Peanut agglutinin-induced structural changes in cowpea Rhizobia as revealed by freeze-etching

1990 ◽  
Vol 21 (4) ◽  
pp. 243-247 ◽  
Author(s):  
Bina Mody ◽  
Rustom Mody ◽  
Vinod Modi
Keyword(s):  
Structural Changes ◽  
Peanut Agglutinin ◽  
Freeze Etching ◽  
Cowpea Rhizobia

scholarly journals A study of rapid mitochondrial structural changes in vitro by spray-freeze-etching

1978 ◽  
Vol 77 (1) ◽  
pp. 134-147 ◽  
Author(s):  
RD Lang ◽  
JR Bronk
Keyword(s):  
Time Course ◽  
Structural Changes ◽  
Rat Liver Mitochondria ◽  
Sampling Device ◽  
Rapid Sampling ◽  
Outer Compartment ◽  
Matrix Volume ◽  
The Matrix ◽  
Freeze Etching

The spray-freeze-etching technique has been used to study energy-linked mitochondrial structural changes in rat liver mitochondria incubated in vitro. The technique involves spraying the suspension of mitochondria into liquid propane at -190 degrees C, and does not require the use of cryoprotectants or chemical fixatives. The results confirmed that freshly isolated mitochondria have a condensed matrix and that this expands at the expense of the outer compartment to give the orthodox configuration when the mitochondria are incubated in a K+ medium in the presence of substrate and phosphate. Addition of adenosine diphosphate (ADP) caused a rapid shrinkage of the matrix compartment, and the time-course and extent of this shrinkage has been measured quantitatively by coupling a rapid sampling device to the spray-freezing apparatus. These data show that for orthodox mitochondria the onset of phosphorylation is accompanied by a reduction of 30% in the matrix volume in 20's, and there is no evidence that the decrease in matrical volume affects the phosphorylation efficiency. These results suggest that natural ionophores in the mitochondrial inner membrane make it permeable enough to permit a rapid readjustment of matrix volume after the addition of ADP, and that the associated ion movement does not cause uncoupling of oxidative phosphorylation.

Freeze-etch study of the cell envelope of cowpea rhizobia: compartmentalization of the periplasmic space in relation to polysaccharide excretion

1990 ◽  
Vol 36 (6) ◽  
pp. 373-383 ◽  
Author(s):  
Bina Mody ◽  
Rustom Mody ◽  
Vinod Modi
Keyword(s):  
Outer Membrane ◽  
Cell Envelope ◽  
Ruthenium Red ◽  
Distinctive Features ◽  
Rhizobium Strains ◽  
Cytoplasmic Membranes ◽  
Rigid Cell Wall ◽  
Freeze Etching ◽  
Ruthenium Red Staining ◽  
Cowpea Rhizobia

Freeze-etching electron microscopy of free-living cowpea rhizobia revealed distinctive features of the cell envelope of this bacteria. The topology of the outer and cytoplasmic membranes was comparable with that described for other Gram-negative bacteria. In cowpea Rhizobium strains JLn(c) and NC-92, the rigid cell wall almost invariably cleaved away from the outer membrane, thus exposing two distinct fracture faces in the outer membrane, which to date have been ill defined in rhizobia. Cross-fractured cells showed enlargement of the periplasmic region as a result of invagination of the cytoplasmic membrane near one end of the cell. This feature was consistent in all exponential and stationary phase cells. Ruthenium red staining of cells in various growth phases showed unipolar initiation of the capsule. A unique structure seen within the enlarged periplasm was the presence of a smooth lipidic vesicle. Key words: Rhizobium, freeze-etch, membrane, polysaccharide, vesicle.

scholarly journals PRESERVATION OF THE ULTRASTRUCTURE OF BACILLUS SUBTILIS BY CHEMICAL FIXATION AS VERIFIED BY FREEZE-ETCHING

1969 ◽  
Vol 42 (3) ◽  
pp. 733-744 ◽  
Author(s):  
N. Nanninga
Keyword(s):  
Bacillus Subtilis ◽  
Plasma Membrane ◽  
Osmium Tetroxide ◽  
Structural Changes ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Fixation Procedure ◽  
Etching Technique ◽  
Freeze Etching

The present study on the ultrastructure of Bacillus subtilis was undertaken in order to examine by means of the freeze-etching technique possible structural changes occurring during the chemical fixation procedure (Ryter-Kellenberger (R-K) fixation). Three stages were followed by freeze-etching, viz.: (a) fixation in osmium tetroxide, (b) fixation in osmium tetroxide and posttreatment with uranyl acetate, and (c) fixation in osmium tetroxide, posttreatment in uranyl acetate, and dehydration in a graded series of acetone. Preparations were made after each stage in the presence of 20% glycerol. Good preservation of ultrastructure was observed, after any of the three treatments, of the outer surface of the plasma membrane, and the inner surface of the plasma membrane. No alteration in fracturing properties could be observed. However, if we are to judge by the results of freeze-etching, any of the successive steps of the chemical fixation procedure achieve strong contrast between the nucleoplasmic region and the cytoplasm. Dependent on the quality of fixation, very delicately preserved DNA fibrils or strongly aggregated ones were seen. It appears that R-K fixation is capable of producing more or less distinctly visible changes in the native state of the nucleoplasm in young cells of B. subtilis.

Fine Structural Changes in the Microvasculature of the Mouse Pinna: Aging and Radiation Injury

1974 ◽  
Vol 32 ◽  
pp. 54-55
Author(s):  
S. Phyllis Steamer ◽  
Rosemarie L. Devine
Keyword(s):  
Structural Changes ◽  
Radiation Treatment ◽  
Arterial Stenosis ◽  
Ultrastructural Changes ◽  
Vascular Effects ◽  
Microvascular Changes ◽  
Surrounding Connective Tissue ◽  
Fission Neutrons ◽  
Total Body

The importance of radiation damage to the skin and its vasculature was recognized by the early radiologists. In more recent studies, vascular effects were shown to involve the endothelium as well as the surrounding connective tissue. Microvascular changes in the mouse pinna were studied in vivo and recorded photographically over a period of 12-18 months. Radiation treatment at 110 days of age was total body exposure to either 240 rad fission neutrons or 855 rad 60Co gamma rays. After in vivo observations in control and irradiated mice, animals were sacrificed for examination of changes in vascular fine structure. Vessels were selected from regions of specific interest that had been identified on photomicrographs. Prominent ultrastructural changes can be attributed to aging as well as to radiation treatment. Of principal concern were determinations of ultrastructural changes associated with venous dilatations, segmental arterial stenosis and tortuosities of both veins and arteries, effects that had been identified on the basis of light microscopic observations. Tortuosities and irregularly dilated vein segments were related to both aging and radiation changes but arterial stenosis was observed only in irradiated animals.

Evaluation of single-electron movies of glutamine synthetase molecules

1983 ◽  
Vol 41 ◽  
pp. 280-281
Author(s):  
W. Kunath ◽  
E. Zeitler ◽  
M. Kessel
Keyword(s):  
Electron Microscope ◽  
Glutamine Synthetase ◽  
Electron Irradiation ◽  
Structural Damage ◽  
Structural Changes ◽  
Single Electron ◽  
Continuous Series ◽  
Digital Recording ◽  
Recording Device ◽  
Low Exposure

The features of digital recording of a continuous series (movie) of singleelectron TV frames are reported. The technique is used to investigate structural changes in negatively stained glutamine synthetase molecules (GS) during electron irradiation and, as an ultimate goal, to look for the molecules' “undamaged” structure, say, after a 1 e/Å2 dose.The TV frame of fig. la shows an image of 5 glutamine synthetase molecules exposed to 1/150 e/Å2. Every single electron is recorded as a unit signal in a 256 ×256 field. The extremely low exposure of a single TV frame as dictated by the single-electron recording device including the electron microscope requires accumulation of 150 TV frames into one frame (fig. lb) thus achieving a reasonable compromise between the conflicting aspects of exposure time per frame of 3 sec. vs. object drift of less than 1 Å, and exposure per frame of 1 e/Å2 vs. rate of structural damage.

Comparison of Acrylamide and Agar Gels by Freeze-Etching

1977 ◽  
Vol 35 ◽  
pp. 606-607
Author(s):  
Russell L. Steere ◽  
Eric F. Erbe
Keyword(s):  
Electron Microscope ◽  
Sodium Hydroxide ◽  
Sodium Hypochlorite ◽  
Chromic Acid ◽  
Distilled Water ◽  
Thin Sheets ◽  
Acid Cleaning ◽  
Agar Gels ◽  
Freeze Etching ◽  
Water Cut

Thin sheets of acrylamide and agar gels of different concentrations were prepared and washed in distilled water, cut into pieces of appropriate size to fit into complementary freeze-etch specimen holders (1) and rapidly frozen. Freeze-etching was accomplished in a modified Denton DFE-2 freeze-etch unit on a DV-503 vacuum evaporator.* All samples were etched for 10 min. at -98°C then re-cooled to -150°C for deposition of Pt-C shadow- and C replica-films. Acrylamide gels were dissolved in Chlorox (5.251 sodium hypochlorite) containing 101 sodium hydroxide, whereas agar gels dissolved rapidly in the commonly used chromic acid cleaning solutions. Replicas were picked up on grids with thin Foimvar support films and stereo electron micrographs were obtained with a JEM-100 B electron microscope equipped with a 60° goniometer stage.Characteristic differences between gels of different concentrations (Figs. 1 and 2) were sufficiently pronounced to convince us that the structures observed are real and not the result of freezing artifacts.

New Dimensions in Freeze-Etching

1969 ◽  
Vol 27 ◽  
pp. 202-203 ◽  
Author(s):  
Russell L. Steere ◽  
Michael Moseley
Keyword(s):  
Fracture Surfaces ◽  
Aluminum Specimen ◽  
Specimen Holder ◽  
Freeze Etching ◽  
The Right

A redesigned specimen holder and cap have made possible the freeze-etching of both fracture surfaces of a frozen fractured specimen. In principal, the procedure involves freezing a specimen between two specimen holders (as shown in A, Fig. 1, and the left side of Fig. 2). The aluminum specimen holders and brass cap are constructed so that the upper specimen holder can be forced loose, turned over, and pressed down firmly against the specimen stage to a position represented by B, Fig. 1, and the right side of Fig. 2.

Fine Structural Changes in the Adenohypophyses of Rats Following Destruction of the Pituitary Stalk

1977 ◽  
Vol 35 ◽  
pp. 496-497
Author(s):  
K. Kovacs ◽  
E. Horvath ◽  
J. M. Bilbao ◽  
F. A. Laszlo ◽  
I. Domokos
Keyword(s):  
Structural Changes ◽  
Tissue Injury ◽  
Anterior Lobe ◽  
Uranyl Acetate ◽  
Male Rats ◽  
Pituitary Stalk ◽  
Sequence Of Events ◽  
Pituitary Glands ◽  
Electrolytic Lesions ◽  
Ultrathin Sections

Electrolytic lesions of the pituitary stalk in rats interrupt adenohypophysial blood flow and result in massive infarction of the anterior lobe. In order to obtain a deeper insight into the morphogenesis of tissue injury and to reveal the sequence of events, a fine structural investigation was undertaken on adenohypophyses of rats at various intervals following destruction of the pituitary stalk.The pituitary stalk was destroyed electrolytically, with a Horsley-Clarke apparatus on 27 male rats of the R-Amsterdam strain, weighing 180-200 g. Thirty minutes, 1,2,4,6 and 24 hours after surgery the animals were perfused with a glutaraldehyde-formalin solution. The skulls were then opened and the pituitary glands removed. The anterior lobes were fixed in glutaraldehyde-formalin solution, postfixed in osmium tetroxide and embedded in Durcupan. Ultrathin sections were stained with uranyl acetate and lead citrate and investigated with a Philips 300 electron microscope.

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