Three-Dimensional Live Imaging of Filamentous Fungi with Light Sheet-Based Fluorescence Microscopy (LSFM).
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.
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe Universität Frankfurt am Main, Max-von-Laue-Str. 15, D-60438, Frankfurt am Main, Germany. francesco.pampaloni@physikalischebiologie.de.
Francesco
F
Pampaloni
Institute of Molecular Biosciences and Cluster of Excellence Frankfurt Macromolecular Complexes, Department of Biosciences, Goethe Universität Frankfurt am Main, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany.
Laura
L
Knuppertz
Institute of Molecular Biosciences and Cluster of Excellence Frankfurt Macromolecular Complexes, Department of Biosciences, Goethe Universität Frankfurt am Main, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany.
Andrea
A
Hamann
Institute of Molecular Biosciences and Cluster of Excellence Frankfurt Macromolecular Complexes, Department of Biosciences, Goethe Universität Frankfurt am Main, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany.
Heinz D
HD
Osiewacz
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe Universität Frankfurt am Main, Max-von-Laue-Str. 15, D-60438, Frankfurt am Main, Germany.
Ernst H K
EHK
Stelzer
D005658:Fungi / Q000166:cytology*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D012984:Software
Advanced light microscopy; Aging model; Ascomycetes; Autophagy; DSLM; LSFM; Light sheet-based fluorescence microscopy; Live cell imaging; Microbiology; Mycology; Podospora anserina; SPIM