A versatile clearing agent for multi-modal brain imaging

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.

Full text

Its licence does not allow us to show the full text. Read it at PubMed Central:

References 30

1. 1.

   Lein E. S. et al. Genome-wide atlas of gene expression in the adult mouse brain. Nature 445, 168–176 (2007 ).

   Not in our library10.1038/nature05453

2. 2.
   A Proposal for a Coordinated Effort for the Determination of Brainwide Neuroanatomical Connectivity in Model Organisms at a Mesoscopic Scale · 2009

   No notice
3. 3.

   Li A. et al. Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain. Science 330, 1404–1408 (2010 ).

   Not in our library10.1126/science.1191776

4. 4.

   Gong H. et al. Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution. Neuroimage 74, 87–98 (2013 ).

   Not in our library10.1016/j.neuroimage.2013.02.005

5. 5.

   Ragan T. et al. Serial two-photon tomography for automated ex vivo mouse brain imaging. Nat Methods 9, 255–258 (2012 ).

   Not in our library10.1038/nmeth.1854

6. 6.

   Huisken J. & Stainier D. Y. Selective plane illumination microscopy techniques in developmental biology. Development 136, 1963–1975 (2009 ).

   Not in our library10.1242/dev.022426

7. 7.

   Dodt H. U. et al. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat Methods 4, 331–336 (2007 ).

   Not in our library10.1038/nmeth1036

8. 8.
   Chemical Clearing and Dehydration of GFP Expressing Mouse Brains · 2012

   Corrected
9. 9.

   Renier N. et al. iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume Imaging. Cell 159, 896–910 (2014 ).

   Not in our library10.1016/j.cell.2014.10.010

10. 10.

    Hama H. et al. Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain. Nat Neurosci 14, 1481–1488 (2011 ).

    Not in our library10.1038/nn.2928

11. 11.

    Ke M. T., Fujimoto S. & Imai T. SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction. Nat Neurosci 16, 1154–1161 (2013 ).

    Not in our library10.1038/nn.3447

12. 12.

    Kuwajima T. et al. ClearT: a detergent- and solvent-free clearing method for neuronal and non-neuronal tissue. Development 140, 1364–1368 (2013 ).

    Not in our library10.1242/dev.091844

13. 13.

    Susaki E. A. et al. Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis. Cell 157, 726–739 (2014 ).

    Not in our library10.1016/j.cell.2014.03.042

14. 14.

    Chung K. et al. Structural and molecular interrogation of intact biological systems. Nature 497, 332–337 (2013 ).

    Not in our library10.1038/nature12107

15. 15.

    Tomer R., Ye L., Hsueh B. & Deisseroth K. Advanced CLARITY for rapid and high-resolution imaging of intact tissues. Nat Protoc 9, 1682–1697 (2014 ).

    Not in our library10.1038/nprot.2014.123

16. 16.

    Staudt T., Lang M. C., Medda R., Engelhardt J. & Hell S. W. 2,2'-thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy. Microsc Res Tech 70, 1–9 (2007 ).

    Not in our library10.1002/jemt.20396

17. 17.

    Appleton P. L., Quyn A. J., Swift S. & Nathke I. Preparation of wholemount mouse intestine for high-resolution three-dimensional imaging using two-photon microscopy. J Microsc 234, 196–204 (2009 ).

    Not in our library10.1111/j.1365-2818.2009.03163.x

18. 18.

    Gonzalez-Bellido P. T. & Wardill T. J. Labeling and confocal imaging of neurons in thick invertebrate tissue samples. Cold Spring Harb Protoc 2012, 969–983 (2012 ).

    Not in our library10.1101/pdb.prot069625

19. 19.

    Frasconi P. et al. Large-scale automated identification of mouse brain cells in confocal light sheet microscopy images. Bioinformatics 30, i587–593 (2014 ).

    Not in our library10.1093/bioinformatics/btu469

20. 20.

    Baek S. T., Gibbs E. M., Gleeson J. G. & Mathern G. W. Hemimegalencephaly, a paradigm for somatic postzygotic neurodevelopmental disorders. Current opinion in neurology 26, 122–127 (2013 ).

    Not in our library10.1097/wco.0b013e32835ef373

21. 21.

    Poguzhelskaya E., Artamonov D., Bolshakova A., Vlasova O. & Bezprozvanny I. Simplified method to perform CLARITY imaging. Molecular neurodegeneration 9, 19 (2014 ).

    Not in our library10.1186/1750-1326-9-19

22. 22.

    Silvestri L., Allegra Mascaro A. L., Costantini I., Sacconi L. & Pavone F. S. Correlative two-photon and light sheet microscopy. Methods 66, 268–272 (2014 ).

    Not in our library10.1016/j.ymeth.2013.06.013

23. 23.

    Jahrling N., Becker K., Schonbauer C., Schnorrer F. & Dodt H. U. Three-dimensional reconstruction and segmentation of intact Drosophila by ultramicroscopy. Front Syst Neurosci 4, 1 (2010 ).

    Not in our library10.3389/neuro.06.001.2010

24. 24.

    Huisken J., Swoger J., Del Bene F., Wittbrodt J. & Stelzer E. H. Optical sectioning deep inside live embryos by selective plane illumination microscopy. Science 305, 1007–1009 (2004 ).

    Not in our library10.1126/science.1100035

25. 25.

    Feng G. et al. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28, 41–51 (2000 ).

    Not in our library10.1016/s0896-6273(00)00084-2

26. 26.

    Taniguchi H. et al. A resource of Cre driver lines for genetic targeting of GABAergic neurons in cerebral cortex. Neuron 71, 995–1013 (2011 ).

    Not in our library10.1016/j.neuron.2011.07.026

27. 27.

    Madisen L. et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13, 133–140 (2010 ).

    Not in our library10.1038/nn.2467

28. 28.

    Knott G. W., Holtmaat A., Trachtenberg J. T., Svoboda K. & Welker E. A protocol for preparing GFP-labeled neurons previously imaged in vivo and in slice preparations for light and electron microscopic analysis. Nat Protoc 4, 1145–1156 (2009 ).

    Not in our library10.1038/nprot.2009.114

29. 29.

    Silvestri L., Bria A., Sacconi L., Iannello G. & Pavone F. S. Confocal light sheet microscopy: micron-scale neuroanatomy of the entire mouse brain. Opt Express 20, 20582–20598 (2012 ).

    Not in our library10.1364/oe.20.020582

30. 30.
    TeraStitcher - A tool for fast automatic 3D-stitching of teravoxel-sized microscopy images · 2012

    No notice

Cited by papers we hold 4

• Optimization of Whole Mount RNA Multiplexed in situ Hybridization Chain Reaction With Immunohistochemistry, Clearing and Imaging to Visualize Octopus Embryonic Neurogenesis · 2022
  Not checked
• Next generation histology methods for three-dimensional imaging of fresh and archival human brain tissues · 2018
  Corrected
• CUBIC pathology: three-dimensional imaging for pathological diagnosis · 2017
  No notice
• ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging · 2016
  Corrected

Reads

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.