Scanning electron microscopy of the respiratory surfaces of Saccobranchus (=Heteropneustes) fossilis (Bloch)

Summary The gill secondary lamellae are generally covered with epithelial cells whose outer surfaces form numerous microvilli. The surface of the primary lamellae is characterised by microridges. A particular type of surface sculpturing seems to be associated with given cell boundaries.Further evidence for the derivation of the air tube and fans which guard its entrance by modification of the basic gill structure has been obtained from both the gross surface architecture and microstructure of the individual cell surfaces. Secondary lamellae are represented by stubby projections which generally have a biserial arrangement. The outer surfaces of the epithelia overlying the capillaries of these respiratory islets are coated with microvilli as in the secondary lamellae. On the other hand, the relatively smooth-surfaced lanes between groups of respiratory islets have a microridged surface similar to that of the primary gill lamellae.It is suggested that previous estimates of surface area, and consequently diffusing capacities of the air-breathing organ, have been low in view of the increased surface, due to both their gross and microstructure. Estimates for gill surface area may need very little correction as the spaces between the microvilli and microridges are probably filled with mucus under normal conditions.We thank Mr. John Clements for his excellent technical assistance and the Department of Botany, Bristol University for the use of their scanning electron microscope     

Scanning electron microscopy of the gills of a freshwater catfish,Rita Rita

Variations in the gross morphology and surface architecture of the gill filaments and secondary lamellae of a freshwater catfish (Rita rita) have been investigated using scanning electron microscopy. Heterogeneity of the gill has been correlated with the distribution of lamellar water-flow at different regions of a gill filament. Higher lamellar water flow (cc/pore/cmH₂O/sec) was estimated for the middle region of the filaments. The filaments are covered with epithelial cells whose surface is provided with well-developed microridges. The lamellae are generally covered with microvillous epithelial cells. The variations in surface architecture of the gill filaments and secondary lamellae have been correlated with their probable functions.     

Scanning electron microscopy of olfactory rosette in sea trout

Summary Scanning electron microscopy has been employed to study the central axis and laminae of the olfactory rosette in adult sea trout (Salmo trutta trutta L.) caught in the River Umeälven when they were homing from sea.—Both flat sides of the primary laminae are secondarily folded all over their surface. In one organ there are about 200 secondary laminae usually arranged in longitudinal, parallel ridges crossing the surface of the primary laminae. Initially they are covered with sensory epithelium, but as the folds grow they become covered with an increasing area of indifferent ciliar epithelium with bushes of cilia separated by microvilli cells and goblet cells. Parts of the central axis and primary laminae have a nonciliar indifferent epithelium. The sensory epithelium has irregularly arranged cilia. Like those of the indifferent epithelium they have uniform thickness and granulated surface. The function of laminae, secretion and cilia is discussed.The author wish to acknowledge the technical facilities and assistance in the use of the scanning electron microscope to Jeolco Stockholm office. This research was supported by grants 2389-10, 2389-11 and 2389-13 from the Swedish Natural Science Research Council.     

Scanning electron microscopy of exposed surfaces of the porcine placenta

The three-dimensional development of the separated parts of the porcine placenta from 9 Danish Landrace sows at gestational stages from 20 to 100 days was studied. After cautious separation of the allantochorion and the endometrium in a 1 mM buffered ethylenediaminetetraacetic acid solution, the separated parts were processed for scanning electron microscopy by routine methods. The macroscopic enlargement resulted from primary and secondary circular folds or plicae, which were permanent on the maternal side, whereas they were mainly non-permanent on the fetal side. The areolar placenta and interareolar placenta with formation of permanent microscopic folding on both sides were described. The observations of the separated parts yielded new information on the development of surface enlargement during gestation and revealed a hitherto unknown regular architecture of the endometrium by the formation of parallel primary ridges or rugae with secondary ridges or rugae at their sides subdividing the maternal troughs or fossae. This configuration on the maternal side explains the transformation of the regular chorionic ridges from the 63-day stage into bulbous protrusions at the 100-day stage. Based on these observations the precise terminology used above was proposed.     

Scanning electron microscopy of dissociated pancreatic acinar cell surfaces

Summary The pancreatic acinar cell surfaces have been studied by SEM with a dissection technique and correlated with results obtained by TEM. The SEM results demonstrate characteristic arrangement of microplicae which in some areas are densely packed.In many areas, the microplicae are distributed in such a manner that they create zones with typical geometrical shapes and show a relatively smooth surface.These smooth areas may coincide, as indicated by correlated TEM results, with the limits of intimate contact between adjacent acinar cells which, in turn, represent part of the junctional complex. Another aspect revealed by these SEM preparations concerns the presence of groups of densely packed microplicae, arranged in regular rows and distributed along some grooves and/or infoldings of the cellular surface. On the basis of SEM and TEM information, it is likely that these structures correspond to intercellular (and possibly, in some cases, intracellular) canaliculi which topographically form a kind of extensive microlabyrinthine arrangement running along all the cell sides.One final point revealed by fractured samples concerns the finding of spherical zymogen droplets within the vesicles of the Golgi complex. Because in many scanning images these vesicles appear connected by small openings, it is suggested that they may represent a system of intercommunicating chambers (vacuoles) through which the zymogen droplets can be continuously accumulated and discharged into the acinar lumen.     

Scanning electron microscopy of the integumental surfaces of Schistosoma haematobium.

The integumental surfaces of critical point dried S. haematobium were studied by scanning electron microscopy at 34 to 8,000 magnifications. There are marked differences between the surface structures of male and female as well as from one part of the same parasite to another. The surface of the male schistosome is moderately rough while that of the female is relatively smooth. SEM reveals certain basic features such as spines in the oral sucker and the acetabulum of both sexes which may facilitate rasping and/or attachment of the parasite for residence in the bloodstream of the definitive host. The lining of the gynecophoral canal is roughened by minute spines. The presence of a gynecophoral fold may enhance anchorage of the female in the grasp of the male. The significance of visualization of surface features by SEM as a means for differentiating species is not yet known.     

Scanning electron microscope studies of some internal surfaces in human skin

Summary The scanning electron microscope has been used to investigate the surfaces of epidermis, hair follicles, fat cells, glands and blood vessels in sections of human skin. The appearances of structures associated with these surfaces have been described. The results demonstrate that this new instrument can become an important tool in the investigation of surface changes that might accompany the application of loads (extensive, compressive or torsional), or physiological or pathological abnormalities.We wish to thank Prof. R. M. Kenedi for his continued encouragement of this work. One of us (I.A.B.) is also grateful to the Science Research Council for the provision of a post-graduate student grant.     

Megalodiscus temperatus: Scanning electron microscopy of the tegumental surfaces

The tegumental surface of Megalodiscus temperatus forms cobblestonelike areas with rows of indentations encircling the worm. This pattern merges in several areas into folds and ridges, some of which represent the musculature of the posterior sucker and genital pore. Papillae surrounding the base of the oral sucker appear as two types: one with a bulb-like base and a short apical knob; the second typified by a hair-like structure (cilium?) of variable length projecting from a pit. From their location on the oral sucker and the resemblance to previously described structures, they are presumed to be sensory receptors. A circle of papillae on the closed posterior sucker was also observed. Rod-shaped bacteria were seen adhering to some of the worms observed but they were not found to be in any special association with the tegument or in any specific areas of the worm's surface.     

Scanning electron microscopic study of the gut mucosa of the rainbow trout Salmo gairdneri Richardson.

The topological characteristics of the entire gut surface of the rainbow trout were investigated utilizing the scanning electron microscope. The mucosa exhibited longitudinal ridges in the oesophagus and stomach, villi in the intestine, fine longitudinal ridges in the caeca and annular folds in the rectum, arranged as a stack of caudally-directed funnels starting from the intestino-rectal valve to the vent. Detailed study revealed taste pores in the anterior oesophagus and the sculpting of the luminal plasmalemma of the surface cells into micro-ridges with complicated patterns. The surfaces of the posterior oesophagus and stomach were demarcated into polygons by rows of stubby microvilli—each polygon representing the luminal surface of an epithelial cell. Each rectal fold consisted of a smooth, caudally-directed apex and a base which was supported by perpendicular buttress-like secondary folds. The functional significance of these features which emphasize the vast difference in the physical length of the gut and the effective surface area is discussed.     

Kinetics of potassium transport across trout gills.

1. Kinetics of potassium transport across trout gills was studied, using an isolated-head preparation. 2. Potassium exchanges were shown to take place across secondary lamellae only. 3. Influx of potassium was saturable and fitted satisfactorily the lineweaver-Burk linear plot. 4. Results suggest that these exchanges occur through potassium channel. 5. Kinetics of potassium exchanges is discussed in relation to the maintenance of the osmoregulation in fish.     

Modifications of mammalian cell surfaces induced by sugars: Scanning electron microscopy

Substitution of galactose, xylose, or mannose for glucose in the growth medium of L cells or the addition of equal concentrations of the alternate sugars to glucose-containing medium results in marked morphologic changes. The changes are revealed by conventional staining for light microscopy and by scanning electron microscopy.L cells grow indefinitely on combinations of equal concentrations of glucose and galactose, xylose, or mannose. There appear to be no significant differences in growth rate on glucose compared to the combinations of sugars cited. Cells subcultured from glucose to the combinations while undergoing rapid multiplication show marked morphologic changes by light and scanning electron microscopy within 36 hr.Of particular note are the loss of microvilli; flattening of the cells; assumption of polygonal shape; prominence of nuclei and nucleoli; and changes in the structure and distribution of filopodia. Virtually all cells in the population exhibit the changes noted.     

Scanning electron microscopy of dairy equipment surfaces contaminated by two milk-borne micro-organisms

Ethanol dehydration followed by argon replacement induced drying (ARID) was found to be a suitable method for the preparation of glass, stainless steel and rubber surfaces which had been in contact with inoculated milk and which were to be examined using scanning electron microscopy (SEM). This technique was used to examine samples of all three materials which had been subjected to both single and repeated inoculation with whole milk containing a Pseudomonas sp. or a Micrococcus sp. and incubated for various periods. Some samples were also prepared for SEM using a cryofixation technique. The Pseudomonas sp. was found to proliferate on glass and stainless steel surfaces but not on rubber. Due to the clumping tendency of the Micrococcus sp. proliferation of this organism was more difficult to assess accurately. In general there was no difference in results obtained between single and repeated inoculation. Various factors which may have aided attachment of micro-organisms to surfaces were identified viz. , surface channels present in stainless steel, milk deposits and the production of extracellular material. The value of using both the cryofixation and chemical preparatory techniques for the identification of artifacts is discussed.     

Scanning electron microscopic evaluation of effects of low pH on gills of Channa punctata (Bloch)

Bioassay of Channa punctata (± 36 g) was carried out in acidic waters of different pH. A 96 h LC₅₀, value was obtained at pH 5.3. Scanning electron microscopy of gills of C. punctata showed fusion of adjacent secondary lamellae. At this low pH, dissociation of epithelium of branchial arches and gill filaments take place. At 360 h the branchial epithelium ruptures developing lesions in the gills exposing the efferent filament vessels. At pH 6.6 the tips of adjacent filaments belonging to both oral and aboral hemibranchs fuse in blocks.     

Fixation of mucus on rainbow trout (Oncorhynchus mykiss Walbaum) gills for light and electron microscopy

Fixation of mucus for the assessment of biofilms and surface associated pathogens often involves complex and expensive techniques. Rainbow trout killed by an overdose of MS 222 had their gills removed and immersion-fixed gently in buffered glutaraldehyde containing 2% Alcian blue. Control tissues consisted of gills fixed in Alcian blue-free fixative. Trout were also killed and directly immersed in liquid nitrogen and the gills freeze-dried then vapour fixed with osmium tetroxide at −50° C. Following fixation gill tissue was processed for light and electron microscopy. A continuous and intact mucous coat was not detected on tissue fixed by conventional methods but the addition of Alcian blue to the fixative preserved an extensive mucous coat trapped between the lamellae and overlying the epithelia. Electron microscopic examination revealed that mucus preservation with the conventional fixative was poor and intermittent whereas the addition of Alcian blue to the fixative greatly enhanced the preservation of the branchial mucous coat. Mucus appeared as interdispersed flocculant material between the epithelial microridges and formed extensive superficial sheets over the epithelium. Freeze-dried/vapour-fixed gill tissue also provides excellent preservation of the integrity of branchial mucous coats, the mucus appearing as a continuous sheet over the filament and secondary lamellae. However, freeze-dried tissue fails to preserve sufficient cellular integrity for this technique to be useful for light or transmission electron microscopy. The potential for use of glutaraldehyde-Alcian blue fixed-gill tissue diagnostically and in research are discussed.     

Endothelin redistributes blood flow through the lamellae of rainbow trout gills

The lamellae of the fish gill are the primary sites for oxygen uptake from the water. Here, only two very thin layers of cells separate the blood from the water. Therefore, energetically costly ion-fluxes will also occur between blood and water, and it has been hypothesised that the blood flow within the lamellae can be regulated through vasoconstriction, but evidence for this has been lacking. Through direct observations of the lamellae of rainbow trout (Oncorhynchus mykiss) in vivo, using epi-illumination microscopy, we show here that an endothelium-derived vasoactive peptide, endothelin-1 (ET-1, 0.2 μg kg⁻¹ or 1.0 μg kg⁻¹), is able to completely constrict the vascular sheet in the lamellae, probably by inducing contraction of pillar cells. This coincided with a dose-dependent increase in ventral aortic blood pressure (rising from 6.6 kPa to 12.0 kPa in response to the high ET-1 dose). However, blood continued to flow through the marginal channel that circumvents each lamella. Thus, ET-1 caused an intralamellar blood shift from the lamellar sheet towards the marginal channels. Vasoconstriction in the lamellae is likely to provide the fish with a mechanism for matching its respiratory surface area with its respiratory needs, thereby minimising ion-fluxes. Accepted: 8 September 1998     

Fine structure of the respiratory lamellae of teleostean gills

Summary The blood-water pathway in respiratory lamellae of teleostean gills consists of an epithelial layer one or two cells thick, a basal lamina and a thin layer of cytoplasm which lines the blood lacunae. This layer of cytoplasm is formed by flange-like extensions of the pillar cells. The resulting location of the pillar cell perikarya between the surfaces of the blood lacunae is probably of paramount importance for maintenance of the flattened form of the lamellae.Collagenous bundles traverse the pillar cells within tubes formed by infolding of the cellular surface. These bundles, which are oriented normal to the flattened aspect of the lamellae, no doubt provide further protection against distension or collapse of the blood spaces. A compartment filled with collagenous tissue is interposed between the basal lamina and the lining layer of the lacunae in some of the species studied.Regulation of blood flow to the respiratory surfaces is thought to result in part from contraction of the pillar cells. This contractility presumably resides in tracts of filaments which course through the cytoplasm of the pillar cells parallel to the collagenous bundles. Since nervous tissue has not been demonstrated within the gill lamellae it is possible that contraction of the pillar cells is under some form of hormonal control, although existence of local control mechanisms (e.g. self-stimulation of the cells as a result of anoxia) is not excluded.Within the limited number of species studied, the structure of the blood-water pathway does not appear to be correlated with the characteristics of the normal habitat of a particular species.This work was performed during the tenure of a post doctoral traineeship under USPHS Grant 5 T 1 GM-136 to the Department of Biological Structure, University of Washington.Particular thanks are due Dr. John H. Luft of the University of Washington for his advice and criticism while this work was in progress and to Drs. Douglas Kelly, James Koehler and Daniel Szollosi for critical assistance with the manuscipt.     

Biochemical responses in gills of rainbow trout exposed to propiconazole

The aim of this study is to investigate the toxic effect of PCZ, a triazole fungicide commonly present in surface and ground water, on the ROS defense system and Na₊-K₊-ATPase in gills of rainbow trout exposed to sublethal concentrations (0.2, 50 and 500 μg L⁻¹) for 7, 20 and 30 days. After prolonged exposure of PCZ at higher test concentrations (50 and 500 μg L⁻¹), oxidative stress was apparent as reflected by the significant higher ROS levels in fish gill, as well as the significant inhibition of SOD and CAT activities. In addition, Na₊-K₊-ATPase activities were significantly lower than those of the control with increasing PCZ concentration and prolonged exposure period. The results of this study indicate that chronic exposure to PCZ has altered multiple physiological indices in fish gill; however, before these parameters are used as unique biomarkers for monitoring residual pharmaceuticals in aquatic environments, more detailed laboratory experiments need to be performed.     

Scanning electron microscopic study of blood cell surfaces of the middle asian tortoise, Testudo horsfieldi

The micromorphology and surface topography of spleen cells of the Middle Asian tortoise, Testudo horsfieldi, have been examined by scanning electron microscopy. Spleen cells comprise a population with surface characteristics similar to counterparts in mammalian spleen except for an, as yet undescribed, lymphocyte which is designated as lamellar type on the basis of its surface contours.