Imaging the molecular constituents of the brain vasculature and lymphatic connectome

PIS：大卫·柯菲德和吴柱豪 项目摘要 脑血管和淋巴管连接体的分子组成成像 延伸至颅骨和脑膜淋巴管的脑血管的复杂连通性 在系统的脑血流循环、免疫反应和废物清除中都是至关重要的。 然而，我们对这些复杂网络的了解仍然有限，这主要是因为 在非均质中以高分辨率成像这些结构的完整三维结构 生物组织。 两个方面的最新技术进展，一个是在分子表征和标记方面，另一个是在组织方面 清除，导致了头骨-脑膜血管连接的发现。然而，波前失真通过 不完全清除的头骨和脑组织严重降低了深度成像的分辨率。在这里我们 建议改进样品制备和成像技术，使其与标记和 减少分辨率方面的损失。我们的光学方法包括非线性成像和使用飞秒 消融术伴有骨和软组织的颞部聚焦。这将使我们能够获得衍射限制的分辨率 头骨/脑膜/大脑过渡区的图像。 这个试点项目将建立一条管道，对整个血管系统和淋巴管进行成像和重建 在小鼠大脑中的细胞分辨率系统。这是成像和成像的长期目标的前奏 为身体所有器官重建相似的地图集。

PIs: David Kleinfeld and Zhuhao Wu Project Summary Imaging the molecular constituents of the brain vasculature and lymphatic connectome The intricate connectivity of the brain vasculature that extends into the skull and the meningeal lymphatic vessels are vital in the systematic cerebral blood flow circulation, immunological responses, and waste product removal. However, our understanding of these complex networks remains limited, largely because of the challenges in imaging the complete three-dimensional structure of these structure at high resolution in heterogeneous biological tissues. Recent technical advances on two fronts, one in molecular characterization and labeling, and the other in tissue clearing, led to the discovery of skull-meninge vascular connections. However, wavefront distortion through imperfectly cleared skull and brain tissue severely deteriorate the resolution of imaging at depth. Here we propose to advance sample preparation and imaging technologies that are compatible with labeling and that mitigate the loss in resolution. Our optical methods involve non-linear imaging as well as the use of femtosecond ablation with temporal focussing of bone and soft tissue. This will enable us to obtain diffraction-limited resolution images at the skull/meningeal/brain transition zone. This pilot project will establish a pipeline to image and reconstruct the entire vasculature system and lymphatic system at cellular resolution in the mouse brain. This is a prelude to the long-term goal of imaging and reconstructing similar atlases for all organs in the body.