Beam damage to organic material is considerably reduced in cryo-electron microscopy

Knapek, E.; Dubochet, Jacques

doi:10.1016/0022-2836(80)90382-4

Cited by 135 publications

(63 citation statements)

References 23 publications

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“…Earlier studies that determined critical exposures did so only for high-resolution Fourier components (6 12 Å) (Taylor and Glaeser, 1976;Hayward and Glaeser, 1979;Unwin and Henderson, 1975;Knapek and Dubochet, 1980) and used electron accelerating voltages (e.g. 100 kV) and temperatures (room temperature or À120°C) no longer common for studying unstained specimens.…”

Section: Resultsmentioning

confidence: 99%

The resolution dependence of optimal exposures in liquid nitrogen temperature electron cryomicroscopy of catalase crystals

Baker

Smith

Bueler

et al. 2010

Journal of Structural Biology

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Section: Resultsmentioning

confidence: 99%

The resolution dependence of optimal exposures in liquid nitrogen temperature electron cryomicroscopy of catalase crystals

Baker

Smith

Bueler

et al. 2010

Journal of Structural Biology

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“…An effective way of decreasing radiation damage is to cool samples to temperatures around 100 K (15)(16)(17)(18)(19)(20), which is routinely done in x-ray crystallography and electron microscopy. At these temperatures, samples can tolerate a significantly higher dose of ionizing radiation, but eventually they show comparable signs of damage as observed at room temperature.…”

mentioning

confidence: 99%

Origin and temperature dependence of radiation damage in biological samples at cryogenic temperatures

Meents

Gutmann

Wagner

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

184

153

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Radiation damage is the major impediment for obtaining structural information from biological samples by using ionizing radiation such as x-rays or electrons. The knowledge of underlying processes especially at cryogenic temperatures is still fragmentary, and a consistent mechanism has not been found yet. By using a combination of single-crystal x-ray diffraction, small-angle scattering, and qualitative and quantitative radiolysis experiments, we show that hydrogen gas, formed inside the sample during irradiation, rather than intramolecular bond cleavage between non-hydrogen atoms, is mainly responsible for the loss of high-resolution information and contrast in diffraction experiments and microscopy. The experiments that are presented in this paper cover a temperature range between 5 and 160 K and reveal that the commonly used temperature in x-ray crystallography of 100 K is not optimal in terms of minimizing radiation damage and thereby increasing the structural information obtainable in a single experiment. At 50 K, specific radiation damage to disulfide bridges is reduced by a factor of 4 compared to 100 K, and samples can tolerate a factor of 2.6 and 3.9 higher dose, as judged by the increase of R free values of elastase and cubic insulin crystals, respectively.

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“…This radiation protection is a direct result of dynamically immobilizing the sample through embedment into vitrified ice (Knapek & Dubochet, 1980). By caging the sample in a frozen environment, free radicals generated from inelastic scattering events are unable to diffuse through the sample and cause secondary damage (Knapek & Dubochet, 1980).…”

Section: Benefits Of Low Temperature Imagingmentioning

confidence: 99%

Low-Dose Imaging Techniques for Transmission Electron Microscopy

Carlson¹,

Evans²

2012

The Transmission Electron Microscope

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Beam damage to organic material is considerably reduced in cryo-electron microscopy

Cited by 135 publications

References 23 publications

The resolution dependence of optimal exposures in liquid nitrogen temperature electron cryomicroscopy of catalase crystals

The resolution dependence of optimal exposures in liquid nitrogen temperature electron cryomicroscopy of catalase crystals

Origin and temperature dependence of radiation damage in biological samples at cryogenic temperatures

Low-Dose Imaging Techniques for Transmission Electron Microscopy

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