Complex Vesicles, Microtubules, and Cytoplasmic Filaments in the Endothelial Cells of Normal Dog Aorta

Author(s):  
C. N. Sun ◽  
J. J. Ghidoni

Endothelial cells in longitudinal and cross sections of aortas from 3 randomly selected “normal” mongrel dogs were studied by electron microscopy. Segments of aorta were distended with cold cacodylate buffered 5% glutaraldehyde for 10 minutes prior to being cut into small, well oriented tissue blocks. After an additional 1-1/2 hour period in glutaraldehyde, the tissue blocks were well rinsed in buffer and post-fixed in OsO4. After dehydration they were embedded in a mixture of Maraglas, D.E.R. 732, and DDSA.Aldehyde fixation preserves the filamentous and tubular structures (300 Å and less) for adequate demonstration and study. The functional significance of filaments and microtubules has been recently discussed by Buckley and Porter; the precise roles of these cytoplasmic components remains problematic. Endothelial cells in canine aortas contained an abundance of both types of structures.

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 710-713 ◽  
Author(s):  
EM Cramer ◽  
D Meyer ◽  
R le Menn ◽  
J Breton-Gorius

Abstract Immunogold staining was used to study the ultrastructural distribution of von Willebrand factor (vWF) in unstimulated platelets. vWF was detected in the alpha-granules with a specific eccentric distribution pattern opposite the nucleoids. Similar findings were obtained with a polyclonal antibody or a pool of monoclonal antibodies to human vWF. This labeling coincided with the presence of tubular structures located at the periphery of the alpha-granules. These structures were better visualized on platelets treated for standard electron microscopy: they formed a group of one to four tubules ranging from 200 A to 250 A in diameter. They closely resembled the internal tubular structures found in Weibel-Palade bodies, which are the storage organelles of vWF in endothelial cells.


1978 ◽  
Vol 26 (5) ◽  
pp. 417-421
Author(s):  
D K Racker

The study of whole negatively stained cells has revealed details of cellular organelles in rat venous endothelial cells. In particular, details of surface membrane organelles and small tubular structures were demonstrated. The surface membrane organelles which appeared "vesicular-like" were found to be connected with small tubular attachments. These findings were correlated with those described by other techniques. It is significant that this simple technique appears to permit the demonstration of fine details of three-dimensional cytoplasmic structures.


Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 710-713 ◽  
Author(s):  
EM Cramer ◽  
D Meyer ◽  
R le Menn ◽  
J Breton-Gorius

Immunogold staining was used to study the ultrastructural distribution of von Willebrand factor (vWF) in unstimulated platelets. vWF was detected in the alpha-granules with a specific eccentric distribution pattern opposite the nucleoids. Similar findings were obtained with a polyclonal antibody or a pool of monoclonal antibodies to human vWF. This labeling coincided with the presence of tubular structures located at the periphery of the alpha-granules. These structures were better visualized on platelets treated for standard electron microscopy: they formed a group of one to four tubules ranging from 200 A to 250 A in diameter. They closely resembled the internal tubular structures found in Weibel-Palade bodies, which are the storage organelles of vWF in endothelial cells.


Author(s):  
R. W. Anderson ◽  
D. L. Senecal

A problem was presented to observe the packing densities of deposits of sub-micron corrosion product particles. The deposits were 5-100 mils thick and had formed on the inside surfaces of 3/8 inch diameter Zircaloy-2 heat exchanger tubes. The particles were iron oxides deposited from flowing water and consequently were only weakly bonded. Particular care was required during handling to preserve the original formations of the deposits. The specimen preparation method described below allowed direct observation of cross sections of the deposit layers by transmission electron microscopy.The specimens were short sections of the tubes (about 3 inches long) that were carefully cut from the systems. The insides of the tube sections were first coated with a thin layer of a fluid epoxy resin by dipping. This coating served to impregnate the deposit layer as well as to protect the layer if subsequent handling were required.


Author(s):  
D. E. Philpott ◽  
W. Sapp ◽  
C. Williams ◽  
Joann Stevenson ◽  
S. Black

The response of spermatogonial cells to X-irradiation is well documented. It has been shown that there is a radiation resistent stem cell (As) which, after irradiation, replenishes the seminiferous epithelium. Most investigations in this area have dealt with radiation dosages of 100R or more. This study was undertaken to observe cellular responses at doses less than 100R of X-irradiation utilizing a system in which the tissue can be used for light and electron microscopy.Brown B6D2F1 mice aged 16 weeks were exposed to X-irradiation (225KeV; 15mA; filter 0.35 Cu; 50-60 R/min). Four mice were irradiated at each dose level between 1 and 100 rads. Testes were removed 3 days post-irradiation, fixed, and embedded. Sections were cut at 2 microns for light microscopy. After staining, surviving spermatogonia were identified and counted in tubule cross sections. The surviving fraction of spermatogonia compared to control, S/S0, was plotted against dose to give the curve shown in Fig. 1.


Author(s):  
M.E. Lee ◽  
A. Moller ◽  
P.S.O. Fouche ◽  
I.G Gaigher

Scanning electron microscopy of fish scales has facilitated the application of micro-structures to systematics. Electron microscopy studies have added more information on the structure of the scale and the associated cells, many problems still remain unsolved, because of our incomplete knowledge of the process of calcification. One of the main purposes of these studies has been to study the histology, histochemistry, and ultrastructure of both calcified and decalcified scales, and associated cells, and to obtain more information on the mechanism of calcification in the scales. The study of a calcified scale with the electron microscope is complicated by the difficulty in sectioning this material because of the close association of very hard tissue with very soft tissues. Sections often shatter and blemishes are difficult to avoid. Therefore the aim of this study is firstly to develop techniques for the preparation of cross sections of fish scales for scanning electron microscopy and secondly the application of these techniques for the determination of the structures and calcification of fish scales.


Author(s):  
Richard Mcintosh ◽  
David Mastronarde ◽  
Kent McDonald ◽  
Rubai Ding

Microtubules (MTs) are cytoplasmic polymers whose dynamics have an influence on cell shape and motility. MTs influence cell behavior both through their growth and disassembly and through the binding of enzymes to their surfaces. In either case, the positions of the MTs change over time as cells grow and develop. We are working on methods to determine where MTs are at different times during either the cell cycle or a morphogenetic event, using thin and thick sections for electron microscopy and computer graphics to model MT distributions.One approach is to track MTs through serial thin sections cut transverse to the MT axis. This work uses a video camera to digitize electron micrographs of cross sections through a MT system and create image files in computer memory. These are aligned and corrected for relative distortions by using the positions of 8 - 10 MTs on adjacent sections to define a general linear transformation that will align and warp adjacent images to an optimum fit. Two hundred MT images are then used to calculate an “average MT”, and this is cross-correlated with each micrograph in the serial set to locate points likely to correspond to MT centers. This set of points is refined through a discriminate analysis that explores each cross correlogram in the neighborhood of every point with a high correlation score.


Author(s):  
J. S. Shah ◽  
R. Durkin ◽  
A. N. Farley

It is now possible to perform High Pressure Scanning Electron Microscopy (HPSEM) in the range 10 to 2000 Pa. Here the effect of scattering on resolution has been evaluated by calculating the profile of the beam in high pressure and assessing its effect on the image contrast . An experimental scheme is presented to show that the effect of the primary beam ionization is to reduce image contrast but this effect can be eliminated by a novel use of specimen current detection in the presence of an electric field. The mechanism of image enhancement is discussed in terms of collection of additional carriers generated by the emissive components.High Pressure SEM (HPSEM) instrumentation is establishing itself as commercially viable. There are now a number of manufacturers, such as JEOL, ABT, ESCAN, DEBEN RESEARCH, selling microscopes and accessories for HPSEM. This is because high pressure techniques have begun to yield high quality micrographs at medium resolution.To study the effect of scattering on the incident electron beam, its profile - in a high pressure environment - was evaluated by calculating the elastic and inelastic scattering cross sections for nitrogen in the energy range 5-25 keV. To assess the effect of the scattered beam on the image contrast, the modification of a sharp step contrast function due to scattering was calculated by single scattering approximation and experimentally confirmed for a 20kV accelerated beam.


1978 ◽  
Vol 39 (03) ◽  
pp. 751-758 ◽  
Author(s):  
B L Sheppard ◽  
J Bonnar

SummaryThe fibrinolytic activity of the intimal cells of decidual spiral arteries and the syncytium of placental villi was studied by electron microscopy in ten normal full-term human pregnancies using a modification of the fibrin slide technique. Endothelial cells lining the intima of the decidual spiral arteries showed a considerably greater fibrinolytic activity than intimal cytotrophoblast and the syncytiotrophoblast showed no activity.The replacement of endothelial cells by an intimal lining of cytotrophoblast, and the presence of cytotrophoblast in the media, appears to play an important role in the reduction of the fibrinolytic activity of the vessel. This inhibition of fibrinolytic activity in the utero-placental arteries may be the physiological mechanism which controls fibrin deposition in these vessels and on the placental villi.


1994 ◽  
Vol 72 (01) ◽  
pp. 140-145 ◽  
Author(s):  
Valeri Kolpakov ◽  
Maria Cristina D'Adamo ◽  
Lorena Salvatore ◽  
Concetta Amore ◽  
Alexander Mironov ◽  
...  

SummaryActivated neutrophils may promote thrombus formation by releasing proteases which may activate platelets, impair the fibrinolytic balance and injure the endothelial monolayer.We have investigated the morphological correlates of damage induced by activated neutrophils on the vascular wall, in particular the vascular injury induced by released cathepsin G in both static and dynamic conditions.Human umbilical vein endothelial cells were studied both in a cell culture system and in a model of perfused umbilical veins. At scanning electron microscopy, progressive alterations of the cell monolayer resulted in cell contraction, disruption of the intercellular contacts, formation of gaps and cell detachment.Contraction was associated with shape change of the endothelial cells, that appeared star-like, while the underlying extracellular matrix, a potentially thrombogenic surface, was exposed. Comparable cellular response was observed in an “in vivo” model of perfused rat arterial segment. Interestingly, cathepsin G was active at lower concentrations in perfused vessels than in culture systems. Restoration of blood flow in the arterial segment previously damaged by cathepsin G caused adhesion and spreading of platelets on the surface of the exposed extracellular matrix. The subsequent deposition of a fibrin network among adherent platelets, could be at least partially ascribed to the inhibition by cathepsin G of the vascular fibrinolytic potential.This study supports the suggestion that the release of cathepsin G by activated neutrophils, f.i. during inflammation, may contribute to thrombus formation by inducing extensive vascular damage.


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