Open AccessThree-dimensional imaging of podocyte ultrastructure using FE-SEM and FIB-SEM tomography
Author(s) -
Takayoshi Miyaki,
Yuto Kawasaki,
Yasue Hosoyamada,
Takashi Amari,
Mui Kinoshita,
Hironori Matsuda,
Soichiro Kakuta,
Tatsuo Sakai,
Koichiro Ichimura
Publication year - 2019
Publication title -
cell and tissue research/cell and tissue research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.64
H-Index - 137
eISSN - 1432-0878
pISSN - 0302-766X
DOI - 10.1007/s00441-019-03118-3
Subject(s) - podocyte , ultrastructure , glomerulus , scanning electron microscope , microscopy , materials science , glomerular basement membrane , focused ion beam , tomography , chemistry , biophysics , anatomy , microbiology and biotechnology , biomedical engineering , biology , pathology , kidney , optics , physics , glomerulonephritis , ion , medicine , proteinuria , composite material , organic chemistry , endocrinology
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, focused-ion 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).