Morphological structure of rat tongue using light and scanning electron microscopy

The rat is one of the most commonly used animals in biological research and experimental investigations in medicine. The ultrafine structural components of the epithelium differ depending on the anatomy of the animal and the papilla type. Animal adaptation to food types and environmental circumstances may also be linked to morphological diversity. In the current study, seven male Wistar rat tongues were investigated. For scanning electron microscope (SEM), two rat tongues were immersed in a 10% formalin solution and the other two rat tongues were immersed in a 2.5% glutaraldehyde solution. The tongues of three rats were fixed for regular histological evaluation using triple staining. The three primary components of the Wistar rat tongue are the apex, body and root. The apex had a rounded and bifurcated shape. Filiform papillae and gustatory papillae were easily identified on the dorsal side of the tongue. There were three forms of gustatory papillae; fungiform papillae, vallate papillae and foliate papillae. The purpose of this study was to expose the tongue morphology of the Wistar rat species, which is widely used in investigations. Also, we wanted to show that formalin fixation can be utilized for morphological research in SEM. Finally, the Wistar rat tongue was thoroughly investigated and compared to those of other species.     

Functional and comparative study of lingual papillae in four species of bear (Ursidae) by scanning electron microscopy

The eight current species of bear (Ursidae) are widely distributed throughout Europe, Asia, and America. They are mainly encountered in the northern hemisphere, except for the spectacled bear and the sun bear, which are also found in the south of the equator. Adaptations of the masticatory apparatus (teeth, tongue, and musculature) to diet are one of the factors that imply the greatest structural changes in the cranium. This diet may be carnivorous, herbivorous, melliferous, or insectivorous, with one type of food predominating according to the time of year. The way in which food is eaten determines the morphology of the lingual surface; generally speaking, all bears put their mouth to the food, which, initially, they lick or they let the food stick to their tongue, as occurs when insects are eaten. As in all mammals, a distinction can be made between mechanical and gustatory papillae and the development and distribution of which depend on the species and their eating habits. In this study of the complete tongues of four species of adult bears, we describe the morphology of the lingual surfaces, the different types of papillae, their characteristics, and topographic distribution. It was seen that there were five main types of lingual papillae: filiform, conical, fungiform, foliate, and vallate. Morphology of the microgrooves and pores was similar to that observed in other mammals. In general, there were no great differences among the four species of bears studied, perhaps due to the similarity in the kind of food they consume in captivity.     

The development of the tongue of the domestic goose from 9th to 25th day of incubation as seen by scanning electron microscopy

The general development of the tongue in birds was described by Lillie ( 1908 ) in chicken. Bryk et al. ( 1992 ) also studied the tongue development in chicken and they observed development of the conical papillae of the body. Our study aims to describe the timing of the development of the tongue morphological features in the domestic goose by using SEM methods. The tongue of the domestic goose is characterized by the widest variety of shape of the particular part of the tongue and mechanical papillae. Results indicated that the formation of the apex, body, lingual prominence, and the root of the tongue take place between the 10th and 19th day of incubation. The tongue elongates rapidly between the 16th and 18th day of incubation. Simultaneously, the median groove appears on the body and the lingual prominence and elongates towards the rostral part of the tongue. The conical papillae of the tongue develop gradually. On the body, the conical papillae develop from the caudal part of the body to the rostral part and on the lingual prominence from the median part of the prominence to the lateral part. Hair‐like papillae at the caudal surface of the body of the tongue remain primordial to the end of the incubation. Our studies on the morphogenesis of the tongue in the domestic goose revealed changes in shape of the particular part of the tongue and rapid pace of the formation of mechanical papillae. The tongue is completely develop before hatching and ready to collect food. © Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

Scanning electron microscopy of the tongue papillae in the pig (Sus scrofa)

A clear understanding of the role of different papillae in the pig may permit the development of a more palatable diet, thus, better utilizing anatomical structures and morphological characteristics to enhance health and productivity in this domestic species. The study used ten healthy market-size, mixed-breed pigs to properly describe the surface structure of the tongue papillae using standard scanning electron microscopic techniques. Two types of sharp and blunt filiform papillae were identified. Fungiform papillae, located on the lateral side of the tongue, were circular and large with surface taste pores. The majority of oval papillae on the dorsal surface of the tongue contained multidirectional surface grooves and lacked taste pores. The foliate papillae, located principally on the caudal third and lateral part of the tongue, were comprised of four to five leaves and they had additional pseudopapillae. The two vallate papillae, found in the caudal third and dorsum of the tongue, had a well-defined vallum and pseudopapillae on their surface. At the root of the tongue, two types of conical papillae with or without apex were observed. The entire area covered by conical papillae was deeply occupied by lingual tonsils. The lingual tonsils were scattered with varying sized lymph nodules in the propria mucosa of the conical papillae deep to epithelium. Functionally, the filiform and conical papillae appeared to have a mechanical role whereas fungiform, foliate, and vallate papillae apparently perform a gustatory role in the pig. The tonsillar location at the root of the tongue provides an ideal experimental model for immunohistological and immunological studies on the lymphoid tissue in man.     

Identification of bacteria by studying one section under light microscopy,scanning, and transmission electron microscopy

A method for bacterial identification has been developed by means of studying the same histological sections through several types of microscopy. With this method, one section was processed and analyzed respectively for light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Sections of gingival biopsies were Gram stained and bacteria tentatively identified by LM. Photographs of the sections were taken and presketched transparent acetate sheets (PTAS) were made from the photos. The same section was later prepared for SEM, areas previously thought to contain bacteria were localized by placing the PTAS onto the SEM monitoring screen. The SEM specimens were subsequently processed for TEM, bacteria were located, and micrographs obtained. The results showed that out of ten diseased gingival biopsies observed under the LM, bacteria were found to be present in all the specimens and were identified as both Gram positive and Gram negative. By transferring the section from LM to SEM, the bacteria could be relocated and their morphotype (cocci, rods, etc.) clearly identified in most of the cases. Since cocci may resemble other biological granular structures under SEM, they require further analysis under TEM for additional positive identification. This study demonstrated that the method described here is a useful tool for assessing the presence and identifying bacteria within the gingival tissues.     

Macroscopic and microscopic analysis of the tongue of the common opossum (Didelphis marsupialis)

We performed a macroscopic and microscopic study of the tongues of common opossums, Didelphis marsupialis, from South America. We studied two males and two females. We collected morphometric data on the tongue with precision calipers. For the light microscopy and scanning electron microscopy analyses, we fixed tissue fragments in 10% formaldehyde and 2.5% glutaraldehyde, respectively. The opossum tongues averaged 5.87 ± 0.20 cm in length, 3.27 ± 0.15 cm in width at the lingual body, and 3.82 ± 0.15 cm in width at the root. The mean thickness of the lingual body was 1.8 ± 0.1 cm, and the thickness of the root was 3.82 ± 0.15 cm. Sharp filiform papillae were scattered across the entire tongue; conical filiform papillae occurred on the lingual body and tongue tip; fungiform papillae were scattered among the filiform papillae on the lingual body and tongue tip; and there were three vallate papillae at the root of the tongue. We found two strands of papillary projections in the tongue root. Despite the low variability observed in the lingual papillae, the morphological data obtained in this study may be related to the opossum's diverse food habits and the extensive geographic distribution of the species throughout America. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Polyethylene glycol (PEG) embedding and subsequent de-embedding as a method for the correlation of light microscopy,scanning electron microscopy,and transmission electron microscopy

The polyethylene glycol (PEG) embedding and subsequent deembedding method was applied to the observation of general tissues in scanning electron microscopy (SEM). Resulting SEM images were of high quality. It was demonstrated that intermicroscopic correlation of images between light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) is easily and reliably done by means of the PEG method. In particular, the exact correlation of immuno-LM with SEM is shown to be of potential value.     

Characterization of an oil-degrading Microcoleus consortium by means of confocal scanning microscopy, scanning electron microscopy and transmission electron microscopy

A consortium of microorganisms with the capacity to degrade crude oil has been characterized by means of confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The analysis using CLSM shows that Microcoleus chthonoplastes is the dominant organism in the consortium. This cyanobacterium forms long filaments that group together in bundles inside a mucopolysaccharide sheath. Scanning electron microscopy and transmission electron microscopy have allowed us to demonstrate that this cyanobacterium forms a consortium primarily with three morphotypes of the heterotrophic microorganisms found in the Microcoleus chthonoplastes sheath. The optimal growth of Microcoleus consortium was obtained in presence of light and crude oil, and under anaerobic conditions. When grown in agar plate, only one type of colony (green and filamentous) was observed.     

Anatomical and scanning electron microscopic investigations of the tongue and laryngeal entrance in the long-legged buzzard (Buteo rufinus, cretzschmar, 1829)

This research aimed to examine the morphological features of the tongue and laryngeal entrance of long‐legged buzzard by macroscopic and scanning electron microscopic methods. Two adult buzzards were used as material. The tongue was fairly elongated and terminated in oval shovel‐like apex. Scale‐like projections were localized on the apex and body of tongue. Both lateral sides of lingual body were contained considerably long thread‐like projections. Many orifices of lingual posterior salivary glands were discerned among scale‐like projections in median sulcus of the lingual body. Papillary crest of sharp conical papilla were observed on the between the body and root of the tongue. No conical papillae or other projections were existent on the root of the tongue, but numerous orifices of posterior salivary gland ducts were detected. In addition, orifices of anterior salivary gland ducts were present on the dorsal and lateral surfaces of the lingual body. Numerous conical papillae were observed on the caudal region of glottis and no conical papillae or any similar projection which were bordered the glottic fissure was noted. Anatomy of these organs in the long‐legged buzzard and white tailed eagle which are the member of the same family, Accipitridae, revealed very high similarity. Microsc. Res. Tech. 75:1245–1252, 2012. © 2012 Wiley Periodicals, Inc.     

Seed morphoanatomy of the genus Passiflora L. (Passifloraceae) by scanning electron microscopy and light microscopy

Morphoanatomical analysis of seeds contributes to knowledge of the development of seedlings and identification of species, as well as supporting conservation studies. The conservation of the species belonging to the Passiflora genus is crucial due to of the threats to the genetic resources of these species. Thus, the objective of this study was to morphoanatomically characterize Passiflora seeds, verify possible injuries to the tissues after cryopreservation and thus contribute to the conservation strategies of the species of this genus. Initially, seeds of Passiflora coccinea, P. edulis, P. gibertii, P. maliformis, P. morifolia, P. setacea, P. suberosa, and P. tenuifila collected from the Passion Fruit Active Germplasm Bank of the Embrapa Cassava and Fruits research unit (Embrapa Mandioca e Fruticultura) were analyzed. Then, their length, width and thickness, shape of the base and tip, and ornamentations present on the body and edge of the seeds were evaluated. The seeds of the species were placed in cryotubes and immersed in liquid nitrogen to assess possible cryoinjuries. The tegument and tissues of the seeds were examined by scanning electron microscopy. The seeds had varied biometric data, with average values of 4.63 mm for length, 3.28 mm for width, and 1.51 mm for thickness. Six ornamentation types were observed: reticulate for the species P. coccinea; finely reticulate for P. edulis; foveolate reticulate for P. gibertii and P. setacea; alveolate reticulate for P. maliformis and P. tenuifila; coarsely reticulate for P. morifolia; and falsifoveolate reticulate for P. suberosa. Some seeds suffered tegument cracks due to the freezing in liquid nitrogen, but without physiological damages to the embryo and endosperm. The cryopreservation of the seeds in the presence of the tegument significantly reduced the cryoinjuries caused to the embryo. Cryopreservation can be promising for long-term conservation of passion fruit seeds.     

3-D morphological characterization of the liver parenchyma by atomic force microscopy and by scanning electron microscopy

A comparative study of atomic force microscopy (AFM) and scanning electron microscopy (SEM) imaging of the healthy human liver parenchyma was carried out to determine the similarities and the differences. In this study, we compared the fine hepatic structures as observed by SEM and AFM. Although AFM revealed such typical hepatic structures as bile canaliculi and hepatocytes, it also showed the location of the nucleus and chromatin granules in rough relief structure, which was not visible by SEM. By contrast, SEM visualized other structures, such as microvilli, the central vein, and collagenous fibers, none of which was visualized by AFM. For better orientation and confirmation of most of the structures imaged by SEM and AFM, Congo Red-stained specimens were also examined. Amyloid deposits in the Disse's spaces were shown especially clearly in these images. The differences between the SEM and AFM images reflected the characteristics of the detection systems and methods used for sample preparation. Our results reveal that more detailed information on hepatic morphology is obtained by exploiting the advantages of both SEM and AFM.     

Morphological investigations on the lips and cheeks of the goat (Capra hircus): A scanning electron and light microscopic study

The current study was done to provide comprehensive information on the anatomical features of the lips and cheeks of the goat by gross examination and morphometric analysis in addition to scanning electron microscope (SEM). Samples from 12 normal healthy adult goat's heads of both sexes were collected directly after slaughtering. The lips and cheeks were dissected, and specimens were collected for both light and SEM. The lips of goat were soft and mobile. The free border of both lips was characterized rostrally by the presence of labial projections. The number, size, and arrangement of labial projections differed in the upper and lower lips. On the other hand, the buccal papillae were arranged into 6–8 longitudinal rows parallel to the cheek teeth. The length of these papillae decreased caudally while they were absent on the most caudal part of the cheek. Presence of several types and shapes of labial projections and papillae, and buccal papillae suggest a high degree of mechanical adaptation of the lips and cheeks of the goat. This study provides baseline data for clinical studies. This study is the first report to shed light on the morphology of the lips and cheeks of the goat by gross and scanning electron microscopy.     

Morpho-anatomical characterization of some members of euglenophycota (algae) of north-east Punjab,Pakistan based on light microscopy (LM) and scanning electron microscopy (SEM)

The study was carried out in four districts, that is, Gujranwala, Gujarat, Narowal and Sialkot of Punjab, Pakistan. The sampling was carried out randomly in different seasons from the water bodies especially from wastewater. Twenty-one species belonging to Euglenophycota were identified using light microscopy and scanning electron microscopy from which 04 species belong to genus Phacus, 02 species belonging to Trachelomonas and Euglena based on light microscopy. It was observed that Euglena was the most diverse genus and it is supposed to be the indicator species of the polluted water. It was observed that E. oblonga was found in maximum pH range, that is, 7.0–11.0. Similarly, E. brevicaudatus was found in maximum EC, that is, 169 ± 1.5 ms/cm these outcomes indicated that for internal examination along with LM, SEM was necessary for correct identification of algal sample up to specie level.     

Image comparisons of snow and ice crystals photographed by light (video) microscopy and low-temperature scanning electron microscopy

Light (video) microscopy and low-temperature scanning electron microscopy (SEM) were used to examine and record images of identical precipitated and metamorphosed snow crystals as well as glacial ice grains. Collection procedures enabled numerous samples from distant locations to be shipped to a laboratory for storage and/or observation. The frozen samples could be imaged with a video microscope in the laboratory at ambient temperatures or with the low-temperature SEM. Stereo images obtained by video microscopy or low-temperature SEM greatly increased the ease of structural interpretations. The preparation procedures that were used for low-temperature SEM did not result in sublimation or melting. However, this technique did provide far greater resolution and depth of focus over that of the video microscope. The advantage of resolution was especially evident when examining the small particles associated with rime and graupel (snow crystals encumbered with frozen water droplets), whereas the greater depth of focus provided clearer photographs of large crystals such as depth hoar, and ice. Because the SEM images contained only surface information while the video images were frequently confounded by surface and internal information, the SEM images also clarified the structural features of depth hoar crystals and ice grains. Low-temperature SEM appears to have considerable promise for future investigations of snow and ice.     

Macro‐ and microscopic study on the tongue and lingual papillae of Bison bonasus hybrid as an interspecific species (Bos taurus × Bison bonasus)

Nowadays the processes of crossing of different mammalian species are well characterized in light of genetics, and possibility of reproduction. Generally, lack detailed studies on changes in anatomy and histology of internal systems of hybrids. Bison bonasus hybrid is an interspecific species between the bull of European bison (Bison bonasus), and female of domesticated cattle (Bos taurus). To study the anatomy of tongue and distribution of lingual papillae on Bison bonasus hybrid tongue, a stereomicroscopic and scanning electron microscopic observations was conducted. We aimed to study the appearance of macro‐ and microscopic traits in relation to parental species and other bovids. Results indicate that Bison bonasus hybrid tongue possesses characteristic traits for parental species and ruminants from Bovidae family and also species‐specific features differ from mentioned animals. Specific new traits in hybrid are: V‐shaped arrangement of lingual papillae on ventral surface of the tongue, rosette arrangement of conical papillae on the top of lingual prominence, bigger number of vallate papillae and closer arrangement of vallate papillae. In hybrid observed also the appearance of combination of parental traits. Similar to European bison are rectangular shape of lingual prominence and absence of filiform papillae on the root of tongue. Presence of lateral processes of filiform papillae on the apex of tongue, and arrangement of vallate papillae into two rows corresponded to cattle. Development of new morphological features of tongue and its mucosal papillae in Bison bonasus hybrid opens new issue for future studies on organs of digestive system in animals after hybridization.     

Study of the crystallographic architecture of corals at the nanoscale by scanning transmission X-ray microscopy and transmission electron microscopy

We have investigated the nanotexture and crystallographic orientation of aragonite in a coral skeleton using synchrotron-based scanning transmission X-ray microscopy (STXM) and transmission electron microscopy (TEM). Polarization-dependent STXM imaging at 40-nm spatial resolution was used to obtain an orientation map of the c-axis of aragonite on a focused ion beam milled ultrathin section of a Porites coral. This imaging showed that one of the basic units of coral skeletons, referred to as the center of calcification (COC), consists of a cluster of 100-nm aragonite globules crystallographically aligned over several micrometers with a fan-like distribution and with the properties of single crystals at the mesoscale. The remainder of the skeleton consists of aragonite single-crystal fibers in crystallographic continuity with the nanoglobules comprising the COC. Our observation provides information on the nm-scale processes that led to biomineral formation in this sample. Importantly, the present study illustrates how the methodology described here, which combines HRTEM and polarization-dependent synchrotron-based STXM imaging, offers an interesting new approach for investigating biomineralizing systems at the nm-scale.     

Taxonomic significance of caryopsis in subfamily Panicoideae (Poaceae) using scanning electron microscopy and light microscopy

In recent study, 13 taxa of subfamily Panicoideae were investigated for morphological characterization of caryopsis. Light microscope (LM) and scanning electron microscope (SEM) were utilized to study macro‐ and micro‐morphological caryopsis features respectively. Caryopsis size in studied taxa was recorded as 1.5–10 mm long and 1–4 mm wide. Caryopsis color was brown, green, yellow, and whitish‐brown. Caryopsis shape studied was obovate, elliptic, linear oblate, and round shallowly obtriangular. Hilum position is grooved and depressed. Caryopsis compression type was lateral and dorsiventral. Major variations among studied taxa were observed in terms of caryopsis surface pattern and epicuticular projection types. Six types of caryopsis surface pattern were observed viz. scabrate, rugose, striate, reticulate, papillate, and scabridulous. Structures such as silica cells, bulges, spines, prickles, granules, and bicellular microhair were studied as epicuticular projections. Major variations were observed among Cenchrus pennisetiformis and Cenchrus ciliaris as both has entirely two different types of surface patterns and epicuticular projections. Anticlinal wall thickness and pattern as well as periclinal wall texture and level were investigated. The present research work emphasized on caryopsis characterization of subfamily Panicoideae and it is recommended to establish phylogeny within subfamily Panicoideae and with other subfamilies of Poaceae.     

A rapid and simple technique for correlating light microscopy,transmission and scanning electron microscopy of fixed tissues in Epon blocks

A simplified and standardized technique for close correlation between light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM) is described. Perfusion and immersion fixed tissue specimens were embedded in Epon 812 and cut for conventional LM and TEM. The Epon blocks with remaining tissue were thereafter treated with epoxy solvent (ethanol-NaOH solution) for partial epoxy resin removal only (dissolving rate approx 33μm/h). The blocks with partially blotted tissue specimens were then critically point dried and gold coated for SEM. This method, in an easy way, allows repeated observations with LM, TEM and SEM with preserved fine structure and exact correlation. Since the technique is so simple and there is no need for special equipment the method can easily be adopted in all laboratories with basic SEM standards.