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Video tracking and analysis of sleep in Drosophila melanogaster

Abstract

In the past decade, Drosophila has emerged as an ideal model organism for studying the genetic components of sleep as well as its regulation and functions. In fruit flies, sleep can be conveniently estimated by measuring the locomotor activity of the flies using techniques and instruments adapted from the field of circadian behavior. However, proper analysis of sleep requires degrees of spatial and temporal resolution higher than is needed by circadian scientists, as well as different algorithms and software for data analysis. Here I describe how to perform sleep experiments in flies using techniques and software (pySolo and pySolo-Video) previously developed in my laboratory. I focus on computer-assisted video tracking to monitor fly activity. I explain how to plan a sleep analysis experiment that covers the basic aspects of sleep, how to prepare the necessary equipment and how to analyze the data. By using this protocol, a typical sleep analysis experiment can be completed in 5–7 d.

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Figure 1: Principles of locomotor detection.
Figure 2: Experimental workflow.
Figure 3: Translating motor activity to sleep.
Figure 4: Components of Drosophila sleep.

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Acknowledgements

I am deeply indebted to A. Keene for his help with a first version of this manuscript. I would like to thank the open source community for creating and sharing valuable software. The work described here was partly funded by a Royal Society Research grant (RG110197) to G.F.G.

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Correspondence to Giorgio F Gilestro.

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The author declares no competing financial interests.

Supplementary information

Supplementary Fig. 1

A. The recording monitor. B. A printed arena. The transparent lid is secured using a rubber band. The middle chamber is filled with food. C. A3D printed arena compared to a glass tube traditionally used for infrared beam-split analysis. The size of the walking chamber is almost identical. (EPS 9269 kb)

Supplementary Table 1

Specialised software for computer assisted behavioural analysis. (DOC 28 kb)

Supplementary Table 2

Comparing costs of video tracking and infrared beam-split. (DOC 18 kb)

Supplementary Video 1

Installing pySolo and pySolo video repositories in Ubuntu Linux A brief screencast guiding the user through the steps required to install the pySolo suite in Ubuntu linux adding custom repositories. The repositories will take automatic care of future updates. (MP4 8097 kb)

Supplementary Video 2

How to build a recording monitor This video shows the basic components of a recording monitor and the criteria for building one of your own. (MP4 36799 kb)

Supplementary Video 3

Preparing the arena for recording How to prepare an arena for recording: from transferring the food to placing the flies. (MP4 50524 kb)

Supplementary Video 4

Configuring the datafetcher script Configuring the datafetcher for automatic fetching and formatting of data. Can be used for pySolo Video or for the original TriKinetics Software. (MP4 21499 kb)

Supplementary Video 5

An introduction to pySolo Video A brief introduction showing how to configure and use pySolo Video. (MP4 18455 kb)

Supplementary Video 6

Using pySolo for data analysis A detailed guide with complete instructions on how to use pySolo for data analysis. (MP4 42015 kb)

Supplementary Data

The zip file contains an electronic drawing of a complete arena, as shown in Supplementary Figure 1b-c. The unzipped file, provided here in STL format, was created using the free software Blender (http://www.blender.org) and can be opened and viewed using the free STL viewer meshlab (http://meshlab.sourceforge.net) (ZIP 64 kb)

Supplementary Manual

Installing, configuring and using PySolo for sleep analysis. (PDF 601 kb)

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Gilestro, G. Video tracking and analysis of sleep in Drosophila melanogaster. Nat Protoc 7, 995–1007 (2012). https://doi.org/10.1038/nprot.2012.041

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