Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus

Koga, Daisuke; Ushiki, Tatsuo; Watanabe, Tsuyoshi

doi:10.1007/s12565-016-0380-8

Cited by 17 publications

(10 citation statements)

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“…3D-EM techniques made data available for the Golgi of plants (Staehelin and Kang 2008) and mammals (Ladinsky et al 1999; Marsh and Pavelka 2013; Koga et al 2017). Telophase is most instructive to understand the metamorphosis of the Golgi.…”

Section: Golgi Transitionmentioning

confidence: 99%

“…The Golgi has been studied extensively in structure and function (Sütterlin and Colanzi 2010 ; Wang and Seemann 2011 ; Yadav and Linstedt 2011 ; Gosavi and Gleeson 2017 ; Wei and Seemann 2017 ). 3D-EM techniques made data available for the Golgi of plants (Staehelin and Kang 2008 ) and mammals (Ladinsky et al 1999 ; Marsh and Pavelka 2013 ; Koga et al 2017 ). Telophase is most instructive to understand the metamorphosis of the Golgi.…”

Section: Golgi Transitionmentioning

confidence: 99%

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Precise and economic FIB/SEM for CLEM: with 2 nm voxels through mitosis

Luckner

Wanner

2018

Histochem Cell Biol

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A portfolio is presented documenting economic, high-resolution correlative focused ion beam scanning electron microscopy (FIB/SEM) in routine, comprising: (i) the use of custom-labeled slides and coverslips, (ii) embedding of cells in thin, or ultra-thin resin layers for correlative light and electron microscopy (CLEM) and (iii) the claim to reach the highest resolution possible with FIB/SEM in xyz. Regions of interest (ROIs) defined in light microscope (LM), can be relocated quickly and precisely in SEM. As proof of principle, HeLa cells were investigated in 3D context at all stages of the cell cycle, documenting ultrastructural changes during mitosis: nuclear envelope breakdown and reassembly, Golgi degradation and reconstitution and the formation of the midzone and midbody.Electronic supplementary materialThe online version of this article (10.1007/s00418-018-1681-x) contains supplementary material, which is available to authorized users.

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Section: Golgi Transitionmentioning

confidence: 99%

Section: Golgi Transitionmentioning

confidence: 99%

Precise and economic FIB/SEM for CLEM: with 2 nm voxels through mitosis

Luckner

Wanner

2018

Histochem Cell Biol

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“…Using the maceration method, the 3D ultrastructure of cell organelles, such as the Golgi apparatus, mitochondria, ER, ribosomes on the rough ER, has been investigated using SEM in various tissues (Tanaka and Naguro, 1981 ; Tanaka and Mitsushima, 1984 ; Tanaka et al, 1986 ; Lea and Hollenberg, 1989 ; Lea et al, 1994 ; Ogata and Yamasaki, 1997 ). We have also applied the maceration method to elucidate the 3D ultrastructural characteristic of the Golgi apparatus and clarify the morphological diversity of this organelle in different types of cells (Koga and Ushiki, 2006 ; Koga et al, 2017a , see also Koga et al, 2017c for a review on this project). Thus, the osmium maceration method has broadened the possibilities for SE-mode SEM in the field of cell biology.…”

Section: Discussionmentioning

confidence: 99%

Applications of Scanning Electron Microscopy Using Secondary and Backscattered Electron Signals in Neural Structure

et al. 2021

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Scanning electron microscopy (SEM) has contributed to elucidating the ultrastructure of bio-specimens in three dimensions. SEM imagery detects several kinds of signals, of which secondary electrons (SEs) and backscattered electrons (BSEs) are the main electrons used in biological and biomedical research. SE and BSE signals provide a three-dimensional (3D) surface topography and information on the composition of specimens, respectively. Among the various sample preparation techniques for SE-mode SEM, the osmium maceration method is the only approach for examining the subcellular structure that does not require any reconstruction processes. The 3D ultrastructure of organelles, such as the Golgi apparatus, mitochondria, and endoplasmic reticulum has been uncovered using high-resolution SEM of osmium-macerated tissues. Recent instrumental advances in scanning electron microscopes have broadened the applications of SEM for examining bio-specimens and enabled imaging of resin-embedded tissue blocks and sections using BSE-mode SEM under low-accelerating voltages; such techniques are fundamental to the 3D-SEM methods that are now known as focused ion-beam SEM, serial block-face SEM, and array tomography (i.e., serial section SEM). This technical breakthrough has allowed us to establish an innovative BSE imaging technique called section-face imaging to acquire ultrathin information from resin-embedded tissue sections. In contrast, serial section SEM is a modern 3D imaging technique for creating 3D surface rendering models of cells and organelles from tomographic BSE images of consecutive ultrathin sections embedded in resin. In this article, we introduce our related SEM techniques that use SE and BSE signals, such as the osmium maceration method, semithin section SEM (section-face imaging of resin-embedded semithin sections), section-face imaging for correlative light and SEM, and serial section SEM, to summarize their applications to neural structure and discuss the future possibilities and directions for these methods.

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“…The method of optical microscopy is based on histological preparations of trabecular bone tissue with post-traumatic changes in structure, studied under an optical microscope. (13) characterized by a decrease in the area of bone beams and a violation of the integrity of bone lacunae [13][14][15].…”

Section: Analysis Of Recent Research and Publicationsmentioning

confidence: 99%

Development of a Method for Analyzing Tomographic Images of Bone Structures

Abramova

Аврунін

2020

Innovative Technologies and Scientific Solutions for Industries

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The subject matter of research in the article is the morphological structure of bone tissue in the lumbar spine, visualized by tomography in the sagittal and axial planes. The goal of the research is to create the most informative investigation method for analyzing the structure of bone tissue, taking into account pathology in the form of metastatic bone lesions. This is justified by the fact that the detection of pathological processes is one of the most important tasks of image processing and analysis; at the same time, early diagnosis of various pathologies, including cancer, significantly increases the chances of patient recovery. Tasks: to consider the existing modern methods for analyzing the structure of bone tissue, to develop and propose a method for detecting bone tissue pathology in multiple myeloma. For the development of methods for the analysis of tomographic images with lesion of the bone structure, one of the fundamental issues is visualization of tomographic data. In this case, it is advisable to provide modules for both two-dimensional and three-dimensional visualization with methods of processing and segmentation of vertebral bodies, as well as correcting the results obtained in an interactive mode. The research uses methods of improving the quality of images, filtering using adaptive local filters, segmentation and stereology methods, cluster analysis method. The result of the work is to obtain a method suitable for use in the analysis of bone tissue with its accompanying pathology in the form of bone metastases. This method will be the basis for the further development of a method aimed at analyzing the microstructure of bone tissue, which will significantly increase the accuracy of calculations. Conclusions. The relevance of the topic under study is of vital importance for a huge number of patients suffering from cancer. In the course of the research, algorithms for processing and analyzing input images were developed, taking into account the modality of the input data. This makes it possible to develop the next stage of analysis aimed at the microstructure of the bone tissue.

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Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus

Cited by 17 publications

References 83 publications

Precise and economic FIB/SEM for CLEM: with 2 nm voxels through mitosis

Precise and economic FIB/SEM for CLEM: with 2 nm voxels through mitosis

Applications of Scanning Electron Microscopy Using Secondary and Backscattered Electron Signals in Neural Structure

Development of a Method for Analyzing Tomographic Images of Bone Structures

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