Fracture, electron microscopy

B. Donnio, D. N. Bruce, H. Delacroix, T. Gulik-Krzywicki. Freeze-fracture electron microscopy of thermotropic cubic and columnar mesophases. Liq... 

The dimer chains of Ca -ATPase can also be observed by freeze-fracture electron microscopy [119,165,166,172-174], forming regular arrays of oblique parallel ridges on the concave P fracture faces of the membrane, with complementary grooves or furrows on the convex E fracture faces. Resolution of the surface projections of individual Ca -ATPase molecules within the crystalline arrays has also been achieved on freeze-dried rotary shadowed preparations of vanadate treated rabbit sarcoplasmic reticulum [163,166,173,175]. The unit cell dimensions derived from these preparations are a = 6.5 nm b = 10.7 nm and 7 = 85.5° [175], in reasonable agreement with earlier estimates on negatively stained preparations [88]. 

Analysis of the lanthanide-induced crystalline arrays by negative staining (Fig. 5) or freeze-fracture electron microscopy reveals obliquely oriented rows of particles, corresponding to individual Ca -ATPase molecules [119]. The unit cell dimensions for the gadolinium-induced Ca -ATPase crystals are a = 6. l A, b = 54.4 A and y = 111°. Similar cell constants were obtained for the crystals induced by lanthanum, praseodymium and calcium. The unit cell dimensions of the Ei crystals are consistent with a single Ca -ATPase monomer per unit cell. The space group of the Eptype crystals is PI [119], while that of the E2 crystals is P2 [88,90]. 

Sternberg, B., Sorgi, F.L., and Huang, L., New structures in complex formation between DNA and cationic liposomes visualized by freeze-fracture electron microscopy, FEBS Letters, 1994, 356, 361-366. 

Freeze fracture electron microscopy has revealed the distribution of the preceding three components as... 

Incorporated amount of PS II and freeze-fracture electron microscopy images of PS II reconstituted vesicles. 

Modification of the ratio of the two phospholipids yielded different structures in freeze-fracture electron microscopy. Without DMPG (molar ratio DMPC/DMPG/AmB 10/0/5), large lamellar structure were observed. The presence of DMPG favored the formation of shorter lamellar structures, both stacked and fused in places or a predominantly disc morphology. 

Copper ions have been reduced in colloidal assemblies differing in their structures (55,56). In all cases, copper metal particles are obtained. Figure 9.3.1 shows the freeze-fracture electron microscopy (FFEM) for the various parts of the phase diagram. Their structures have been determined by SAXS, conductivity, FFEM, and by predictions of microstructures that require only notions of local curvature and local and global packing constraints. 

The extensive studies of the behavior of mixed monolayers or bilayers of di-acetylenic lipids and other amphiphiles parallel to some degree the studies of dienoyl-substituted amphiphiles. Since the dienoyl lipids do not contain a rigid diacetylenic group in the middle of the hydrophobic chains, they tend to be miscible with other lipids over a wide range of temperatures and compositions. In order to decrease the lipid miscibility of certain dienoyl amphiphiles, Ringsdorf and coworkers utilized the well-known insolubility of hydrocarbons and fluorocarbons. Thus two amphiphiles were prepared, one with hydrocarbon chains and the other with fluorocarbon chains, in order to reduce their ability to mix with one another in the bilayer. Of course it is necessary to demonstrate that the lipids form a mixed lipid bilayer rather than independent structures. Elbert et al. used freeze fracture electron microscopy to demonstrate that a molar mixture of 95% DM PC and 5% of a fluorinated amphiphile formed phase-separated mixed bilayers [39]. Electron micrographs showed extensive regions of the ripple phase (Pb phase) of the DM PC and occasional smooth patches that were attributed to the fluorinated lipid. In some instances it is possible to... 

Figure 18-2 (A) Schematic diagram of mitochondrial structure. (B) Model showing organization of particles in mitochondrial membranes revealed by freeze-fracture electron microscopy. The characteristic structural features seen in the four half-membrane faces (EF and PF) that arise as a result of fracturing of the outer and inner membranes are shown. The four smooth membrane surfaces (ES and PS) are revealed by etching. From Packer.8...

Miscibility of a natural lipid (DMPC) and the monomeric and polymeric lecithin analogue (26) was studied in large unilamellar vesicles using freeze-fracture electron microscopy and photobleaching by H. Gaub 100>. Before polymerization the two lipids appear miscible at all compositions in the fluid state and at DMPC concentrations at or below 50 mol/o in the solid state. After polymerization a two-dimensional solution of the polymer in DMPC is obtained at T > T (T phase transition temperature of polymeric 26) while lateral phase segregation into DMPC-rich domains and patches of the polymer is observed T < T. The diameter of the polymerized lipid domains was found to average 400 A. 

Phase-separated monolayers and liposomes were characterized by R. Elbert1011 who synthesized saturated and polymerizable fluorocarbon amphiphiles (59, 60, 61) and investigated their mixing behavior with CH2-analogues and natural lipids. In these systems the fluorocarbon compounds are incompatible with hydrocarbon lipids in a wide range of compositions and tend to form domains of pure fluorocarbon and hydrocarbon amphiphiles. The domains can be visualized by freeze-fracture electron microscopy. 

Freeze-fracture electron microscopy, (a) When struck with a sharp knife, membranes embedded in ice usually fracture between the monolayer leaflets of the lipid bilayer, (b) Freeze-fracture electron micrograph of the plasma membrane of Streptococcus faecalis, showing a large number of protrusions (presumably proteins) on the outer fracture face and the relative lack of such particles on the inner... 

Pasquali-Ronchetti, I., Fornieri, C., Baccarani Contri, M., and Volpin, D. (1979). The ultrastructure of elastin revealed by freeze-fracture electron microscopy. Micron. 10, 89-99. 

There are several genetic skin diseases with known defects in the lipid metabolism. Atopic dermatitis, lamellar ichthyosis, and psoriasis have been the most widely studied with respect to epidermal barrier function and alterations in the lipid profile. Deviations in the lipid profile have been linked with an impaired stratum corneum barrier function. Atopic dermatitis is characterized by inflammatory, dry and easily irritable skin, and overall reduced ceramide levels in the stratum corneum [58-60]. In particular a significant decrease in the ceramide 1 level is observed, whereas the levels of oleate that is esterified to ceramide 1 are elevated [59]. Both aberrations may be responsible for the reduced order of the lamellar phases as observed with freeze fracture electron microscopy [61]. It has further been established that, in comparison to healthy stratum corneum, the fraction of lipids forming a hexagonal packing is increased [61]. A recent study reveals that the level of free fatty acids... 

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