Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy
Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy
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DOI：
10.1038/s41467-019-09552-9
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发表时间：
2019-04-10
影响因子：
16.6
通讯作者：
Beaurepaire, Emmanuel
中科院分区：
文献类型：
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作者：
Abdeladim, Lamiae;Matho, Katherine S.;Beaurepaire, Emmanuel
大规模显微镜方法正在改变脑成像，但目前缺乏有效的多色对比模式。介绍了彩色多光子序列显微技术（ChroMS），这是一种将单次多色多光子激发与连续块面图像采集相结合的方法。该方法在整个成像体积上提供了具有恒定微米级分辨率和亚微米通道配准的光谱不同荧光蛋白和无标记非线性信号的器官尺度微米成像。我们展示了几立方毫米的三维（3D）多色成像以及全脑连续二维多通道成像。我们通过对小鼠大脑皮层星形胶质细胞形态和接触的基于颜色的3D分析，在密集的脑弓标记组织中追踪单个锥体神经元，以及用光谱不同的示踪剂标记的轴突投影的多路全脑映射来说明这种方法的优势。ChroMS将是在神经科学和超越的多尺度和系统级研究的资产。
Large-scale microscopy approaches are transforming brain imaging, but currently lack efficient multicolor contrast modalities. We introduce chromatic multiphoton serial (ChroMS) microscopy, a method integrating one-shot multicolor multiphoton excitation through wavelength mixing and serial block-face image acquisition. This approach provides organ-scale micrometric imaging of spectrally distinct fluorescent proteins and label-free nonlinear signals with constant micrometer-scale resolution and sub-micron channel registration over the entire imaged volume. We demonstrate tridimensional (3D) multicolor imaging over several cubic millimeters as well as brain-wide serial 2D multichannel imaging. We illustrate the strengths of this method through color-based 3D analysis of astrocyte morphology and contacts in the mouse cerebral cortex, tracing of individual pyramidal neurons within densely Brainbow-labeled tissue, and multiplexed whole-brain mapping of axonal projections labeled with spectrally distinct tracers. ChroMS will be an asset for multiscale and system-level studies in neuroscience and beyond.