Glass-forming microemulsions: vitrification of simple liquids and electron microscope probing of droplet-packing modes

Dubochet, Jacques; Adrian, Marc; Teixeira, J.; Alba, C.; Kadiyala, R. K.; MacFarlane, Douglas R.; Angell, C. Austen

doi:10.1021/j150670a042

Cited by 77 publications

(54 citation statements)

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“…They characterised vitreous, cubic and hexagonal forms of ice, and developed a simple plunge-freezing method for preparing grids Dubochet et al 1982Dubochet et al , 1984. Their work depended on the development of the first ultra-high vacuum specimen environment in the Philips 400 electron microscope, which had been required by the materials science community to avoid contamination of their clean metallic specimens.…”

Section: The Path To Cryoemmentioning

confidence: 99%

“…Following the preparation of samples for cryoEM using the method of Dubochet et al (1982Dubochet et al ( ,1984, we typically apply a small drop (3 µl) of a solution of the biological structure of interest to a holey carbon film on a metal EM grid. This is then blotted for a few seconds (using filter paper) and rapidly frozen by plunging the grid into liquid ethane cooled by liquid nitrogen to a temperature just above the freezing point of liquid ethane (−183°C).…”

Section: Beam-induced Specimen Movementmentioning

confidence: 99%

See 1 more Smart Citation

Single particle electron cryomicroscopy: trends, issues and future perspective

Vinothkumar

Henderson

2016

Quart. Rev. Biophys.

173

115

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Abstract. There has been enormous progress during the last few years in the determination of three-dimensional biological structures by single particle electron cryomicroscopy (cryoEM), allowing maps to be obtained with higher resolution and from fewer images than required previously. This is due principally to the introduction of a new type of direct electron detector that has 2-to 3-fold higher detective quantum efficiency than available previously, and to the improvement of the computational algorithms for image processing. In spite of the great strides that have been made, quantitative analysis shows that there are still significant gains to be made provided that the problems associated with image degradation can be solved, possibly by minimising beam-induced specimen movement and charge build up during imaging. If this can be achieved, it should be possible to obtain near atomic resolution structures of smaller single particles, using fewer images and resolving more conformational states than at present, thus realising the full potential of the method. The recent popularity of cryoEM for molecular structure determination also highlights the need for lower cost microscopes, so we encourage development of an inexpensive, 100 keV electron cryomicroscope with a high-brightness field emission gun to make the method accessible to individual groups or institutions that cannot afford the investment and running costs of a state-of-the-art 300 keV installation. A key requisite for successful high-resolution structure determination by cryoEM includes interpretation of images and optimising the biochemistry and grid preparation to obtain nicely distributed macromolecules of interest. We thus include in this review a gallery of cryoEM micrographs that shows illustrative examples of single particle images of large and small macromolecular complexes.

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Section: The Path To Cryoemmentioning

confidence: 99%

Section: Beam-induced Specimen Movementmentioning

confidence: 99%

Single particle electron cryomicroscopy: trends, issues and future perspective

Vinothkumar

Henderson

2016

Quart. Rev. Biophys.

173

115

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“…25 Although we did not take into account the possible dependence of v on concen- Glass-transition temperature for pure benzene is from ref. 27 . The dashed line on the top represents the FH prediction for melting temperature of polyisoprene in the mixture based on eq 2; the dotted line at the bottom is the estimation for glass-transition temperature of the mixture based on the Fox equation and the horizontal dashed line refers to the theoretical prediction for the eutectic point in this system.…”

Section: Solvent Melting In Uncrosslinked Polyisoprenementioning

confidence: 99%

Melting of solvents nanoconfined by polymers and networks

Qin

McKenna

2006

J Polym Sci B Polym Phys

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Thermoporosimetry is becoming increasingly used to study nanoscale heterogeneity and structure in polymer networks. The starting point for thermoporosimetry is the Gibbs-Thomson (GT) relation between melting point and inverse crystal size. In the case of polymers, the Flory-Huggins (FH) model also predicts that there is a depression of the melting point because of the mixing of the polymer and the solvent molecules, and this needs to be taken into account. The first step in analysis of size heterogeneity using thermoporosimetry and the GT equation requires that there be quantitative agreement between the FH theory and the melting points of the diluent in the uncrosslinked rubber. We find that both benzene and hexadecane exhibit excessive melting point depressions in uncrosslinked polyisoprene. This may imply that the uncrosslinked polymer is divided into 'nanoheterogeneities'. We further find that the heat of fusion decreases as polymer concentration increases for the benzene, but not for the hexadecane. Finally, we compare pore size distributions obtained for a crosslinked polyisoprene as determined from the melting behavior of n-hexadecane as a diluent, using different references for how the uncrosslinked polymer behaves. While number average distribution is not very different between the different analyses, the weight average distribution is.

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“…In other films of simple organic molecules, T c F of amorphous naphthalene [6] is 105 K and that of amorphous anthracene [7] is 200 K. In a bulk state, the molecular reorientation of benzene molecules was observed at 90 K by the NMR experiments [8]. In contrast, glass transition temperature (T g ) of the bulk benzene was estimated to be around 130 K [9]. Relating to amorphous, it is reported that the complex relaxation process in the supercooled liquid of benzene is detected by time-resolved optical Kerr effect experiments [10].…”

Section: Introductionmentioning

confidence: 99%