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STED Imaging in Drosophila Brain Slices

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Light Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1563))

Abstract

Super-resolution microscopy is a very powerful tool to investigate fine cellular structures and molecular arrangements in biological systems. For instance, stimulated emission depletion (STED) microscopy has been successfully used in recent years to investigate the arrangement and colocalization of different protein species in cells in culture and on the surface of specimens. However, because of its extreme sensitivity to light scattering, super-resolution imaging deep inside tissues remains a challenge. Here, we describe the preparation of thin slices from the fruit fly (Drosophila melanogaster) brain, subsequent immunolabeling and imaging with STED microscopy. This protocol allowed us to image small dendritic branches from neurons located deep in the fly brain with improved resolution compared with conventional light microscopy.

*Author contributed equally with other authors.

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Acknowledgments

We are indebted to H. Leonhardt and the BioImaging Network Munich for generous support. We thank Marianne Braun and Ursula Weber for excellent help with technical procedures, and Aljoscha Nern, Gerald M. Rubin, and Barry Dickson for providing transgenic flies.

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    Authors and Affiliations

    1. Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152, Martinsried, Munich, Germany

      Sandra Fendl, Jesús Pujol-Martí, Alexander Borst & Robert Kasper

    2. LMU Munich, Biocenter Martinsried, Grosshadernerstr. 2, 82152, Martinsried, Munich, Germany

      Joel Ryan

    Authors
    1. Sandra Fendl
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    2. Jesús Pujol-Martí
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    3. Joel Ryan
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    4. Alexander Borst
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    5. Robert Kasper
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    Corresponding author

    Correspondence to Robert Kasper .

    Editor information

    Editors and Affiliations

    1. Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, California, USA

      Yolanda Markaki

    2. Biozentrum der LMU Munchen, Planegg-Martinsried, Germany

      Hartmann Harz

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    Fendl, S., Pujol-Martí, J., Ryan, J., Borst, A., Kasper, R. (2017). STED Imaging in Drosophila Brain Slices. In: Markaki, Y., Harz, H. (eds) Light Microscopy. Methods in Molecular Biology, vol 1563. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6810-7_10

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    • DOI: https://doi.org/10.1007/978-1-4939-6810-7_10

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-6808-4

    • Online ISBN: 978-1-4939-6810-7

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