The X-CLARITY™ systems and reagents are engineered to standardize, simplify, and expedite each phase of the tissue clearing process, providing a streamlined approach to preparing tissues for detailed imaging. At the heart of this process is the CLARITY method, which involves embedding preserved tissues in a hydrogel matrix, followed by the active extraction of lipids through electrophoresis. This results in a stable, optically transparent tissue-hydrogel hybrid that remains chemically accessible for repeated rounds of antibody labeling and imaging. Crucially, the native cytoarchitecture of the tissue is preserved, and endogenous fluorescence proteins remain intact, enabling robust fluorescence imaging downstream.
The process begins with hydrogel infusion using the X-CLARITY™ Hydrogel Solution Kit (C1310X). This pre-tested kit provides a uniform and consistent approach to tissue-hydrogel hybridization. The kit includes the X-CLARITY™ Hydrogel Solution, an acrylamide-based solution used to create polyacrylamide, and the X-CLARITY™ Polymerization Initiator, a thermal free radical initiator known as VA-044. Together, these components ensure that the hydrogel matrix is uniformly integrated into the tissue, setting the stage for effective clearing.
Following hydrogel infusion, tissue-hydrogel hybridization is carried out using the X-CLARITY™ Polymerization System (C20001). This standalone, automated system simplifies the critical process of hybridization, allowing for efficient and high-throughput sample processing. Multiple samples can be processed simultaneously in multi-well plates or conical tubes. The system offers precise control over polymerization parameters, including vacuum strength, temperature, and timing, all managed through an intuitive touchscreen interface. This automation streamlines the hybridization process, ensuring consistency and reliability in sample preparation.
The next phase involves lipid removal, which is efficiently managed by the X-CLARITY™ Tissue Clearing System II (C30001). This all-in-one system is designed for rapid and effective electrophoretic tissue clearing. The system’s advanced design accelerates lipid extraction while preserving the structural integrity of the tissue. For instance, it can clear an entire mouse brain in just six hours, a remarkable eight times faster than previous technologies. The clearing process is controlled via a user-friendly touchscreen interface, and in ETC (electrophoretic tissue clearing) mode, platinum-plated electrodes generate an electric field to enhance lipid removal. The system features built-in temperature control to maintain consistent buffer temperatures, and a continuously circulated buffer ensures consistent buffering capacity and removal of clearing byproducts. This advanced setup guarantees efficient, rapid, and uniform tissue clearing, significantly improving the overall workflow for high-resolution imaging studies.
