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Genetically targeted 3D visualisation of Drosophila neurons under Electron Microscopy and X-Ray Microscopy using miniSOG

Julian Ng, Alyssa Browning, Lorenz Lechner, Masako Terada, Gillian Howard, Gregory S. X. E. Jefferis
doi: https://doi.org/10.1101/070755
Julian Ng
1Department of Zoology, Downing Street, Cambridge, CB2 3EJ, United Kingdom
4Neurobiology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, CB2 0-H, United Kingdom
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Alyssa Browning
2Carl Zeiss X-ray Microscopy Inc., 4385 Hopyard Rd., Suite 100, Pleasanton, CA 94588, USA
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Lorenz Lechner
2Carl Zeiss X-ray Microscopy Inc., 4385 Hopyard Rd., Suite 100, Pleasanton, CA 94588, USA
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Masako Terada
2Carl Zeiss X-ray Microscopy Inc., 4385 Hopyard Rd., Suite 100, Pleasanton, CA 94588, USA
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Gillian Howard
3Cell Biology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, CB2 0-H, United Kingdom
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Gregory S. X. E. Jefferis
1Department of Zoology, Downing Street, Cambridge, CB2 3EJ, United Kingdom
4Neurobiology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, CB2 0-H, United Kingdom
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Abstract

Large dimension, high-resolution imaging is important for neural circuit visualisation as neurons have both long- and short-range patterns: from axons and dendrites to the numerous synapses at their endings. Electron Microscopy (EM) is the favoured approach for synaptic resolution imaging but how such structures can be segmented from high-density images within large volume datasets remains challenging.

Fluorescent probes are widely used to localise synapses, identify cell-types and in tracing studies. The equivalent EM approach would benefit visualising such labelled structures from within sub-cellular, cellular, tissue and neuroanatomical contexts.

Here we developed genetically-encoded, electron-dense markers using miniSOG. We demonstrate their ability in 1) labelling cellular sub-compartments of genetically-targeted neurons, 2) generating contrast under different EM modalities, and 3) segmenting labelled structures from EM volumes using computer-assisted strategies. We also tested non-destructive X-ray imaging on whole Drosophila brains to evaluate contrast staining. This enables us to target specific regions for EM volume acquisition.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted August 22, 2016.
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Genetically targeted 3D visualisation of Drosophila neurons under Electron Microscopy and X-Ray Microscopy using miniSOG
Julian Ng, Alyssa Browning, Lorenz Lechner, Masako Terada, Gillian Howard, Gregory S. X. E. Jefferis
bioRxiv 070755; doi: https://doi.org/10.1101/070755
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Genetically targeted 3D visualisation of Drosophila neurons under Electron Microscopy and X-Ray Microscopy using miniSOG
Julian Ng, Alyssa Browning, Lorenz Lechner, Masako Terada, Gillian Howard, Gregory S. X. E. Jefferis
bioRxiv 070755; doi: https://doi.org/10.1101/070755

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