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- Evaluate imaging and reconstruction time tradeoffs. Imaging at a higher resolution could improve downstream automation and proofreading, which is currently the bottleneck. With the recent reliability improvements to FIB-SEM, we can now more thoroughly examine this tradeoff.
- Start imaging hot-knife sections from the whole fly brain. We need to evaluate the hot-knife imaging methodology over large brain regions to enable analysis of important biological circuits.
- Improve tissue preparation to reduce membrane holes. During our sample preparation, we often discover broken membranes that hinder automatic segmentation algorithms. We aim to find preparation protocols that greatly reduce this problem.
- Label gap junctions in dataset. Electrical coupling of neurons through gap junctions is not evident at the production resolution of FIB-SEM (8nm x 8nm x 8nm). We aim to explore several strategies to identify gap junctions including extra-cellular sample preparation, fluorescent labeling and correlating LM with EM, very high-resolution EM imaging, and electron dense labeling of gap junctions.
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