. This software generates tracking data, which can be exported in a programmable-language-compatible format, such as the MATLAB format. From the data, calculate the distance traveled per frame, the mean walking distance for each fly, and the average distance traveled per fly. For statistical significance, expect to test about 100 flies per treatment group.

tracking-freely-walking-flies-method-to-assess-locomotion

Research

JoVE Encyclopedia of Experiments

Biology

Drosophila melanogaster (fruit fly)

Adult

2.3K Views. - Use a covered arena-- for example, a Petri dish filled with clear silicone-- to ensure flies can ...

Tracking Freely Walking Flies: A Method to Assess Locomotion in Drosophila

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Abstract

Protocol

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Tags

Name	Company	Catalog Number	Comments
60 mm Petri Dishes	Fisher Scientific	FB0875713A	Used as a tracking arenas
Flow Regulator	Genesee Scientific	59-122WC
Fine Brush	Genesee Scientific	59-204
Flypad	Genesee Scientific	59-114
Sylgard Silicone Elastomer	Dow Corning	4019862
CCD Camera	Microsoft	HD-5000
Ctrax Walking Fly Tracker	Caltech	Ctrax 0.2.11
MATLAB Image Processing Toolbox	MATLAB	R2015b