Principles and Practice of Variable Pressure/Environmental Scanning Electron Microscopy (VP‐ESEM)

Stokes, D. J.

doi:10.1002/9780470758731

Cited by 213 publications

(196 citation statements)

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“…We observed that areas of higher water content such as cytoplasm or extracellular space show a brighter grey level in the in-lens image than lipid-rich structures such as membranes and myelin. This effect is probably due to differences in charge dissipation which cause negative (SE1-release promoting) or positive (SE1-release suppressing) surface potentials as discussed (see chapter 5 (Stokes, 2008)). Surface potentials can be also influenced by Ga + implantation and loss of charged milling products during FIB milling.…”

Section: Resultsmentioning

confidence: 99%

Cryo FIB-SEM: Volume imaging of cellular ultrastructure in native frozen specimens

Schertel

Snaidero

Hong-mei

et al. 2013

Journal of Structural Biology

170

113

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Volume microscopy at high resolution is increasingly required to better understand cellular functions in the context of three-dimensional assemblies. Focused ion beam (FIB) milling for serial block face imaging in the scanning electron microscope (SEM)is an efficient and fast method to generate such volume data for 3D analysis. Here, we apply this technique at cryo-conditions to image fully hydrated frozen specimen of mouse optic nerves and Bacillus subtilis spores obtained by high-pressure freezing (HPF). We established imaging conditions to directly visualize the ultrastructure in the block face at −150°C by using an in-lens secondary electron (SE) detector. By serial sectioning with a focused ion beam and block face imaging of the optic nerve we obtained a volume as large as X=7.72 µm, Y=5.79 µm and Z=3.81 µm with a lateral pixel size of 7.5 nm and a slice thickness of 30 nm in Z. The intrinsic contrast of membranes was sufficient to distinguish structures like Golgi cisternae, vesicles, endoplasmic reticulum and cristae within mitochondria and allowed for a threedimensional reconstruction of different types of mitochondria within an oligodendrocyte and an astrocytic process. Applying this technique to dormant Bacillus subtilis spores we obtained volumes containing numerous spores and discovered a bright signal in the core, which can not be related to any known structure so far. In summary, we describe the use of cryo FIB-SEM as a tool for direct and fast 3D cryo-imaging of large native frozen samples including tissues.

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Section: Resultsmentioning

confidence: 99%

Cryo FIB-SEM: Volume imaging of cellular ultrastructure in native frozen specimens

Schertel

Snaidero

Hong-mei

et al. 2013

Journal of Structural Biology

170

113

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“…It may be due to relying more on unconfirmed assumptions than on rigorous quantitative analysis of the system. The present findings suggest that previous works expounding on the purpose and capabilities of ESEM (Cameron & Donald 1994;Stokes et al 2004;Stokes 2006;Griffin 2007;Stokes & Baken 2007;Toth et al 2007;Stokes 2008;Stokes et al 2008) should be looked at in a new light, and also in view of related prior works by the present author (Danilatos & Robinson, 1979;Danilatos, 1980Danilatos, , 1988Danilatos, , 1990Danilatos, , 1992Danilatos, , 1993aDanilatos, ,b, 2004.…”

Section: Discussionmentioning

confidence: 97%

“…Actually, the field of view needs not be sacrificed, if the instrument were to incorporate the relocation of the rocking point of the beam close to or at the PLA1, which would allow a very large field of view even with the use of very small PLAs including sizes of 100 µm and even 50 µm (Danilatos, 2004). Instead, the minimum size of a rather thick PLA appears invariable by the industry to only 0.5 mm diameter (Stokes, 2008) for a long time, for some hitherto arbitrary reason.…”

Section: Discussionmentioning

confidence: 99%

Figure of merit for environmental SEM and its implications

Danilatos¹

2011

Journal of Microscopy

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Summary A recently introduced figure of merit for environmental and low vacuum scanning electron microscopes has now been computed in the full operational pressure range for one commercial instrument. The direct simulation Monte Carlo method has been used in lieu of experimental measurements. The theory of this figure of merit is further consolidated. It is shown that a thin pressure limiting aperture can indeed be used as an optimum reference system for all instruments employing differential pumping in the transfer of an electron beam from high vacuum to high pressure. The implications of the results obtained are discussed both in relation to existing commercial instruments and associated literature to pave the way for future progress in the field.

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“…SEM can achieve resolution better than 1 nanometer. Specimens can be observed in high vacuum in conventional SEM, or in low vacuum or wet conditions in variable pressure or environmental SEM, and at a wide range of cryogenic or elevated temperatures with specialized instruments [7] The mechanical phases of PBDP were obtained from triaxle tester.…”

Section: Testingmentioning

confidence: 99%

The contribution agriculture waste reinforcement to the floor topping of renovation building architectures: compressive strength test by trial the percentage of the pseudostem of banana dried powder

et al. 2018

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Abstract. The growing demand of construction around the world has led to an increased usage of concrete. However, convention concrete making materials are not entirely environmental friendly and this has enthused research on seeking greener alternative for concrete production. Agriculture industries are one of the major industries globally that harvested products such as food and biomass for organism around the world. After harvesting products, there are effluence of agriculture waste as left, straw and pseudostem. This research focusing on mechanical properties as compressive Strength Test of mortar cement adding with the pseudostem of banana dried powder (PBDP) by trial the percentage of PBDP and chemical properties as using scanning electron microscope (SEM). These findings indicate that 10% of PBDP replacement is the best ratio and element of PBDP which C was the most element in PBDP as 59.96% of mass and 71.49 % of atomic.

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Principles and Practice of Variable Pressure/Environmental Scanning Electron Microscopy (VP‐ESEM)

Cited by 213 publications

References 0 publications

Cryo FIB-SEM: Volume imaging of cellular ultrastructure in native frozen specimens

Cryo FIB-SEM: Volume imaging of cellular ultrastructure in native frozen specimens

Figure of merit for environmental SEM and its implications

The contribution agriculture waste reinforcement to the floor topping of renovation building architectures: compressive strength test by trial the percentage of the pseudostem of banana dried powder

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