ELECTRON MICROSCOPY OF ISOLATED PLANT MITOCHONDRIA AND PLASTIDS USING BOTH THE THIN-SECTION AND NEGATIVE-STAINING TECHNIQUES

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.

Author(s):  
J. C. Russ ◽  
E. McNatt

In order to study the retention of copper in cirrhotic liver, rats were made cirrhotic by carbon tetrachloride inhalation twice weekly for three months and fed 0.2% copper acetate ad libidum in drinking water for one month. The liver tissue was fixed in osmium, sectioned approximately 2000 Å thick, and stained with lead citrate. The section was examined in a scanning electron microscope (JEOLCO JSM-2) in the transmission electron mode.Figure 1 shows a typical area that includes a red blood cell in a sinusoid, a disse, and a portion of the cytoplasm of a hepatocyte which contains several mitochondria, peribiliary dense bodies, glycogen granules, and endoplasmic reticulum.


1961 ◽  
Vol 9 (3) ◽  
pp. 609-617 ◽  
Author(s):  
M. Zalokar

Normal and centrifuged hyphae of Neurospora were studied with the electron microscope. The following cell structures could be identified: nuclei with nucleoli, mitochondria, endoplasmic reticulum, ribosomes, glycogen, fat bodies, vacuoles, and vesicles with an inner canalicular system, of unknown nature. In centrifuged hyphae, the glycogen layer appeared as a light area, with a slight indication of granular structure. The ribosome layer consisted of densely packed ribosomes without any membranes. The mitochondrial layer contained spaces filled with ribosomes. The nuclei were loosely packed, with endoplasmic reticulum between them. The "enchylema" layer was composed of vesicles belonging to the endoplasmic reticulum. The vacuolar layer was poorly preserved and consisted of double-walled vesicles. Fat appeared as stellate osmiophilic droplets. These observations were compared with previous observations under the optical microscope and their meaning for cell physiology was discussed.


1984 ◽  
Vol 99 (3) ◽  
pp. 875-885 ◽  
Author(s):  
A Saito ◽  
S Seiler ◽  
A Chu ◽  
S Fleischer

We have developed a procedure to isolate, from skeletal muscle, enriched terminal cisternae of sarcoplasmic reticulum (SR), which retain morphologically intact junctional "feet" structures similar to those observed in situ. The fraction is largely devoid of transverse tubule, plasma membrane, mitochondria, triads (transverse tubules junctionally associated with terminal cisternae), and longitudinal cisternae, as shown by thin-section electron microscopy of representative samples. The terminal cisternae vesicles have distinctive morphological characteristics that differ from the isolated longitudinal cisternae (light SR) obtained from the same gradient. The terminal cisternae consist of two distinct types of membranes, i.e., the junctional face membrane and the Ca2+ pump protein-containing membrane, whereas the longitudinal cisternae contain only the Ca2+ pump protein-containing membrane. The junctional face membrane of the terminal cisternae contains feet structures that extend approximately 12 nm from the membrane surface and can be clearly visualized in thin section through using tannic acid enhancement, by negative staining and by freeze-fracture electron microscopy. Sections of the terminal cisternae, cut tangential to and intersecting the plane of the junctional face, reveal a checkerboardlike lattice of alternating, square-shaped feet structures and spaces each 20 nm square. Structures characteristic of the Ca2+ pump protein are not observed between the feet at the junctional face membrane, either in thin section or by negative staining, even though the Ca2+ pump protein is observed in the nonjunctional membrane on the remainder of the same vesicle. Likewise, freeze-fracture replicas reveal regions of the P face containing ropelike strands instead of the high density of the 7-8-nm particles referable to the Ca2+ pump protein. The intravesicular content of the terminal cisternae, mostly Ca2+-binding protein (calsequestrin), is organized in the form of strands, sometimes appearing paracrystalline, and attached to the inner face of the membrane in the vicinity of the junctional feet. The terminal cisternae preparation is distinct from previously described heavy SR fractions in that it contains the highest percentage of junctional face membrane with morphologically well-preserved junctional feet structures.


1983 ◽  
Vol 31 (6) ◽  
pp. 755-764 ◽  
Author(s):  
P Liesi

Laminin was localized in cultured mouse C1300 neuroblastoma cells by applying the peroxidase-antiperoxidase technique in preembedding electron microscopy. The results were compared to those obtained by indirect immunofluorescence and by the colloidal gold second antibody method on Epon-embedded ultrathin sections. Laminin was found in the cell membranes and within the rough endoplasmic reticulum as well as in intracytoplasmic vacuoles. Plasma membranes of the neuroblastoma cells showed a patchy localization of laminin that was apparently involved in cell-to-substrate attachment and in gap junction-like intercellular connections. Under normal conditions, the Golgi cisternae contained no laminin. Pretreatment of cells with micromolar concentrations of monensin, however, lead to an accumulation of laminin within the Golgi cisternae. These results support a role for laminin as an adhesion protein in cultured neuroblastoma cells and indicate that laminin is transported through the Golgi complex.


Author(s):  
Ronald H. Jones ◽  
Zensaku Yoshii ◽  
Oscar J. Carver

Four strains (Reiter, Kazan 2,4,5) of treponemata were cultivated at 37° C for 3-7 days in standard media containing complement inactivated rabbit serum (Cannefax, Spirolate) and in a defined medium devised by Johnson and his associates (unpublished data). Electron microscope preparations included cells which were unwashed (harvested), washed up to seven times, autolysed, or ruptured by explosive decompression. Each preparation was fixed with gluteraldehyde or left unfixed. Negative staining was performed using phosphotungstic acid (PTA - pH 7.2) or ammonium molybdate (AM - pH 7.0-7.2). Electron microscopy was done with a JEM 6A model at 80kV using a magnification range of 10,000-50,000X.The autolytic and disruption techniques were most effective for visualizing the ultrastructure of the more internal cell layers or moieties. The number of axial fibers (AF) arising from each cell pole varied from 4-7, six being the most common.


Blood ◽  
1964 ◽  
Vol 23 (3) ◽  
pp. 300-320 ◽  
Author(s):  
ROBERT J. CAPONE ◽  
EVA LURIE WEINREB ◽  
GEORGE B. CHAPMAN

Abstract The development of representative myeloid elements is traced by correlated light and electron microscopy. Cytoplasmic changes during maturation of granulocytes from the myeloblast include loss of basophilia, development of the endoplasmic reticulum complex, decrease in number of mitochondria, and granule formation. The endoplasmic reticulum vesicles increase in size and number during the promyelocyte and myelocyte stages, accompanied by the appearance of non-specific and specific granules, and decrease again during the cytosomal maturation of the metamyelocyte. A reduction in number of mitochondria is noted through the metamyelocyte stage. The apparent continuity of the limiting membranes of both the granules and mitochondria with those of the cisternae of endoplasmic reticulum suggests a direct connection among cytosomal organelles. The role of the endoplasmic reticulum in granulogenesis is discussed. Maturation of the nucleus involves a loss of nucleolar differentiation by a loosening of the compact fibrillar aggregates, and progressive chromatin condensation.


1964 ◽  
Vol 20 (1) ◽  
pp. 175-183 ◽  
Author(s):  
G. G. Selman ◽  
A. Jurand

Ultrasonic treatment of the tails of Triturus alpestris tadpoles, at intensities of 8 to 15 watts/cm2, at 1 megacycle/sec., for 5 minutes, disrupted the epidermis and caused pycnosis in individual cells of the muscle and neural tube, but caused no damage to the notochord that could be detected by light microscopy. Electron microscopy showed that this ultrasonic treatment disordered nearly all the endoplasmic reticulum (ER) of the notochord cells into irregularly rounded vesicles, but within 3 hours after treatment some parallel arrays of normal endoplasmic reticulum were seen near, and continuous with, the outer nuclear membrane. In addition, a re-ordering of the previously disordered ER took place throughout the cytoplasm, in some cases. A classification was made of the state of the ER as shown in electron micrographs of material fixed immediately, 3, and 24 hours after treatment. This showed that more than half the total endoplasmic reticulum in notochord cells was normal again by 24 hours after treatment.


1984 ◽  
Vol 32 (8) ◽  
pp. 849-861 ◽  
Author(s):  
R Davis ◽  
G B Koelle ◽  
U J Sanville

Ciliary ganglia (CG) of cats were stained for acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) by the bis-(thioacetoxy) aurate (I), or Au(TA)2, method for examination by electron microscopy. Acetylcholinesterase was localized along the axolemmas of the preganglionic fibers and their terminals and on the plasmalemmas of the perikarya and dendrites of the ganglion cells, as in the cat superior cervical ganglion (SCG). In contrast to the SCG, AChE was also found in significant amounts in the rough endoplasmic reticulum of the CG cells and dendrites, and in varying but high concentrations in channels of extracellular space in the complex capsular region surrounding the perikarya and dendrites. Butyrylcholinesterase was confined chiefly to the dendritic and perikaryonal plasma membranes of the ganglion cells, as in the SCG. Lysosomes and mitochondria were stained chiefly for non-cholinesterase enzymes, as indicated by the physostigmine-treated controls. The significance of these distributions is discussed.


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.


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