title: Genetically targeted 3D visualisation of Drosophila neurons under Electron Microscopy and X-Ray Microscopy using miniSOG.
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 terminal 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 enabled us to target specific regions for EM volume acquisition.
authors.affiliation: Department of Zoology, Downing Street, Cambridge, CB2 3EJ, United Kingdom.|Neurobiology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, CB2 0QH, United Kingdom.
authors.forename: Julian
authors.initials: J
authors.lastname: Ng
authors.affiliation: Carl Zeiss X-ray Microscopy Inc., 4385 Hopyard Rd., Suite 100, Pleasanton, CA 94588, USA.
authors.forename: Alyssa
authors.initials: A
authors.lastname: Browning
authors.affiliation: Carl Zeiss X-ray Microscopy Inc., 4385 Hopyard Rd., Suite 100, Pleasanton, CA 94588, USA.
authors.forename: Lorenz
authors.initials: L
authors.lastname: Lechner
authors.affiliation: Carl Zeiss X-ray Microscopy Inc., 4385 Hopyard Rd., Suite 100, Pleasanton, CA 94588, USA.
authors.forename: Masako
authors.initials: M
authors.lastname: Terada
authors.affiliation: Cell Biology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, CB2 0QH, United Kingdom.
authors.forename: Gillian
authors.initials: G
authors.lastname: Howard
authors.affiliation: Department of Zoology, Downing Street, Cambridge, CB2 3EJ, United Kingdom.|Neurobiology Division, MRC Laboratory of Molecular Biology, Francis Crick Avenue, CB2 0QH, United Kingdom.
authors.forename: Gregory S X E
authors.initials: GSXE
authors.lastname: Jefferis
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001369:Axons / Q000648:ultrastructure; D003712:Dendrites / Q000648:ultrastructure; D004330:Drosophila / Q000235:genetics* / Q000648:ultrastructure; D007089:Image Enhancement; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D009474:Neurons / Q000648:ultrastructure*; D014965:X-Rays