Morphometry characterisation of European eel spermatozoa with computer-assisted spermatozoa analysis and scanning electron microscopy

2006 ◽  
Vol 65 (7) ◽  
pp. 1302-1310 ◽  
Author(s):  
F. Marco-Jiménez ◽  
L. Pérez ◽  
M.P. Viudes de Castro ◽  
D.L. Garzón ◽  
D.S. Peñaranda ◽  
...  
Author(s):  
G. Pereira

Previous electron microscopic observations of the spleen have revealed the white pulp to be completely separated from the extravasated blood in the surrounding marginal zone by a strategically-located, double layer of reticular cells ensheathing a coarse reticular fiber. Similarly, a single reticular cell layer has been observed to form a continuous investment for all white pulp capillaries. To test the significance of this apparent isolation of the splenic white pulp from the blood, the distribution and composition of silver deposits in the spleen of argyric rats were determined by transmission and scanning electron microscopy coupled with computer-assisted x-ray analysis.Young male albino rats were made argyric by supplying them for many months with drinking water to which 1.5gm per liter of silver nitrate had been added. Specimens from the spleens of control and argyric animals were prepared for conventional transmission electron microscopy by glutaraldehyde-osmium fixation. For scanning electron microscopy, other specimens were fixed in buffered glutaraldehyde, freeze-dried in vacuo, coated with a thin film of gold- palladium and examined in a Cambridge Stereoscan Mark II.


Author(s):  
P.J. Lea ◽  
R.J. Temkin ◽  
T. Banoub ◽  
M. Silverman ◽  
M.J. Hollenberg

The principal applications of scanning electron microscopy (SEM) to renal ultrastructure have been in the study of the surface topography of various kidney cell types and their orientation and distribution in both health and disease. SEM study, however, has been limited in a major way by a lack of resolution sufficient to readily examine in detail and in three dimensions such features as glomerular basement membrane substructure and the structural organization at high magnification of the glomerular, capillary endothelium. Consequently, most of our current information about the 3D ultrastructure of rat kidney glomerulus has been obtained from transmission electron (TEM) micrographs obtained from thin sections cut at various planes followed by computer assisted, serial reconstructions. Recent advances in specimen preparation techniques and scanning electron microscope design have permitted ultrastructural examination of the glomerular capillary wall in three dimensions using high resolution scanning electron microscopy (HRSEM). Specimens in which the cytosol and cytoskeleton have been extracted, but cell membranes nuclear structures and organelles left in place, were studied using a Hitachi SEM with a resolution of approximately 3 nm.


1992 ◽  
Vol 263 (2) ◽  
pp. E231-E238
Author(s):  
C. Penel ◽  
J. Mauchamp

Pseudopods at the apical pole of porcine thyroid monolayers in culture were considered as reflecting individual thyrocyte responses to thyrotropin (TSH) stimulation. Scanning electron microscopy and computer-assisted image analysis showed that whatever TSH stimulation was used, the pseudopods were characterized by two populations: P1 with small diameters (2 microns) and P2 with greater diameters (5 microns). The density of P1 rapidly increased to reach a plateau, while P2 continuously increased during stimulation. Two-dimensional pseudopod patterns were compared with random point distributions by means of two topographical parameters: the interpseudopod distances and angles. A factorial analysis of experimental distribution of pseudopods obtained after increasing stimulation times displayed a shift from a nonrandom (10-20 min) to a random (60-90 min) distribution. Clusters of three pseudopods characterized by short distances (6-9 microns) and equilateral organization (angles 40-60 degrees) were observed after a 10-min stimulation. These results suggested that early thyrocyte response to TSH stimulation is characterized by interrelations between three adjoining cells, with the thyrocyte response later appearing as random.


Author(s):  
P.S. Porter ◽  
T. Aoyagi ◽  
R. Matta

Using standard techniques of scanning electron microscopy (SEM), over 1000 human hair defects have been studied. In several of the defects, the pathogenesis of the abnormality has been clarified using these techniques. It is the purpose of this paper to present several distinct morphologic abnormalities of hair and to discuss their pathogenesis as elucidated through techniques of scanning electron microscopy.


Author(s):  
P.J. Dailey

The structure of insect salivary glands has been extensively investigated during the past decade; however, none have attempted scanning electron microscopy (SEM) in ultrastructural examinations of these secretory organs. This study correlates fine structure by means of SEM cryofractography with that of thin-sectioned epoxy embedded material observed by means of transmission electron microscopy (TEM).Salivary glands of Gromphadorhina portentosa were excised and immediately submerged in cold (4°C) paraformaldehyde-glutaraldehyde fixative1 for 2 hr, washed and post-fixed in 1 per cent 0s04 in phosphosphate buffer (4°C for 2 hr). After ethanolic dehydration half of the samples were embedded in Epon 812 for TEM and half cryofractured and subsequently critical point dried for SEM. Dried specimens were mounted on aluminum stubs and coated with approximately 150 Å of gold in a cold sputtering apparatus.Figure 1 shows a cryofractured plane through a salivary acinus revealing topographical relief of secretory vesicles.


Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).


Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.


Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).


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