Nature Methods
- 4, 331 - 336 (2007)
Published online: 25 March 2007; | doi:10.1038/nmeth1036
Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brainHans-Ulrich Dodt1, 3, Ulrich Leischner1, Anja Schierloh1, Nina Jährling1, 3, Christoph Peter Mauch1, Katrin Deininger2, Jan Michael Deussing1, Matthias Eder1, Walter Zieglgänsberger1 & Klaus Becker1, 31
Max Planck Institute of Psychiatry, Kraepelinstr. 2, 80804 Munich, Germany. 2
Department of Molecular Neurobiology, Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Martinsried, Germany. 3
Present address: Department of Bioelectronics, Institute of Solid State Electronics, Vienna University of Technology, Floragasse 7, 1040 Vienna, Austria.
Correspondence should be addressed to Hans-Ulrich Dodt dodt@mpipsykl.mpg.de 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.
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