Neuroanatomic analysis depends on the reconstruction of complete cell shapes. High-throughput reconstruction of neural circuits, or connectomics, using volume electron microscopy requires dense staining of all cells, which leads even experts to make annotation errors. Currently, reconstruction speed rather than acquisition speed limits the determination of neural wiring diagrams. We developed a method for fast and reliable reconstruction of densely labeled data sets. Our approach, based on manually skeletonizing each neurite redundantly (multiple times) with a visualization-annotation software tool called KNOSSOS, is ~50-fold faster than volume labeling. Errors are detected and eliminated by a redundant-skeleton consensus procedure (RESCOP), which uses a statistical model of how true neurite connectivity is transformed into annotation decisions. RESCOP also estimates the reliability of consensus skeletons. Focused reannotation of difficult locations promises a rather steep increase of reliability as a function of the average skeleton redundancy and thus the nearly error-free analysis of large neuroanatomical datasets.
At a glance
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- Supplementary Text and Figures (594K)
Supplementary Figures 1–5
- Supplementary Movie 1 (68M)
Skeleton tracing with KNOSSOS. 3 orthogonal views are displayed (xy, top left, yz, top right, xz, bottom left) and a 3-dimensional view of the dataset bounding box and the skeleton being created (bottom right). The 19MB version requires a video player compatible with DivX-encoded files. The 68MB version can be viewed with a generic player but is shorter.