High-Resolution 3D Mapping of Human Cortical Vasculature

Abstract

A quantitative description of the three-dimensional organization of the human cortical vasculature at micron scale is critical for understanding the influence of the vascular architecture on cerebral blood flow, neurovascular coupling, neurological disease, and human neuroimaging signals. However, high-resolution volumetric vascular data from human cortex are scarce, as the acquisition relies on postmortem microscopy, and tissue-clearing and whole-mount immunolabeling approaches developed for rodent tissues are less effective in heavily crosslinked, pigment rich fixed human cortex. Here we introduce h-iDISCO+, a workflow that enables whole-mount immunolabeling and high-resolution light-sheet imaging of arteries, capillaries, and veins in long-term formalin fixed human brain tissue. By integrating extended oxidative and photobleaching prior to tissue staining, we achieve uniform sample transparency and full-depth antibody penetration. Applying this workflow to tissue samples from human primary visual cortex, we quantified vessel geometrical properties and vascular density across cortical depth. This approach allows quantitative reconstruction of human cortical vascular networks at micron resolution, enabling volumetric datasets of the human cortical vasculature that were previously unavailable.