Kidneys
(mouse)

Movie 1: Basic kidney anatomy

Tissue/pathology:

Young adult male C57BL/6J control mouse kidney

Antibodies:

488 autofluorescence

Movie description:

This movie is based on 488 tissue autofluorescence. The basic anatomy of the kidney is demonstrated with horizontal and coronal optical slicing.

Tissue/pathology:

Young adult male C57BL/6J control mouse kidney

Antibodies:

488 autofluorescence (gray)
aquaporin 2 (Aq2, green)
nephrin (red)

Movie description:

AF (gray) = autofluorescence, depicting the shape of the organ, MIP 3D rendering.

Nephrin (red) is a protein that depicts the shape of the glomeruli. Nephrin is a transmembrane protein that localizes in the tips of the podocytes in the Bowman capsule. Bowman capsule and the glomerulus together form the filtration unit in the kidney. 

Aquaporin2 (green) is a protein that localizes in the kidney collective duct apical membrane. This depicts all collective ducts that link the nephrons to the calyx/renal pelvis. Collective ducts are important in the reabsorption of water, before the final urine formation. 

The movie first shows the autofluorescence shadow of the organ. Then, overview and zoom to nephrin+Aq2 volume co-staining. Then, zoom to Aq2 staining in coronal orientation: here we see how the collective ducts (which collect and modify the created urine) target the pelvis. Then, we see the nephrin channel with all glomeruli-Bowmann capsules. Between 2:39 -3:36 the movie shows a quantification: each glomerulus is indicated by a whitish dot (‘spot detection’ function of Imaris software). Finally, we see some turn again for nephrin+Aq2 double staining.

Movie 3: A piece of renal cortex, higher resolution

Tissue/pathology:

Young adult male C57BL/6J control mouse kidney

Antibodies:

Aquaporin 2 (green)
Nephrin (red)

Movie description:

Close up to the ‘mess’ of glomeruli and collective ducts in various perspectives with both channels or in a single channel.

Movie 4: This movie is about the same blocks as MOVIE 3, but with ‘blend’ 3D rendering mode

Tissue/pathology:

Young adult male C57BL/6J control mouse kidney

Antibodies:

Aquaporin 2 (green)
Nephrin (red)

Movie description:

Notably, the nephrin volume staining has a penetration gradient issue (the glomeruli closer to the organ surface are stronger in staining intensity), and the ‘blend’ mode attenuates this effect.