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A protocol for dissecting Drosophila melanogaster brains for live imaging or immunostaining

Nature Protocols volume 1, pages 2110–2115 (2006)Cite this article

Abstract

This protocol describes a basic method for dissection and immunofluorescence staining of the Drosophila brain at various developmental stages. The Drosophila brain has become increasingly useful for studies of neuronal wiring and morphogenesis in combination with techniques such as the 'mosaic analysis with a repressible cell marker' (MARCM) system, where single neurons can be followed in live and fixed tissues for high-resolution analysis of wild-type or genetically manipulated cells. Such high-resolution anatomical study of the brain is also important in characterizing the organization of neural circuits using genetic tools such as GAL4 enhancer trap lines, as Drosophila has been intensively used for studying the neural basis of behavior. Advantages of fluorescence immunostaining include compatibility with multicolor labeling and confocal or multiphoton imaging. This brain dissection and immunofluorescence staining protocol requires approximately 2 to 6 d to complete.

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Figure 1: Visualizing neurons in the Drosophila brain.

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Acknowledgements

We thank members of our laboratory for their helpful comments on this protocol. Research in our lab has been generously supported by grants from the US National Institutes of Health, and more recently from the Howard Hughes Medical Institute, for which L.L. is an investigator.

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Authors and Affiliations

  1. Department of Biological Sciences, Howard Hughes Medical Institute, Neurosciences Program, Stanford University, Stanford, 94305-5020, California, USA

    Joy S Wu & Liqun Luo

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  1. Joy S Wu
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Correspondence to Liqun Luo.

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Wu, J., Luo, L. A protocol for dissecting Drosophila melanogaster brains for live imaging or immunostaining. Nat Protoc 1, 2110–2115 (2006). https://doi.org/10.1038/nprot.2006.336

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