Large-volume en-bloc staining for electron microscopy-based connectomics

Nat Commun. 2015 Aug 3:6:7923. doi: 10.1038/ncomms8923.


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.

MeSH terms

  • Animals
  • Connectome / methods*
  • Imaging, Three-Dimensional / methods*
  • Mice
  • Microscopy, Electron / methods*
  • Neocortex / ultrastructure*
  • Neurites / ultrastructure*
  • Neuroglia / ultrastructure*
  • Neurons / ultrastructure*
  • Staining and Labeling / methods*