The structure of the mitochondrial cloud of Xenopus laevis oocytes

Development ◽  
1976 ◽  
Vol 36 (3) ◽  
pp. 697-710
Author(s):  
F. S. Billett ◽  
Elizabeth Adam

The ultrastructure of the mitochondrial cloud (Balbiani body) of the pre-vitellogenic oocytes of Xenopus laevis has been examined using transmission and stereoscan electron microscopy. Examination of conventional thin sections confirm previous observations which suggest that the cloud consists essentially of many thousands of mitochondria and numerous small vesicles; larger clouds, in oocytes greater than 200µm in diameter, contain relatively more vesicles. Using a standard electron microscope at 100 kV very long and coursing arrays of mitochondrial profiles can be detected. The presence of very long mitochondrial lements has been confirmed using a high voltage microscope operating at 500–1000 kV. Stereoscan preparations, isolated from pre-vitellogenic oocytes, lend some support to the view that the mitochondrial cloud may consist of a mass of long filamentous mitochondria and the possibility that there are large continuous regions of mitochondrial material cannot be ruled out.

Author(s):  
J. Temple Black

Since its introduction by Fernandez-Moran, the diamond knife has gained wide spread usage as a common material for cutting of thin sections of biological and metallic materials into thin films for examination in the transmission electron microscope. With the development of high voltage E.M. and scanning transmission E.M., microtomy applications will become increasingly important in the preparation of specimens. For those who can afford it, the diamond knife will thus continue to be an important tool to accomplish this effort until a cheaper but equally strong and sharp tool is found to replace the diamond, glass not withstanding.In Figs. 1 thru 3, a first attempt was made to examine the edge of a used (β=45°) diamond knife by means of the scanning electron microscope. Because diamond is conductive, first examination was tried without any coating of the diamond. However, the contamination at the edge caused severe charging during imaging. Next, a thin layer of carbon was deposited but charging was still extensive at high magnification - high voltage settings. Finally, the knife was given a light coating of gold-palladium which eliminated the charging and allowed high magnification micrographs to be made with reasonable resolution.


Author(s):  
William H. Massover

Stereoscopic examination of thick sections of fixed and embedded biological tissues by high voltage electron microscopy has been shown to allow direct visualization of three-dimensional fine structure. The present report will consider the occurrence of some new technical problems in specimen preparation and Image interpretation that are not common during lower voltage studies of thin sections.Thick Sectioning and Tissue Coloration - Epon sections of 0.5 μm or more that are cut with glass knives do not have a uniform thickness as Judged by their interference colors; these colors change with time during their flotation on the knife bath, and again when drying onto the specimen support. Quoted thicknesses thus must be considered only as rough estimates unless measured in specific regions by other methods. Chloroform vapors do not always result in good spreading of thick sections; however, they will spread spontaneously to large degrees after resting on the flotation bath for several minutes. Ribbons of thick sections have been almost impossible to obtain.


Author(s):  
Brenda R. Eisenberg ◽  
Lee D. Peachey

Analysis of the electrical properties of the t-system requires knowledge of the geometry of the t-system network. It is now possible to determine the network parameters experimentally by use of high voltage electron microscopy. The t-system was marked with exogenous peroxidase. Conventional methods of electron microscopy were used to fix and embed the sartorius muscle from four frogs. Transverse slices 0.5-1.0 μm thick were viewed at an accelerating voltage of 1000 kV using the JEM-1000 high voltage electron microscope at Boulder, Colorado and prints at x5000 were used for analysis.The length of a t-branch (t) from node to node (Fig. 1a) was measured with a magnifier; at least 150 t-branches around 30 myofibrils were measured from each frog. The mean length of t is 0.90 ± 0.11 μm and the number of branches per myofibril is 5.4 ± 0.2 (mean ± SD, n = 4 frogs).


Author(s):  
M.A. O’Keefe ◽  
J. Taylor ◽  
D. Owen ◽  
B. Crowley ◽  
K.H. Westmacott ◽  
...  

Remote on-line electron microscopy is rapidly becoming more available as improvements continue to be developed in the software and hardware of interfaces and networks. Scanning electron microscopes have been driven remotely across both wide and local area networks. Initial implementations with transmission electron microscopes have targeted unique facilities like an advanced analytical electron microscope, a biological 3-D IVEM and a HVEM capable of in situ materials science applications. As implementations of on-line transmission electron microscopy become more widespread, it is essential that suitable standards be developed and followed. Two such standards have been proposed for a high-level protocol language for on-line access, and we have proposed a rational graphical user interface. The user interface we present here is based on experience gained with a full-function materials science application providing users of the National Center for Electron Microscopy with remote on-line access to a 1.5MeV Kratos EM-1500 in situ high-voltage transmission electron microscope via existing wide area networks. We have developed and implemented, and are continuing to refine, a set of tools, protocols, and interfaces to run the Kratos EM-1500 on-line for collaborative research. Computer tools for capturing and manipulating real-time video signals are integrated into a standardized user interface that may be used for remote access to any transmission electron microscope equipped with a suitable control computer.


1979 ◽  
Vol 35 (1) ◽  
pp. 403-415
Author(s):  
N. Chegini ◽  
V. Aleporou ◽  
G. Bell ◽  
V.A. Hilder ◽  
N. Maclean

Adult Xenopus laevis, rendered anaemic by phenylhydrazine injection, have been studied during the recovery from such anaemia. Electron microscopy of liver and spleen sections indicates that both of these organs are active in the phagocytosis and destruction of the old damaged red blood cells. May-Grunwald and Giemsa staining of liver and spleen cells following anaemia has been used to show that erythropoiesis also occurs in both liver and spleen, and this has been confirmed by electron-microscope studies of these organs. Cell counting and radiolabelling of the new population of circulating erythroid cells in the period following phenylhydrazine injection suggests that a sudden release of basophilic erythroblasts from liver and spleen is followed by mitosis of this new cell population in circulation, and that no further release of erythroid cells from these organs is likely until complete recovery has occurred.


2020 ◽  
Vol 68 (6) ◽  
pp. 389-402
Author(s):  
Lars Möller ◽  
Gudrun Holland ◽  
Michael Laue

Diagnostic electron microscopy is a useful technique for the identification of viruses associated with human, animal, or plant diseases. The size of virus structures requires a high optical resolution (i.e., about 1 nm), which, for a long time, was only provided by transmission electron microscopes operated at 60 kV and above. During the last decade, low-voltage electron microscopy has been improved and potentially provides an alternative to the use of high-voltage electron microscopy for diagnostic electron microscopy of viruses. Therefore, we have compared the imaging capabilities of three low-voltage electron microscopes, a scanning electron microscope equipped with a scanning transmission detector and two low-voltage transmission electron microscopes, operated at 25 kV, with the imaging capabilities of a high-voltage transmission electron microscope using different viruses in samples prepared by negative staining and ultrathin sectioning. All of the microscopes provided sufficient optical resolution for a recognition of the viruses tested. In ultrathin sections, ultrastructural details of virus genesis could be revealed. Speed of imaging was fast enough to allow rapid screening of diagnostic samples at a reasonable throughput. In summary, the results suggest that low-voltage microscopes are a suitable alternative to high-voltage transmission electron microscopes for diagnostic electron microscopy of viruses.


1975 ◽  
Vol 66 (2) ◽  
pp. 404-413 ◽  
Author(s):  
J J Paulin

The unitary nature of the chondriome of two species of trypanosomatids, Blastocrithidia culicis and Trypanosoma cruzi, has been demonstrated by utilizing serial thick-sectioning techniques combined with high voltage electron microscopy. Profiles of mitochondrial elements seen in thin sections and suspected to be parts of a continuum were confirmed by serial thick sectioning (0.25-0.50 mum thick) and stereopair analysis to be parts of the same mitochondrion. Three-dimensional models obtained from tracings of mitochondrial profiles on cellulose acetate reveal the mitochondrion of B. culicis to consist of a posterior mass with six tubular extensions extending upward and terminating in the anterior apex. The kinetoplast was found suspended between two of the tubular extensions, or less frequently, protuding as a nodule from one of the extensions. A bifurcation of one of the extensions was found in some specimens. The mitochondrion of T. cruzi consists of a triangular-shaped convoluted tubule, the base being the kinetoplast portion while the apex is directed posteriorly. The mitochondrion bifurcates behind the flagellar pocket, lateral to the kinetoplast, sending two entwined extensions into the tenuous anterior apex. Whether the mitochondrion of T. cruzi is unitary in the trypomastigote form was not determined in this study, since only epimastigote forms were used.


2009 ◽  
Vol 610-613 ◽  
pp. 591-597
Author(s):  
Zhi Wei Du ◽  
Z.M. Sun ◽  
B.L. Shao ◽  
A.S. Liu

Cu99.8Y0.2, Cu99.2Y0.8 and Cu98Y2 alloy ribbons were prepared by single roller melt spinning. The microstructure was studied by X-ray diffraction (XRD), transmission electron microscopy (TEM), high voltage electron microscope (HVEM) and high resolution electron microscopy (HREM). The results showed that α-Cu was the dominative phase in the rapid solidification ribbons of three alloys. A secondary phase Cu4Y was detected in the Cu98Y2 ribbon by XRD. The grain size was in a range of 50-200 nm in the Cu99.2Y0.8 and Cu98Y2 ribbons. Many nano-scale twins and some dislocations existed inside the larger grains. However, the grains in Cu99.8Y0.2 ribbon were in the size of microns and the sub-grains with small misorentations were in 100-200 nm. To understand the deformation mechanism, in situ tensile test were carried out at a High Voltage Electron Microscope (HVEM). The results showed that the deformation is predominated by the dislocation slip in larger grains. To accommodate the deformation, elastic deformation occured in the small grains in the initial stage of the deformation. Meanwhile, some small grains maybe deform by grain rotations. With strain increasing, some fractures generated and propagated along the grain boundaries or across the grains.


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