A critical overview of ESEM applications in the biological field

Muscariello, Livio; Rosso, Francesco; Marino, G.; Giordano, Antonio; Barbarisi, Manlio; Cafiero, Gioacchino; Barbarisi, Alfonso

doi:10.1002/jcp.20444

Cited by 187 publications

(134 citation statements)

References 32 publications

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“…After fixation, the samples were rinsed twice with distilled water to avoid salt precipitation, and observed using a FEI-Quanta 200 Scanning Electron Microscope (Hillsboro, OR, USA) equipped with a gaseous secondary electron detector (GSED) under specific conditions: 12.5 kV, a spot size of 5-6, 5 torr vacuum, 2500× magnification and a 6 mm working distance. To avoid the collapse of cells, a pump-down routine of four cycles of 5/10 torr, finishing at 5 torr, was carried out for a specimen cooled to 3 • C (Muscariello et al, 2005). Image analysis was performed with ImageJ 1.42 (NIH, Bethesda, MD, USA), and the following parameters were evaluated: cellular shape was determined by area measurement and maximum Feret's diameter (maximum caliper length); the solidity of cells was estimated by the ratio of the area of a cell to the area of its convex hull (Peterbauer et al, 2011).…”

Section: Environmental Scanning Electron Microscopy (E-sem)mentioning

confidence: 99%

Morphological changes induced by advanced glycation endproducts in osteoblastic cells: Effects of co-incubation with alendronate

et al. 2013

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a b s t r a c tAdvanced glycation endproducts (AGEs) accumulate with age in various tissues, and are further increased in patients with Diabetes mellitus, in which they are believed to contribute to the development and progression of chronic complications that include a decrease in bone quality. Bisphosphonates are antiosteoporotic drugs that have been used for the treatment of patients with diabetic bone alterations, although with contradictory results. In the present study, we have evaluated the in vitro alterations on osteoblastic morphology by environmental scanning electron microscopy, in actin cytoskeleton and apoptosis induced by AGEs, as well as the modulation of these effects by alendronate (an N-containing bisphosphonate). Our present results provide evidence for disruption induced by AGEs of the osteoblastic actin cytoskeleton (geodesic domes) and significant alterations in cell morphology with a decrease in cellsubstratum interactions leading to an increase in apoptosis of osteoblasts and a decrease in osteoblastic proliferation. High concentrations of alendronate (10 −5 M, such as could be expected in an osteoclastic lacuna) further increase osteoblastic morphological and cytoskeletal alterations. However, low doses of alendronate (10 −8 M, compatible with extracellular fluid levels to which an osteoblast could be exposed for most of its life cycle) do not affect cell morphology, and in addition are able to prevent AGEs-induced alterations and consequently apoptosis of osteoblasts.

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Section: Environmental Scanning Electron Microscopy (E-sem)mentioning

confidence: 99%

Morphological changes induced by advanced glycation endproducts in osteoblastic cells: Effects of co-incubation with alendronate

et al. 2013

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“…This technique is particularly well adapted for the observation or experimentation on biological samples (Muscariello et al, 2005). Images of small and highly hydrated samples such as liposomes have been obtained by several authors (Perrie et al, 2007 ;Ruozi et al;) without any particular sample preparation.…”

Section: Biology and Soft Matter Applicationsmentioning

confidence: 99%

In Situ Experiments in the Scanning Electron Microscope Chamber

Podor¹,

Ravaux²,

Brau³

2012

Scanning Electron Microscopy

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“…A second report showed that it is possible to view unfixed, fully hydrated and uncoated cells with no dehydration effects for extended periods (in some cases even hours) but the user must have good knowledge of their sample. The osmotic pressure and rate of water loss from the specimen needs to be known to achieve this task [75,92].…”

Section: Advances In Sample Preparation Techniquesmentioning

confidence: 99%

Towards native-state imaging in biological context in the electron microscope

2009

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Modern cell biology is reliant on light and fluorescence microscopy for analysis of cells, tissues and protein localisation. However, these powerful techniques are ultimately limited in resolution by the wavelength of light. Electron microscopes offer much greater resolution due to the shorter effective wavelength of electrons, allowing direct imaging of sub-cellular architecture. The harsh environment of the electron microscope chamber and the properties of the electron beam have led to complex chemical and mechanical preparation techniques, which distance biological samples from their native state and complicate data interpretation. Here we describe recent advances in sample preparation and instrumentation, which push the boundaries of high-resolution imaging. Cryopreparation, cryoelectron microscopy and environmental scanning electron microscopy strive to image samples in near native state. Advances in correlative microscopy and markers enable high-resolution localisation of proteins. Innovation in microscope design has pushed the boundaries of resolution to atomic scale, whilst automatic acquisition of high-resolution electron microscopy data through large volumes is finally able to place ultrastructure in biological context.

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A critical overview of ESEM applications in the biological field

Cited by 187 publications

References 32 publications

Morphological changes induced by advanced glycation endproducts in osteoblastic cells: Effects of co-incubation with alendronate

Morphological changes induced by advanced glycation endproducts in osteoblastic cells: Effects of co-incubation with alendronate

In Situ Experiments in the Scanning Electron Microscope Chamber

Towards native-state imaging in biological context in the electron microscope

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