A versatile clearing agent for multi-modal brain imaging

Year: 2015

Authors: Costantini I., Ghobril J.P., Di Giovanna A.P., Allegra Mascaro A. L., Silvestri L., Müllenbroich M.C., Onofri L., Conti V., Vanzi F., Sacconi L., Guerrini R., Markram H., Iannello G., Pavone F.S.

Autors Affiliation: Univ Florence, European Lab Nonlinear Spect, I-50019 Sesto Fiorentino, Italy; CNR, Natl Opt Inst, I-50125 Florence, Italy; Univ Florence, Dept Phys & Astron, I-50019 Sesto Fiorentino, Italy; Ecole Polytech Fed Lausanne, Brain Mind Inst, Lab Neural Microcircuitry, CH-1015 Lausanne, Switzerland; Univ Campus Biomed Rome, Dept Engn, I-00128 Rome, Italy; Univ Florence, A Meyer Childrens Hosp, Dept Neurosci Pharmacol & Child Hlth, Pediat Neurol & Neurogenet Unit, I-50139 Florence, Italy; Univ Florence, A Meyer Childrens Hosp, Dept Neurosci Pharmacol & Child Hlth, Labs, I-50139 Florence, Italy; Univ Florence, Dept Biol, I-50125 Florence, 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.

Journal/Review: SCIENTIFIC REPORTS

Volume: 5      Pages from: 9808-1  to: 9808-9

More Information: The research leading to these results has received funding from the European Union Seventh Framework Program (FP7/2007-2013) under grant agreements no. 604102 (Human Brain Project) and nu 284464 (LASERLAB-EUROPE). The research has also been supported by the Italian Ministry for Education, University and Research in the framework of the Flagship Project NANOMAX, by Ente Cassa di Risparmio di Firenze (private foundation) and by Regione Toscana (grant number: POR-CreO 2007-2013). We thank Prof. Giovanni Delfino for discussions on the electron microscopy data and Marcel van’t Hoff for his help on LabVIEW code programming.
KeyWords: Mouse; 2-photon; Light; Reconstruction; Tomography; Neurons; Clarity; Scale; Mice; Vivo
DOI: 10.1038/srep09808

Citations: 190
data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2024-11-17
References taken from IsiWeb of Knowledge: (subscribers only)
Connecting to view paper tab on IsiWeb: Click here
Connecting to view citations from IsiWeb: Click here