scholarly journals 'Chlorobium-type' Vesicles of Photosynthetically-grown Chloroflexus aurantiacus Observed Using Negative Staining Techniques

1977 ◽  
Vol 102 (2) ◽  
pp. 279-285 ◽  
Author(s):  
M. T. MADIGAN ◽  
T. D. BROCK
Author(s):  
A. C. Reimschuessel ◽  
V. Kramer

Staining techniques can be used for either the identification of different polymers or for the differentiation of specific morphological domains within a given polymer. To reveal morphological features in nylon 6, we choose a technique based upon diffusion of the staining agent into accessible regions of the polymer.When a crystallizable polymer - such as nylon 6 - is cooled from the melt, lamellae form by chainfolding of the crystallizing long chain macromolecules. The regions between adjacent lamellae represent the less ordered amorphous domains into which stain can diffuse. In this process the lamellae will be “outlined” by the dense stain, giving rise to contrast comparable to that obtained by “negative” staining techniques.If the cooling of the polymer melt proceeds relatively slowly - as in molding operations - the lamellae are usually arranged in a radial manner. This morphology is referred to as spherulitic.


1967 ◽  
Vol 2 (2) ◽  
pp. 169-192
Author(s):  
O. BEHNKE ◽  
A. FORER

Experiments were performed on crane-fly spermatids (Nephrotoma suturalis Loew), ratsperm, and rat tracheal cilia to test whether all microtubules respond in the same way to different treatments. Crane-fly spermatids contain cytoplasmic microtubules, accessory tubules, and the 9 + 2 complex of tubules; rat sperm and rat tracheal cilia contain only the 9 + 2 tubules. Crane-fly spermatid tubules responded to the experimental treatments as follows. After colchicine treatment, or storage at 0 °C, the cytoplasmic microtubules disappeared, while the 9 + 2 tubules were normal. After storage at 50 °C the cytoplasmic microtubules disappeared, and then the 9 + 2 tubules were affected: first the central tubules and B-tubules were affected, and later the A-tubules. After brief pepsin treatment, the 9 doublet tubules disappeared, while the other tubules appeared normal; after prolonged pepsin treatment the accessory, central, and cytoplasmic tubules disappeared. After negative staining at pH 7, the cytoplasmic microtubules were never seen, the central tubules were only sometimes seen, the B-tubules were sometimes fragmented, and the A-tubules were intact. On the basis of these responses, it was concluded that there are 4 classes of tubules in crane-fly spermatids, namely cytoplasmic microtubules; accessory tubules and central tubules (of the 9 + 2 complex); B-tubules (of the 9 + 2 complex); and A-tubules (of the 9 + 2 complex). At least some of the different responses appeared to be due to intrinsic physical and/or chemical differences between the tubules themselves. Pepsin digestion and negative staining of rat sperm tails gave results similar to those with crane-fly spermatids. In addition, the 9 + 2 tubules responded differently to pepsin digestion at different points along their length. This gradient of sensitivity was attributed to synthesis of new tubules occurring at one end of the sperm tail. Pepsin digestion and negative staining of rat tracheal cilia gave results similar to those with crane-fly spermatids and rat sperm tails. All the tubules had a similar substructure, as revealed by negative-staining techniques. It was concluded that microtubules are proteinaceous, at least in part, and that microtubules are different in composition from membranes. It is suggested that the walls of the B-tubules are composed of two materials--(1) the portions adjacent to the A-tubules, and (2) the remaining portion.


1970 ◽  
Vol 45 (2) ◽  
pp. 272-290 ◽  
Author(s):  
Daniel A. Goodenough ◽  
Jean Paul Revel

Zonulae occludentes and gap junctions were examined both in the intact mouse liver and in a junction-rich membrane fraction from homogenized mouse liver. These preparations were visualized with the techniques of uranyl acetate staining en bloc, staining with colloidal lanthanum, negative staining with phosphotungstate, and freeze-cleaving. The zonula occludens is arranged as a meshwork of branching and anastomosing threadlike contacts sealing the lumen of the bile canaliculus from the liver intercellular space. The gap junction is characterized in section by a 20 A gap between the apposed junctional membrane outer leaflets, and permeation of this space with lanthanum or phosphotungstate reveals a polygonal lattice of subunits with a center-to-center spacing of 90–100 A. Freeze-cleaved gap junctions show a similar lattice. Extraction of junction-rich fractions with 60% aqueous acetone results in a disappearance of the 20 A gap in sectioned pellets and an inability to demonstrate the polygonal lattice with either the freeze-cleave or negative staining techniques. Extraction of the membranes with 50% acetone does not produce this effect. Thin-layer chromatography of the acetone extracts reveals a group of phospholipids in the 60% extract that are not detectable in the 50% extract. Acetone does not cause any detectable change in the structure of the zonula occludens, but the occluding junction becomes leaky to lanthanum following acetone treatment. The effects of other reagents on the junctions are reported.


Author(s):  
M. Ohtsuki ◽  
A. V. Crewe

Extracellular hemoglobins of annelids consist of 12 subunits with appearance of superposed hexagons. So far, electron microscopy using negative staining techniques indicated that only the hemoglobins of Nephtys and Oenone contain an additional central subunit.


1965 ◽  
Vol 43 (6) ◽  
pp. 647-655 ◽  
Author(s):  
D. F. Parsons ◽  
W. D. Bonner Jr. ◽  
J. G. Verboon

Six types of plant mitochondria have been isolated by improved techniques and examined with the electron microscope by the thin-section and negative-staining techniques. In general, the mitochondria appeared well preserved. There was minimal contamination by endoplasmic reticulum but some plastids were present. The morphology of the plant mitochondria is compared with that of animal cell mitochondria. Negative staining showed that the inner membranes were covered with projecting knob-like subunits, as previously described for animal and insect cell mitochondria. The outer membrane showed a characteristic pitted appearance that was apparently due to 28 Å holes in the membrane. A preliminary report is also given of the appearance of negatively stained membranes of chloroplasts.


1969 ◽  
Vol 42 (1) ◽  
pp. 202-220 ◽  
Author(s):  
Toku Kanaseki ◽  
Ken Kadota

Five points are discussed regarding the vesicular structure isolated by fractionation techniques from the brain and liver of the guinea pig. 1. One type of vesicle, fixed by OsO4 and shown in thin sections, is identified with the coated vesicle that has been observed in many varieties of tissues. 2. The vesicle contained in a spherical polygonal "basketwork" shown by the negative-staining techniques is identical with the coated vesicle shown in sections. 3. Despite minute observation of this basketwork we could not confirm the existence of "hairlike projections" extending from the convex cytoplasmic surface of the vesicle. We are inclined to believe, therefore, that the hairlike projections are actually the superimposed visual images of the regular hexagons and pentagons of the network composing the basketwork. 4. We repeat the hypothesis originally advanced by Roth and Porter (1) that the "coating" of the coated vesicle plays a role in the mechanism of the infolding and fission of the membrane; we suggest that these events are caused by the transformation of the regular hexagons (of the coating) into regular pentagons. 5. Finally, we make a suggestion as to the nature of those vesicles which have on their surface subparticles which look like "elementary particles (2)."


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