Multiplex Immunolabeling and Imaging of Functionally Essential Kidney Structures in X-CLARITY-Cleared Tissue

Tissue clearing techniques render tissues transparent and allow for 3D analysis of anatomical structure in intact volumes of tissues. CLARITY-based techniques embed tissues in a porous hydrogel matrix that preserves tissue architecture during clearing and facilitates effective immunostaining in thick tissue volumes [1]. Furthermore, hydrogel-embedded tissue is highly amenable to multiple rounds of immunostaining and imaging without significant tissue loss [2]. In this study, we utilize the X-CLARITY tissue clearing system (Logos Biosystems) to clear kidney tissue sections of varying thickness and demonstrate techniques for antibody labeling, stripping, and reprobing of cleared tissue sections. This allows for the imaging and mapping of multiple structurally and functionally important components of the kidney in 3D within the same tissue.