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Three-dimensional architecture of podocytes revealed by block-face scanning electron microscopy.

Ichimura K, Miyazaki N, Sadayama S, Murata K, Koike M, Nakamura K, Ohta K, Sakai T - Sci Rep (2015)

Bottom Line: Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process.The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process.In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Life Structure, Juntendo University Graduate School of Medicine, Tokyo, Japan.

ABSTRACT
Block-face imaging is a scanning electron microscopic technique which enables easier acquisition of serial ultrastructural images directly from the surface of resin-embedded biological samples with a similar quality to transmission electron micrographs. In the present study, we analyzed the three-dimensional architecture of podocytes using serial block-face imaging. It was previously believed that podocytes are divided into three kinds of subcellular compartment: cell body, primary process, and foot process, which are simply aligned in this order. When the reconstructed podocytes were viewed from their basal side, the foot processes were branched from a ridge-like prominence, which was formed on the basal surface of the primary process and was similar to the usual foot processes in structure. Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process. The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process. In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

No MeSH data available.


Related in: MedlinePlus

Comparison with block-face SEM and conventional TEM images of podocytes.The block-face images obtained with SBF-SEM (a1, a2) and FIB/SEM (b1, b2) are similar to the conventional TEM images (c1, c2). Due to the combinatorial heavy metal en bloc staining, the Golgi apparatus and endoplasmic reticulum in the cell bodies of podocytes are electron-densely depicted (pink arrowheads in a1, b1, c1). The electron-dense actin bundles, which are visualized at the luminal cytoplasm of the foot processes in the conventional TEM image (yellow arrowheads in c2), are difficult to see in the FIB/SEM and SBF-SEM images. Cap, capillary lumen. Bar scales, 500 nm in (a1), (b1), (c1); 200 nm in (a2), (b2), (c2).
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f2: Comparison with block-face SEM and conventional TEM images of podocytes.The block-face images obtained with SBF-SEM (a1, a2) and FIB/SEM (b1, b2) are similar to the conventional TEM images (c1, c2). Due to the combinatorial heavy metal en bloc staining, the Golgi apparatus and endoplasmic reticulum in the cell bodies of podocytes are electron-densely depicted (pink arrowheads in a1, b1, c1). The electron-dense actin bundles, which are visualized at the luminal cytoplasm of the foot processes in the conventional TEM image (yellow arrowheads in c2), are difficult to see in the FIB/SEM and SBF-SEM images. Cap, capillary lumen. Bar scales, 500 nm in (a1), (b1), (c1); 200 nm in (a2), (b2), (c2).

Mentions: We obtained serial block-face images of podocytes from a perfusion-fixed rat kidney using SBF-SEM and FIB/SEM. Contrast-inverted block-face images achieved a quality comparable to conventional TEM images (Supplementary Fig. S1). However, several differences were found between conventional TEM and contrast-inverted block-face images (Fig. 2).


Three-dimensional architecture of podocytes revealed by block-face scanning electron microscopy.

Ichimura K, Miyazaki N, Sadayama S, Murata K, Koike M, Nakamura K, Ohta K, Sakai T - Sci Rep (2015)

Comparison with block-face SEM and conventional TEM images of podocytes.The block-face images obtained with SBF-SEM (a1, a2) and FIB/SEM (b1, b2) are similar to the conventional TEM images (c1, c2). Due to the combinatorial heavy metal en bloc staining, the Golgi apparatus and endoplasmic reticulum in the cell bodies of podocytes are electron-densely depicted (pink arrowheads in a1, b1, c1). The electron-dense actin bundles, which are visualized at the luminal cytoplasm of the foot processes in the conventional TEM image (yellow arrowheads in c2), are difficult to see in the FIB/SEM and SBF-SEM images. Cap, capillary lumen. Bar scales, 500 nm in (a1), (b1), (c1); 200 nm in (a2), (b2), (c2).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4355681&req=5

f2: Comparison with block-face SEM and conventional TEM images of podocytes.The block-face images obtained with SBF-SEM (a1, a2) and FIB/SEM (b1, b2) are similar to the conventional TEM images (c1, c2). Due to the combinatorial heavy metal en bloc staining, the Golgi apparatus and endoplasmic reticulum in the cell bodies of podocytes are electron-densely depicted (pink arrowheads in a1, b1, c1). The electron-dense actin bundles, which are visualized at the luminal cytoplasm of the foot processes in the conventional TEM image (yellow arrowheads in c2), are difficult to see in the FIB/SEM and SBF-SEM images. Cap, capillary lumen. Bar scales, 500 nm in (a1), (b1), (c1); 200 nm in (a2), (b2), (c2).
Mentions: We obtained serial block-face images of podocytes from a perfusion-fixed rat kidney using SBF-SEM and FIB/SEM. Contrast-inverted block-face images achieved a quality comparable to conventional TEM images (Supplementary Fig. S1). However, several differences were found between conventional TEM and contrast-inverted block-face images (Fig. 2).

Bottom Line: Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process.The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process.In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Life Structure, Juntendo University Graduate School of Medicine, Tokyo, Japan.

ABSTRACT
Block-face imaging is a scanning electron microscopic technique which enables easier acquisition of serial ultrastructural images directly from the surface of resin-embedded biological samples with a similar quality to transmission electron micrographs. In the present study, we analyzed the three-dimensional architecture of podocytes using serial block-face imaging. It was previously believed that podocytes are divided into three kinds of subcellular compartment: cell body, primary process, and foot process, which are simply aligned in this order. When the reconstructed podocytes were viewed from their basal side, the foot processes were branched from a ridge-like prominence, which was formed on the basal surface of the primary process and was similar to the usual foot processes in structure. Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process. The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process. In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

No MeSH data available.


Related in: MedlinePlus