Scanning electron microscopy of intestinal microvilli

Millington, P. F.; Critchley, David R.; Tovell, P. W. A.; Pearson, Roger

doi:10.1111/j.1365-2818.1969.tb00681.x

Cited by 22 publications

(9 citation statements)

References 8 publications

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“…In these four preparations the microvilli were morphologically identical, and in addition, were similar to those seen in intact mucosa from human and rat jejunum (Swift and Marsh, 1968;Marsh and Swift, 1969). It is therefore apparent that the demonstration of microvilli is not due to artefacts in preparation, as suggested by Millington, Critchley, Tovell, and Pearson (1969).…”

Section: Discussionsupporting

confidence: 67%

Observations of isolated enterocytes and of their subcellular components using transmission and scanning electron microscopy

Marsh¹,

Peters²,

Brown³

1971

Gut

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SUMMARY Isolated epithelial cells and their subcellular organelles were examined both by transmission and by scanning electron microscopy. It was shown that scanning electron microscopy is particularly useful for displaying the three-dimensional structure of the brush border. Furthermore, it has been possible to observe the basal aspect of separated epithelium for the first time. Studies on the scanning electron microscopic appearance of isolated nuclei, mitochondria, and lysosomes did not reveal any evidence of surface structure.The use of the scanning electron microscope for studying intestinal epithelium has already been described in this journal (Marsh and Swift, 1969). Several techniques are now available which may be used to prepare isolated intestinal epithelial cells (enterocytes), and subcellular fractions of these cells may also be obtained in relatively pure form. In this paper the appearances of some of these fractions have been studied using scanning electron microscopy and compared with those observed through the transmission electron microscope. The results of these investigations are described in relation to existing knowledge of the structure of the intestinal epithelium. Materials and MethodsIsolated epithelial cells were prepared from the jejunum of adult rats and guinea pigs. Pieces of small intestinal mucosa were incubated either in Ca2+-and Mg2+-free Tyrode's solution containing 0-1 ,g/100 ml phospholipase C or in buffered 10 mM EDTA (Evans, Wrigglesworth, Burdett, and Pover, personal communication). The cells were harvested by gentle centrifugation, and were washed and resuspended in buffered 0-2 M mannitol.Brush borders were prepared by the methods of Eichholz and Crane (1965) and of Hubscher, West,

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Section: Discussionsupporting

confidence: 67%

Observations of isolated enterocytes and of their subcellular components using transmission and scanning electron microscopy

Marsh¹,

Peters²,

Brown³

1971

Gut

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“…Previously, AFM studies have shown that it is possible to track protein motion 9 and capture cell surface receptor interactions 10 at room-temperature in aqueous media, which is an experimentally demanding environment when compared to cryogenic AFM measurements conducted in an ultraclean and thermally stable experimental platform. Although it is imperative to study biomolecular dynamics under physiological conditions, it has been shown that the intricate structures of fixed cells (in a dry state) can be better resolved with electron microscopy operating in vaccum 11 than with AFM in water 12 . During electron microscopy studies of fixed cells the samples are protected from ambient contamination but at the cost of losing soluble cell contents 11 .…”

Section: Introductionmentioning

confidence: 99%

“…Although it is imperative to study biomolecular dynamics under physiological conditions, it has been shown that the intricate structures of fixed cells (in a dry state) can be better resolved with electron microscopy operating in vaccum 11 than with AFM in water 12 . During electron microscopy studies of fixed cells the samples are protected from ambient contamination but at the cost of losing soluble cell contents 11 . Consequently, an alternative method is required, which leverages recent advances in instrumentation, preserves cellular morphology and simultaneously detects subcellular topological features without the requirement for multistep sample preparation procedures as in cryosectioning for AFM based cellular imaging 13 .…”

Section: Introductionmentioning

confidence: 99%

Subcellular Imaging of Liquid Silicone Coated-Intestinal Epithelial Cells

Nirmalraj

Lehner

Thompson

et al. 2018

Sci Rep

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Surface contamination and the formation of water bridge at the nanoscopic contact between an atomic force microscope tip and cell surface limits the maximum achievable spatial resolution on cells under ambient conditions. Structural information from fixed intestinal epithelial cell membrane is enhanced by fabricating a silicone liquid membrane that prevents ambient contaminants and accumulation of water at the interface between the cell membrane and the tip of an atomic force microscope. The clean and stable experimental platform permits the visualisation of the structure and orientation of microvilli present at the apical cell membrane under standard laboratory conditions together with registering topographical features within a microvillus. The method developed here can be implemented for preserving and imaging contaminant-free morphology of fixed cells which is central for both fundamental studies in cell biology and in the emerging field of digital pathology.

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“…Electron microscopy (EM), with its ability to provide information about ultrastructural details, was the key method for microvilli characterization in the pioneering works 41 . However, EM cannot visualize specific proteins efficiently, since the labelling densities are limited by ligand/antigen accessibility, steric hindrance and electron repulsion [42][43][44] .…”

Section: Introductionmentioning

confidence: 99%

Unraveling nanotopography of cell surface receptors

Franke

Chum

Kvíčalová

et al. 2019

Preprint

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Cells communicate with their environment via surface receptors, but receptor organization with respect to complex cell surface morphology remains unclear. This is mainly due to a lack of accessible, robust and high-resolution methods. Here, we present an approach for mapping the topography of receptors at the cell surface with nanometer precision. The method involves coating glass coverslips with a glycine hydrogel, which preserves the fine membrane morphology while allowing immobilized cells to be positioned close to the optical surface. We developed an advanced and simplified algorithm for the analysis of single-molecule localization data acquired in a biplane detection scheme. These advancements enable direct and quantitative mapping of protein distribution on ruffled plasma membranes with near isotropic 3D nanometer resolution. Tested successfully on CD4 and CD45, the described workflow is a simple and straightforward technique to study molecules and their interactions at the complex surface nanomorphology of differentiated metazoan cells.

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Scanning electron microscopy of intestinal microvilli

Cited by 22 publications

References 8 publications

Observations of isolated enterocytes and of their subcellular components using transmission and scanning electron microscopy

Observations of isolated enterocytes and of their subcellular components using transmission and scanning electron microscopy

Subcellular Imaging of Liquid Silicone Coated-Intestinal Epithelial Cells

Unraveling nanotopography of cell surface receptors

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