Vesicles electron microscopy

Cationic quaternary ammonium compounds such as distearyldimethylammonium-chloride (DSDMAC) used as a softener and as an antistatic, form hydrated particles in a dispersed phase having a similar structure to that of the multilayered liposomes or vesicles of phospholipids 77,79). This liposome-like structure could be made visible by electron microscopy using the freeze-fracture replica technique as shown by Okumura et al. 79). The concentric circles observed should be bimolecular lamellar layers with the sandwiched parts being the entrapped water. In addition, the longest spacings of the small angle X-ray diffraction pattern can be attributed to the inter-lamellar distances. These liposome structures are formed by the hydrated detergent not only in the gel state but also at relatively low concentrations. 

With the freeze-fracture technique, the fracture plane passes through liposomes which are randomly positioned in the frozen sample. Some liposomes will be cut far from their midplane sections, others through their midplane section. Therefore, the analysis of freeze-fracture pictures requires corrections for nonequatorial fracture. Besides, corrections have to be made for the size-dependent probability of a vesicle being in the fracture plane (Jousma et al., 1987 Guiot et al., 1980). Recently, results with a new technique based on electron microscopy was discussed this technique allows analysis not only of liposome size, but also of the number of bilayers (Lauten-schlager et al., 1988). 

Scattering experiments can be performed to help determine the size and shape of the vesicles without the need for the extensive sample preparation required for electron microscopy and AFM. Dynamic (DLS) and static light scattering (SLS) are widely used to determine the size and possible shape of vesicle systems [40,42,48,49,51,... 

Hinton, DP Johnson, CS, Diffusion Coefficients, Electrophoretic Mobilities, and Morphologies of Charged Phospholipid Vesicles by Pulsed Field Gradient NMR and Electron Microscopy, Journal of Colloid and Interface Science 173, 364, 1995. 

Vesicles lie at the heart of intracellular transport of many proteins. Recently, significant progress has been made in understanding the events involved in vesicle formation and transport. This has transpired because of the use of a number of approaches. These include establishment of cell-free systems with which to study vesicle formation. For instance, it is possible to observe, by electron microscopy, budding of vesicles from Golgi preparations incubated with cytosol and ATP. The development of genetic approaches for studying vesicles in yeast has also been crucial. The piemre is complex, with its own nomenclamre (Table 46-7), and involves a variety of cytosolic and membrane proteins, GTP, ATP, and accessory factors. 

Two separate lines of research led to the proposal that transmitter released in response to neuronal excitation is derived from a vesicle-bound pool rather than from the neuronal cytoplasm. One piece of evidence came from electron microscopy which... 

Of the 20 residues that react with A-ethylmaleimide in the non-reduced denatured Ca -ATPase at least 15 are available for reaction with various SH reagents in the native enzyme [75,239,310]. These residues are all exposed on the cytoplasmic surface. After reaction of these SH groups with Hg-phenyl azoferritin, tightly packed ferritin particles can be seen by electron microscopy only on the outer surface of the sarcoplasmic reticulum vesicles [143,311-314]. Even after the vesicles were ruptured by sonication, aging, or exposure to distilled water, alkaline solutions or oleate, the asymmetric localization of the ferritin particles on the outer surface was preserved [311,313,314]. 

Crawford AR, Smith AJ, Hatch VC, Oude Elferink RP, Borst P, Crawford JM. Hepatic secretion of phospholipid vesicles in the mouse critically depends on mdr2 or MDR3 P-glycoprotein expression. Visualization by electron microscopy. J Clin Invest 1997 100(10) 2562-2567. 

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

Analysis of SFV entry has thus shown that the virus binds to receptors on the cell surface and moves by lateral diffusion into coated pits to be internalized by coated vesicles. The endocytosed virus is delivered into endosomes. Here presumably, the viral envelope is activated by the low pH prevailing in this compartment to fuse with the vacuolar membrane. This results in the release of the viral nucleocapsid into the cytoplasm. During normal infection, the virus might not enter into lysosomes although SFV particles have been identified in this compartment using the large loads of virus needed to visualize the entry process by electron microscopy. Even if this were to happen normally, the viral nucleocapsid would escape destruction because of the rapidity of the fusion mechanism. 

Turner TJ (2004) Nicotine enhancement of dopamine release by a calcium-dependent increase in the size of the readily releasable pool of synaptic vesicles. J Neurosci 24 11328-11336 Unwin N (1995) Acetylcholine receptor channel imaged in the open state. Nature 373 37-43 Unwin N (2003) Structure and action of the nicotinic acetylcholine receptor explored by electron microscopy. FEBS Lett 555 91-95... 

Figures 9d,e show aqueous dispersions of vesicles. The smaller the vesicles, the less probable is an upcoming cross-fracture. Thus the question whether the vesicle is uni- or multilamellar can hardly be answered. At least for fluid vesicle dispersions it is possible to solve the problem with the help of cryotransmission electron microscopy.

Our electron microscopy observations have revealed some of the roles of cell organellae involved in biosynthesis of cell wall components (i) the plasma membrane is the site of cellulose synthesis. This supports the proposal that terminal and rosette complexes at the plasma membrane are responsible for cellulose synthesis, (ii) The Golgi-bodies and small circular vesicles derived from the r-ER s are involved in the biosynthesis and/or transport of the hemicelluloses. Our investigations, however, could not distinguish between what type of cell organellae contained what kind of hemicelluloses, and how these polymers were processed in the organellae. 

Fig. 4. Experimental study of RMN3-silicone oil mixture in rabbits (electron microscopy after 1-month internal tamponade with heavy silicone oil) (a) moderate intercellular oedema between photoreceptor external articles note the presence of a mundane vesicle (b) normal aspect of retina internal layers and (c) normal aspect of internal limiting layer and vitreous cavity.

---