Enhancement of outer cell layers of a methanotrophic bacterium using uranyl acetate en bloc

1988 ◽  
Vol 46 ◽  
pp. 74-75
Author(s):  
K. Putzer ◽  
T.A. Fassel ◽  
C.C. Remsen
Keyword(s):  
Uranyl Acetate ◽  
Outer Cell ◽  
Type I ◽  
Type Ii ◽  
En Bloc ◽  
Wall Structures ◽  
Intracytoplasmic Membranes ◽  
Cell Layers ◽  
Ethanol Series ◽  
Autotrophic Microorganisms

Obligate methanotrophic bacteria are autotrophic microorganisms capable of oxidizing methane to supply both their carbon and energy requirements. Methanotrophs are placed in one of two groups based on certain biochemical features and the arrangement of intracytoplasmic membranes (ICM). Type I methanotrophs contain ICM arranged as bundles of disc-like vessicles in the interior of the cell, while Type II methanotrophs display paired peripheral ICM. The organism used in this study, Methylosinus trichosporium OB3b, is a Type II methanotroph and has a capsular structure surrounding the cell as well as several complex cell wall structures described as cup-like, filamentous, and spike-shaped. This study examines the external surface of M. trichosporium OB3b following en bloc staining with uranyl acetate.Cells were harvested in mid-log phase, fixed in 0.5% glutaraldehyde and postfixed in 1% OsO4. Cells were enrobed in 4% agar prior to dehydration in a graded ethanol series.

Labelling of non-A, non-B hepatitis infected chimpanzee hepatocytes with nuclease-gold conjugates

1984 ◽  
Vol 42 ◽  
pp. 250-251
Author(s):  
G. D. Gagne ◽  
M. F. Miller ◽  
D. A. Peterson
Keyword(s):  
Endoplasmic Reticulum ◽  
Experimental Infection ◽  
Uranyl Acetate ◽  
Type I ◽  
Cellular Injury ◽  
Type Ii ◽  
Tubular Structures ◽  
Type Iii ◽  
Type Iv ◽  
Infected Chimpanzee

Experimental infection of chimpanzees with non-A, non-B hepatitis (NANB) or with delta agent hepatitis results in the appearance of characteristic cytoplasmic alterations in the hepatocytes. These alterations include spongelike inclusions (Type I), attached convoluted membranes (Type II), tubular structures (Type III), and microtubular aggregates (Type IV) (Fig. 1). Type I, II and III structures are, by association, believed to be derived from endoplasmic reticulum and may be morphogenetically related. Type IV structures are generally observed free in the cytoplasm but sometimes in the vicinity of type III structures. It is not known whether these structures are somehow involved in the replication and/or assembly of the putative NANB virus or whether they are simply nonspecific responses to cellular injury. When treated with uranyl acetate, type I, II and III structures stain intensely as if they might contain nucleic acids. If these structures do correspond to intermediates in the replication of a virus, one might expect them to contain DNA or RNA and the present study was undertaken to explore this possibility.

Ruthenium red and alcian blue effects on outer structures of methanotrophic bacteria

1988 ◽  
Vol 46 ◽  
pp. 72-73
Author(s):  
T.A. Fassel ◽  
M.J. Schaller ◽  
C.C. Remsen
Keyword(s):  
Research Effort ◽  
Ruthenium Red ◽  
Outer Cell ◽  
Methanotrophic Bacteria ◽  
En Bloc ◽  
Methylosinus Trichosporium ◽  
Wall Structures ◽  
Methylomonas Albus ◽  
Type Strains ◽  
Outer Cell Wall

Methane, a contributor to the “greenhouse effect”, is oxidized in the natural environment by methanotrophic bacteria. As part of a comprehensive research effort, we have been examining the ultrastructure of methanotrophs. These microorganisms have complex outer cell wall structures similar to those frequently found in other chemol itho- trophic bacteria. (1,2)In our work, we have focused on the “type” strains of Methylomonas albus BG8 and Methylosinus trichosporium OB3b. Between Spurr and LR White embedding resins, we found a difference 1n the preservation of an outer cup layer of BG8 external to the peripheral membranes. Cells from the same sample embedded in Spurr consistently lacked this feature (FIG. 1). This effect was overcome by an en bloc ruthenium red (RR) protocol that resulted in successful retention of the cup layer in Spurr resin (FIG. 2). For OB3b cells, the en bloc RR protocol resulted in an exterior bead feature distinguishable in thin section (FIG. 4) that previously was seen only by SEM.

Comparative surface and ultrastructural views of methylotrophic bacteria by TEM, STEM, SE, and freeze-etch electron microscopy.

1987 ◽  
Vol 45 ◽  
pp. 792-793
Author(s):  
T.A. Fassel ◽  
M.J. Schaller ◽  
M.E. Lidstrom ◽  
C.C. Remsen
Keyword(s):  
Cytoplasmic Membrane ◽  
Structural Features ◽  
Methylotrophic Bacteria ◽  
Type I ◽  
Type Ii ◽  
Membrane Complex ◽  
Oxidation Of Methane ◽  
Methane Oxidizing Bacteria ◽  
Wall Structures ◽  
Methylomonas Albus

Methylotrophic bacteria play an Important role in the environment in the oxidation of methane and methanol. Extensive intracytoplasmic membranes (ICM) have been associated with the oxidation processes in methylotrophs and chemolithotrophic bacteria. Classification on the basis of ICM arrangement distinguishes 2 types of methylotrophs. Bundles or vesicular stacks of ICM located away from the cytoplasmic membrane and extending into the cytoplasm are present in Type I methylotrophs. In Type II methylotrophs, the ICM form pairs of peripheral membranes located parallel to the cytoplasmic membrane. Complex cell wall structures of tightly packed cup-shaped subunits have been described in strains of marine and freshwater phototrophic sulfur bacteria and several strains of methane oxidizing bacteria. We examined the ultrastructure of the methylotrophs with particular view of the ICM and surface structural features, between representatives of the Type I Methylomonas albus (BG8), and Type II Methylosinus trichosporium (OB-36).

scholarly journals Co-culture of type I and type II pneumocytes as a model of alveolar epithelium

2021 ◽  
Author(s):  
Oliver Brookes ◽  
Sonja Boland ◽  
René Lai Kuen ◽  
Armelle Baeza-Squiban
Keyword(s):  
Cell Lines ◽  
Barrier Function ◽  
Single Cells ◽  
Light And Electron Microscopy ◽  
Alveolar Epithelium ◽  
Type I ◽  
Apical Surface ◽  
Type Ii ◽  
Cell Layers ◽  
Distal Lung

AbstractThe epithelial tissues of the distal lung are continuously exposed to inhaled air, and are of research interest in studying respiratory exposure to both hazardous and therapeutic materials. Pharmaco-toxicological research depends on the development of sophisticated models of the alveolar epithelium, which better represent the different cell types present in the native lung and interactions between them.We developed an air-liquid interface (ALI) model of the alveolar epithelium which incorporates cell lines representative of both type I (NCI-H441) and type II (hAELVi) epithelial cells. We compared morphology of single cells and the structure of cell layers of the two lines using light and electron microscopy. Working both in monotypic cultures and cocultures, we measured barrier function by trans-epithelial electrical resistance (TEER), and demonstrated that barrier properties can be maintained for 30 days. We created a mathematical model of TEER development over time based on these data in order to make inferences about the interactions occurring in these culture systems. We assessed expression of a panel of relevant genes that play important roles in barrier function and differentiation.The coculture model was observed to form a stable barrier akin to that seen in hAELVi, while expressing surfactant protein C, and having a profile of expression of claudins and aquaporins appropriate for the distal lung. We described cavities which arise within stratified cell layers in NCI-H441 and cocultured cells, and present evidence that these cavities represent an aberrant apical surface. In summary, our results support the coculture of these two cell lines to produce a model which better represents the breadth of functions seen in native alveolar epithelium.

The anatomy of two functional types of mechanoreceptive 'free' nerve-ending in the head skin of Xenopus embryos

1983 ◽  
Vol 218 (1210) ◽  
pp. 61-76 ◽  
Keyword(s):  
Nerve Ending ◽  
Type I ◽  
Type Ii ◽  
Free Nerve Ending ◽  
Skin Cells ◽  
Skin Cell ◽  
Functional Types ◽  
Superficial Skin ◽  
Cell Layers ◽  
Movement Detectors

The structure of the two functional types of ‘free’ nerve-ending in the head skin of late Xenopus embryos has been examined by horseradish peroxidase staining through their cells in the trigeminal ganglion and by electron microscopy. Type I neurites are identified as the ‘movement’ detectors by their purely homolateral innervation. They have many fine branches between the superficial skin cells, bearing numerous large varicosities. Type II neurites cross the midline to innervate both sides of the head as do the ‘rapid transient’ detectors found by physiology. They have a few fairly straight branches between the skin cell layers with few elongated varicosities.

scholarly journals Co-culture of type I and type II pneumocytes as a model of alveolar epithelium

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0248798
Author(s):  
Oliver Brookes ◽  
Sonja Boland ◽  
René Lai Kuen ◽  
Dorian Miremont ◽  
Jamileh Movassat ◽  
...  
Keyword(s):  
Cell Lines ◽  
Barrier Function ◽  
Single Cells ◽  
Light And Electron Microscopy ◽  
Alveolar Epithelium ◽  
Type I ◽  
Apical Surface ◽  
Type Ii ◽  
Cell Layers ◽  
Distal Lung

The epithelial tissues of the distal lung are continuously exposed to inhaled air, and are of research interest in studying respiratory exposure to both hazardous and therapeutic materials. Pharmaco-toxicological research depends on the development of sophisticated models of the alveolar epithelium, which better represent the different cell types present in the native lung and interactions between them. We developed an air-liquid interface (ALI) model of the alveolar epithelium which incorporates cell lines which bear features of type I (hAELVi) and type II (NCI-H441) epithelial cells. We compared morphology of single cells and the structure of cell layers of the two lines using light and electron microscopy. Working both in monotypic cultures and cocultures, we measured barrier function by trans-epithelial electrical resistance (TEER), and demonstrated that barrier properties can be maintained for 30 days. We created a mathematical model of TEER development over time based on these data in order to make inferences about the interactions occurring in these culture systems. We assessed expression of a panel of relevant genes that play important roles in barrier function and differentiation. The coculture model was observed to form a stable barrier akin to that seen in hAELVi, while expressing surfactant protein C, and having a profile of expression of claudins and aquaporins appropriate for the distal lung. We described cavities which arise within stratified cell layers in NCI-H441 and cocultured cells, and present evidence that these cavities represent an aberrant apical surface. In summary, our results support the coculture of these two cell lines to produce a model which better represents the breadth of functions seen in native alveolar epithelium.

scholarly journals INTRAMITOCHONDRIAL FILAMENTOUS BODIES IN THE THICK LIMB OF HENLE OF THE RAT KIDNEY

1967 ◽  
Vol 33 (3) ◽  
pp. 605-623 ◽  
Author(s):  
Teruo Suzuki ◽  
F. K. Mostofi
Keyword(s):  
Structural Characteristics ◽  
Rat Kidney ◽  
Uranyl Acetate ◽  
Type I ◽  
Type Ii ◽  
Type Iii ◽  
Tubular System ◽  
The Matrix ◽  
Developmental Form ◽  
Filamentous Inclusions

"Intramitochondrial filamentous bodies" (IMFB) were occasionally found within the matrix of some mitochondria of the thick limb of Henle of the rat kidney, but not elsewhere in the tubular system. Three types were recognized: type I, an accumulation of filaments 55 A thick; type II, a bundle of parallel filaments having the same thickness as those of type I and regular spacing, 87 A apart, from center to center; and type III, consisting of type II with regular light bands of 280 A periodicity and a helical border of prismatic tubular cristae. In addition to these, electron-opaque masses showing variable and faint substructures were found in the matrix of mitochondria. It is suggested that all these IMFB may originate from mitochondrial cristae and that type II IMFB may be an intermediate developmental form between type I and type III. After uranyl acetate staining, IMFB and the membranes of prismatic tubular cristae showed highly increased electron opacity. The literature has been reviewed for reports of intramitochondrial filamentous inclusions in various types of cells. These inclusions have been classified according to their structural characteristics and the localization in the mitochondria and compared with IMFB reported herein.

Potassium Permanganate Staining Allows For The Visualization Of An Outer Surface Component Of Selected Ehrlichiae

1999 ◽  
Vol 5 (S2) ◽  
pp. 1318-1319
Author(s):  
S.F. Hayes ◽  
U. GMunderloh ◽  
J.L. Goodman ◽  
K.M. Kocan
Keyword(s):  
Potassium Permanganate ◽  
Sodium Citrate ◽  
Uranyl Acetate ◽  
Human Beings ◽  
En Bloc ◽  
Resin Sample ◽  
Granulocytic Ehrlichiosis ◽  
Human Granulocytic Ehrlichiosis ◽  
Staining Protocol ◽  
Ethanol Series

The Ehrlichia spp. are tick-borne pathogens that infect leucocytes and erythrocytes of human beings and animals (1-4). Three ehrlichial organisms, E. chaffeensis, E. equi, HGE (human granulocytic ehrlichiosis agent, equivalent to E. phagocytophila and closely related to E. equi), and Anaplasmamarginale were studied with respect to their morphology after using a combined fixative/potassium permanganate staining protocol (5). Cell cultures infected with the respective organisms were fixed and treated as follows: Tissues were initially fixed with Karnovsky's fixative [glutaraldehydeparaformaldehyde, 2.5/4.0% respectively], containing 0.1% tannic acid, in a Sorenson's phosphate buffer with 0.1M sucrose. Post fixation with OsO4 or without it occurred in some samples. En bloc staining with 1% aqueous uranyl acetate was always done. Dehydration was in a graded ethanol series followed by embedding in Spurr's™ resin. Sample sections were stained for 30-60 minutes at RT with 1% potassium permanganate. These were then washed for 5-15 seconds with 0.025-0.05% sodium citrate, jet washed with dH20, dried and examined (6).

Platinum Compounds as Stains for Electron Microscopy

1974 ◽  
Vol 32 ◽  
pp. 230-231
Author(s):  
S. K. Aggarwal ◽  
P. McAllister ◽  
R. W. Wagner ◽  
B. Rosenberg
Keyword(s):  
Electron Microscopy ◽  
Hela Cells ◽  
Uranyl Acetate ◽  
Sarcoma 180 ◽  
Platinum Compounds ◽  
Kb Cells ◽  
En Bloc ◽  
Human Lymphoma ◽  
Ultrathin Sections ◽  
Blue Coloration

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.

Cellular Contacts Within the Interhemal Membrane of the Rat Placenta at Term

1974 ◽  
Vol 32 ◽  
pp. 146-147
Author(s):  
William P. Jollie
Keyword(s):  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Capillary Endothelium ◽  
En Bloc ◽  
Aldehyde Fixation ◽  
Chorioallantoic Placenta

By routine EM preparative techniques, the tissues which, collectively, separate maternal and fetal bloods in the fully formed chorioallantoic placenta of the rat have been shown to consist of three chorionic layers, or trophoblast, and a layer of allantoic capillary endothelium [Fig. 1]. Relationships between these layers are best demonstrated by special techniques, viz., cacodylate-buffered aldehyde fixation, collidine-buffered osmium tetroxide postfixation, and en bloc staining with uranyl acetate. By using this method on placentas at term, the cells of the outermost chorionic layer (Trophoblast 1) appear to be attached to each other by means of maculae adherentes which sometimes occur in clusters [Fig. 2].

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