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Abstract Serial section transmission electron microscopy (TEM) has proven to be one of the leading methods for millimeter-scale 3D imaging of brain tissues at nanoscale resolution. It is important to further improve imaging efficiency to acquire larger and more brain volumes. We report here a threefold increase in the speed of TEM by using a beam deflecting mechanism to enable highly efficient acquisition of multiple image tiles (nine) for each motion of the mechanical stage. For millimeter-scale areas, the duty cycle of imaging doubles to more than 30%, yielding a net average imaging rate of 0.3 gigapixels per second. If fully utilized, an array of four beam deflection TEMs should be capable of imaging a dataset of cubic millimeter scale in five weeks. 

Abstract Understanding the brain requires understanding neurons’ functional responses to the circuit architecture shaping them. Here we introduce the MICrONS functional connectomics dataset with dense calcium imaging of around 75,000 neurons in primary visual cortex (VISp) and higher visual areas (VISrl, VISal and VISlm) in an awake mouse that is viewing natural and synthetic stimuli. These data are co-registered with an electron microscopy reconstruction containing more than 200,000 cells and 0.5 billion synapses. Proofreading of a subset of neurons yielded reconstructions that include complete dendritic trees as well the local and inter-areal axonal projections that map up to thousands of cell-to-cell connections per neuron. Released as an open-access resource, this dataset includes the tools for data retrieval and analysis^1,2. Accompanying studies describe its use for comprehensive characterization of cell types^3–6, a synaptic level connectivity diagram of a cortical column⁴, and uncovering cell-type-specific inhibitory connectivity that can be linked to gene expression data^4,7. Functionally, we identify new computational principles of how information is integrated across visual space⁸, characterize novel types of neuronal invariances⁹and bring structure and function together to uncover a general principle for connectivity between excitatory neurons within and across areas^10,11.