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Sci Rep. 2015 May 7;5:9808. doi: 10.1038/srep09808.

A versatile clearing agent for multi-modal brain imaging.

Author information

1
European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy.
2
Laboratory of Neural Microcircuitry, Brain Mind Institute, EPFL, Station 15, CH-1015 Lausanne, Switzerland.
3
Pediatric Neurology and Neurogenetics Unit and Laboratories, Department of Neuroscience, Pharmacology and Child Health, A. Meyer Children's Hospital - University of Florence, Viale Pieraccini 24, 50139 Florence, Italy.
4
1] European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy [2] Department of Biology, University of Florence, Via Romana 17, 50125 Florence, Italy.
5
1] National Institute of Optics, National Research Council, Largo Fermi 6, 50125 Florence, Italy [2] European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy.
6
Department of Engineering, University Campus Bio-Medico of Rome, Via Alvaro del Portillo 21, 00128 Roma, Italy.
7
1] European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy [2] National Institute of Optics, National Research Council, Largo Fermi 6, 50125 Florence, Italy [3] Department of Physics and Astronomy, University of Florence, Via Sansone 1, 50019 Sesto Fiorentino, Italy.

Abstract

Extensive mapping of neuronal connections in the central nervous system requires high-throughput µm-scale imaging of large volumes. In recent years, different approaches have been developed to overcome the limitations due to tissue light scattering. These methods are generally developed to improve the performance of a specific imaging modality, thus limiting comprehensive neuroanatomical exploration by multi-modal optical techniques. Here, we introduce a versatile brain clearing agent (2,2'-thiodiethanol; TDE) suitable for various applications and imaging techniques. TDE is cost-efficient, water-soluble and low-viscous and, more importantly, it preserves fluorescence, is compatible with immunostaining and does not cause deformations at sub-cellular level. We demonstrate the effectiveness of this method in different applications: in fixed samples by imaging a whole mouse hippocampus with serial two-photon tomography; in combination with CLARITY by reconstructing an entire mouse brain with light sheet microscopy and in translational research by imaging immunostained human dysplastic brain tissue.

PMID:
25950610
PMCID:
PMC4423470
DOI:
10.1038/srep09808
[Indexed for MEDLINE]
Free PMC Article

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