An Accessible Optical Toolbox for Saturated Nanoscale Analysis of Neural Architecture

对神经元完整结构的纳米级重建提供了 对正常和疾病大脑中神经元回路研究的潜在革命性进展 但这一承诺远未实现，因为更大的神经科学界无法 访问提供大容量和纳米级重建的技术 分析它们所需的计算基础设施。因此，神经解剖学在 纳米尺度仅限于较小体积的一次性研究，很少延伸到过去 单个动物的小体积重建(n&lt；=1)，只有符合以下条件的实验室才能访问 能够负担得起所需的非凡的时间投入、劳动力和费用 即使是简单的神经元回路也能重建。在这里，我们提出了一种新的方法： 开发越来越复杂的硬件来进行纳米级的重建 大脑，我们建议开发一个工具箱，包括可访问的分子、化学和 大体积物体纳米尺度精确重建的计算方法 脑靶向荧光显微镜和传统的共聚焦显微镜- 神经科学武器库中使用最广泛的两种工具。通过提供一个 可接近的管道重建脑细胞之间的自然形状和空间 在活体大脑中发现了广泛的实验条件，我们将帮助交付 获取更广泛的神经科学的全面纳米级神经解剖学 社区。

Nanoscale-resolution reconstructions of the complete architectures of neurons offer the potential to revolutionize the study of neuronal circuitry in normal and diseased brains but this promise is far from realized because the larger neuroscience community cannot access the technologies that provide nanoscale reconstructions over large volumes and the computational infrastructure required to analyze them. As a result, neuroanatomy at the nanoscale is restricted to one-off studies of smaller volumes, rarely extending past small volume reconstructions in single animals (n<=1), and accessible only to labs that can afford the extraordinary time commitment, labor, and expense required to reconstruct even simple neuronal circuits. Here we propose a novel approach: rather than developing more and more sophisticated hardware for nanoscale reconstructions of the brain, we propose to develop a toolbox of accessible molecular, chemical, and computational approaches for accurate nanoscale reconstructions of large volumes of the brain targeting fluorescence microscopy and the conventional confocal microscope – two of the most widely accessed tools in the neuroscience arsenal. By providing an accessible pipeline to reconstructing the natural shape and space between brain cells found in the living brain over wide range of experimental conditions, we will help deliver access to comprehensive nanoscale neuroanatomy to the broader neuroscience community.