A versatile new technique to clear mouse and human brain

Irene Costantini; Antonino Paolo Di Giovanna; Anna Letizia Allegra Mascaro; Ludovico Silvestri; Marie Caroline Müllenbroich; Leonardo Sacconi; Francesco  S. Pavone

doi:10.1117/12.2184289

14 July 2015 A versatile new technique to clear mouse and human brain

Irene Costantini, Antonino Paolo Di Giovanna, Anna Letizia Allegra Mascaro, Ludovico Silvestri, Marie Caroline Müllenbroich, Leonardo Sacconi, Francesco S. Pavone

Author Affiliations +

Proceedings Volume 9536, Advanced Microscopy Techniques IV; and Neurophotonics II; 95360X (2015) https://doi.org/10.1117/12.2184289
Event: European Conferences on Biomedical Optics, 2015, Munich, Germany

Abstract

Large volumes imaging with microscopic resolution is limited by light scattering. In the last few years based on refractive index matching, different clearing approaches have been developed. Organic solvents and water-based optical clearing agents have been used for optical clearing of entire mouse brain. Although these methods guarantee high transparency and preservation of the fluorescence, though present other non-negligible limitations. Tissue transformation by CLARITY allows high transparency, whole brain immunolabelling and structural and molecular preservation. This method however requires a highly expensive refractive index matching solution limiting practical applicability. In this work we investigate the effectiveness of a water-soluble clearing agent, the 2,2'-thiodiethanol (TDE) to clear mouse and human brain. TDE does not quench the fluorescence signal, is compatible with immunostaining and does not introduce any deformation at sub-cellular level. The not viscous nature of the TDE make it a suitable agent to perform brain slicing during serial two-photon (STP) tomography. In fact, by improving penetration depth it reduces tissue slicing, decreasing the acquisition time and cutting artefacts. TDE can also be used as a refractive index medium for CLARITY. The potential of this method has been explored by imaging a whole transgenic mouse brain with the light sheet microscope. Moreover we apply this technique also on blocks of dysplastic human brain tissue transformed with CLARITY and labeled with different antibody. This clearing approach significantly expands the application of single and two-photon imaging, providing a new useful method for quantitative morphological analysis of structure in mouse and human brain.

Citation Download Citation

Irene Costantini, Antonino Paolo Di Giovanna, Anna Letizia Allegra Mascaro, Ludovico Silvestri, Marie Caroline Müllenbroich, Leonardo Sacconi, and Francesco S. Pavone "A versatile new technique to clear mouse and human brain", Proc. SPIE 9536, Advanced Microscopy Techniques IV; and Neurophotonics II, 95360X (14 July 2015); https://doi.org/10.1117/12.2184289

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
6 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Brain

Tissues

Refractive index

Luminescence

Neuroimaging

Microscopy

Optical clearing

Show All Keywords

Keywords/Phrases

Search In:

Publication Years