Lens fiber organization in four avian species: a scanning electron microscopic study

The three-dimensional organization of the eye lenses of the chicken, the canary, the song-thrush and the kestrel was studied using light and scanning electron microscopy. The lenses of birds are characterized by the presence of two distinct compartments: the annular pad and the main lens body, separated by a cavum lenticuli. The annular pad fibers had a hexagonal circumference all contained a round nucleus and except for the canary were smooth-surfaced and lacking anchoring devices. In the canary, however, the annular pad fibers were studded with edge protrusions and ball-and-socket junctions. The semicircular main lens body fibers of all four species were studded with ball-and-socket junctions and edge protrusions. In contrast with mammals these anchoring devices were present throughout the lens up to the embryonal nucleus. Superficially the main lens body fibers were extremely flat. Additionally membrane elevations and depressions and globular elements were found on these central fibers in three species, the kestrel being the exception. At the transition between annular pad and main lens body the fibers turned their course and the nuclei became oval and disappeared in the deeper aspect of the main lens body. The cavum lenticuli was filled with globules tied off from the annular pad fibers. It seems attractive to assume that the presence of a separated annular pad, a cavum lenticuli filled with globular elements, the extreme flatness of the superficial central fibers and the studding of these central fibers with anchoring devices up to the embryonal nucleus are morphological expressions of the mouldability of the bird's eye lenses and consequently would explain their efficient accommodative mechanism including formation of a lenticonus. The presence of nuclei in the annular pad fibers and their typical change at the transitional zone between annular pad and main lens body are suggestive for a two-phased differentiation in bird's lens fibers: differentiation of the germinative epithelial cells to annular pad fibers which migrate to the main lens body after which they differentiate further to main lens body fibers.     

Human chromatin from interphase to metaphase: a scanning electron microscopic study

An improved technique for microinjecting individual paramecium cells with fluid, making possible injection of 30 to 40 cells/h, is described. This technique has been used to show that cytoplasmically determined erythromycin resistance in Paramecium aurelia may be transferred to sensitive cells by the injection of mitochondria prepared from resistant clones. The ability of these mitochondria to transmit erythromycin resistance is unaffected by DNAse or RNAse but is completely removed by low concentrations of a non-ionic detergent. This evidence strongly suggests that the erythromycin resistance determinants are located in the mitochondria, probably in the mitochondrial DNA.     

Nasal pit formation in the hamster: a transmission and scanning electron microscopic study

The surface morphology of cells comprising the nasal placode and adjacent body surface was examined by transmission and scanning electron microscopy during nasal pit formation in hamster embryos ranging in age from $\text{8}\text{1}\text{4}$ to $\text{9}\text{1}\text{2}$ days post coitum. A sharp distinction between the apical surface appearance of cells of the nasal epithelium and cells of the surrounding periderm develops at the periphery of the nasal placode. Periderm cells increase in surface area, exhibit a change in the distribution of surface microvilli, and many acquire a single cilium per cell. Cells of the nasal placode retain a dense surface coat of microvilli and exhibit relatively smaller apical surface areas. Olfactory rods can be positively identified on the basis of their ultrastructure at the nasal groove stage. The ventral margin of the nasal groove does not initially depend on the maxillary process, but is bounded by the lateral and medial nasal processes. From their earliest development the oral and nasal cavities appear to be separated in this species.     

Rhinoglena frontalis (Rotifera,Monogononta): a scanning electron microscopic study

Females and males of Rhinoglena frontalis (Monogononta, Epiphanidae) are observed by SEM and their external morphologies are compared. The two sexes differ in size and shape of the body. The female body is fusiform with a short, conical foot, while the male body is more slender and has a rather long foot. The rotatory apparatus (or corona) of both sexes is similar with only minor differences and consists of rows and tufts of cilia arranged around the mouth opening. The corona is made of two paired lobes lateral to the mouth and of a third prominent dorsal lobe, usually called proboscis. The three lobes are lined externally by dense rows of cilia, which constitute the cingulum, used for swimming. The central surface of the proboscis is covered with numerous longitudinal rows of cilia bent towards the mouth. The lateral lobes show, on their central surfaces, two concentric arcs of cirri (made of tightly packed cilia) bent towards the mouth. The similar organization of the rotatory apparatus of both sexes is related to the fact that the male, in this species, is able to feed and has a developed mastax and digestive system. The trophi of both sexes are illustrated and compared.     

Subchondral bone microarchitecture and failure mechanism under compression: A finite element study

Subchondral bone (SCB) microdamage is commonly observed in traumatic joint injuries and has been strongly associated with post-traumatic osteoarthritis (PTOA). Knowledge of the three-dimensional stress and strain distribution within the SCB tissue helps to understand the mechanism of SCB failure, and may lead to an improved understanding of mechanisms of PTOA initiation, prevention and treatment. In this study, we used high-resolution micro-computed tomography (µCT)-based finite element (FE) modelling of cartilage-bone to evaluate the failure mechanism and the locations of SCB tissue at high-risk of initial failure under compression. The µCT images of five cartilage-bone specimens with an average SCB thickness of 1.23 ± 0.20 mm were used to develop five µCT-based FE models. The FE models were analysed under axial compressions of approximately 30 MPa applied to the cartilage surface while the bone edges were constrained. Strain and stress-based failure criteria were then applied to evaluate the failure mechanism of the SCB tissue under excessive compression through articular cartilage. µCT-based FE models predicted two locations in the SCB at high-risk of initial failure: (1) the interface of the calcified-uncalcified cartilage due to excessive tension, and (2) the trabecular bone beneath the subchondral plate due to excessive compression. µCT-based FE models of cartilage-bone enabled us to quantify the distribution of the applied compression which was transferred through the articular cartilage to its underlying SCB, and to investigate the mechanism and the mode of SCB tissue failure. Ultimately, the results will help to understand the mechanism of injury formation in relation to PTOA.     

Interpretation of scanning electron microscopic images of abraded forming bone surfaces

The experimental abrasion of forming bone surfaces was conducted so that such surfaces could be characterized. This is particularly important to bone remodeling studies utilizing scanning electron microscope (SEM) imaging of archeological material. Forming surfaces derived from subadult macaque cranial bone were treated by particle abrasion, water abrasion, sliding abrasion, brushing, manual rubbing, weight, exfoliation, chipping and replication. Acetic acid treatments were also performed. The effects of abrasive agents are specific but generally fall into rough (particle and water abrasion) and smooth (sliding abrasion, brushing, rubbing and weight) categories. Protohistoric human and Plio-Pleistocene hominid subadult craniofacial remains were observed with the SEM for comparison with experimental data. The more recent material appeared smooth, probably as a result of specimen preparation procedures using brushes. Surfaces were still interpretable as forming, however, using a more abrasion-resistant feature called intervascular ridging (IVR) described in this study. The IVR pattern is also recognized on the hominid sample, confirming the possibility of performing remodeling studies on abraded fossil material. The abrasion characteristics are somewhat more difficult to classify, however. Abrasion is defined and discussed relative to remodeling studies and taphonomy. The usefulness of the experimental data reported here, however, in paleoenvironmental reconstruction, has yet to be fully realized. Acid and mechanical preparation techniques are briefly addressed. It is concluded that it is possible to characterize a forming surface as abraded according to the findings of this study and that acid, if handled with care, will more likely preserve microanatomical surface detail. It would also be in everyone's interest to employ a less abrasive cleaning regime on archeological specimens.     

A scanning electron microscopic study of the luteo-follicular complex

Summary As observed by SEM, the repair of an ovulated mammalian follicle is accompanied by a sequence of morphogenetic processes. In the initial phase, a mass of cells and coagulated fluids forms at the site of rupture. Shortly thereafter, connective cells, recruited from the adjacent and subjacent connective tissue stroma begin to proliferate and to migrate over this mass such that in the rabbit, the entire site of disruption is covered by a layer of connective cells by approximately 2 days following ovulation. Coincident with the migration of the connective tissue, superficial cells from undisturbed lateral and basal areas of an ovulated follicle also proliferate and begin to migrate over the newly established connective tissue matrix. By approximately 4 days following ovulation in the rabbit, the surface of an ovulated follicle is repopulated by elements of the superficial epithelium. The formation of the underlying corpus luteum (corpora luted) involves characteristic morphological changes as granulosa cells transform into steroid secreting luteal cells. The luteal cells become organized into cords of cells which usually surround capillary vessels. When examined by SEM, the smooth-surfaced endoplasmic reticulum of the luteal cell is quite apparent and is observed to form a three-dimension network of anastomosing tubules which are continuous with the nuclear membrane. Variations in the appearance of the surface of the ovary which directly overlies corpora lutea were observed when the mouse, rat and rabbit were compared. The regression of corpora lutea involves the infiltration of the luteal mass by connective tissue and both degeneration and vacuolization of the luteal cells. The regressing corpus luteum is a honey-comb-like structure in which each space is occupied by a degenerating luteal cell.This work was supported by grants from the National Institutes of Health, Public Health Service (to J.V.B., no. HD-04274), and from Consiglio Nationale delle Ricerche (C.N.R., contracts nos. CT 76.01288.04 and CT 77.01921.4)     

Brown adipose tissue: a scanning electron microscopic study of tissue and cultured adipocytes

The ultrastructure of rat brown adipose tissue (BAT) and of adipocytes cultured from BAT were studied by scanning electron microscopy (SEM). Brown adipocytes in the intact tissue were arranged in lobules with bundles of collagen among them; within each lobule 20- to 40-microns-large adipocytes were packed together. Fibers of reticular collagen enveloped each adipocyte and also connected each cell to vessels and nerves. At the adipocyte surface rounded protrusions were present, which corresponded to the BAT-typical multivacuolar lipid depot. Gradual digestion of the stroma with collagenase disclosed a more delicate, felt-like cover surrounding each adipocyte, probably representing the external lamina of the cell. Complete digestion of the stroma showed a smooth plasmalemma with occasional roundish blebs which varied in size. Cultured (24 h) brown adipocytes from the stromal-vascular fraction of BAT were elongated or polygonal in shape, with a flattened, central nucleus and a number of spherical cytoplasmatic inclusions which have the same dimension and location as lipid droplets. These inclusions were arranged either at cellular poles or around the nucleus; this suggests that brown adipocytes with mature features were present in the culture. Pictures suggesting the detachment of lipid droplets from the cell body were also visible. Lipid droplet extrusion could be a complementary mechanism which might explain the rapid delipidation of brown adipocytes in culture.     

Tumor-basophil interactions in vitro--a scanning and transmission electron microscopic study.

Purified guinea pig basophils, or basophils either specifically degranulated with antigen or nonspecifically degranulated with lectin, were cultured with guinea pig line 1 hepatoma cells for 1 to 24 hr and studied ultrastructurally. As early as 1 hr of culture, degranulated or nongranulated basophils and tumor cells formed close contacts by mutually intertwined elongated cell processes and also in cultures containing degranulated basophils, extruded membrane-free basophil cytoplasmic granules became firmly attached to tumor cells. At later intervals, some tumor cells cultured with basophils exhibited cytostatic and cytopathic changes, including dense mitochondria, centralization of organelles, dilated perinuclear and rough endoplasmic cisternae, cell swelling and cytoplasmic lucency, disrupted cytoplasmic organelle and plasma membranes, nuclear pyknosis and fragmentation. Some tumor cell specialized surface attachments were either disrupted or damaged at points of basophil or basophil granule adhesion. Tumor damage was most extensive in cultures containing degranulated basophils, although only a minority of tumor cells (less than 10%) was affected. Tumor injury was seen much less frequently in the presence of nondegranulated basophils, and was absent in control cultures of tumor alone. The occasional viable tumor cells that phagocytosed basophil granules were apparently unharmed, suggesting that internalization of basophil granules by tumor cells is not cytotoxic.     

A scanning electron microscopic study of rat Masugi nephritis.

Changes in the glomerular capillaries in the first phase of rat Masugi nephritis were studied by scanning electron microscopy. The changes developed immediately after the injection of nephrotoxic rabbit IgG and early endothelial lesions (2 to 6 h) were characterized by an increase in microvilli and a decrease in endothelial pores. The microvilli were fused and produced abundant pored projections (cytofolds). The peripheral endothelium was then lifted off from the glomerular basement membrane (GBM), leaving scattered endothelial fragments on the GBM. The denuded GBM exhibited a rather uniform, thick carpet-like appearance with occasional crater formation. Depositon of fibrin strands was seen associated with endothelial exfoliation. These later dissolved and were converted to a fibrinoid material, consisting of a complex of fragmented, thin fibrils. A parallel study using the electron microscope revealed that the fibrinoid material was removed by emigrating monocytic macrophages. At the stage of resolution (24 to 72 h), the denuded GBM was covered mostly with a regenerating endothelial layer. A possible process of reorganization of the endothelial pores is discussed.     

The morphology of the esophageal mucosa of the rat: a scanning electron microscopic study

The morphology of the oesophageal mucosa of adult male and female albino rats (Rattus norvegicus) is studied by means of scanning electron microscopy of several levels. A precise surface analysis of the upper, middle, and lower portion of the esophagus is done and the results are compared with each other.     

A scanning electron microscopic study of the rat liver sinusoid

The surface ultrastructure of Kupffer cells in the rat liver has been studied by scanning electron microscopy (SEM). The results demonstrate that Kupffer cells are both significantly different and clearly distinct from endothelial cells. Kupffer cells have neither pores (and/or "sieve plates") nor fenestrations, all of which are present in endothelial cells. They possess a stellate shape, and only indirectly, with slender and irregular evaginations, contribute to the lining of the sinusoidal wall. Furthermore, the luminal surface in some areas contains a large population of short microvilli, microphicae and invaginations. These elements form a kind of microlabyrinth which may correpond to the "worm-like" structures described by transmission electron microscopy (TEM). In the present study, transition forms between endothelial and Kupffer cells were never found. On the contrary, considering the highly fenestrated nature of the endothelial cells, the Kupffer cells may, by ameboid movements, easily cross the overlapping barrier of the sinusoid and protrude into the lumen. Thus, acting as activated macrophages, the Kupffer cells might function to prevent the entrance of foreign material into the tissues of the liver through the fragile and highly fenestrated endothelium. Finally, the topographical reconstruction of the sinusoid by correlated SEM and TEM studies demonstrates the Kupffer cells, with their protruding cytoplasm and ability to extend into the lumen of the sinusoid, may actually change the caliber of the vessel, and thus function as a "sphincter" which causes a temporary arrest of the blood flow when the diameter of the sinusoidal lumen is reduced.     

The vasculature of the rat penis: a scanning electron microscopic and histologic study.

As a basis for understanding the mechanism of erection in an animal model frequently used in research in reproductive biology, the angioarchitecture of the penis of the rat has been described using scanning electron microscopy. Study of the penile vasculature of the rat indicates that the corpora cavernosa penis and the corpus spongiosum are independent erectile tissues, each with its own arterial and venous vessels. The large vascular spaces and abundant smooth muscle of the penile crura are compatible with its role in regulating blood flow to more distal penile tissues. Helicine arteries of the crura, but not the parent deep penile artery or arteries elsewhere, have muscular cushions in their walls. The venous drainage of the penile crura is via subtunical veins which are thought to be compressed during erection to elevate pressure within the penis. Large, paired cavernous veins drain the shaft of the penis. A unique method for inhibiting blood flow from the penis is indicated by the division of the cavernous veins into smaller channels prior to joining the subtunical venous plexus. Erectile tissue in the bifid origins of the corpus spongiosum has abundant cavernous muscle, while in the remainder of the corpus spongiosum little smooth muscle lines the cavernous spaces. The cavernous spaces on either side of the urethra coalesce to form vessels, each of which communicates with cavernous spaces in the glans. In addition, a bypass of the glans is effected by communication of these vessels directly with the deep dorsal vein. The apparent absence of muscular pads in vessels of the spongiosum, the relative paucity of cavernous smooth muscle, and the ample venous drainage provided by the deep dorsal vein may account for the lack of a venous occlusive mechanism similar to that of the corpora cavernosa penis.