ELECTRON MICROSCOPE STUDIES ON THE SURFACE COAT OF THE NEPHRON

Attempts to make visible the carbohydrate coat at the free cell surface of glomeruli as well as the tubules of rabbit kidney were undertaken. The ruthenium red procedure was performed, according to Luft, at various pH values. Moreover, the colloidal iron and the colloidal thorium methods were used. Neuraminidase digestion was also performed. In the ruthenium red procedure the luminal face of the epithelial cells of the nephron was coated distinctly with reaction product. The results obtained revealed that some of the differences at various levels of the nephron depended on the pH values. In glomeruli and proximal convoluted tubules the optimum pH value was 7.4; in the ascending limb of Henle loops and distal convoluted tubules the optimum pH value was 6.8. The ruthenium red-positive surface coat was either closely connected with, or appeared as a part of, the outer leaflet of the unit membrane. The slit pores of glomeruli were also covered by a coat continuous with the surface coat of the adjacent foot processes. The coat lining the microvilli of proximal convoluted tubules completely filled the intervillous spaces. Also, both the colloidal iron method and the colloidal thorium method evidenced the presence of surface coat. Pre-treatment with neuraminidase abolished the effect of the Hale reaction. These results may indicate that the surface coat of the epithelia of the nephron shows the presence of glycoproteins containing siliac acid residues.     

ELECTRON MICROSCOPE STUDIES ON THE HAEMOGLOBIN MOLECULES

Haemoglobin molecules isolated from normal human subjects have been directly micrographed under the electron microscope following in general Hall's technique. The average height (h) and the widths along (w₁₁) and perpendicular (w) to the shadow direction of the molecules have been measured as 56.5 ± 6.6 A, 122.7 ± 15 A, and 120.9 ± 20 A, respectively. The exaggeration in the molecular widths due to the deposition of metal cap ranges between 60 to 70 A. The probable resolution of the substructure of the molecule, e.g., presence of "holes" and dimples, in the present electron microscopic evidence has been discussed. The electron microscopic results on the size of the individual haemoglobin molecules are in satisfactory agreement with the recent x-ray diffraction model of Perutz and his associates for horse haemoglobin.     

ELECTRON MICROSCOPE STUDIES OF THE MICROVILLI OF HELA CELLS

Microvilli of HeLa cells cultured in vitro were preserved for electron microscopic examination at different stages of routine cultivation procedures. By a double-embedding technique, vertical sectioning for electron microscopy was possible. It revealed that, although the microvilli were present on all sides of the cell in the dispersed stage and in the attached stage, they were not present on the bottom of the cell when it was stretched on the surface of the dish. When the cells were grown in dense colonies, they were found on top of each other, and microvilli were present on all sides, except on the bottom surface of those cells in contact with the dish. We achieved a more dramatic demonstration of the microvilli by developing a surface-replica technique which retains their spatial arrangement and permits characterization of the distribution of their number, length, and diameter.     

ELECTRON MICROSCOPE STUDIES ON THE ORIGIN OF ANNULATE LAMELLAE IN OOCYTES OF NECTURUS

Developing oocytes, ranging from approximately 0.1 to 1.0 mm in diameter, in Necturus were studied with the electron microscope. The outer layer of the nuclear envelope is actively engaged in the formation of vesicular elements along most of its surface, especially in smaller oocytes. Groups of vesicles appear to be released into the ooplasm at about the same time, resulting in long chains of individual vesicles immediately adjacent to the nuclear membrane. This process is repeated so that chains of vesicles grouped in rather ordered ranks extend progressively into the surrounding cytoplasm. Eventually, the cytoplasm becomes more concentrated with chains of vesicles and the distance between the individual rows becomes less. Very soon after a chain of vesicles has been budded off from the nuclear membrane, fine intervesicular connections appear between certain of the vesicles comprising the rows. Several of the vesicles in a row may then fuse, forming short, flattened cisternae. Fusion of vesicles continues, individual rows of vesicles become more closely packed and, finally, regions appear in the cytoplasm which have the appearance of annulate lamellae. Further growth of the lamellae appears to occur by the progressive fusion of vesicles at the ends of those lamellae already present, as well as by the addition of other fusing rows of vesicles.     

ELECTRON MICROSCOPE STUDIES ON THE ACTIVE ACCUMULATION OF SR++ BY RAT-LIVER MITOCHONDRIA

Electron-opaque granules are deposited in isolated rat-liver mitochondria concomitant with the energy-linked accumulation of Sr⁺⁺ by these organelles. High temperature microincineration (600°C) of thin sections of mitochondria containing different amounts of Sr⁺⁺ shows that a clear qualitative correlation exists between the number of inorganic residues remaining after incineration and the amount of Sr⁺⁺ translocated into the mitochondria. By loading the mitochondria with consecutive pulses of small amounts of Sr⁺⁺ ("multiple-pulse" loading), very early stages of granule formation can be detected; the first detectable deposits are seen closely associated with the cristae. The evidence presented supports the hypothesis that mineral deposition following or during the in vitro accumulation of ions by mitochondria occurs, at least initially, at sites on these membranes and not as nonspecific precipitates in the mitochondrial matrix. The large number of electron-opaque deposits (100 to 200) seen in single thin sections of individual mitochondria having accumulated intermediate levels of Sr⁺⁺ clearly exceeds the number of normal dense granules in rat-liver mitochondria, indicating that the normal matrix granules per se do not constitute sites essential for deposition. At the highest levels of Sr⁺⁺ uptake studied in the multiple-pulse loading experiments, needlelike deposits are seen, a result which suggests that the structural form ("crystallinity") of the mineral deposits may be determined by the rate of accumulation.     

ELECTRON MICROSCOPE STUDIES ON THE DICTYOSOMES AND ACROBLASTS IN THE MALE GERM CELLS OF THE CRICKET

The dictyosome (Golgi body) in the secondary spermatocyte of the cricket appears in electron micrographs as a duplex structure composed of (a) a group of parallel double-membraned lamellae and (b) a group of associated vacuoles arranged along the compact lamellae in a chain-like fashion. This arrangement of ultramicroscopic structure for the dictyosomes is strikingly comparable to that described for the Golgi apparatus of vertebrates. Accordingly, the two are considered homologous structures. Associated with the duplex structure of the dictyosomes is a differentiated region composed of small vacuoles. This is thought to represent the pro-acrosome region described in light microscope preparations. In the spermatid the dictyosomes fuse, giving rise to the acroblast. Like the dictyosomes, the acroblasts are made up of double-membraned lamellae and associated vacuoles. In addition, a differentiated acrosome region is present which, in some preparations, may display the acrosome vacuole and granule. Both the dictyosomes and acroblasts are distinct from mitochondria.     

MOTION PICTURE AND ELECTRON MICROSCOPE STUDIES ON THE EMBRYONIC AVIAN OSTEOCLAST

Time-lapse motion picture studies were carried out on isolated fowl embryo osteoclasts in vitro, the cells have an extremely active ruffled border, and show vigorous pinocytotic activity. Electron microscope studies on osmium-fixed cells showed that the pinocytotic vacuoles contained bone salt crystals (as well as material which could not be identified on morphological grounds), and that the folds of the ruffled border enclosed crystals and collagen fibrils. Changes were seen in the matrix beneath the ruffled border. Initially, the collagen fibres became separated from each other and at the same time bone salt crystals became detached from them. Later, as crystals and ground substance disappeared, the outline and cross-striation of the collagen became distinct. The implications of these findings are discussed with respect to the mechanism of bone erosion.     

LIGHT AND ELECTRON MICROSCOPE STUDIES OF THE PRIMARY XYLEM OF RICINUS COMMUNIS

Scott , Flora Murray , Virginia Sjaholm, and Edwin Bowler (U. California, Los Angeles.) Light and electron microscope studies of the primary xylem of Ricinus communis. Amer. Jour. Bot. 47(3) : 162-173. Illus. 1960.–The development of annular and spiral vessels in Ricinus communis has been examined under light and electron miscroscopes. Under the light microscope it is seen that spiral elements make up the bulk of the primary xylem. Pits and plasmodesmata are ubiquitous and are demonstrable in vertical and end walls. Plasmodesmata are evident in spiral thickenings. During tissue growth, intercellular spaces are formed between surrounding cells and developing vessels. These circum-vessel spaces are first lined with and later occluded by suberinlike substances. Traces of a material similar in microchemical reaction are laid down in the middle lamella. A suberin-like lining, termed in this paper the lipid lining, stainable with dimethylaminoazobenzene, occurs in mature living vessel elements. Innumerable minute fat-like droplets, refringent, and stainable with Sudan III, Sudan Black and also with osmic acid, occur in the outer cytoplasm and appear to be attached to the vessel lining by fine protoplasmic strands. They presumably are the source of the wall deposits. After the death of the protoplast, the vessel walls appear completely suberized. When contiguous cells are removed by treatment with I₂ki-H₂SO₄, their site and the site of the intercellular spaces remain marked by linear suberized ridges on the vessel wall. Annular and spiral thickenings arise as cellulose strands and begin to lignify only when the vessel reaches maximum diameter. In transverse section, the broken end of an extracted spiral thickening appears stratified. Under the electron microscope, pits and plasmodesmata are evident in procambial and in differentiating xylem elements in all walls. Annular and spiral thickening are distinguishable first as closely woven microfibrillar cellulose bands. As lignin is deposited in the microfibrillar mesh, the thickenings become dense to the electron beam. Irradiation with the full strength of the electron beam distinguishes between spiral thickenings in younger and older vessels. Older spirals remain apparently unchanged. Younger spirals instantly swell, volatilize in part, and assume a moniliform outline. The bead-like swellings consist of a matrix partly transparent to the electron beam and an internal framework of a material comparatively dense to the electron beam. Similar intense irradiation differentiates between younger and older vessel linings. Older linings appear unchanged, while the younger react violently, volatilize in part and stabilize as an irregular coagulum set in a basic mesh. The volatilized substances appear as granules on lining surface or on substrate. The changing microfibrillar pattern of the cell wall is observed from the procambial stage to the final deposition of the lipid vessel lining.     

ELECTRON MICROSCOPE STUDIES ON DEVELOPING CRAYFISH OOCYTES WITH SPECIAL REFERENCE TO THE ORIGIN OF YOLK

The endoplasmic reticulum is composed, in places, of stacks of parallel cisternae which are limited by membranes having great numbers of ribosomes attached to their outer surface. These are connected with other cisternae of similar structure but with fewer ribosomes and without preferred orientation. The latter extend in all directions from the stacked cisternae, branching and anastomosing freely so that the entire system of membrane-limited cisternae appears interconnected; a morphological condition suitable to serve as the basis for an active transport system. Within the stacked cisternae appear granules about 40 to 60 mµ in diameter. These are thought to represent the precursors of proteinaceous yolk, and the hypothesis is advanced that most of the intracisternal granules are synthesized here, possibly under the influence of the ribosomes. They then "flow" into and along the unoriented cisternae to regions where they collect, expand the cisternae, and undergo transformation into finely granular, relatively large proteinaceous yolk bodies. The mitochondria are somewhat pleomorphic, often show atypical cristae, and frequently contain a few dense granules. Lipid is abundant. Other cytoplasmic components are illustrated.     

多甲藻属扫描电镜样品制备方法的比较研究

用扫描电镜(SEM)观察藻类细胞的表面形态特征,是研究藻类的系统分类、形态和生态学的重要手段之一。