Dodt Hans-Ulrich, Leischner Ulrich, Schierloh Anja, Jährling Nina, Mauch Christoph Peter, Deininger Katrin, Deussing Jan Michael, Eder Matthias, Zieglgänsberger Walter, Becker Klaus
Max Planck Institute of Psychiatry, Kraepelinstr. 2, 80804 Munich, Germany.
Nat Methods. 2007 Apr;4(4):331-6. doi: 10.1038/nmeth1036. Epub 2007 Mar 25.
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.
迄今为止，在完整大脑中可视化整个神经元网络以进行分析是不可能的。计算机断层扫描或磁共振成像（MRI）等技术无法提供细胞分辨率，而机械切片程序不足以实现三维高分辨率重建。在此，我们提出一种能够对整个固定小鼠大脑进行成像的方法。我们通过将“超显微镜”与一种特殊的组织透明化程序相结合对其进行了改进。我们表明，这项新技术能够以细胞分辨率对固定小鼠大脑进行光学切片，并可用于检测切除的小鼠海马体中单个绿色荧光蛋白（GFP）标记的神经元。我们获得了分离海马体中CA1神经元群体的树突棘和树突的三维（3D）图像。同样，在果蝇和小鼠胚胎中，我们能够通过对自发荧光成像来观察解剖细节。我们的方法非常适合对转基因小鼠进行高通量表型筛选，因此将有助于疾病模型的研究。
