Tissue Clearing, the X-CLARITY™ Systems and Reagents
Overview
Tissues are inherently three dimensional in nature, which makes imaging intact tissues a necessity for a more
complete study into the relationship between structure and function and the system-level study of cellular
mechanisms.
Tissue clearing has become an important step for imaging tissues in 3D at single-cell resolution.
The X-CLARITY™ is a collection of systems and ready-to-use reagents to standardize, simplify, and accelerate
each step of the tissue clearing process.
X-CLARITY™ is based on the CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining /in situ-hybridization-compatible Tissue hYdrogel) method. With CLARITY, preserved tissues are embedded in a hydrogel matrix and lipids are actively extracted through electrophoresis to create a stable and optically transparent tissue-hydrogel hybrid that is chemically accessible for multiple rounds of antibody labeling and imaging.
A tissue sample is incubated in the X-CLARITY™ Hydrogel Solution Kit to allow hydrogel monomers to diffuse uniformly throughout the sample. The sample is then placed in the X-CLARITY™ Polymerization System. Monomers polymerize in the anaerobic environment, linking the biomolecules to a hydrogel network, which preserves molecular information and structural integrity
Once the tissue-hydrogel hybrid has been formed, the hybrid is cleared in the X-CLARITY™ Tissue Clearing System II using the ready-to-use Electrophoretic Tissue Clearing Solution. Lipids are extracted actively through electrophoresis
or passively, leaving behind a stable and transparent tissue-hydrogel hybrid that is chemically accessible for molecular phenotyping
The transparent sample is then labeled with antibodies using the DeepLabel™ Antibody Staining Kit, which enhances antibody penetration into clarified tissues. Optimized for clarified tissue samples, DeepLabel™ can be used with tissues
cleared with various clearing methods such as passive or active CLARITY, DISCO, 3DISCO, uDISCO, Visikol or CUBIC.
Prior to imaging, the tissue-hydrogel hybrid is placed in X-CLARITY™ Mounting Solution, a refractive index matching solution (RIMS), to homogenize the environment within the tissue to the solution. This reduces light scatter, which in turn increases optical transparency and consequently increases image quality and imaging depth.
Features of the X-CLARITY™ Systems & Reagents for Tissue Clearing:
A collection of systems and ready to use reagents to make electrophoretic tissue clearing simple, rapid and efficient
Preserved tissues are embedded in a hydrogel matrix
Lipids are actively extracted through electrophoresis to create a stable transparent tissue-hydrogel hybrid
Chemically accessible for multiple rounds of antibody labelling and imaging
Cytoarchitecture remains intact
Endogenous fluorescence proteins are preserved for fluorescence imaging
Features of the X-CLARITY™ Hydrogel Solution Kit:
Pre-tested for uniform and consistent tissue-hydrogel hybridization
Kit consists of hydrogel solution and a polymerization initiator
X-CLARITY™ Hydrogel Solution is acrylamide based solution to create polyacrylamide
