Imaging a 1 mm 3 Volume of Rat Cortex Using a MultiBeam SEM
Imaging a 1 mm 3 Volume of Rat Cortex Using a MultiBeam SEM
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使用多束 SEM 对 1 mm 3 体积的大鼠皮层进行成像
DOI：
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发表时间：
2016
影响因子：
2.8
通讯作者：
J. Lichtman
中科院分区：
文献类型：
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作者：
R. Schalek;Dongil Lee;N. Kasthuri;A. Peleg;T. Jones;V. Kaynig;D. Haehn;Hanspeter Pfister;D. Cox;J. Lichtman
啮齿动物的大脑是由厘米到纳米的长度尺度组成的--6个数量级[1]。在厘米尺度上，大脑由皮质叶、小脑、脑干和脊髓组成。毫米尺度的神经元排列成列、层或以其他方式聚集。最近的成像技术进步允许生成跨越纳米到数百微米空间范围的神经元数据集[2，3]。使用最快的信号束SEM以4 nm x-y分辨率收集1 mm体积的脑组织数据集将需要约6年。要达到神经元回路的下一个长度和体积规模，需要几项技术进步。多束扫描电子显微镜（mSEM）代表了一种变革性的成像技术，使神经科学家能够解决毫米级的皮层电路问题。在这项工作中，我们描述了从组织采集到成像的工作流程，该工作流程将在不到6个月的时间内生成大鼠视觉皮层的2 PB数据集（> 300，000，000张图像），该数据集以4 nm x 4 nm x-y（膜的奈奎斯特采样）和30 nm切片厚度成像。
The rodent brain is organized with length scales spanning centimeters to nanometers --6 orders of magnitude [1]. At the centimeter scale, the brain consist of lobes of cortex, the cerebellum, the brainstem and the spinal cord. The millimeter scale have neurons arranged in columns, layers, or otherwise clustered. Recent technological imaging advances allow the generation of neuronal datasets spanning the spatial range from nanometers to 100s of microns [2,3]. Collecting a 1 mm volume dataset of brain tissue at 4 nm x-y resolution using the fastest signal-beam SEM would require ~6 years. To move to the next length and volume scale of neuronal circuits requires several technological advances. The multibeam scanning electron microscope (mSEM) represents a transformative imaging technology that enables neuroscientists to tackle millimeter scale cortical circuit problems. In this work we describe a workflow from tissue harvest to imaging that will generate a 2 petabyte dataset (> 300,000,000 images) of rat visual cortex imaged at a 4nm x 4nm x-y (Nyquist sampling of membranes) and 30nm section thickness in less than 6 months.