A 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.
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.
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.
“[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.”
“Thanks to X-CLARITY, we now have access to the threedimensions of the whole heart organ.”