title: Large-volume en-bloc staining for electron microscopy-based connectomics.
abstract: Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic.
authors.affiliation: Department of Connectomics, Max Planck Institute of Brain Research, Max-von-Laue-Strasse 4, D-60438 Frankfurt, Germany.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Hua
authors.affiliation: Department of Connectomics, Max Planck Institute of Brain Research, Max-von-Laue-Strasse 4, D-60438 Frankfurt, Germany.
authors.forename: Philip
authors.initials: P
authors.lastname: Laserstein
authors.affiliation: Department of Connectomics, Max Planck Institute of Brain Research, Max-von-Laue-Strasse 4, D-60438 Frankfurt, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Helmstaedter
mesh_terms: D000818:Animals; D063132:Connectome / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008854:Microscopy, Electron / Q000379:methods*; D019579:Neocortex / Q000648:ultrastructure*; D016501:Neurites / Q000648:ultrastructure*; D009457:Neuroglia / Q000648:ultrastructure*; D009474:Neurons / Q000648:ultrastructure*; D013194:Staining and Labeling / Q000379:methods*