Scanning electron microscopy: an introduction

Vernon–Parry, K.D.

doi:10.1016/s0961-1290(00)80006-x

Cited by 163 publications

(128 citation statements)

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“…TEM specimen supporters with fine controlled porous structures were selected as the modular substrate and fabricated in 3D CCISs. Micrographs of the exemplary HDFs cultured on the thin modular substrate were captured at multiple‐ and single‐cell levels using optical microscopes during cell culture; in situ analysis was also conducted at micro‐ and nanoscales after cell culture via SEM and TEM with higher resolutions (Clarke, ; Vernon‐Parry, ). Our research demonstrated that the scale‐down study design is an effective approach to systematically evaluate individual elementary structural components of complex 3D matrices.…”

Section: Discussionmentioning

confidence: 99%

A novel scale‐down cell culture and imaging design for the mechanistic insight of cell colonisation within porous substrate

Gabbott

Zhou

Han

et al. 2017

Journal of Microscopy

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SummaryAt the core of translational challenges in tissue engineering is the mechanistic understanding of the underpinning biological processes and the complex relationships among components at different levels, which is a challenging task due to the limitations of current tissue culture and assessment methodologies. Therefore, we proposed a novel scale-down strategy to deconstruct complex biomatrices into elementary building blocks, which were resembled by thin modular substrate and then evaluated separately in miniaturised bioreactors using various conventional microscopes. In order to investigate cell colonisation within porous substrate in this proof-of-concept study, TEM specimen supporters (10-30 μm thick) with fine controlled open pores (100ß600 μm) were selected as the modular porous substrate and suspended in 3D printed bioreactor systems. Noninvasive imaging of human dermal fibroblasts cultured on these free-standing substrate using optical microscopes illustrated the complicated dynamic processes used by both individual and coordinated cells to bridge and segment porous structures. Further in situ analysis via SEM and TEM provided high-quality micrographs of cell-cell and cell-scaffold interactions at microscale, depicted cytoskeletal structures in stretched and relaxed areas at nanoscale. Thus this novel scaled-down design was able to improve our mechanistic understanding of tissue formation not only at singleand multiple-cell levels, but also at micro-and nanoscales, which could be difficult to obtain using other methods.

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Section: Discussionmentioning

confidence: 99%

A novel scale‐down cell culture and imaging design for the mechanistic insight of cell colonisation within porous substrate

Gabbott

Zhou

Han

et al. 2017

Journal of Microscopy

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“…After high-energy electron beams have been applied to a specimen, EM instruments collect signals from transmitted electrons (transmission EM) (9), reflected electrons (reflection EM) (10), or secondary electrons (scanning EM) (11). The resolution of EM is determined by the wavelength of the incident electron beam.…”

Section: Strategies For Peptidoglycan Imagingmentioning

confidence: 99%

Imaging Bacterial Cell Wall Biosynthesis

Radkov

Hsu

Booher

et al. 2018

Annu. Rev. Biochem.

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Peptidoglycan is an essential component of the cell wall that protects bacteria from environmental stress. A carefully coordinated biosynthesis of peptidoglycan during cell elongation and division is required for cell viability. This biosynthesis involves sophisticated enzyme machineries that dynamically synthesize, remodel, and degrade peptidoglycan. However, when and where bacteria build peptidoglycan, and how this is coordinated with cell growth, have been long-standing questions in the field. The improvement of microscopy techniques has provided powerful approaches to study peptidoglycan biosynthesis with high spatiotemporal resolution. Recent development of molecular probes further accelerated the growth of the field, which has advanced our knowledge of peptidoglycan biosynthesis dynamics and mechanisms. Here, we review the technologies for imaging the bacterial cell wall and its biosynthesis activity. We focus on the applications of fluorescent d-amino acids, a newly developed type of probe, to visualize and study peptidoglycan synthesis and dynamics, and we provide direction for prospective research.

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“…Also an independent determination of the density and the thickness is not possible. Techniques based on electrons [27] like Auger electron spectroscopy (AES) or X-ray photoelectron spectroscopy (XPS) are surface-sensitive and give, in contrast to ellipsometry [28], scanning electron microscopy (SEM) [29] or scanning tunneling microscopy (STM) [30], a direct information on the elemental and chemical composition. However, the mean penetration range of electrons severely limits the thickness of the sample which can be analyzed if this technique is not combined with sputtering.…”

Section: Introductionmentioning

confidence: 99%

High-energy-resolution grazing emission X-ray fluorescence applied to the characterization of thin Al films on Si

Kayser

Szlachetko

Banaś

et al. 2013

Spectrochimica Acta Part B: Atomic Spectroscopy

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The grazing emission X-ray fluorescence (GEXRF) technique was applied to the analysis of different Al films, with nominal thicknesses in the range of 1 nm to 150 nm, on Si wafers. In GEXRF the sample volume from which the fluorescence intensity is detected is restricted to a near-surface region whose thickness can be tuned by varying the observation angle. This is possible because of the refraction of the fluorescence X-rays and the quite long emission paths within the probed sample. By recording the X-ray fluorescence signal for different shallow emission angles, defined relatively to the flat, smooth sample surface, the deposited Al surface layers of the different samples could be well characterized in terms of layer thickness, layer density, oxidation and surface roughness. The advantages offered by synchrotron radiation and the employed wavelength-dispersive detection setup were profited from. The GEXRF results retrieved were confirmed by complementary measurements. The experimental setup, the principles and advantages of GEXRF and the analysis of the recorded angular intensity profiles will be discussed in details.

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Scanning electron microscopy: an introduction

Cited by 163 publications

References 0 publications

A novel scale‐down cell culture and imaging design for the mechanistic insight of cell colonisation within porous substrate

A novel scale‐down cell culture and imaging design for the mechanistic insight of cell colonisation within porous substrate

Imaging Bacterial Cell Wall Biosynthesis

High-energy-resolution grazing emission X-ray fluorescence applied to the characterization of thin Al films on Si

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