Journal
ANNUAL REVIEW OF BIOCHEMISTRY, VOL 90, 2021
Volume 90, Issue -, Pages 451-474Publisher
ANNUAL REVIEWS
DOI: 10.1146/annurev-biochem-072020-020231
Keywords
cryo-EM; grid preparation; air-water interface; protein denaturation; affinity grids
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Funding
- National Institutes of Health [GM051487, GM135666]
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Transmission electron microscopy poses extreme risk of damaging biological macromolecules due to interactions with the air-water interface. Current research is focused on finding methods to substantially reduce this hazard, primarily through immobilizing particles and reducing interaction time. There is also a growing focus on gaining more effective control of sample thickness before vitrification.
resolution electron microscopy, poses extreme risk of damaging biological macromolecules due to interactions with the air-water interface. Although the rapid increase in the number of published structures initially gave little indication that this was a problem, the search for methods that substantially mitigate this hazard is now intensifying. The two main approaches under investigation are (a) immobilizing particles onto structure-friendly support films and (b) reducing the length of time during which such interactions may occur. While there is little possibility of outrunning diffusion to the interface, intentional passivation of the interface may slow the process of adsorption and denaturation. In addition, growing attention is being given to gaining more effective control of the thickness of the sample prior to vitrification.
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