Safety Rules for Cryopreparation

Sitte, Hellmuth; Neumann, Klaus; Edelmann, L

doi:10.1007/978-3-642-72815-0_16

Cited by 7 publications

(2 citation statements)

References 3 publications

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“…Before starting any experimental work involving cryogenic temperatures, all experimenters must become familiar with the potential hazards and the actions to be taken if an accident occurs. Some basic facts are given here and more information is given in work by Sitte, Neumann & Edelmann (1987) and Ryan & Liddicoat (1987) and in the references therein.…”

Section: Safetymentioning

confidence: 99%

Macromolecular Cryocrystallography

Garman¹,

Schneider²

1997

J Appl Crystallogr

344

289

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The current techniques for X-ray-diffraction data collection from macromolecular crystals at cryogenic temperatures are reviewed. The development of the experimental methods is outlined and the basic concepts pertaining to radiation damage and cryoprotection are summarized. Emphasis is placed on the practical aspects important to the success of the techniques, and a detailed account of these is presented.

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Section: Safetymentioning

confidence: 99%

Macromolecular Cryocrystallography

Garman¹,

Schneider²

1997

J Appl Crystallogr

344

289

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“…After pelleting the melittincardiolipin complexes, they were quenched by plunge freezing with the Reichert Jung KF 80 [46] without the use of cryoprotectants. The freezefracture replicas were analysed with a Philips 420 microscope.…”

Section: Freeze-fracture Electron Microscopymentioning

confidence: 99%

Melittin induces HII phase formation in cardiolipin model membranes

Batenburg

Hibbeln

Verkleij

et al. 1987

Biochimica et Biophysica Acta (BBA) - Biomembranes

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The interaction of melittin with bovine heart cardiolipin model membranes was investigated via binding assays, 3t P-NMR, freeze-fracture electron microscopy, small angle X-ray diffraction and fluorescence based fusion assays. A strong binding (K d < 10-7 M) appeared to be accompanied by the formation of large structures, resulting from a fusion process of extremely fast initial rate. As the melittin content is increased, bilayer structure is gradually lost and from a cardiolipin to melittin ratio of about 6 the lipid starts to organize itself in an hexagonal H ii phase. At lower temperatures (T < 40 ° C) the coexistence of another structure is observed, characterized by a broad isotropic 31p-NMR signal and giving rise to sharp X-ray reflections, most probably a cubic phase, as suggested also by freeze-fracture images, showing orderly stacked particles. The results are discussed in relation to contrasting observations on the structural changes induced by melittin in the zwitterionic phospholipid system of dipalmitoylphosphatidylcholine (Dufourcq, J. et al. (1986) Biochim. Biophys. Acta 859, 33-48). The biological relevance of the observations with respect to the process of protein insertion into membranes is indicated.

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A simple pneumatic device for plunge‐freezing cells grown on electron microscopy grids

Cole

Matuszek

See

et al. 1990

J. Elec. Microsc. Tech.

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A detailed design for a simple and inexpensive variable-speed (1.0-5.8 m s-1) pneumatic plunge-freezing device is presented. Cultured cells, grown on Formvar-coated 75-mesh gold finder grids, are pneumatically driven into a stirring mixture of propane/isopentane (3:1) cooled by liquid nitrogen (LN2). Premature freezing of the sample in the cryogenic vapors above the cryogen is prevented by plunging through an entry tube into an insulating box, to which a partial vacuum is applied. The cryogenic vapors are drafted into the box at the level of the liquid cryogen by the vacuum, thereby preventing a layer of cold gas from collecting above the cryogen. To prevent the sample from thawing during transfer from the cryogen to the substitution medium, the box top is removed and compressed air is forced through a corrugated tube running the length of the box. The resulting boiling LN2 creates an atmosphere below -120 degrees C in which the transfer can be accomplished.

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Safety Rules for Cryopreparation

Cited by 7 publications

References 3 publications

Macromolecular Cryocrystallography

Macromolecular Cryocrystallography

Melittin induces HII phase formation in cardiolipin model membranes

A simple pneumatic device for plunge‐freezing cells grown on electron microscopy grids

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