Correlation of organelle dynamics between light microscopic live imaging and electron microscopic 3D architecture using FIB-SEM

Ohta, Keisuke; Hirashima, Shingo; Miyazono, Yoshihiro; Togo, Akinobu; Nakamura, Kei‐ichiro

doi:10.1093/jmicro/dfaa071

Cited by 14 publications

(8 citation statements)

References 37 publications

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“…Secondly, it is difficult to accurately integrate immunohistochemical characteristics with 3D observations of the IC ultrastructure using the presented method. This limitation can be overcome using 3D correlative light-electron microscopy 34 – 36 . Finally, the major limitation of our study is that only a small number of samples could be analyzed by 3D ultrastructure reconstruction, due to the time-consuming manual segmentation.…”

Section: Discussionmentioning

confidence: 99%

Three-dimensional analysis of interstitial cells in the lamina propria of the murine vas deferens by confocal laser scanning microscopy and FIB/SEM

Hiroshige

Uemura

Hirashima

et al. 2022

Sci Rep

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The present study aimed to explore the three-dimensional (3D) ultrastructure of interstitial cells (ICs) within the lamina propria of the murine vas deferens and the spatial relationships between epithelial cells and surrounding cells. Focused ion beam scanning electron microscopy and confocal laser scanning microscopy were performed. ICs within the lamina propria had a flat, sheet-like structure of cytoplasm with multiple cellular processes. In addition, two types of 3D structures that comprised cell processes of flat, sheet-like ICs were observed: one was an accordion fold-like structure and the other was a rod-shaped structure. ICs were located parallel to the epithelium and were connected to each other via gap junctions or adherens junctions. Moreover, multiple sphere-shaped extracellular vesicle-like structures were frequently observed around the ICs. The ICs formed a complex 3D network comprising sheet-like cytoplasm and multiple cell processes with different 3D structures. From this morphological study, we noted that ICs within the lamina propria of murine vas deferens may be involved in signal transmission between the epithelium and smooth muscle cells by physical interaction and by exchanging extracellular vesicles.

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Section: Discussionmentioning

confidence: 99%

Three-dimensional analysis of interstitial cells in the lamina propria of the murine vas deferens by confocal laser scanning microscopy and FIB/SEM

Hiroshige

Uemura

Hirashima

et al. 2022

Sci Rep

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“…Indeed, even if the ribbon organization appears to be mediated by the formation and fusion of tubules connecting the stacks (Feinstein and Linstedt, 2008), the few ultrastructural studies conducted so far indicate that the non-compact zones have the form of tubular-saccular membranes, suggesting that these membranes have a structure more complex than that of simple tubules (Saraste and Prydz, 2019). Therefore, we hope for a revived interest in this topic thanks to the development of electron microcopy (EM) approaches characterized by improved resolution and automatization (FIB-SEM, STEM, Array Tomography) (Ferguson et al, 2017;Koga et al, 2017;Ohta et al, 2021) that now also allow a faster examination of the detailed ultrastructure of larger and deeper Golgi areas.…”

Section: The Ribbon Is Actively Separated Into Stacks During G2mentioning

confidence: 99%

Structural Organization and Function of the Golgi Ribbon During Cell Division

Ayala

Colanzi²

2022

Front. Cell Dev. Biol.

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The Golgi complex has a central role in the secretory traffic. In vertebrate cells it is generally organized in polarized stacks of cisternae that are laterally connected by membranous tubules, forming a structure known as Golgi ribbon. The steady state ribbon arrangement results from a dynamic equilibrium between formation and cleavage of the membrane tubules connecting the stacks. This balance is of great physiological relevance as the unlinking of the ribbon during G2 is required for mitotic entry. A block of this process induces a potent G2 arrest of the cell cycle, indicating that a mitotic “Golgi checkpoint” controls the correct pre-mitotic segregation of the Golgi ribbon. Then, after mitosis onset, the Golgi stacks undergo an extensive disassembly, which is necessary for proper spindle formation. Notably, several Golgi-associated proteins acquire new roles in spindle formation and mitotic progression during mitosis. Here we summarize the current knowledge about the basic principle of the Golgi architecture and its functional relationship with cell division to highlight crucial aspects that need to be addressed to help us understand the physiological significance of the ribbon and the pathological implications of alterations of this organization.

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“…However, a focused ion beam (FIB) with a gallium ion source allows precise milling. Therefore, the tomography technique using FIB-SEM is used for the 3D reconstruction of cell organelles 11,12 . However, FIB can process a small area, limiting the processing of large samples 13 .…”

Section: Introductionmentioning

confidence: 99%

Intact observation of secondary xylem and parenchyma cells in woody plants using a combination of scanning electron microscopy and broad argon ion beam milling

Hatano

Nakaba

Horikawa

et al. 2022

Preprint

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The secondary xylem of woody plants consists of fragile living cells and rigid cell walls. However, xylem structures are easily damaged during mechanical cross-sectioning for electron microscopy analysis. A broad argon ion beam (BIB) milling technique is commonly employed for analysis using scanning electron microscopy (SEM) in material science to generate a large and distortion-free cross-section. However, BIB milling is rarely used in plant science. In this study, SEM combined with the BIB milling technique was validated as an accurate tool for structural observation of xylem tissues of two woody plants, Pinus densiflora (Japanese red pine) and Quercus phillyraeoides (Ubame oak), in comparison to cutting with the classical microtome. The BIB milling method does not require epoxy resin embedding because of prior chemical fixation and critical point drying of the sample, thus producing a three-dimensional image. The results showed that xylem structures were well-preserved in their natural state in the BIB-milled cross-section compared with the microtome cross-section. The observation using SEM with BIB milling was found to be useful for imaging hard and soft plant tissues, particularly the ultrastructure of fragile secondary xylem cells. Thus, we propose a new imaging method for detailed observation of plant tissue structures.

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Correlation of organelle dynamics between light microscopic live imaging and electron microscopic 3D architecture using FIB-SEM

Cited by 14 publications

References 37 publications

Three-dimensional analysis of interstitial cells in the lamina propria of the murine vas deferens by confocal laser scanning microscopy and FIB/SEM

Three-dimensional analysis of interstitial cells in the lamina propria of the murine vas deferens by confocal laser scanning microscopy and FIB/SEM

Structural Organization and Function of the Golgi Ribbon During Cell Division

Intact observation of secondary xylem and parenchyma cells in woody plants using a combination of scanning electron microscopy and broad argon ion beam milling

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