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Nat Commun. 2018 Nov 9;9(1):4731. doi: 10.1038/s41467-018-07192-z.

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

Author information

1
Department for Bioelectronics, FKE, Vienna University of Technology, Gußhausstraße 25-25A, Building CH, 1040, Vienna, Austria. marko.pende@tuwien.ac.at.
2
Section of Bioelectronics, Center for Brain Research, Medical University of Vienna, Spitalgasse 4, 1090, Vienna, Austria. marko.pende@tuwien.ac.at.
3
Department for Bioelectronics, FKE, Vienna University of Technology, Gußhausstraße 25-25A, Building CH, 1040, Vienna, Austria.
4
Section of Bioelectronics, Center for Brain Research, Medical University of Vienna, Spitalgasse 4, 1090, Vienna, Austria.
5
Department of Neurobiology, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
6
Department of Ecogenomics and Systems Biology, Archaeal Biology and Ecogenomics Division, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
7
Department for Bioelectronics, FKE, Vienna University of Technology, Gußhausstraße 25-25A, Building CH, 1040, Vienna, Austria. hans.ulrich.dodt@tuwien.ac.at.
8
Section of Bioelectronics, Center for Brain Research, Medical University of Vienna, Spitalgasse 4, 1090, Vienna, Austria. hans.ulrich.dodt@tuwien.ac.at.

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.

PMID:
30413688
PMCID:
PMC6226481
DOI:
10.1038/s41467-018-07192-z
[Indexed for MEDLINE]
Free PMC Article

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