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STED Imaging in Drosophila Brain Slices

  • Sandra Fendl
  • Jesús Pujol-Martí
  • Joel Ryan
  • Alexander Borst
  • Robert KasperEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

STED Drosophila melanogaster Immunofluorescence Cryostat sectioning Brain slice 

Notes

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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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Sandra Fendl
    • 1
  • Jesús Pujol-Martí
    • 1
  • Joel Ryan
    • 2
  • Alexander Borst
    • 1
  • Robert Kasper
    • 1
    Email author
  1. 1.Max Planck Institute of NeurobiologyMartinsried, MunichGermany
  2. 2.LMU Munich, Biocenter MartinsriedMartinsried, MunichGermany

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