Whole-Kidney Three-Dimensional Staining with CUBIC

This protocol allows a visualization of three-dimensional structures in full kidney, which can lead to innovative discoveries in kidney research by providing a macroscopic perspective. Whole-mount organ staining is challenging because of the difficulty in antibody penetration. This protocol has demonstrated the high quality, whole-mount staining of the kidneys by the antibodies.

To begin, perform the immersion fixation immediately after the perfusion fixation and kidney sampling, by immersing the kidney in 4%paraformaldehyde at four degrees Celsius for 16 hours. Give three washes using PBS for two hours each. Perform the decolorization and delipidation process by immersing the fixed kidney in seven millimeters of 50%CUBIC-L in a 14 milliliter round-bottom tube, with gentle shaking at room temperature.