Limits...
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

Luminal surface of reconstructed podocytes based on serial FIB/SEM images.Individual podocytes are represented with different colors. (a) The reconstructed image is quite similar to the conventional SEM images as shown in Figure 1a. (b, b′) These two reconstructed images are shown in the same region, and the green-colored podocyte is displayed semi-transparently in b′. The distal ends of foot processes are situated under the primary process (asterisks in b′), and both lateral surfaces of the ridge face the distal ends of foot processes. Bar scales, 1 μm in (a), 200 nm in (b′). These three-dimensionally reconstructed podocytes can be observed from any directions on the reconstruction software. CB, cell body of podocyte; P, primary process. (See also Supplementary Movies S1, S2).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4355681&req=5

f3: Luminal surface of reconstructed podocytes based on serial FIB/SEM images.Individual podocytes are represented with different colors. (a) The reconstructed image is quite similar to the conventional SEM images as shown in Figure 1a. (b, b′) These two reconstructed images are shown in the same region, and the green-colored podocyte is displayed semi-transparently in b′. The distal ends of foot processes are situated under the primary process (asterisks in b′), and both lateral surfaces of the ridge face the distal ends of foot processes. Bar scales, 1 μm in (a), 200 nm in (b′). These three-dimensionally reconstructed podocytes can be observed from any directions on the reconstruction software. CB, cell body of podocyte; P, primary process. (See also Supplementary Movies S1, S2).

Mentions: We segmented individual podocytes on serial block-face images obtained by FIB/SEM and SBF-SEM, and then reconstructed the segmented podocytes using an AMIRA 5.1 image analysis and reconstruction software. The three-dimensional reconstruction images based on the serial FIB/SEM and SBF-SEM images appeared quite similar to conventional SEM images (Figs. 3, 4, Supplementary Movie S1). However, the surface of podocytes appeared smoother in the FIB/SEM-based reconstruction than in the SBF-SEM-based one, because the electrical distortion of each block-face image due to surface charging was less in the FIB/SEM images.


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)

Luminal surface of reconstructed podocytes based on serial FIB/SEM images.Individual podocytes are represented with different colors. (a) The reconstructed image is quite similar to the conventional SEM images as shown in Figure 1a. (b, b′) These two reconstructed images are shown in the same region, and the green-colored podocyte is displayed semi-transparently in b′. The distal ends of foot processes are situated under the primary process (asterisks in b′), and both lateral surfaces of the ridge face the distal ends of foot processes. Bar scales, 1 μm in (a), 200 nm in (b′). These three-dimensionally reconstructed podocytes can be observed from any directions on the reconstruction software. CB, cell body of podocyte; P, primary process. (See also Supplementary Movies S1, S2).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Luminal surface of reconstructed podocytes based on serial FIB/SEM images.Individual podocytes are represented with different colors. (a) The reconstructed image is quite similar to the conventional SEM images as shown in Figure 1a. (b, b′) These two reconstructed images are shown in the same region, and the green-colored podocyte is displayed semi-transparently in b′. The distal ends of foot processes are situated under the primary process (asterisks in b′), and both lateral surfaces of the ridge face the distal ends of foot processes. Bar scales, 1 μm in (a), 200 nm in (b′). These three-dimensionally reconstructed podocytes can be observed from any directions on the reconstruction software. CB, cell body of podocyte; P, primary process. (See also Supplementary Movies S1, S2).
Mentions: We segmented individual podocytes on serial block-face images obtained by FIB/SEM and SBF-SEM, and then reconstructed the segmented podocytes using an AMIRA 5.1 image analysis and reconstruction software. The three-dimensional reconstruction images based on the serial FIB/SEM and SBF-SEM images appeared quite similar to conventional SEM images (Figs. 3, 4, Supplementary Movie S1). However, the surface of podocytes appeared smoother in the FIB/SEM-based reconstruction than in the SBF-SEM-based one, because the electrical distortion of each block-face image due to surface charging was less in the FIB/SEM images.

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