A Device for Ribbon Collection for Array Tomography with Scanning Electron Microscopy

Koike, Taro; Kataoka, Yosky; Maeda, Masayuki; Hasebe, Yuji; Yamaguchi, Yuuki; Suga, Mitsuo; Saitô, Akira; Yamada, Hisao

doi:10.1267/ahc.17013

Cited by 19 publications

(14 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, it is impossible to obtain serial FIB/SEM images that contain the entire volume of one nephrocyte, podocyte, or glomerulus. Array tomography, which is used to obtain serial images from serial ultrathin sections mounted on silicon or glass bases, is a potential solution to this problem (Koga et al 2018;Koike et al 2017;Micheva and Smith 2007;Wacker and Schroeder 2013). This method enables serial images of larger volumes than FIB/SEM tomography to be obtained, which may enable the entire volume of the murine glomerular capillary to be reconstructed by array tomography (Dittmayer et al 2018).…”

Section: Limitations Of Fib/sem Tomographymentioning

confidence: 99%

Three-dimensional architecture of pericardial nephrocytes in Drosophila melanogaster revealed by FIB/SEM tomography

et al. 2019

View full text Add to dashboard Cite

Nephrocytes are similar in structure to podocytes and play a role in the isolation of toxic substances from hemolymph in insects. Drosophila melanogaster nephrocytes have recently been used to study podocyte function and disease. However, the threedimensional ultrastructure of nephrocytes is not clearly understood because their surrounding basement membrane makes it difficult to observe using conventional scanning electron microscopy. We reconstructed the three-dimensional ultrastructure of Drosophila pericardial nephrocytes using serial focused-ion beam/scanning electron microscopy (FIB/SEM) images. The basal surfaces were occupied by foot processes and slit-like spaces between them. The slit-like spaces corresponded to the podocyte filtration slits and were formed by longitudinal infolding/invagination of the basal plasma membrane. The basal surface between the slit-like spaces became the foot processes, which ran almost linearly, and had a Bwashboard-like^appearance. Both ends of the foot processes were usually anastomosed to neighboring foot processes and thus free ends were rarely observed. We demonstrated that FIB/SEM is a powerful tool to better understand the three-dimensional architecture of nephrocytes.

show abstract

Section: Limitations Of Fib/sem Tomographymentioning

confidence: 99%

Three-dimensional architecture of pericardial nephrocytes in Drosophila melanogaster revealed by FIB/SEM tomography

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Array tomography is a potential solution to this problem, and it was used to obtain serial images from serial ultrathin sections mounted on bases, such as silicon wafer and glass plate (Figure 9). [28][29][30][31] These serial ultrathin sections are physically stable and repeatedly observable. For long-term future studies, we aim to establish a "serial ultrathin section library" of renal biopsy specimens and to analyze them using array tomography.…”

Section: Protein Leakage Pathways In Pan Nephrosismentioning

confidence: 99%

Morphological Processes of Foot Process Effacement in Puromycin Aminonucleoside Nephrosis Revealed by FIB/SEM Tomography

Ichimura

Miyaki²,

Kawasaki³

et al. 2018

JASN

View full text Add to dashboard Cite

Background Foot process effacement is one of the pathologic indicators of podocyte injury. However, the morphologic changes associated with it remain unclear.Methods To clarify the developmental process, we analyzed puromycin nephrotic podocytes reconstructed from serial focused-ion beam/scanning electron microscopy (FIB/SEM) images.Results Intact podocytes consisted of four subcellular compartments: cell body, primary process, ridgelike prominence (RLP), and foot process. The RLP, a longitudinal protrusion from the basal surface of the cell body and primary process, served as an adhesive apparatus for the cell body and primary process to attach to the glomerular basement membrane. Foot processes protruded from both sides of the RLP. In puromycin nephrotic podocytes, foot process effacement occurred in two ways: by type-1 retraction, where the foot processes retracted while maintaining their rounded tips; or type-2 retraction, where they narrowed across their entire lengths, tapering toward the tips. Puromycin nephrotic podocytes also exhibited several alterations associated with foot process effacement, such as deformation of the cell body, retraction of RLPs, and cytoplasmic fragmentation. Finally, podocytes were reorganized into a broad, flattened shape. ConclusionsThe three-dimensional reconstruction of podocytes by serial FIB/SEM images revealed the morphologic changes involved in foot process effacement in greater detail than previously described.

show abstract

“…Array tomography is a potential solution to this problem. It has been used to obtain serial images from a series of ultrathin sections mounted on silicon wafer and glass plate substrates (Koga et al 2018;Koike et al 2017). These ultrathin sections have been shown to be physically stable and repeatable.…”

Section: Fib-sem Tomographymentioning

confidence: 99%

Three-dimensional imaging of podocyte ultrastructure using FE-SEM and FIB-SEM tomography

et al. 2019

View full text Add to dashboard Cite

Podocytes are specialized epithelial cells used for glomerular filtration in the kidney. They can be divided into the cell body, primary process and foot process. Here, we describe two useful methods for the three-dimensional(3D) visualization of these subcellular compartments in rodent podocytes. The first method, field-emission scanning electron microscopy (FE-SEM) with conductive staining, is used to visualize the luminal surface of numerous podocytes simultaneously. The second method, focusedion beam SEM (FIB-SEM) tomography, allows the user to obtain serial images from different depths of field, or Z-stacks, of the glomerulus. This allows for the 3D reconstruction of podocyte ultrastructure, which can be viewed from all angles, from a single image set. This is not possible with conventional FE-SEM. The different advantages and disadvantages of FE-SEM and FIB-SEM tomography compensate for the weaknesses of the other. The combination renders a powerful approach for the 3D analysis of podocyte ultrastructure. As a result, we were able to identify a new subcellular compartment of podocytes, "ridge-like prominences" (RLPs).

show abstract

A Device for Ribbon Collection for Array Tomography with Scanning Electron Microscopy

Cited by 19 publications

References 12 publications

Three-dimensional architecture of pericardial nephrocytes in Drosophila melanogaster revealed by FIB/SEM tomography

Three-dimensional architecture of pericardial nephrocytes in Drosophila melanogaster revealed by FIB/SEM tomography

Morphological Processes of Foot Process Effacement in Puromycin Aminonucleoside Nephrosis Revealed by FIB/SEM Tomography

Three-dimensional imaging of podocyte ultrastructure using FE-SEM and FIB-SEM tomography

Contact Info

Product

Resources

About