Ultramicroscopy:: three-dimensional visualization of neuronal networks in the whole mouse brain
Ultramicroscopy:: three-dimensional visualization of neuronal networks in the whole mouse brain
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DOI：
10.1038/nmeth1036
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发表时间：
2007-04-01
期刊：
影响因子：
48
通讯作者：
Becker, Klaus
中科院分区：
文献类型：
--
作者：
Dodt, Hans-Ulrich;Leischner, Ulrich;Becker, Klaus
可视化整个神经元网络以进行完整的大脑分析，到目前为止一直不可能。计算机断层扫描或磁共振成像（MRI）等技术不会产生细胞分辨率，机械切片程序不足以在三个维度上实现高分辨率重建。在这里，我们提出了一种允许对整个固定小鼠大脑进行成像的方法。我们通过将其与清除组织的特殊程序结合使用来改变“超显微镜检查”。我们表明，这种新技术允许使用细胞分辨率的固定小鼠大脑进行光学切片，可用于检测切除的小鼠海马中的单个GFP标记的神经元。我们获得了孤立海马中Ca1神经元种群的树突树和棘突的三维（3D）图像。同样在果蝇和小鼠胚胎中，我们能够通过成像自动荧光来可视化解剖学的细节。我们的方法非常适合转基因小鼠的高通量表型筛查，因此将有益于疾病模型的研究。
Visualizing entire neuronal networks for analysis in the intact brain has been impossible up to now. Techniques like computer tomography or magnetic resonance imaging (MRI) do not yield cellular resolution, and mechanical slicing procedures are insufficient to achieve high-resolution reconstructions in three dimensions. Here we present an approach that allows imaging of whole fixed mouse brains. We modified 'ultramicroscopy' by combining it with a special procedure to clear tissue. We show that this new technique allows optical sectioning of fixed mouse brains with cellular resolution and can be used to detect single GFP-labeled neurons in excised mouse hippocampi. We obtained three-dimensional (3D) images of dendritic trees and spines of populations of CA1 neurons in isolated hippocampi. Also in fruit flies and in mouse embryos, we were able to visualize details of the anatomy by imaging autofluorescence. Our method is ideally suited for high-throughput phenotype screening of transgenic mice and thus will benefit the investigation of disease models.