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.     

Light microscopy of Pakistani Berberis leaf cuticles and its taxonomic implications

Taxonomy of the genus Berberis is quite complex, due to overlapping morphological characters, making it very difficult to differentiate the species within the genus. In order to resolve this taxonomic complexity, the foliar anatomy of 10 Berberis L. species was carried out, for the first time from Pakistan, using light microscopy (LM). Significant variation in terms of epidermal cells shape, size, cell wall pattern, and stomata type was observed. B. baluchistanica has the largest epidermal cells, Adaxial: length = 45–(53.9 ± 3.6)–62.5 μm; and width = 22.5–(26.3 ± 1.3)–30 μm; Abaxial: length = 37.5–(43.25 ± 2.5)–50 μm; and width = 20–(22.6 ± 0.8)–25. The highest number of stomata was observed in B. glaucocarpa as 62 on the abaxial surface while the lowest number of stomata was recorded in B. baluchistanica as 8 on the adaxial surface. Of 10 investigated species, 6 possess anomocytic type stomata, while 2 species that is, B. aitchisonii and B. parkeriana have both anomocytic and anisocytic stomata while B. baluchistanica and B. calliobotrys have only paracytic type stomata. The highest number of cells per unit area was present on the adaxial surface of B. calliobotrys ranging from 245–(252.4)–260 followed by B. parkeriana with 209–(227.8)–250 on the abaxial surface. Stomatal index (SI) also varied considerably and was the lowest (2.6) percentage in B. baluchistanica and highest (31.9) percentage in B. kunawurensis. A taxonomic key based on micro‐morphological characters is provided for species identification.     

Local burn versus local cold induced acute effects on in vivo microcirculation and histomorphology of the human skin

Background: The impact of burns and colds on human skin microcirculation and histomorphology has not been compared as yet. Reflectance confocal microscopy (RCM) enables in vivo insight in human skin on cellular and subcellular levels. We evaluated analogies and differences of thermal injuries on microcirculation and histomorphology in vivo using RCM. Methods: Local superficial burn (6 female, 4 male; aged 28.4 ± 2.9 years, burn group) versus superficial cold (4 female, 6 male; aged 30.4 ± 5.2 years, cold group) was induced on the dorsum of the hand in an experimental immersion hand model. In vivo RCM was performed prior (control), immediately (t1) and 15 minutes (t2) following thermal injury to evaluate: Individual blood cell flow (IBCF), functional capillary density (FCD), epidermal thickness (ET), and granular cell size (GCS). Results: In the burn group, IBCF was increased at t1 (78.02 ± 2.60/min) and remained elevated at t2 (84.16 ± 3.04/min). In the cold group, IBCF decreased at t1 (12.62 ± 2.12 min) and increased at t2 (74.24 ± 3.14/min, P < 0.05) compared to the controls (58.23 ± 3.21/min). FCD was 6.74 ± 0.52/mm² in controls and increased at both t1 (7.82 ± 0.72/mm²) and t2 (8.02 ± 0.81/mm²) in the burn group. In the cold group, FCD decreased at t1 (2.60 ± 0.42/mm²) and increased at t2 (7.92 ± 0.44/mm², P < 0.05). ET increased at both t1 (43.12 ± 4.08 μm, P > 0.05) and t2 (47.26 ± 4.72 μm, P < 0.05) in the burn group. In the cold group, ET decreased at t1 (39.92 ± 3.14 μm, P > 0.05) and increased at t2 (44.72 ± 4.06 μm, P < 0.05) compared to the controls (41.26 ± 3.82 μm). Control GCS was 726.9 ± 59.4 μm² and increased at both t1 (739.8 ± 69.8 μm², P > 0.05) and t2 (762.6 ± 71.4 μm², P < 0.05) in the burn group. In the cold group, GCS decreased at t1 (712.4 ± 53.8 μm², P > 0.05) and increased at t2 (742.6 ± 64.8 μm², P < 0.05). Conclusions: Superficial burn induces more cellular destruction and cold leads to huge fluctuation in tissue perfusion, however, with moderate impact on histomorphology. The effect on dermal capillaries suggests a selective neural control and cold injuries might down‐regulate this system, much more than burns can activate it. Microsc. Res. Tech., 2011. © 2011 Wiley‐Liss, Inc.     

Enameloid microstructure of the serrated cutting edges in certain fossil carcharhiniform and lamniform sharks

The triple‐layered enameloid organization of neoselachian teeth has proven to be a reliable systematic character of the group. This study uses scanning electron microscopy to investigate the orientation of the parallel enameloid bundles in the area of the serrated cutting edges in certain fossil elasmobranchs. The examined teeth come from two Upper Cretaceous Squalicorax species and the Upper Miocene carcharhiniforms Galeocerdo sp., Carcharhinus sp., and Hemipristis serra. The parallel bundles are revealed by surface etching, which removes the superficial shiny‐layered enameloid. In the teeth of Squalicorax, the bundles around the cutting edge bend once, before they reach the serrations. The studied carcharhiniform species show a more complicated pattern with a change of parallel bundle course inside the serrations. H. serra teeth do not display the first bending of the bundles, whereas it was present in the other two carcharhiniforms. The course of the crystalline bundles in both Squalicorax species is not affected by the presence of the serrations, regardless of the twofold difference in tooth size between them. In the carcharhiniform species, the bended bundles occur within the primary and secondary serrations and are always associated with them. This feature might have functional significance by strengthening the cutting edge or could simply develop as a consequence of the enameloid mineralization around the individual serrae. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Reflectance confocal‐laser‐scanning microscopy in vivo assessments of cigarette‐induced dynamic alterations of cutaneous microcirculation on histomorphological level

Objective: Until now, high resolution reflectance confocal‐laser‐scanning microscopy (CLSM) was used for observation of cutaneous morphology in vivo and in real time. We hypothesized that CLSM also allows observation of dynamic processes of cutaneous microcirculation. Methods: Reflectance CLSM (Vivascope1500; Lucid, Rochester, NY) was performed in 24 young male habitual smokers (23 years, range: 19–26, body mass index 23.9 ± 4.04) with relatively limited cigarette exposure (mean: 3.1 ± 2.4 pack‐years). Eight matched nonsmokers served as controls. The quantitative blood cell flow and the diameter of capillary loops were determined prior (baseline), during, as well as 5 and 10 min after smoking. Results: Baseline value for blood cell flow was 55.50 ± 2.33 cells/min, and decreased over 45% during smoking (30.43 ± 3.76/min; P = 0.02). They were still 22% lower (43.33 ± 2.45/min; P = 0.01) 5 min after smoking and exceeded baseline values 10 min after smoking by 13% (63.00 ± 3.10/min; P > 0.05). The baseline values for capillary loop diameter (9.03 ± 0.22 μm) decreased by 21% (7.18 ± 0.28 μm; P = 0.03) during smoking, remained about 9% (8.23 ± 0.18 μm; P = 0.01) lower 5 min after smoking and exceeded baseline values insignificantly by 4% (9.38 ± 0.28 μm; P > 0.05) 10 min after smoking. There were no significant differences to the controls. Conclusion: Reflectance CLSM enables qualitative and quantitative observation of dynamic processes of cutaneous microcirculation on histomorphological level. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.     

Characterization of Albanian water frog,Pelophylax shqipericus,sperm traits and morphology,by using phase contrast microscopy

Mature spermatozoa traits and morphology of endangered Albanian water frog, Pelophylax shqipericus, have been characterized for the first time through phase contrast microscopy, as part of successful implementation of in vitro fertilization technique for this species. The basic morphology of P. shqipericus spermatozoa consists of an elongated, thick, smooth‐edged, and solid‐staining head, continuing with a thin and long tail which usually extends 2.48 times the head length. The acrosome was not clearly discernible so the measurements were done on the head as a whole, while the middle section was better visible. Average length of head, including the acrosome and midsection was estimated to be 11.78 μm ± 0.32, while the tail length resulted 29.24 ± 1.75 μm. The average thickness of the head was shown to be 3.45 μm. The total sperm length resulted to be 41.02 ± 1.83 μm. The average sperm concentration was estimated of 25.5 × 10⁶/ml. Sperm amount, survival rate and motility were also measured. The sperm survival rate was maximal immediately after preparation of the suspension and tended to decrease over time of storage, reaching 50% after 72 hr. Decreased sperm motility seemed to follow the same trend as sperm viability. Sperm traits resulted to be very similar both in size and in shape with those of “Lessonae” frog group, one of the lineages of Western Palearctic species complex, suggesting a strong phylogenetic relationship among these species.     

Palyno‐morphological investigations of subtropical endangered flora of Capparidaceae through light and scanning electron microscopy

The research was performed to investigate pollen morphology of endangered species of Capparidaceae in subtropical regions of Pakistan. The distinguishing characters were investigated by using light microscope and scanning electron microscope. Palynological study is comprised of pollen shape, pollen type, exine sculpturing, polar and equatorial diameter, length and width of colpi, mesocolpium, and exine thickness. In polar view, Cleome viscosa exhibited the highest pollen size 26.4 (32.7–24.5 μm) ±0.776 whereas Capparis spinosa appeared to be the lowest 12.6 (14.5?10.7 μm) ±0.400. In equatorial view, Cleome viscosa had the largest pollen size 17.1 (20.0–15.0 μm) ±0.606 and Capparis spinosa had the smallest pollen size 9.7 (12.50–8.00 μm) ±0.394. The maximum fertility percentage has been observed in Capparis spinosa, that is, 98.96% and minimum in Cleome viscosa, that is, 82.93%. Diagnostic key has been constructed to state the essential diagnostic features by means of which the taxa can be identified. Remarkable variations have been observed in pollen size, shape, and exine sculpturing. All the selected species were tricolporate. Prolate to subprolate pollen were observed. There is a great variation existed in exine sculpturing such as in Capparis decidua and C. sp. nova sculpturing is reticulate, in Capparis himalayensis sculpturing is Scabrate granulate, in Capparis spinosa sculpturing is Psilate, in Cleome viscosa sculpturing is regulate‐reticulate, in Dipterygium glaucum sculpturing is regulate and in Gynandropsis gynandra sculpturing is striate‐regulate. On the basis of overall characteristics of pollen it seems that palynology of this family is helpful at the generic and specific level.     

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.     

Observations on the oral organelle,cytopharynx, and subpellicular structure of a Dileptus sp.

We used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to observe the oral organelle, cytopharynx, and subpellicular structure of a Dileptus sp. The main results were as follows: (a) the cytostome was located on the ventral surface of the base of the beak, surrounded by a periportal matrix that integrated 135 microtube bundles. When these microtube bundles contract, radially arranged into a disk, the cytostome was closed. When these microtube bundles were stretch, they fell into the cytostome and opens. The diameter of the cytostome was about 16 μm regardless of its closure or opening, indicating that the contraction or elongation of these microtube bundles did not change the size of the cytostome, which was only related to whether it blocked the cytostome, thus determining the opening and closing of the cytostome. There were many microtube bundles on two sides of the feeding trough, which could widen or narrow the feeding trough and facilitate beak feeding. (b) The cytopharynx was basket‐like without a bottom with a diameter of about 6 μm and was woven from two kind fibers about 0.08 and 0.19 μm. (c) There were two types of extrusomes under the pellicle. Using transmission electron microscopy，the Type I extrusomes showed narrow and long egg shape, its cross section was circular which is composed by various electronic density of concentric. Using the scanning electron microscope, they were two slightly thin clavate, the length was about 5 μm, the diameter of the middle section was about 0.75 μm, and the diameter of the two ends was about 0.32 μm, they were distributed abundantly between the microtubule fasciculi which were located on both sides of the gap on the feeding groove. Using transmission electron microscopy, the Type II extrusomes showed egg shape. Using the scanning electron microscopy, they were about 1.6 × 0.8 μm in size, they were distributed abundantly under the body pellicle while rarely the proboscis. In addition, many different of developmental stages two types of extrusomes could be also seen in the cytoplasm. (d) There were very well‐developed fibrous systems under the pellicle that were woven from fibers about 0.14 μm in diameter that attached to the pellicle and bound some organelles in the cytoplasm (e.g., mitochondria, extrusomes) and other structures to the cytoplasm and maintained cell morphology. The results of this study not only supplement and enrich the morphological contents of the Dileptus sp., but also provide the basis for the study of the taxonomy of the Dileptus sp. It also provides a new method for researchers to explore the morphology and structure of ciliate cells under the cortex by SEM.     

Pollen morphological variation of Berberis L. from Pakistan and its systematic importance

Due to overlapping and diverse morphological characters, Berberis is among the most taxonomically complex genera. Palynology is one of the taxonomic tools for delimitation and identification of complex species. In this study, pollens of 10 Berberis species were analyzed through light and scanning electron microscopy. Qualitative as well as quantitative features (pollen shape, size, presence or absence of colpi, colpi length and width, exine thickness, ornamentation, pollen class, aperture, and polar–equatorial ratio) were measured. Five species were observed to have colpate (pantocolpate) with elongated ends, radially symmetrical, isopolar, monads, and psilate‐regulate pollens. In polar view, six pollen were spheroidal, two were ovoid, one spherical, and one oblate. Similarly, variation in pollen length was prominent and the largest pollen on polar view was recorded for B. psodoumbellata 60–65 μm (62.4 ± 0.9), while the smallest one was observed for B. lycium 29–35 μm (32.2 ± 1). The observed variation in both quantitative and qualitative features were important in taxonomic identification. This shows that palynological characters are helpful in identification of Berberis genus at the species level.     

Microscopic characters of the leaf and stem of Lavandula dentata L. (Lamiaceae)

Lavandula dentata L. is an aromatic plant used in folk medicine for different purposes and, for this reason, phytochemical surveys have been carried out in the search for bioactive substances aiming to support its uses. Since there is little knowledge on the structural aspects of L. dentata, this work has studied the anatomical characters of the leaf and stem using light and scanning electron microscopy, in order to assist the species identification. As a result, there are different types of trichomes: capitate glandular with uni‐ or bicellular head, peltate glandular with multicellular head, and branched non‐glandular. The leaf is hypostomatic showing diacytic stomata. The epidermis is uniseriate and coated with striate cuticle. The mesophyll is dorsiventral and the midrib is concave–convex and traversed by a single collateral vascular bundle. The stem is quadrangular and has alternating strands of collenchyma and cortical parenchyma as well as a typical endodermis in the cortex. The phloem and xylem cylinders are traversed by narrow rays and there is an incomplete sclerenchymatic sheath adjoining the phloem. These results are a novelty for the species and contribute to distinguish it from other lavenders. Microsc. Res. Tech. 77:647–652, 2014. © 2014 Wiley Periodicals, Inc.     

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.     

Microscopic investigation of palyno‐morphological features of melliferous flora of Lakki Marwat district,Khyber Pakhtunkhwa,Pakistan

The present study was carried out to investigate the polliniferous bee flora of Lakki Marwat district, Khyber Pakhtunkhwa, Pakistan. A total of 18 species, belonging to 13 different families were identified for the foraging activities of honeybees, namely, Cenchrus biflorus, Cestrum nocturnum, Citrus limon, Combretum indicum, Datura innoxia, Duranta erecta, Hamelia patens, Helianthus annuus, Ipomoea cairica, Luffa aegyptiaca, Nerium oleander, Ocimum basilicum, Parthenium hysterophorus, Pennisetum typhoides, Prosopis cineraria, Prosopis juliflora, Saccharum spontaneum, and Ziziphus jujuba. The identified melliferous flora was collected for the morpho‐palynological investigation. Pollen were acetolyzed, measured, and described qualitatively using light microscopy. The pollen types varied from psilate to echinate and colpi from tricolpate to hexacolpate. Quantitative characteristics including polar diameter, equatorial diameter, polar to equatorial ratio, colpi length, colpi width, spine length, spine width, and exine thickness of the pollen were determined and analyzed statistically using software IBM SPSS Statistics 20. The results of the present study highlighted the significance of pollen morphology of the bee flora of the area which may lead to the identification of the potential and useful botanical sources for beekeepers and to check the honey quality marketed from the study area.     

Morphological and microscopic identification of three major medicinal Dendrobium species in Ta‐pieh Mountains area

Dendrobium is an important medicinal material in China. It has the effect of nourishing the stomach, nourishing yin and clearing heat. In China, there are many types of Dendrobium, and different Dendrobium species have different efficacy. The present study is aimed at distinguishing three major Dendrobium species from morphological and microscopic identification in Ta‐pieh mountains area. In this article, the roots, stems and leaves of Dendrobium huoshanense, Dendrobium officinale, and Dendrobium moniliforme are used as materials to compare the differences of tissues of these three Dendrobium species by morphological indexes and microscopic identification of different Dendrobium. The stem morphology of these three Dendrobium species was significantly different except for stem internodes number and the middle part of the stem diameter by measuring the stems of Dendrobium. To ensure the safe use of Dendrobium, we built a fast and convenient method combining normal and fluorescence microscopy was applied in the present study to distinguish D. huoshanense, D. officinale, and D. moniliforme. The microscopic results show that different types of Dendrobium exhibit different states that can be distinguished under normal light normal and fluorescence microscopy. This comparative study of morphology and microscopy contributes to the development of identification and quality evaluation of Dendrobium.     

Microscopic evaluation of vesicles shed by erythrocytes at elevated temperatures

The images of human erythrocytes and vesicles were analyzed by a light microscopy system with spatial resolution of better than 90 nm. The samples were observed in an aqueous environment and required no freezing, dehydration, staining, shadowing, marking, or any other manipulation. Temperature elevation resulted in significant concentration increase of structurally transformed erythrocytes (echinocytes) and vesicles in the blood. The process of vesicle separation from spiculated erythrocytes was video recorded in real time. At a temperature of 37°C, mean vesicle concentrations and diameters were found to be 1.50 ± 0.35 × 10⁶ vesicles per microliter and 0.365 ± 0.065 μm, respectively. The vesicle concentration increased approximately threefold as the temperature increased from 37 to 40°C. It was estimated that 80% of all vesicles found in the blood are smaller than 0.4 μm. Accurate account of vesicle numbers and dimensions suggest that 86% of the lost erythrocyte material is lost not by vesiculation but by another, as yet, unknown mechanism. Microsc. Res. Tech. 76:1163–1170, 2013. © 2013 Wiley Periodicals, Inc.     

Identification of Daqingye and Banlangen including crude drugs and decoction dregs from three plant species by normal light and fluorescence microscopy

Daqingye and Banlangen are commonly used Chinese medicinal materials derived from the leaves and roots of Isatis indigotica Fort., respectively, which clinical effects have been confirmed by many studies in recent years. However, many problems have arisen concerning the quality and identity of materials sold in the market under these two names. Thus, the identification of Daqingye and Banlangen has drawn public attention. In this work, transverse sections of Daqingye and Banlangen from I. indigotica Fort. and two easily confused species, namely Baphicacanthus cusia (Nees) Bremek. and Clerodendrum cyrtophyllum Turcz., were investigated with normal light and fluorescence microscopy. The distinguishing features were 7–9 vascular bundles, cystoliths and nonglandular hairs in the leaves of I. indigotica, B. cusia, and C. cyrtophyllum, respectively. The Banlangen could be distinguished according to the characteristics of parenchymous cells, cystoliths, and stone cells. Moreover, the fluorescence features of Daqingye and Banlangen investigated in this study can provide direct points for differentiating those samples. Importantly, whether the crude drugs were decocted could be easily identified by their different fluorescence features, which can ensure their quality in clinical application. This is the first report to distinguish the three species that are commonly found in the market sold as Daqingye and Banlangen by normal light and fluorescence microscopy. This work indicates that the combination of normal light and fluorescence microscopy could be powerful, convenient, and economical for authenticating Daqingye and Banlangen from the three species, including crude drugs and decoction dregs. Microsc. Res. Tech. 76:774–782, 2013. © 2013 Wiley Periodicals, Inc.     

Three-dimensional morphometrical study of dendritic spines of the granule cell in the rat dentate gyrus with HVEM stereo images

Number, length, and diameters of dendritic spines of the granule cell in the dorsal leaf of the rat dentate gyrus were measured by using high-voltage electron microscope stereo images of 5-m?m-thick Golgi preparations with the aid of a three-dimensional image analyzer system. Spine densities of 2.02 ± 0.28, 2.28 ± 0.33, and 3.36± 0.35 per 1 μm at distal, middle, and proximal portions of the dendrite were obtained. These values were about 1.6-fold of the previous light microscopical report. Mean three-dimensional spine length were 1.244 ± 0.506 μm, 1.262 ± 0.563 μm, and 1.254 ± 0.584 μm at distal, middle, and proximal portions, respectively, which were about 1.4 times longer than those measured in two dimensions. By using measured morphometrical parameters of spines such as lengths, diameters, and population densities, total spine surface areas of 2.401 μm², 2.806 μm², and 4.180 μm² per 1 μm of the dendrite at distal, middle, and proximal portions, respectively, were obtained. The total surface area of dendrite was about doubled by the addition of the spines at each dendritic portion. The advantageous features and the problems of the present method are discussed.