Vitrification of cryoelectron microscopy specimens revealed by high‐speed photographic imaging

@article{Kasas2003VitrificationOC,
  title={Vitrification of cryoelectron microscopy specimens revealed by high‐speed photographic imaging},
  author={Sandor Kasas and Guy Dumas and Giovanni Dietler and Stefano Catsicas and Marc Adrian},
  journal={Journal of Microscopy},
  year={2003},
  volume={211}
}
Cryoelectron microsopy is a widely used technique to observe biological material in an almost physiological, fully hydrated state. The sample is prepared for electron microsopy observation by quickly reducing its temperature to −180 °C. The high‐speed cooling induces the formation of vitreous water, which preserves the sample conformation. However, the way vitrification occurs is still poorly understood. In order to better understand the phenomenon, we have used a stroboscopic device to… Expand
Cryo-EM structures from sub-nl volumes using pin-printing and jet vitrification
TLDR
The authors present VitroJet, a single device, where sub-nl volumes of samples are deposited by pin printing thus eliminating the need for sample blotting, which is followed by jet vitrification, and they show that high-resolution structures can be obtained using four standard proteins. Expand
Understanding the invisible hands of sample preparation for cryo-EM.
TLDR
This Review aims to categorize and explain the principles behind various techniques in the preparation of vitrified samples for the electron microscope, and to give guidelines and posit future directions for improvement. Expand
Imaging nanostructured fluids using cryo-TEM
Cryogenic transmission electron microscopy (cryo-TEM) has gained increasing popularity among researchers in the field of amphiphilic self-assembly as an experimental method of choice because of itsExpand
HPM Live μ for a full CLEM workflow
TLDR
This chapter introduces a novel HPF live μ instrument with a focus on its coupling to a light microscope, elaborate on the optimization of sample preservation and the time needed to capture a biological event, going from live imaging to cryo-arrest using HPF, and addresses the adaptation of HPF to novel correlation workflows related to the forthcoming transition from imaging 2D to 3D samples. Expand
HPM live μ for a full CLEM workflow.
TLDR
The optimization of sample preservation and the time needed to capture a biological event, going from live imaging to cryo-arrest using HPF is addressed, and the HPF core technology has been revisited to allow easy modification of the environmental parameters during vitrification. Expand
Microstructure of milk gel and cheese curd observed using cryo scanning electron microscopy and confocal microscopy
Cryo scanning electron microscopy (cryo SEM) and confocal laser scanning microscopy (CLSM) were used to visualise changes in the microstructure of milk, rennet-induced gel and curd during theExpand
Nanofluidic chips for cryo-EM structure determination from picoliter sample volumes
TLDR
It is shown that a nanofluidic sample support with well-defined geometry can be used to prepare cryo-EM specimens with reproducible ice thickness from picoliter sample volumes and to perform high-resolution structure determination with three standard protein specimens. Expand
New controlled environment vitrification system for preparing wet samples for cryo‐SEM
TLDR
Complementary cryo‐SEM images revealed that the freezing rate and surface shearing of a sample need to be balanced by adjusting the plunging attitude, and a new hypothesis about the width of the band of extremely rapid cooling by deeply subcooled nucleate boiling below the line of entry was interpreted. Expand
Effects of cryo-EM cooling on structural ensembles
TLDR
This work combined continuum model calculations of the temperature drop, molecular dynamics simulations of a ribosome complex before and during cooling with kinetic models, and the quantification of the heterogeneity of room-temperature ensembles from cryo-EM structures to suggest three effects markedly contribute to the narrowing of the structural ensemble. Expand
Molecular dynamics of protein complexes from four-dimensional cryo-electron microscopy.
TLDR
The strategy employed in a recent analysis of herpes simplex virus procapsid maturation is described, and various factors that impinge on the feasibility of performing similar experimental analyses of molecular dynamics in the general case are discussed. Expand
...
1
2
3
...

References

SHOWING 1-10 OF 16 REFERENCES
Cryo-electron microscopy of vitrified specimens.
Water is the most abundant component of biological material, but it is systematically excluded from conventional electron microscopy. This is because water evaporates rapidly under the vacuumExpand
Vitrification of aqueous suspensions from a controlled environment for electron microscopy: An improved plunge‐cooling device
In the process of vitrifying aqueous suspensions for cryotransmission electron microscopy, water is solidified without crystallization. Vitrification can be achieved by rapidly plunging an aqueousExpand
Concentration of solutes during preparation of aqueous suspensions for cryo‐electron microscopy
TLDR
It is demonstrated that solutes are likely to be significantly concentrated under conditions commonly used to prepare vitrified aqueous suspensions for transmission electron microscopy, probably due to an increase in salt concentration caused by evaporation of water immediately prior to freezing. Expand
New controlled environment vitrification system for cryo‐transmission electron microscopy: design and application to surfactant solutions
TLDR
These experiments demonstrate that cryo TEM experiments not only provide direct structural information, but furthermore permit a quantitative determination of micellar properties, which very critically depends on the composition of the sample. Expand
Evaporation during preparation of unsupported thin vitrified aqueous layers for cryo-electron microscopy.
TLDR
The drying effect is generally negligible when specimens are prepared on a hydrophilic perforated support but it is frequently important when hydrophobic films are used instead and this effect is quantitatively studied by measuring the contrast of polystyrene spheres in a metrizamide solution. Expand
Transmission Electron Microscopy of Complex Fluids: The State of the Art
Transmission electron microscopy (TEM) is potentially one of the most useful techniques in the study of micro-structured fluid systems. It is the one technique that provides high resolution directExpand
Cryo-electron microscopy of viruses
TLDR
Cryo-electron microscopy of vitrified specimens offers possibilities for high resolution observations that compare favourably with any other electron microscopical method. Expand
Controlled environment vitrification system: an improved sample preparation technique.
TLDR
The controlled environment vitrification system (CEVS) is described and the value of the technique is illustrated with TEM micrographs of surfactant microstructures in which specimen preparation artifacts were avoided. Expand
Fast-freezing devices for cryo-electron-microscopy
Abstract The design, construction and performance of two single fast-freezing devices are described. The devices are driven pneumatically at a controlled speed of up to 100msec -1 . They are suitableExpand
Fast processes imaging device
Reference LPMV-ARTICLE-2003-011doi:10.1119/1.1523074View record in Web of Science Record created on 2007-06-20, modified on 2017-05-12
...
1
2
...