High-resolution ultramicroscopy of the developing and adult nervous system in optically cleared Drosophila melanogaster

Marko Pende (), Klaus Becker, Martina Wanis, Saiedeh Saghafi, Rashmit Kaur, Christian Hahn, Nika Pende, Massih Foroughipour, Thomas Hummel and Hans-Ulrich Dodt ()
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Marko Pende: Vienna University of Technology
Klaus Becker: Vienna University of Technology
Martina Wanis: Vienna University of Technology
Saiedeh Saghafi: Vienna University of Technology
Rashmit Kaur: University of Vienna
Christian Hahn: Vienna University of Technology
Nika Pende: University of Vienna
Massih Foroughipour: Medical University of Vienna
Thomas Hummel: University of Vienna
Hans-Ulrich Dodt: Vienna University of Technology

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: Abstract The fruit fly, Drosophila melanogaster, is an important experimental model to address central questions in neuroscience at an organismic level. However, imaging of neural circuits in intact fruit flies is limited due to structural properties of the cuticle. Here we present a novel approach combining tissue clearing, ultramicroscopy, and data analysis that enables the visualisation of neuronal networks with single-cell resolution from the larval stage up to the adult Drosophila. FlyClear, the signal preserving clearing technique we developed, stabilises tissue integrity and fluorescence signal intensity for over a month and efficiently removes the overall pigmentation. An aspheric ultramicroscope set-up utilising an improved light-sheet generator allows us to visualise long-range connections of peripheral sensory and central neurons in the visual and olfactory system. High-resolution 3D reconstructions with isotropic resolution from entire GFP-expressing flies are obtained by applying image fusion from orthogonal directions. This methodological integration of novel chemical, optical, and computational techniques allows a major advance in the analysis of global neural circuit organisation.

Date: 2018
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DOI: 10.1038/s41467-018-07192-z

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