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 CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining /
in situ-hybridization-compatible Tissue hYdrogel), a tissue clearing method developed by the Deisseroth lab at Stanford.
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. Native cytoarchitecture remains intact and even endogenous fluorescence proteins are preserved for robust fluorescence imaging downstream.

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  STEP 1. Tissue-Hydrogel Hybridization

  A fixed tissue sample is incubated in the X-CLARITY™ Hydrogel Solution Kit to allow hydrogel monomers to diffuse uniformly throughout the sample. Once the solution has permeated the tissue, the sample is placed in the X-CLARITY™ Polymerization System. Monomers polymerize in the anaerobic environment, covalently linking the biomolecules to a sturdy hydrogel network, preserving molecular information and structural integrity.

  X-CLARITY™ Hydrogel Solution Kit   >
  X-CLARITY™ Polymerization System   >

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  STEP 2. Tissue Clearing

  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.

  X-CLARITY™ Tissue Clearing System II   >
  Electrophoretic Tissue Clearing Solution   >

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  STEP 3. Labeling

  The transparent sample is then labled 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, iDISCO, 3DISCO, uDISCO, Visikol, or CUBIC.

  DeepLabel Antibody Staining Kit   >

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  STEP 4. Imaging

  Prior to imaging, the tissue-hydrogel hybrid is placed in a refractive index (RI) matching solution in order 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.

  X-CLARITY™ Mounting Solution   >