Palyno-morphological attributes of some selected plant species of family Asteraceae from district Dera Ismail Khan,KPK, Pakistan

The current study aimed to investigate the palyno-morphological features of Asteraceous species from District Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan. A total of 13 species were collected, pressed, identified, and examined by using light and scanning electron microscopy. Both qualitative and quantitative pollen characters were observed, that is, equatorial and polar diameter, number of pores and colpi, exine thickness, exine sculpturing, pollen shape, and P/E ratio. Pollen shapes observed in studied taxa were spheroidal, suboblate, oblate-spheroidal, and lophate-spheroidal. The aperturation patterns of pollens vary from tricolporate to tetracolporate. The maximum polar and equatorial diameter of 75–100 (87.5 ± 13.6) μm and 87.5–117.5 (102.5 ± 16.4) μm, respectively, was recorded in Echinops echinatus, while minimum polar and equatorial diameter of 20–22.5 (21 ± 1.36) μm and 22.5–25 (23.5 ± 1.36) μm, respectively, was recorded in Aster subulatus. Six types of exine sculpturing patterns were observed; echinate, micro-echinate, echinate-microreticulate, microreticulate, echinate-perforate, and scabrate. Maximum exine thickness of 8.75 μm was recorded in E. echinatus and minimum of 1.25 μm in Launaea mucronata. The pollen morphology has valuable significance in understanding the taxonomy of different plant groups and plays a central role in the correct identification and classification of Asteraceous flora at species, generic, and tribe levels.     

Bundles of hexagonally arranged tubules in timothy grass pollen: Detection of a novel pollen component using anhydrous fixation and image analysis techniques in transmission electron microscopy

Pollen from timothy grass (Phleum pratense L.) was subjected to various aqueous and non‐aqueous fixation and preparation protocols for transmission electron microscopy. Only in the cytoplasm of anhydrously prepared pollen grains were conspicuous inclusions observed that range in size from less than 1 μm up to 8 or 10 μm. These bodies have so far not been described in the literature. Higher magnifications show that these inclusions consist of bundles of hexagonally arranged small tubules. In order to obtain details of the ultrastructure of this novel pollen component, TEM micrographs of ultrathin sections of hexagonally arranged tubules were analyzed using Fourier transform techniques of image analysis. It was found that the tubules form groups with quasi‐periodic hexagonal arrangement, with an average centre‐to‐centre spacing between the neighbouring tubules of approximately 42 nm. Individual tubules are formed by 12 or 13 particles. The outer diameter of the tubules ranges between 22 and 24 nm. From our experiments, we conclude that the quasi‐periodic hexagonally arranged tubules forming conspicuous cytoplasmic inclusions in dry timothy grass pollen grains are structurally similar to microtubules.     

Measurement of the width of thin,cylindrical, transparent objects by phase contrast light microscopy6

Measurement of the width of a thin, cylindrical, transparent object by phase contrast light microscopy has been frustrated by the absence of an established relationship between the true width of the object and its apparent width in the phase contrast image. We have solved this problem by devising a simple method by which individual glass fibres may be measured using both phase contrast light microscopy and electron microscopy. Using this method we have constructed calibration curves relating the diameter measured by phase contrast microscopy to the real diameter of the fibres. These curves are linear in the range 0.10-2.5 μm real diameter, with slopes close to unity and intercepts of about 0.2 μm. Thus widths of such objects are overestimated. The precise value of the intercept is related to the overall numerical aperture of the optical system. Each calibration curve permits the true width of a cylindrical object to be estimated by phase contrast microscopy with an accuracy of better than ±0.05 μm. We have found that greater precision is obtained by taking measurements of light micrographs subjectively using a microcomparator rather than objectively using a microdensitometer.     

Palynological investigation of lactiferous flora (Apocynaceae) of District Rawalpindi,Pakistan, using light and scanning electron microscopy

This was the first comprehensive pollen micromorphological investigation of lactiferous flora (Apocynaceae) of District Rawalpindi, Pakistan. The pollen morphology of 10 species of the family Apocynaceae was observed and documented using light microscopy and scanning electron microscopy. Pollen was found subspheroidal in shape in most of the species, however peroblate shape was noted in Vinca major. Exine sculpturing patterns (psilate, rugulate, scabrate, and microreticulate) were observed. The result indicated that exine ornamentation of Apocynaceae taxa is systematically informative at generic and species levels. Most of the species have tricolporate type pollen but tetraporate pollen was also observed in Trachelospermum jasminoides and tetracolpate in V. major. Minimum equatorial diameter was noted in Carissa edulis (27.13 μm) and maximum in V. major (108.25 μm). Similarly, maximum exine thickness was found in Cascabela thevetia (9.5 μm). In the present findings, the pollen morphological data are compared with available other pollen studies to evaluate the taxonomic value of pollen traits in Apocynaceae taxa by using multiple microscopic techniques. Furthermore, molecular and phylogenetic studies were recommended to strengthen the systematics of Apocynaceae taxa.     

Taxonomic importance of spore morphology in Thelypteridaceae from Northern Pakistan

Spore morphology of Thelypteridaceae species growing in Malakand Division, Northern Pakistan, was studied using both light microscopy and scanning electron microscopy. The taxa are Christella dentata and Glaphyropteridopsis erubescens in the subfamily Thelypteridoideae, and Phegopteris connectilis, Pseudophegopteris pyrrhorhachis, and Pseudophegopteris levingei in the subfamily Phegopteridoideae. The studied species exhibit differences in spore size, exospore thickness, color, and ornamentation. Spores of the studied species are monolete and medium‐sized, and shape is ellipsoidal in both polar and equatorial views. The average measurement of the polar diameter ranges from 27 μm to 31 μm, whereas in the equatorial direction it varied from 20 μm to 40 μm. The exospore thickness ranges from 1.2 μm to 2.4 μm. Reticulate, laevigate with microgranules, cristate, and coarsely echinate surface ornamentation are observed among the species. Multivariate analysis including unweighted pair group method with arithmetic mean and principal component analysis was used for the grouping and discrimination of species and genera.     

Pollen morphology and its taxonomic potential in some selected taxa of Caesalpiniaceae observed under light microscopy and scanning electron microscopy

Pollen micromorphology is not only used to check the functional and structural evolution in plants but also to solve the taxonomic problem related to the classification of plants. Therefore, keeping in view the significance of pollen traits, selected taxa of the subfamily Caesalpiniaceae was collected from different geographical regions of Pakistan. The species were then analyzed under both light microscopy and scanning electron microscopy techniques to investigate the importance of micromorphological characters of pollen in the identification and classification of species. Great variation was recorded in equatorial shape, surface ornamentation, tectum, polar diameter, equatorial diameter, and exine thickness. However, little variation was observed in pollen type, polar shape, and fertility of pollen. The equatorial shape of five types was observed: prolate, prolate-spheroidal, spheroidal-subprolate, subspheroidal-prolate, and subspheroidal. Four types of surface ornamentation, psilate, granulate, clavate, and perforate, were recorded. Tectum of five types, intactate, reticulate regulate, medium reticulate, tactate, and striate, was observed. Sexine was thicker than nexine in all studied species. The largest polar diameter was observed in Caesalpinia pulcherrima 64.1 μm while the smallest in Parkinsonia aculeata 26.1 μm. The largest equatorial diameter was found in C. pulcherrima of 70.25 μm whereas the smallest in P. aculeata 27.57 μm. All the pollens analyzed were tricolporate. All studied species have a fertility ratio of more than 90%. A taxonomic key was developed to show the variation in pollen features and delimit species for the correct identification. In conclusion, the pollen traits were found useful to define species boundaries at various taxonomic ranks and will strengthen the taxonomy of this subfamily. Besides, this study also explored the palynological traits and their implication in the taxonomy of the subfamily Caesalpiniaceae.     

Effect of the Pitot microtube diameter on pressure measurement in plane supersonic microjets

The article reports on results of estimating the Pitot tube diameter effect on the streamwise and lateral pressure distributions in plane supersonic air microjets. The study is aimed at obtaining reliable information on the microjet structure. The nozzles with the size of 22.3 × 2593 μm and 83.3 × 3823 μm are used in experiments. The supersonic flow from that nozzles is investigated by glass Pitot tubes in inner/outer diameters of 24/70 and 16/42 μm. Additional measurements are performed by a Pitot microtube 8 μm in outer diameter and ≈0.1 μm wall thickness. The value ranges for accuracy determination of supersonic jets main characteristics (shock cell sizes and supersonic core length) have been found.     

Ultrastructural aspects of Callithrix penicillata lingual papillae

Callithrix penicillata belongs to the family Callitrichidae, Callithrix genus. They are basically insectivorous, but they consume fruits. The mucosa of the tongue is composed of some papillary types, revealing different levels of expertise. The present study attempted to describe the morphological and ultrastructural aspects of the dorsal surface of the C. penicillata, describing the characteristics and distribution of papillae found. Five tongues of C. penicillata (two females and three males), obtained from breeding colonies of CENP-Ananindeua-PA, died from natural causes. The material was fixed partly in a buffer solution paraformaldehyde 10% and partly in modified Karnovsky solution, divided into apex, body, and root, and then the fragments were used in light microscopy and scanning electron microscopy. The average length of the tongue of the females was 22 mm and for males 20.5 mm. Three types of papillae were described: filiform (along all tissue extension with 154 μm of diameter), fungiform (along all tissue extension with 275 μm of diameter), and vallate (just three units in caudal (dorsal) portion with 672 μm of diameter). Data analysis indicates that the distribution and ultrastructural morphology of the C. penicillata lingual papillae are some similar to other primates.     

A novel application of microsphere perfusion and scanning electron microscopy to the identification of pulmonary arterioles in guinea-pig and rabbit lungs

In arterioles of the lung the intravascular blood pressures are lower than in comparable vessels in the systemic circulation and the arteriole walls are thinner. Therefore, it is very difficult to distinguish between arterioles and venules of the same size using scanning electron microscopy. This study describes a novel application of latex microsphere perfusion and scanning electron microscopy which distinguishes between pulmonary arterioles and venules on the basis of endothelial cell morphology. Microspheres, 90 and 45 μm in diameter, were perfused into the arterial side of the pulmonary circulation of guinea-pig and rabbit lungs. Scanning electron microscopy of the arterioles on both sides of the lodged microspheres indicated that the endothelial cells are spindle shaped. In contrast, the endothelial cells of equal diameter venules are polygonal. Furthermore, the nuclei of the arteriolar endothelial cells were significantly (P = 0·019) narrower than those of endothelial cells in venules of equal diameter. Finally, it was observed that the differences between arteriole and venule endothelial cells persisted distally to the capillaries.     

Comparative foliar anatomical and pollen morphological studies of Acanthaceae using light microscope and scanning electron microscope for effective microteaching in community

In this study, foliar anatomy and pollen morphology of 10 species of Acanthaceae has been investigated using light and scanning electron microscopy. The study was aimed to highlight the role of microscopy in microteaching at community for proper characterization of plants using palyno‐anatomical characters including pollen type, exine sculpturing, shape of epidermal cells, pattern of anticlinal wall, type and size of stomata, and trichome. Most of the species have polygonal cell shapes but some species have irregular, tetragonal, and pentagonal shape of epidermal cells. The largest epidermal cell length on adaxial and abaxial surface were observed in Asystasia gangetica 66.95 and 87.40 μm whereas least was observed on adaxial surface in Justicia adhatoda 36.9 μm and on abaxial surface in Barleria cristata 35.65 μm. In anatomy, species have diacytic type of stomata, whereas stomata of paracytic type observed in two species, while in A. gangetica cyclocytic type of stomata are present. Quantitively on abaxial surface, largest stomata length 29.9 μm and width 24.30 μm was noted in B. cristata. While shortest stomata length was observed in Ruellia prostrata 25.95 μm whereas minimum width of stomata was examined in Barleria acanthoides 2.05 μm. The diversity of trichomes are present in all species except in Ruellia brittoniana. Acanthaceae can be characterized by exhibiting different pollen morphology having five types of pollen shapes, prolate, spheroidal, perprolate, subprolate, and oblate spheroidal. Exine peculiarities showing variations such as reticulate, granulate, coarsely reticulate, lophoreticulate, perforate tectate, and granulate surface were examined.     

Erosion of copper single crystals under conditions of 30° incidence

High purity copper single crystals were eroded at 30° to a (111) face using glass spheres 210 μm in diameter travelling at a velocity of 133 m s^?1. Detailed scanning electron microscopy of both eroded surfaces and metallographic sections has been performed together with transmission electron microscopy of erosion debris. It was observed that there was extensive surface shear that led to ripple formation, folding of the surface, subsurface crack propagation and material loss by flaking similar to delamination wear. Direct evidence was obtained for recrystallization of the surface layers.     

Surface structure in an abraded titanium alloy

The surface of a hexagonal close-packed titanium alloy Ti-6%Al-4%V which was abraded by extrusion honing has been examined by X-ray diffraction and by transmission electron microscopy. A deformed layer about 10 μm thick under a compressive stress of 420 MPa was observed. Warren-Averbach analysis of the chromium Kβ diffraction pattern, monochromated with a Si(Li) detector, showed negligible microstrain and a mean coherent domain size of 12 nm with a half-width of 6 nm. Electron diffraction shows nearly single crystal regions, corresponding to the original undeformed grains, with tangential streaking due to the presence of subgrains misoriented by a few degrees. Dark field images revealed these subgrains to be typically some 50 nm in diameter. Below about 10 μm from the surface the sample appeared undeformed. These observations are interpreted as showing that the abraded surface consists of nearly dislocation-free subgrains about 12 nm thick and 50 nm in diameter. Such a mosaic structure is consistent with the laminated structure described in Suh's delamination theory of wear.     

Mechanisms of solid particle erosive wear for 90° impact on copper and iron

High purity copper single crystals and fully annealed polycrystalline α-Fe were eroded by glass spheres 70 μm in diameter suspended in an argon gas stream and travelling normal to the target surface at 122 m s^?1. Detailed scanning electron microscopy of eroded surfaces, together with transmission electron microscopy of erosion debris, was performed. It was observed that the surfaces developed a stable hill-and-valley topology not reported previously. The mechanism of material removal was plowing on the hillsides and flaking due to subsurface crack propagation in the valleys. Subsurface voids were created independently of second-phase particles and evidence existed for a local temperature rise at the surface.     

Taxonomic study of subfamily Nepetoideae (Lamiaceae) by polynomorphological approach

The present study is insight into pollen morphology for characterizing species and their utility in the taxonomic separation of certain taxa of subfamily Nepetoideae (Lamiaceae) from Pakistan. The pollen micromorphology of 11 species of the Nepetoideae was analyzed and documented using light microscopy and scanning electron microscopy (SEM) for both qualitative and quantitative characteristics. Most species have hexazonocolpate pollen grains but trizonocolpate and tetrazonocolpate pollen with circular and oval amb were also rarely observed in Mentha spicata. The basic pollen shape in most of the studied species was subspheroidal but prolate grains were also observed in M. spicata, S. coccinea, and S. plebeia. The exine sculpturing of Nepetoideae pollen was taxonomically very informative particularly at subfamily level. Observations of exine sculpturing with SEM revealed various types of pollen grains: reticulate, bireticulate, microreticulate, perforate, aerolate, and gammate. The bireticulate type further subdivided into three subtypes based on the number of secondary lumina in each primary lumen and is characterized by varying characteristics of the secondary reticulum and primary muri. A significant variation was observed in colpus surface ornamentation. The maximum polar diameter was found in O. americanum (58 ± 5.8 μm) and the maximum equatorial diameter observed in O. basilicum (50.25 ± 1.37 μm). Pollen features of the studied species were discussed and compared based on the current taxonomical concepts. The results showed that pollen traits of the subfamily Nepetoideae was found significant to classify the taxa. Furthermore, pollen features provide additional evidence to distinguish macromorphologically similar taxa from each other.     

Palyno-anatomical characters and their systematic significance in the family Apiaceae from Chitral,eastern Hindu Kush,Pakistan

The present study was performed to provide a detailed explanation of leaf epidermal anatomy and pollen micromorphological features of selected species of family Apiaceae from Chitral, eastern Hindu Kush region as the basis of forthcoming studies. In the present article pollen morphology of eight species and foliar epidermal of seven species of family Apiaceae have been examined through microscopic techniques. In results two types of pollen prolate (five species) and perprolate (three species) with three colpi have been recorded. The exine ornamentation was found to be regulate, striate, and cerebroid. Largest pollen was found in Heracleum leucocarpum with the polar diameter of 43.25 μm and equatorial diameter of 21.6 μm. Smallest pollen was observed in Elaeosticta chitralica with the polar diameter of 18.4 μm. The P/E ratio varied from 1.59 to 2.16. Regarding to foliar epidermal anatomy, three types of epidermal cells including rectangular, irregular, and polygonal with variation in anticlinal wall pattern were determined. In the selected species three kinds of stomata comprising anisocytic, anomocytic, and paracytic type were reported in the current research. The size of epidermal cells ranged from 106 × 42.50 μm in Bupleurum falcatum subsp. cernuum and 77.25 × 26.35 μm in Prangos pabularia in adaxial surface. Largest stomatal complex was found in Prangos pabularia both in adaxial 33.55 × 20.05 μm and abaxial 50.25 × 39.40 μm. All the observed quantitative and qualitative features of the species were proved to be useful in the delimitation of species at generic and species level.     

Identification of fenestrated capillary segments in microvascular corrosion casts of the rat exocrine pancreas

The capillary bed of the rat exocrine pancreas was studied by scanning and transmission electron microscopy of corrosion casts and tissue sections. Two types of capillaries were distinguished in corrosion casts. First, there were straight capillaries of relatively constant width (mean diameter 4.79±0.87 μm), which were characterized by numerous circular constrictions on their surface. About 37% of the capillaries belonged to this type. Second, there were undulating capillaries which showed smooth surfaced eccentric dilatations defined by similar surface constrictions. The bulging areas measured 8.43 ± 1.33 μm, the constrictions next to the bulges figured for 6.45 ± 1.53 μm. About 63% of the capillaries belonged to the second type. Two types of capillaries were also identified in tissue sections. First, there were capillaries with continuous endothelial lining (26% of capillary profiles; mean diameter 5.48 ± 1.67 μm); 27% of their endothelial lining was provided with underlying pericytes. Second, there were capillaries with fenestrated endothelium (64% of capillary profiles; mean diameter 6.16 ± 1.75 μm); 12% of their endothelial lining was accompanied by pericytes. According to frequency and dimension of these two types of capillaries, we conclude that bulged and undulating capillary casts correspond to fenestrated capillaries and straight capillary casts of constant width correspond to nonfenestrated capillaries. The frequency of crests on the surface of capillary casts correlates with the different frequency of pericyte processes on fenestrated and nonfenestrated capillaries. It is concluded that pericyte processes beneath the endothelium hold resistance against luminal pressure. Bulging areas of capillary casts correlate with fenestrated areas of endothelial lining, that is, areas which are not reinforced by pericyte processes.     

Ultrastructure and distribution of superficial neuromasts of blind cavefish,Phreatichthys andruzzii,juveniles

Transmission and scanning electron microscopy (TEM, SEM) were used to study the ultrastructure of superficial neuromasts in 15 six-month old blind cavefish juveniles, Phreatichthys andruzzii (Cyprinidae). In five specimens examined with SEM, the number of superficial neuromasts over the fish body (480–538) was recorded. They were localized mainly on the head (362–410), including the dorsal surface, the mentomandibular region, and laterally from the mouth to the posterior edge of the operculum. Neuromasts were also present laterally on the trunk and near the caudal fin (116–140). A significantly higher number of neuromasts were present on the head compared to the trunk (t-test, P < 0.05). Superficial neuromasts of the head and those along the trunk were similar in ultrastructure. Each neuromast comprised sensory hair cells surrounded by nonsensory support cells (mantle cells and supporting basal cells) with the whole covered by a cupula. Each hair cell was pear-shaped, 15–21 μm high and 4–6 μm in diameter, with a single long kinocilium and several short stereocilia. Most support cells were elongated, with nuclei occupying a large portion of the cytoplasm. In the margin of the neuromast, mantle cells were particularly narrow. Both types of support cells had well-developed Golgi apparatus and rough endoplasmic reticulum. The number of hair cells and nonsensory support cells of the anterior lateral line (head) did not differ significantly from those of the posterior lateral line (trunk) (t-test, P > 0.05). Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc.     

流动沸腾条件下微通道壁面温度的红外特征

设计并搭建了沸腾换热试验台,采用TH5104红外热像仪测量微通道壁面温度来研究混合制冷工质在微通道内的沸腾换热特性.测量试件是一外径为1.22 mm,内径为0.86 mm,长为200 mm的不锈钢单圆管.实验利用红外热像仪测量并记录下质量流量为1 726～8 635 kg/m2·s,热流密度为65～231 kW/m2时壁面温度的变化情况.实验分析和讨论结果显示:微通道壁面的温度分布沿着轴向变化有明显的规律性;水平微尺度通道内流动沸腾过程中,试件前后段有较大的温差效应,温差的正负与热流密度的大小有关;壁面温度的变化与热流密度、管内工质的流型和换热形式关系密切,流型越复杂,壁面温度变化越剧烈.     

Integration of a high‐NA light microscope in a scanning electron microscope

We present an integrated light‐electron microscope in which an inverted high‐NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high‐resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub‐10 μm alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum‐compatible immersion oil. For a 40‐nm‐diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry.     

Sephadex globules placed as landmarks in combined stereo microscopy,scanning and transmission electron microscopy

Regions of specific interest in tissue blocks were localized in a stereo microscope and landmarked with Sephadex spheres (10–40 μm in diameter). This procedure made it possible to recognize these regions easily and quickly in the scanning electron microscope. When the tissue was later embedded in Epon the spheres facilitated orientation when sectioning. Thus it was possible with great certainty to determine the level at which ultrathin sections should be cut for transmission electron microscopy to attain an exact correspondence between scanning and transmission electron microscopic observations. The procedure is described and an example of its application is shown in a study of experimental hypertensive endothelial changes.