Publication

Methods to overcome preferred orientation in cryo-samples for single particle analysis

Marcel Drabbels
2025

Abstract

A method for reducing a preferred orientation of particles in a cryo-sample includes receiving the cryo-sample in a vacuum chamber, depositing amorphous ice onto the cryo-sample while the cryo-sample is in the vacuum chamber, delivering an energy pulse to the cryo-sample in the vacuum chamber to temporarily melt the amorphous ice on the cryo-sample to liquid water to promote movement of the particles in the cryo-sample to a changed orientation different from the preferred orientation, and allowing the liquid water to revitrify to amorphous ice upon cessation of the energy pulse to fix or trap the particles in the cryo-sample in the changed orientation.

Official source

https://infoscience.epfl.ch/entities/publication/25b03f45-322c-4599-98fb-d5041a38331a

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Marcel Drabbels
2025

Abstract

Official source

https://infoscience.epfl.ch/entities/publication/25b03f45-322c-4599-98fb-d5041a38331a

About this result

Related concepts (21)

Single particle analysis

Single particle analysis is a group of related computerized image processing techniques used to analyze images from transmission electron microscopy (TEM). These methods were developed to improve and extend the information obtainable from TEM images of particulate samples, typically proteins or other large biological entities such as viruses. Individual images of stained or unstained particles are very noisy, and so hard to interpret. Combining several digitized images of similar particles together gives an image with stronger and more easily interpretable features.

Cryogenic electron microscopy

Cryogenic electron microscopy (cryo-EM) is a cryomicroscopy technique applied on samples cooled to cryogenic temperatures. For biological specimens, the structure is preserved by embedding in an environment of vitreous ice. An aqueous sample solution is applied to a grid-mesh and plunge-frozen in liquid ethane or a mixture of liquid ethane and propane. While development of the technique began in the 1970s, recent advances in detector technology and software algorithms have allowed for the determination of biomolecular structures at near-atomic resolution.

Liquid

A liquid is a nearly incompressible fluid that conforms to the shape of its container but retains a nearly constant volume independent of pressure. It is one of the four fundamental states of matter (the others being solid, gas, and plasma), and is the only state with a definite volume but no fixed shape. The density of a liquid is usually close to that of a solid, and much higher than that of a gas. Therefore, liquid and solid are both termed condensed matter.

Related publications (1)

Methods to overcome preferred orientation in cryo-samples for single particle analysis

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Laser flash melting cryo-EM samples to overcome preferred orientation