Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain

ABSTRACT 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. Access through your institution Buy or

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references ACKNOWLEDGEMENTS We thank G. Ryseck for help with initial experiments and S. Espinoza, L. Luo, E. Kramer and C. Wotjak for specimens. This work was supported by grants of the

Hertie foundation and the SFB391. AUTHOR INFORMATION Author notes * Hans-Ulrich Dodt, Nina Jährling & Klaus Becker Present address: Present address: Department of Bioelectronics,

Institute of Solid State Electronics, Vienna University of Technology, Floragasse 7, 1040 Vienna, Austria., AUTHORS AND AFFILIATIONS * Max Planck Institute of Psychiatry, Kraepelinstr. 2,

Munich, 80804, Germany Hans-Ulrich Dodt, Ulrich Leischner, Anja Schierloh, Nina Jährling, Christoph Peter Mauch, Jan Michael Deussing, Matthias Eder, Walter Zieglgänsberger & Klaus

Becker * Department of Molecular Neurobiology, Max Planck Institute of Neurobiology, Am Klopferspitz 18, Martinsried, 82152, Germany Katrin Deininger Authors * Hans-Ulrich Dodt View author

publications You can also search for this author inPubMed Google Scholar * Ulrich Leischner View author publications You can also search for this author inPubMed Google Scholar * Anja

Schierloh View author publications You can also search for this author inPubMed Google Scholar * Nina Jährling View author publications You can also search for this author inPubMed Google

Scholar * Christoph Peter Mauch View author publications You can also search for this author inPubMed Google Scholar * Katrin Deininger View author publications You can also search for this

author inPubMed Google Scholar * Jan Michael Deussing View author publications You can also search for this author inPubMed Google Scholar * Matthias Eder View author publications You can

also search for this author inPubMed Google Scholar * Walter Zieglgänsberger View author publications You can also search for this author inPubMed Google Scholar * Klaus Becker View author

publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Hans-Ulrich Dodt. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare

no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY VIDEO 1 whole mouse brain reconstructed from 550 optical sections. (MOV 2686 kb) SUPPLEMENTARY VIDEO 2 Granule cells

with dendrites in the hippocampus of a thy-1 GFP mouse. (MOV 2732 kb) SUPPLEMENTARY VIDEO 3 Excised whole hippocampus. (MOV 1016 kb) SUPPLEMENTARY VIDEO 4 Optical sectioning of a whole

hippocampus. (MOV 1521 kb) SUPPLEMENTARY VIDEO 5 3D-reconstruction and animation of a part of a whole hippocampus. (MOV 2262 kb) SUPPLEMENTARY VIDEO 6 3D reconstruction and animation of

axonal bundles in the hippocampal alveus and dendritic spines of CA1 pyramidal neurons. (MOV 2134 kb) SUPPLEMENTARY VIDEO 7 Primary and secondary barrel field made visible by excitation of

autofluorescence in the whole brain of a 10 day old mouse. (MOV 1040 kb) SUPPLEMENTARY VIDEO 8 Optical sectioning of a mouse brain imaged by detection of scattered light. Note the appearance

of fibre tracts during the movement of the optical sectioning plane through the brain. (MOV 2115 kb) SUPPLEMENTARY METHODS (PDF 10 KB) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT

THIS ARTICLE CITE THIS ARTICLE Dodt, HU., Leischner, U., Schierloh, A. _et al._ Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. _Nat Methods_

4, 331–336 (2007). https://doi.org/10.1038/nmeth1036 Download citation * Received: 22 December 2006 * Accepted: 28 February 2007 * Published: 25 March 2007 * Issue Date: April 2007 * DOI:

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