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.
STEP 1. Tissue ClearingA fixed tissue sample is incubated in the X-CLARITY™ Hydrogel Solution Kit to allow hydrogel monomers to diffuse uniformly throughout the sample and then placed in the X-CLARITY™ Polymerization System. Monomers polymerize in the anaerobic environment, linking the biomolecules to a sturdy hydrogel network, preserving molecular information and structural integrity. The tissue-hydrogel 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.
STEP 2.Antibody LabelingThe 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.
STEP 3. RI Matching for ImagingPrior 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.
MAKE TISSUE CLEARING SIMPLE, RAPID, AND
REPRODUCIBLE WITH THE X-CLARITY
Clear a wide variety of tissue types and sizes
Achieve exceptional transparency
Preserve endogenous fluorescent proteins long-term
Label cleared tissues with antibodies
“[The X-CLARITY] has actually been fundamental to the Data Brain Project because without being able to clear tissues of the lipids, we wouldn’t be able to image deep into brains. So we wouldn’t be able to extract that structural information that is so important to understand the function of mammalian brains. It’s been a huge advantage and a great leap forward in imaging.”
Arti Ahluwalia, PhD
Centro E. Piaggio | University of Pisa
“The X-CLARITY saves a lot of time. We were able to image an entire set of brain and spinal cords in a relatively short amount of time. This allowed us to clearly see the benefit of the approaches we’re having on spinal cord networks following an injury. It's a great device to save time for your research.”
Quentin Barraud, PhD