Whole-brain vasculature reconstruction at the single capillary level

The distinct organization of the brain’s vascular network ensures that it is adequately supplied with oxygen and nutrients. However, despite this fundamental role, a detailed reconstruction of the brain-wide vasculature at the capillary level remains elusive, due to insufficient image quality using the best available techniques. Here, we demonstrate a novel approach that improves vascular demarcation by combining CLARITY with a vascular staining approach that can fill the entire blood vessel lumen and imaging with light-sheet fluorescence microscopy. This method significantly improves image contrast, particularly in depth, thereby allowing reliable application of automatic segmentation algorithms, which play an increasingly important role in high-throughput imaging of the terabyte-sized datasets now routinely produced. Furthermore, our novel method is compatible with endogenous fluorescence, thus allowing simultaneous investigations of vasculature and genetically targeted neurons. We believe our new method will be valuable for future brain-wide investigations of the capillary network

Semantic Segmentation of Neuronal Bodies in Fluorescence Microscopy Using a 2D+3D CNN Training Strategy with Sparsely Annotated Data

Semantic segmentation of neuronal structures in 3D high-resolution fluorescence microscopy imaging of the human brain cortex can take advantage of bidimensional CNNs, which yield good results in neuron localization but lead to inaccurate surface reconstruction. 3D CNNs, on the other hand, would require manually annotated volumetric data on a large scale and hence considerable human effort. Semi-supervised alternative strategies which make use only of sparse annotations suffer from longer training times and achieved models tend to have increased capacity compared to 2D CNNs, needing more ground truth data to attain similar results. To overcome these issues we propose a two-phase strategy for training native 3D CNN models on sparse 2D annotations where missing labels are inferred by a 2D CNN model and combined with manual annotations in a weighted manner during loss calculation