Scanning electron microscopy as a new tool for diagnostic pathology and cell biology

Hyams, Tzipi Cohen; Mam, Keriya; Killingsworth, Murray C.

doi:10.1016/j.micron.2019.102797

Cited by 27 publications

(16 citation statements)

References 20 publications

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“…In addition to working with tissues, ultra-thin epoxy resin embedding has also been applied to the preparation of cell cultures for scanning electron microscopy to image individual cells and cell-cell interactions on planar and threedimensional substrates in preference to critical point drying, but this technique was not optimised for BSEM to visualise intracellular compartments (70). Importantly, high-resolution images have been obtained only in studies examining semi-thin and ultra-thin sections (58,59), where staining with uranyl acetate and lead citrate enabled the reaching of a BSEM resolution comparable to that of transmission electron microscopy because such samples can be fully penetrated by an electron beam. However, this method demands tissue sectioning, which rarely allows one to keep the integrity of the samples including mineral deposits or metal implants.…”

Section: Discussionmentioning

confidence: 99%

“…In biomedicine, BSEM is broadly used for the imaging of unstained, freeze-dried cell cultures ( 54 ), wet calcified atherosclerotic lesions ( 55 ), implant-bone interface ( 56 ), cartilage and bone tissue ( 57 ), and a diversity of other tissues ( 58 ). Despite high-resolution BSEM and field emission SEM having been earlier employed for the multiscale visualisation of semi-thin and ultra-thin sections, respectively ( 58 , 59 ), these sample preparation approaches are inconvenient when working with mineralised tissues or specimens with incorporated solid implants. The techniques of correlative histology [i.e., a time-resolved combination of imaging modalities from lower to higher magnifications ( 1 , 60 )], such as the three-sectioning method, where epoxy resin-embedded tissue is sequentially cut into thick (≈300 μm), semi-thin (1–3 μm), and ultra-thin (60–90 nm) sections to specify regions of interest, have been proposed for working with hard tissues ( 61 ).…”

Section: Discussionmentioning

confidence: 99%

“…Importantly, high-resolution images have been obtained only in studies examining semi-thin and ultra-thin sections ( 58 , 59 ), where staining with uranyl acetate and lead citrate enabled the reaching of a BSEM resolution comparable to that of transmission electron microscopy because such samples can be fully penetrated by an electron beam. However, this method demands tissue sectioning, which rarely allows one to keep the integrity of the samples including mineral deposits or metal implants.…”

Section: Discussionmentioning

confidence: 99%

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EMbedding and Backscattered Scanning Electron Microscopy: A Detailed Protocol for the Whole-Specimen, High-Resolution Analysis of Cardiovascular Tissues

Mukhamadiyarov

Богданов

Глушкова

et al. 2021

Front. Cardiovasc. Med.

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Currently, an ultrastructural analysis of cardiovascular tissues is significantly complicated. Routine histopathological examinations and immunohistochemical staining suffer from a relatively low resolution of light microscopy, whereas the fluorescence imaging of plaques and bioprosthetic heart valves yields considerable background noise from the convoluted extracellular matrix that often results in a low signal-to-noise ratio. Besides, the sectioning of calcified or stent-expanded blood vessels or mineralised heart valves leads to a critical loss of their integrity, demanding other methods to be developed. Here, we designed a conceptually novel approach that combines conventional formalin fixation, sequential incubation in heavy metal solutions (osmium tetroxide, uranyl acetate or lanthanides, and lead citrate), and the embedding of the whole specimen into epoxy resin to retain its integrity while accessing the region of interest by grinding and polishing. Upon carbon sputtering, the sample is visualised by means of backscattered scanning electron microscopy. The technique fully preserves calcified and stent-expanded tissues, permits a detailed analysis of vascular and valvular composition and architecture, enables discrimination between multiple cell types (including endothelial cells, vascular smooth muscle cells, fibroblasts, adipocytes, mast cells, foam cells, foreign-body giant cells, canonical macrophages, neutrophils, and lymphocytes) and microvascular identities (arterioles, venules, and capillaries), and gives a technical possibility for quantitating the number, area, and density of the blood vessels. Hence, we suggest that our approach is capable of providing a pathophysiological insight into cardiovascular disease development. The protocol does not require specific expertise and can be employed in virtually any laboratory that has a scanning electron microscope.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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EMbedding and Backscattered Scanning Electron Microscopy: A Detailed Protocol for the Whole-Specimen, High-Resolution Analysis of Cardiovascular Tissues

Mukhamadiyarov

Богданов

Глушкова

et al. 2021

Front. Cardiovasc. Med.

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“…Scanning electron microscopy (SEM) has traditionally been used in the biomedical sciences to characterize the topography of the cell and tissue surfaces [ 42 ]. In addition, although several techniques are available to measure the size and shape of particles, those based on electron microscopy are often considered to be the preferred methods for characterizing their dimensional properties [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

In Silico Study, Physicochemical, and In Vitro Lipase Inhibitory Activity of α,β-Amyrenone Inclusion Complexes with Cyclodextrins

Oliveira

Menezes

Silva

et al. 2021

IJMS

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α,β-amyrenone (ABAME) is a triterpene derivative with many biological activities; however, its potential pharmacological use is hindered by its low solubility in water. In this context, the present work aimed to develop inclusion complexes (ICs) of ABAME with γ- and β-cyclodextrins (CD), which were systematically characterized through molecular modeling studies as well as FTIR, XRD, DSC, TGA, and SEM analyses. In vitro analyses of lipase activity were performed to evaluate possible anti-obesity properties. Molecular modeling studies indicated that the CD:ABAME ICs prepared at a 2:1 molar ratio would be more stable to the complexation process than those prepared at a 1:1 molar ratio. The physicochemical characterization showed strong evidence that corroborates with the in silico results, and the formation of ICs with CD was capable of inducing changes in ABAME physicochemical properties. ICs was shown to be a stronger inhibitor of lipase activity than Orlistat and to potentiate the inhibitory effects of ABAME on porcine pancreatic enzymes. In conclusion, a new pharmaceutical preparation with potentially improved physicochemical characteristics and inhibitory activity toward lipases was developed in this study, which could prove to be a promising ingredient for future formulations.

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“…Soft X-ray contact microscopy (SXCM) is a potent technique enabling imaging with a nanometric resolution of specific features in cells [1,2]. Most structures of mammalian cells have already been visualized by electron and fluorescent microscopy, commonly used in biological research [3,4]. However, these methods were biased by protocols for consecutive sample preparation and had insufficient resolution to analyse the structures inside the cell [5,6].…”

Section: Introductionmentioning

confidence: 99%

Imaging of Cell Structures Using Optimized Soft X-ray Contact Microscopy

et al. 2020

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This work is to study the relationship between the exposure conditions and the quality of cell imaging with soft X-ray contact microscopy (SXCM). It is a crucial step in the efficient visualization of cell structures. Three different human cell lines: DU145 prostate carcinoma cells, HCC38 breast cancer cells, and Poietics mesenchymal stem cells were used to establish the optimal exposure conditions in SXCM. The image quality depended on the soft X-ray (SXR) absorbed energy and photoresist development conditions. At lower SXR energy (200 or 400 SXR pulses), sharp cell edges, membrane projections, and cell–cell connections were visible. In contrast, higher energy (600 or 800 SXR pulses) allowed observation of the cytoskeleton and the nucleus in a cell type-dependent manner (the influence of cell thickness and internal complexity was noted).

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Scanning electron microscopy as a new tool for diagnostic pathology and cell biology

Cited by 27 publications

References 20 publications

EMbedding and Backscattered Scanning Electron Microscopy: A Detailed Protocol for the Whole-Specimen, High-Resolution Analysis of Cardiovascular Tissues

EMbedding and Backscattered Scanning Electron Microscopy: A Detailed Protocol for the Whole-Specimen, High-Resolution Analysis of Cardiovascular Tissues

In Silico Study, Physicochemical, and In Vitro Lipase Inhibitory Activity of α,β-Amyrenone Inclusion Complexes with Cyclodextrins

Imaging of Cell Structures Using Optimized Soft X-ray Contact Microscopy

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