Radiation damage of water in environmental scanning electron microscopy

Royall, C. Patrick; Thiel, B. L.; Donald, Athene M.

doi:10.1046/j.1365-2818.2001.00948.x

Cited by 107 publications

(98 citation statements)

References 17 publications

(40 reference statements)

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“…However, application of a coating of course requires that the specimen be in the solid state (either dried or frozen)-a step that was avoided for the purposes of this investigation. Finally, it should be noted that although there is a possibility of radiation damage to uncoated specimens in the water vapor environment of an ESEM (Jenkins and Donald 1997, Kitching and Donald 1998, Royall et al 2001, this was not found to be significant under the conditions used for this work.…”

Section: Resultsmentioning

confidence: 84%

Electron microscopy of mammalian cells in the absence of fixing, freezing, dehydration, or specimen coating

et al. 2003

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Summary: Human osteoblast-like (bone-forming) cells were imaged using environmental scanning electron microscopy (ESEM). The cells were hydrated, unfrozen, and uncoated. Specimens were cooled to 3°C and imaged in water vapor, with partial pressures varying from saturated conditions to a humidity of approximately 50%, relative to pure water. The ESEM images show the presence of cell nuclei, nucleoli, and cytoplasmic membranes. Comparisons between chemically fixed and unfixed specimens (neither dried nor coated) show that cell morphologies are similar in both cases. These results are compared with a fixed, dried, carbon-coated specimen. Thermodynamic and kinetic arguments are used to show that humidities significantly lower than 100% correspond to metastable states suitable for stabilizing hydrated biological tissues and cells. The ability to perform observations with minimal specimen preparation is potentially useful for studying interactions between mammalian cells and biomaterials that are developed for tissue engineering. The methods employed are equally applicable to the study of specimens in the biological, materials, and physical sciences where careful control over specimen stability is required.

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Section: Resultsmentioning

confidence: 84%

Electron microscopy of mammalian cells in the absence of fixing, freezing, dehydration, or specimen coating

et al. 2003

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“…For further details concerning radiation damage of water and of wet samples see e.g. Kitching and Donald (1998) or Royall et al (2001). To minimize radiation the acceleration voltage was kept low (8 kV) and the beam exposure time short.…”

Section: Resultsmentioning

confidence: 99%

The fracture behaviour of single wood fibres is governed by geometrical constraints: in situ ESEM studies on three fibre types

2008

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In situ tensile tests were performed in an environmental scanning electron microscope (ESEM) on earlywood, transition wood and latewood cells of Norway spruce (Picea abies [L.] Karst.). In order to examine the single wood fibres in a wet state, a specially designed tensile testing stage with a cooling device was built. The fracture behaviour of the cell types was studied at high resolution while straining. Different failure mechanisms were observed for the three tissue types. The thin-walled earlywood fibres showed tension buckling which gave rise to crack initiation and resulted in low tensile strength, whereas thick-walled latewood fibres predominately failed by transverse crack propagation without fibre folding.

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“…This implies that the beam damage to yeast cells occurred significantly faster under the low vacuum mode than the high vacuum. The result is expected as the water molecules present under the low vacuum mode can accelerate specimen degradation (Royall et al 2001;Kitching and Donald 1998). …”

Section: Calibration Of the Moving Speed Of The Nanomanipulatormentioning

confidence: 99%

Investigation of the morphology, viability and mechanical properties of yeast cells in environmental SEM

2008

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The mechanical properties of biological cells at nanoscale may be characterized using an environmental scanning electron microscopy (ESEM) combined with a force measurement device. However, the electron beam radiation in an ESEM may damage a specimen. So far, little is known about the radiation damage to biological cells. In this work, single yeast cells were imaged using an ESEM under both high and low vacuum modes. The changes in their morphology and viability were monitored as a function of radiation time for a given beam current of 538 pA corresponding to 10 kV accelerating voltage and spot size 4. Under the two modes, the radiation damage to the morphology of yeast cells became evident after an exposure time of 3 min, but under the low vacuum mode, the damage to their morphology was more severe. However, all cells lost their viability after 5 min under the high vacuum mode with the electron beam off from an initial viability of 95+/-1%. In contrast, the viability of cells under the low vacuum mode was found to be approximately 20% after 20 min. In addition, a newly developed ESEM-based nanomanipulation technique was applied to measure the force imposed on single yeast cells and their deformation, including contact diameter and central lateral diameter for the compression of single yeast cells to a given displacement within a time frame of 1 min, and the data obtained may be used to validate mathematical modeling of the stress-strain relationship for the compression of cells in order to determine their intrinsic mechanical property parameters.

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Radiation damage of water in environmental scanning electron microscopy

Cited by 107 publications

References 17 publications

Electron microscopy of mammalian cells in the absence of fixing, freezing, dehydration, or specimen coating

Electron microscopy of mammalian cells in the absence of fixing, freezing, dehydration, or specimen coating

The fracture behaviour of single wood fibres is governed by geometrical constraints: in situ ESEM studies on three fibre types

Investigation of the morphology, viability and mechanical properties of yeast cells in environmental SEM

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