The Surface Structure of Cultured Rabbit Kidney Cells as Revealed by Electron Microscopy

1970 ◽  
Vol 7 (3) ◽  
pp. 719-737
Author(s):  
ELIZABETH DIMMOCK
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Cell Surface ◽  
Blood Group ◽  
Potassium Permanganate ◽  
Ruthenium Red ◽  
Rabbit Kidney ◽  
Dense Layer ◽  
Kidney Cells ◽  
Thin Sections

An epithelioid line of rabbit kidney cells (RK 13), in which the distribution of blood group antigen A had previously been investigated by mixed agglutination, was chosen for studies of the structure of the cell surface. Cells were grown in monolayer culture, and thin sections were examined by electron microscopy after fixation in glutaraldehyde and osmium tetroxide and staining of the sections with lead, or uranyl and lead. Cells were also treated with ruthenium red incorporated in the osmium fixative. Other cells were fixed in lanthanum or potassium permanganate, and both stained and unstained sections were examined. The morphology of RK 13 cell surfaces is described. There is apparently no great degree of cellular specialization. The treatment with ruthenium red resulted in dense staining of a layer of the cell surface that is not visible in conventional preparations, and sometimes in staining of the surfaces of intracellular organelles. Lanthanum permanganate fixation also revealed a dense layer on the surface of the plasma membrane; a less dense surface layer was distinguished in many cells fixed in potassium permanganate. The reaction of ruthenium red with the cell surface is probably due to the presence of acidic glycoproteins, but the chemical specificity of the staining method is not yet clear. The nature of the material revealed by lanthanum and potassium permanaganates is also undefined. However, these staining methods reveal that the cell surface is more complex than is apparent in cells prepared by conventional techniques. The additional surface layer is probably the site of many blood group substances and other compounds involved in the physiological reactions of the cell surface.

The Location of Blood Group Antigen a on Cultured Rabbit Kidney Cells as Revealed by Ferritin-Labelled Antibody

1972 ◽  
Vol 10 (2) ◽  
pp. 525-533
Author(s):  
ELIZABETH DIMMOCK ◽  
D. FRANKS ◽  
AUDREY M. GLAUERT
Keyword(s):  
High Resolution ◽  
Cell Surface ◽  
Blood Group ◽  
Blood Group Antigen ◽  
Rabbit Kidney ◽  
Kidney Cells ◽  
Specific Staining ◽  
Antigen A

Fluorescein- and ferritin-labelled antibodies were used to locate blood group antigen A on the surfaces of cultured rabbit kidney cells (RK 13). Fluorescent anti-A revealed the antigen on some cells, but not on others. Controls indicated that specific staining could be obtained. Comparable observations were made with ferritin-labelled antibody, and the comparatively high resolution of this method allowed individual antigen molecules to be located. Measurements were made of the distances between the centres of the ferritin molecules and the cell surface, and the position of the blood group antigen relative to the visible components of the membrane is discussed in relation to this.

Ultrastructural differences in the exosporium of the Sterne and Vollum strains of Bacillus anthracis

1968 ◽  
Vol 14 (12) ◽  
pp. 1297-1299 ◽  
Author(s):  
M. J. Kramer ◽  
Ivan L. Roth
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Bacillus Anthracis ◽  
Potassium Permanganate ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Single Application ◽  
Thin Sections ◽  
Acetate Lead ◽  
Different Strains

Spores from three different strains of Bacillus anthracis were examined by electron microscopy for the presence of a hair-like nap previously reported to be present on the exosporium of spores of the Sterne strain (avirulent). In addition to that strain, the Vollum strain (virulent) and a rough, avimlent variant of the Vollum were utilized in the current studies.Spores were fixed with Kellenberger's standard OsO4 fixative and embedded in Maraglas. Thin sections were poststained with various combinations of the following: potassium permanganate, uranyl acetate, lead citrate. The nap on the exosporium of spores of the Sterne strain was revealed most clearly when thin sections were poststained with all of the aforementioned stains. Post-staining by a single application of any of the three reagents resulted in a nap that was barely perceptible.The surface of the exosporium of spores from the Vollum strain and the rough, avirulent variant was found to be quite different from that of the Sterne strain. On the two former, the surface layer is approximately one-third as thick as the layer of hairs in the nap on the latter.

scholarly journals Further Observations on the Fine Structure of Chrysochromulina Ericina Parke & Manton

1961 ◽  
Vol 41 (1) ◽  
pp. 145-155 ◽  
Author(s):  
I. Manton ◽  
G. F. Leedale
Keyword(s):  
Electron Microscopy ◽  
Fine Structure ◽  
Cell Surface ◽  
Light Microscopy ◽  
Flat Plate ◽  
Outer Layer ◽  
Living Cells ◽  
Layered Plates ◽  
Thin Sections ◽  
Micro Anatomy

C. ericina Parke & Manton has been re-investigated to add salient features of micro-anatomy from the electron microscopy of thin sections and also to add photographs of living cells taken with anoptral contrast light microscopy.The most important new observations concern the scales which are shown to be essentially two-layered plates in which the layers in the very large spined scales have become separated except at their edges, with the outer layer greatly hypertrophied to produce a hollow spine with a flared base closed at the bottom by a flat plate. The patterns of external marking on the two layers are very similar in both plate-scales and spines in this species and the orientation of both with respect to the cell surface has been demonstrated by a section of the scales in situ.

Ultrastructure of the in situ adherence of Mobiluncus to vaginal epithelial cells

1988 ◽  
Vol 34 (6) ◽  
pp. 757-766 ◽  
Author(s):  
Jan M. De Boer ◽  
Friso H. F. Plantema
Keyword(s):  
Electron Microscopy ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Bacterial Vaginosis ◽  
Vaginal Fluid ◽  
Thin Sections ◽  
Epithelial Cell Surface ◽  
Curved Rods

From patients with bacterial vaginosis motile, anaerobic, comma-shaped bacteria can be isolated, which have recently been placed into the new genus Mobiluncus. In this study, electron microscopy was used to examine the in situ adherence of these motile curved rods to detached epithelial cells (comma cells) in vaginal fluid from two patients with bacterial vaginosis. Thin sections showed that the curved rods attached both directly to the epithelial cell surface and at various distances from it. It is concluded that after initial attachment these motile bacteria can grow at the epithelial cell surface in sessile microcolonies. Ruthenium red staining demonstrated a coating of precipitated glycocalyx material both on the surface of the curved rods and on their flagella. This may indicate that in situ the adherent curved rods were enclosed in a very hydrated matrix of exopolysaccharides. Conspicuous was the ability of the curved rods to attach to the epithelial cell surface via their cell tips. However, in situ no specialized bacterial cell surface structures were seen that might explain this polar attachment. Electron microscopy of pure cultures demonstrated that both Mobiluncus curtisii subsp. curtisii and Mobiluncus mulieris can produce a glycocalyx in vitro.

AEM study of reacted surface layer of borosilicate nuclear waste glasses

1992 ◽  
Vol 50 (1) ◽  
pp. 352-353
Author(s):  
L.M. Wang ◽  
S.A. Kaser ◽  
R.C. Ewing ◽  
J.K. Bates
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Nuclear Waste ◽  
Reaction Pathway ◽  
Sample Surface ◽  
Carbon Films ◽  
Waste Glass ◽  
Thin Sections ◽  
Nuclear Waste Glass

Analysis of the reacted surface layer of borosilicate glass is important to the understanding of the long term nuclear waste glass reaction process. The objective is to assess the simulated nuclear waste glass/water reaction pathway by identifying new crystalline phases that appear on the glass surfaces during the reaction. The results can be used to validate models generated to predict long-term performance of the nuclear waste glass under a range of conditions.In this study, extensive scanning electron microscopy (SEM) with qualitative energy dispersive x-ray spectroscopy (EDS) analysis, quantitative analytical and high resolution transmission electron microscopy (AEM and HRTEM) have been performed on two 202U glasses which were reacted in saturated water vapor for 14 and 23 days, respectively. In order to study the microchemical and microstructural changes with increasing depth from the reaction surface, TEM specimens were prepared in cross-section using the ultramicrotomy slicing technique similar to that described by Bates et al. In this process, small chunks containing the reacted surface layer and a thin layer of glass were first broken off from the sample surface and each of these chunks was then embedded in resin to form a block. Finally, thin sections, approximately 90 nm thick, were microtomed from these blocks and were transfered to copper mesh grids covered by holey carbon films for observation. AEM and HRTEM analyses were accomplished using a JEOL JEM-2000FX microscope attached with a Noran/TN-5500 EDS system at the University of New Mexico.

Microscopic detection of adventitious viruses in cell cultures

1972 ◽  
Vol 18 (3) ◽  
pp. 299-304 ◽  
Author(s):  
Nan Anderson ◽  
Frances W. Doane
Keyword(s):  
Electron Microscopy ◽  
Foamy Virus ◽  
Light And Electron Microscopy ◽  
Monkey Kidney ◽  
Contamination Rate ◽  
Kidney Cells ◽  
African Green Monkey Kidney ◽  
Thin Sections ◽  
Tumor Viruses ◽  
Viral Contaminants

Sixty-five batches of three types of monkey kidney cells were examined by light and electron microscopy for the presence of simian viruses. Cynomolgus cells showed the highest contamination rate (67%), followed by rhesus (18%). Foamy virus was the most common contaminant, being detected in 13 batches, twice in concert with a paramyxovirus. Six batches contained a paramyxovirus; one batch contained a papovavirus. Of the 22 batches of African green monkey kidney cells examined, none showed evidence of virus. "Healthy" established cell lines from three laboratories were also checked by electron microscopy, and several contaminating viruses were found. These were apparently either accidentally introduced during handling, as in the case of SV5, reovirus, and parvovirus, or were latent viruses (hamster and mouse) derived from the host tissue. While negative staining provided the simplest technique for detection of the viral contaminants, thin sectioning was required to locate foamy virus and the latent "tumor" viruses. A rapid method for the preparation of thin sections is described.

Mesophyll cell walls in hemlock, Tsuga canadensis

1977 ◽  
Vol 55 (11) ◽  
pp. 1510-1515
Author(s):  
R. Fischer ◽  
N. G. Dengler
Keyword(s):  
Cell Surface ◽  
Cell Walls ◽  
Mesophyll Cell ◽  
Tsuga Canadensis ◽  
Structural Features ◽  
Ruthenium Red ◽  
Intercellular Spaces ◽  
Mesophyll Cells ◽  
Thin Sections ◽  
Histochemical Tests

Observations have been made on the walls of the mesophyll cells in the leaves of hemlock, Tsuga canadensis, to determine if there were any structural features which would account for the relatively high resistance measured at the mesophyll cell surface in infiltrated leaves by Tyree et al. (1975). There was no indication of an internal cuticle based on histochemical tests of fresh sections and the appearance of the wall in electron micrographs. Examination of Alcian-blue-stained fresh sections and thin sections of material treated with ruthenium red indicated that mesophyl) cell walls may be covered with a mucilaginous layer that retards the evaporation of water to intercellular spaces.

scholarly journals Adenosinetriphosphatase and 5-Nucleotidase Activities in the Plasma Membrane of Liver Cells as Revealed by Electron Microscopy

1958 ◽  
Vol 4 (6) ◽  
pp. 711-716 ◽  
Author(s):  
Edward Essner ◽  
Alex B. Novikoff ◽  
Bertha Masek
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Cell Surface ◽  
Cell Function ◽  
Molecular Transport ◽  
Enzyme Reaction ◽  
Bile Canaliculus ◽  
Nucleotidase Activity ◽  
Reaction Products ◽  
Thin Sections

The sites of reaction product resulting from ATPase and 5-nucleotidase activities remaining in parenchymatous cells of osmium-fixed rat liver were studied by electron microscopy of thin sections. These indicate that both ATPase and 5-nucleotidase activities are localized in the plasma membrane where it folds to form the microvilli of the bile canaliculus, and that 5-nucleotidase activity is also present in the microvilli at the sinusoidal aspects of the cells. It is suggested that these enzymes, particularly ATPase, may play a role in molecular transport or in some kind of membrane activity at the cell surface. Of special interest is the apparent differential localization of these enzymes at the absorptive and secretory regions of the plasma membrane of the cell. It may be of interest to study changes in these enzyme localizations in pathologic states, as a sign of changed cell function. Some of the difficulties in the interpretation of enzyme reaction products seen in electron micrographs are discussed.

Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

1978 ◽  
Vol 36 (2) ◽  
pp. 82-83 ◽  
Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Light Microscopy ◽  
Thin Sections ◽  
Transmission Electron ◽  
Identical Cell ◽  
Electron Microscopes ◽  
Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

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