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Three-Dimensional Live Imaging of Filamentous Fungi with Light Sheet-Based Fluorescence Microscopy (LSFM)

  • Francesco PampaloniEmail author
  • Laura Knuppertz
  • Andrea Hamann
  • Heinz D. Osiewacz
  • Ernst H. K. Stelzer
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1563)

Abstract

We describe a method for the three-dimensional live imaging of filamentous fungi with light sheet-based fluorescence microscopy (LSFM). LSFM provides completely new opportunities to investigate the biology of fungal cells and other microorganisms with high spatial and temporal resolution. As an example, we study the established aging model Podospora anserina. The protocol explains the mounting of the live fungi for the light sheet imaging, the imaging procedure and illustrates basic image processing of data.

Key words

Advanced light microscopy Light sheet-based fluorescence microscopy Live cell imaging LSFM SPIM DSLM Podospora anserina Ascomycetes Mycology Microbiology Aging model Autophagy 

Notes

Acknowledgments

The authors thank the LOEWE initiative of the Federal State of Hessen (Ub-Net, speaker Ivan Dikic) and the Deutsche Forschungsgemeinschaft (DFG, Os75/15-1, and Excellenzcluster für Makromolekulare Komplexe, CEF-MC II, EXC 115, speaker Volker Doetsch) for financial support.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Francesco Pampaloni
    • 1
    Email author
  • Laura Knuppertz
    • 2
  • Andrea Hamann
    • 2
  • Heinz D. Osiewacz
    • 2
  • Ernst H. K. Stelzer
    • 1
  1. 1.Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS)Goethe Universität Frankfurt am MainFrankfurt am MainGermany
  2. 2.Institute of Molecular Biosciences and Cluster of Excellence Frankfurt Macromolecular Complexes, Department of BiosciencesGoethe Universität Frankfurt am MainFrankfurt am MainGermany

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