scholarly journals STUDIES ON NUCLEI USING CORRELATED CYTOCHEMICAL, LIGHT, AND ELECTRON MICROSCOPE TECHNIQUES

1956 ◽  
Vol 2 (4) ◽  
pp. 397-406 ◽  
Author(s):  
Montrose J. Moses

In this paper, a procedure for correlating electron microscope and light microscope cytochemical studies using immediately adjacent serial thin and thick sections has been described and discussed. This technique, combined with the Feulgen reaction for DNA, has been of particular value in framing and answering both general and specific questions about the nucleus. The results may be summarized as follows:— Apparent nuclear homogeneity in the electron microscope is not due to loss of DNA as evidenced by positive Feulgen reactions in such nuclei. Arrangement of Feulgen-positive material in chromosomes, heterochromatin, perinuclear and perinucleolar chromatin, etc., is similar to that customarily observed in the light microscope but this is not necessarily reflected in a cursory survey of the electron image. Careful comparison of light and electron images shows that fine differences in structure are associated with chromatin localization. Primary spermatocyte prophase chromosomes of crayfish have been positively identified by their Feulgen-positive nature. Core-like axial structures in such chromosomes have been observed (9) and are described further. A remarkable feature of spermiogenesis in the crayfish is an elaboration of the nuclear envelope of the spermatid accompanying the formation of what becomes a mass of convoluted membranes in the sperm. In the spermatid, perinuclear chromatin follows outpocketings of the nuclear envelope into the cytoplasm. In the early sperm, on the other hand, although the nuclear envelope is continuous with the system of convoluted membranes, the chromatin is distinct from it and is retained in the nucleus proper by some mechanism independent of the nuclear envelope. None of the above observations was apparent from the electron microscope images alone; they were possible only by virtue of the correlated cytochemical and electron microscope study of adjacent sections. The successful use of other cytochemical tests, such as the PAS reaction for certain carbohydrates, in such correlated studies is also described.

1973 ◽  
Vol 51 (11) ◽  
pp. 2065-2074 ◽  
Author(s):  
Rand McNitt

Mitosis in zoosporangia of the chytrid Phlyctochytrium irregulare is described from electron microscope observations and also from light microscope observations of both living and haematoxylin-stained thalli. At the onset of prophase the centriole complex replicates, and the complexes migrate to polar positions. The semi-persistent nucleolus is appressed to the nuclear envelope as the nuclear pockets invaginate, finally rupturing to create polar fenestrae, through which spindle microtubules penetrate the nucleus from the region of the centrioles at prometaphase. Metaphase chromosomes form an equatorial plate. Initial separation at anaphase seems to be accomplished mainly by shortening of chromosome-to-pole microtubules; additional anaphase and telophase separation is accomplished by elongation of the nucleus. A system of perinuclear endoplasmic reticulum is formed during prophase and is completed by metaphase. It persists during all division stages after its formation. Features of this mitotic apparatus are discussed with reference to earlier light microscope studies of chytrid mitosis. The ultrastructure of P. irregulare's mitotic apparatus is similar to that of certain unicellular green algae.


Author(s):  
S. Takashima ◽  
H. Hashimoto ◽  
S. Kimoto

The resolution of a conventional transmission electron microscope (TEM) deteriorates as the specimen thickness increases, because chromatic aberration of the objective lens is caused by the energy loss of electrons). In the case of a scanning electron microscope (SEM), chromatic aberration does not exist as the restrictive factor for the resolution of the transmitted electron image, for the SEM has no imageforming lens. It is not sure, however, that the equal resolution to the probe diameter can be obtained in the case of a thick specimen. To study the relation between the specimen thickness and the resolution of the trans-mitted electron image obtained by the SEM, the following experiment was carried out.


Author(s):  
Oliver C. Wells

The low-loss electron (LLE) image in the scanning electron microscope (SEM) is useful for the study of uncoated photoresist and some other poorly conducting specimens because it is less sensitive to specimen charging than is the secondary electron (SE) image. A second advantage can arise from a significant reduction in the width of the “penetration fringe” close to a sharp edge. Although both of these problems can also be solved by operating with a beam energy of about 1 keV, the LLE image has the advantage that it permits the use of a higher beam energy and therefore (for a given SEM) a smaller beam diameter. It is an additional attraction of the LLE image that it can be obtained simultaneously with the SE image, and this gives additional information in many cases. This paper shows the reduction in penetration effects given by the use of the LLE image.


Author(s):  
J. H. Luft

Ruthenium red is one of the few completely inorganic dyes used to stain tissues for light microscopy. This novelty is enhanced by ignorance regarding its staining mechanism. However, its continued usefulness in botany for demonstrating pectic substances attests to selectivity of some sort. Whether understood or not, histochemists continue to be grateful for small favors.Ruthenium red can also be used with the electron microscope. If single cells are exposed to ruthenium red solution, sufficient mass can be bound to produce observable density in the electron microscope. Generally, this effect is not useful with solid tissues because the contrast is wasted on the damaged cells at the block surface, with little dye diffusing more than 25-50 μ into the interior. Although these traces of ruthenium red which penetrate between and around cells are visible in the light microscope, they produce negligible contrast in the electron microscope. However, its presence can be amplified by a reaction with osmium tetroxide, probably catalytically, to be easily visible by EM. Now the density is clearly seen to be extracellular and closely associated with collagen fibers (Fig. 1).


Author(s):  
P. Bonhomme ◽  
A. Beorchia

We have already described (1.2.3) a device using a pockel's effect light valve as a microscopical electron image converter. This converter can be read out with incoherent or coherent light. In the last case we can set in line with the converter an optical diffractometer. Now, electron microscopy developments have pointed out different advantages of diffractometry. Indeed diffractogram of an image of a thin amorphous part of a specimen gives information about electron transfer function and a single look at a diffractogram informs on focus, drift, residual astigmatism, and after standardizing, on periods resolved (4.5.6). These informations are obvious from diffractogram but are usualy obtained from a micrograph, so that a correction of electron microscope parameters cannot be realized before recording the micrograph. Diffractometer allows also processing of images by setting spatial filters in diffractogram plane (7) or by reconstruction of Fraunhofer image (8). Using Electrotitus read out with coherent light and fitted to a diffractometer; all these possibilities may be realized in pseudoreal time, so that working parameters may be optimally adjusted before recording a micrograph or before processing an image.


Author(s):  
T. Guha ◽  
A. Q. Siddiqui ◽  
P. F. Prentis

The Primary Spermatocytes represent a stage in spermatogenesis when the first meiotic cell division occurs. They are derived from Spermatogonium or Stem cell through mitotic division. At the zygotene phase of meiotic prophase the Synaptonemal complex appears in these cells in the space between the paired homologous chromosomes. Spermatogenesis and sperm structure in fish have been studied at the electron microscope level in a few species? However, no work has yet been reported on ultrastructure of tilapia, O. niloticus, spermatozoa and spermatogenetic process. In this short communication we are reporting the Ultrastructure of Primary Spermatocytes in tilapia, O. niloticus, and the fine structure of synaptonemal complexes seen in the spermatocyte nuclei.


1956 ◽  
Vol 2 (4) ◽  
pp. 123-128 ◽  
Author(s):  
H. W. Beams ◽  
T. N. Tahmisian ◽  
R. L. Devine ◽  
Everett Anderson

The dictyosome (Golgi body) in the secondary spermatocyte of the cricket appears in electron micrographs as a duplex structure composed of (a) a group of parallel double-membraned lamellae and (b) a group of associated vacuoles arranged along the compact lamellae in a chain-like fashion. This arrangement of ultramicroscopic structure for the dictyosomes is strikingly comparable to that described for the Golgi apparatus of vertebrates. Accordingly, the two are considered homologous structures. Associated with the duplex structure of the dictyosomes is a differentiated region composed of small vacuoles. This is thought to represent the pro-acrosome region described in light microscope preparations. In the spermatid the dictyosomes fuse, giving rise to the acroblast. Like the dictyosomes, the acroblasts are made up of double-membraned lamellae and associated vacuoles. In addition, a differentiated acrosome region is present which, in some preparations, may display the acrosome vacuole and granule. Both the dictyosomes and acroblasts are distinct from mitochondria.


1960 ◽  
Vol 7 (3) ◽  
pp. 433-441 ◽  
Author(s):  
Susumu Ito

Spermatogenic cells of Drosophila virilis were studied by light and electron microscopy. The persistence of a "nuclear wall" during the meiotic divisions has been reported by a number of early cytologists, but this interpretation has been a subject of debate. Electron micrographs of dividing spermatocytes reveal the presence of multiple layers of paired membranes surrounding the nuclear region. These lamellar membrane systems are not typical of the nuclear envelope, but were interpreted as such by light microscopists. The membranes constituting a pair are separated by an interspace of ∼ 100 A and successive pairs are 200 to 400 A apart. These spacings are similar but not identical to those found in the lamellar systems of the Golgi complex. The cisternae of the endoplasmic reticulum in this material are devoid of attached ribonucleoprotein particles, are more precisely ordered than in vertebrate cells, and show a uniform, narrow intracisternal space of ∼ 100 A. The conspicuous asters appear to be made up of similar paired membranes radiating from the centriolar region. The primary spermatocyte has numerous dictyosomes and a well developed endoplasmic reticulum in cisternal form, but no typical Golgi complex or endoplasmic reticulum is found during the meiotic division stages of metaphase to telophase. Evidence is presented that these cytoplasmic organelles contribute to the formation of the extensive lamellar systems that appear during meiosis. The results of the Golgi silver staining methods and staining tests for phospholipids, basophilia, and the PAS reaction, indicate that the lamellar arrays of membranes present during meiosis are indistinguishable from the Golgi complex in their tinctorial properties.


1985 ◽  
Vol 63 (3) ◽  
pp. 611-615 ◽  
Author(s):  
Kevin D. Hyde ◽  
E. B. Gareth Jones

Collections of filamentous marine fungi in Seychelles included Nimbospora effusa Koch and an undescribed species, Nimbospora bipolaris Hyde & Jones sp. nov. Tha latter differs from N. effusa in the size of the ascospores and in ascospore appendage morphology. Both species are illustrated by light microscope and scanning electron microscope micrographs.


1975 ◽  
Vol 18 (2) ◽  
pp. 327-346
Author(s):  
F.O. Perkins

The fine structure of the haplosporidan mitotic apparatus is described from observations of plasmodial nuclei of Minchinia nelsoni, M. costalis, Minchinia sp., and Urosporidium crescens. The apparatus, which is the Kernstab of light-microscope studies, consists of a bundle of microtubules terminating in a spindle pole body (SPB) at each end of the bundle. A few microtubules extend from SPB to SPB, but most either extend from an SPB and terminate in the nucleoplasm or lie in the nucleoplasm, free of either SPB. The bundle lengthens during mitosis, increasing the SPB-to-SPB distance by a factor of 2 to 3 as compared to interphase nuclei. SPBs are not in contact with the nuclear envelope, being found always in the nucleoplasm which is delimited by the nuclear envelope throughout mitosis. The mitotic apparatus is persistent through interphase, at least in a form which is not significantly different from that found in mitotic nuclei.


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