Quantifying Podocyte Number in a Small Sample Size of Glomeruli with CUBIC to Evaluate Podocyte Depletion of db/db Mice

Figure 1

Optical clearing, light-sheet microscope, and 3D rendering for whole glomeruli. (a) 2 mm thick kidney tissue from C57 mice was analyzed by optical clearing using CUBIC method. Red arrow displayed the location of the translucent tissue. (b) Double immunofluorescence staining with podocyte cytoplasmic marker synaptopodin (green) and podocyte nuclear marker WT-1 (red) identified glomeruli and podocytes with light-sheet microscope by using the CUBIC method (screenshots from three-dimensional video). (c) Podocyte nuclei stained with WT-1 (red) of a single whole glomerulus and the linked 3D reconstruction image are shown, which depicted the practicality of counting podocyte number with the CUBIC method. Shown are representative images from five C57 mice with similar results (scale bar: 50 μm in all images of (b); 400 μm in all images of (c)).