The sub-membrane reticulum of the human erythrocyte: A scanning electron microscope study

A web-like reticulum underlying the human erythrocyte membrane was studied at a resolution of 5–10 nm by means of a scanning electron microscope. The network was visualized in isolated membranes (ghosts) torn open to reveal their interior space and in residues derived from ghosts extracted with Triton X-100. It formed a continuous (rather than patchy) cover over the entire cytoplasmic surface, except where lifted off or torn away. Filaments (5–40 nm in diameter), annular figures (40–60 nm in diameter), and nodes (30–100 nm in diameter) were prominent in different networks. The dimensions of the filaments and the interstices in the reticulum varied with conditions, suggesting that the network has elastic properties. This reticulum is probably related to the erythrocyte membrane proteins spectrin and actin.     

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.     

A scanning electron microscope study of the papilla basilaris of Gekko gecko

Summary The sensory hair cells of the ventral 2/3 of the papilla basilaris of Gekko gecko are divided into anterior (pre-axial) and posterior (post-axial) portions by a mid-axial gap or hiatus where there are no hair cells. There is no separation of the hair cells in the dorsal third of the papilla. There are three tectorial membrane modifications: an attached thickened membrane covering the pre-axial hair cells, sallets covering the post-axial hair cells, and an attached filamentous membrane covering the dorsal hair cells. The number of hair cells is greatest ventrally and decreases dorsally. There are approximately 2000 to 2100 hair cells. The kinocilia of the hair cells of the anterior halves of both the pre- and the post-axial vertical hair-cell rows are oriented posteriorly, while the kinocilia of the posterior halves are oriented anteriorly. The kinocilia of the hair cells of the dorsal third of the papilla are mostly oriented posteriorly. Thus, kinocilial orientation of the ventral 2/3 of the papilla is doubly bidirectional, and the dorsal 1/3, largely unidirectional.I would like to thank Ms. Maria Maglio for her skill in handling the technical aspects of the scanning electron microscopy as well as her artistry in achieving photographic excellence on the scope, David Akers for expert photographic assistance, and Wayne Emery for the drawings. Research sponsored by United States Public Health Service Grant NS-09231.     

Immunospecific labeling of mouse lymphocytes in the scanning electron microscope

Bone marrow-derived (B) and thymus-derived (T) Balb/c mouse lymphocytes were identified in the scanning electron microscope (SEM) by the immunospecific attachment of one of several kinds of large-molecular-weight markers distinguishable in SEM. These markers (tobacco mosaic virus, keyhole limpet hemocyanin, bushy stunt virus, and bacteriophage T4) could be modified with hapten groups and linked with anti-hapten antibody, in an indirect (sandwich) scheme, to hapten-modified anti-cell-surface antibody bound to the cell surface. Hapten-modified antibodies to B cell antigens (goat anti-mouse-immunoglobulin) or to T cell antigens (rabbit anti-mouse brain) were employed to identify these two lymphoid cell types in unfractionated spleen, mesenteric lymph node, bone marrow, and thymus cell populations. The topography of B cells was always indistinguishable from that of T cells. No surface features were found to be unique to either cell type. In suspension, the majority of B and T cells had one or no microvilli regardless of the tissue source of the labeled cells. Cells in suspension that had microvilli (usually 10% of the total cell population) were always unlabeled. However, after cell contact with a glass surface, approximately half of both the B and T cell populations had a villous topography.     

Light and scanning electron microscopic study of cerebellar cortex of teleost fishes

Summary The teleostean cerebellar cortex has been studied with respect to its cytoarchitectonic arrangement and intracortical neuronal circuits. Samples of fish cerebellum were fixed either by immersion or vascular perfusion in 5% glutaraldehyde solution and processed for light and scanning electron microscopy. The cerebellar cortex shows four distinct layers: granular; fibrous stratum; Purkinje cell; and molecular layers. In the granular layer, mossy and climbing fiber glomeruli were characterized. The mossy glomerular region appeared as polygonal, round or ovoid clews formed by the convergence of up to 17 dendritic profiles upon a thick mossy fiber branch. The en passant nature of mossy fiber-granule cell dendrite synaptic relationship was clearly appreciated. The climbing fibers showed tendril and glomerular collaterals. The latter form thin, elongated glomeruli. Remnants of a neuroglial envelope were observed in the mossy fiber glomeruli but are apparently absent from the climbing fiber glomeruli. The beaded-shape Golgi cell axonal ramifications were observed participating in the formation of both glomerular types. Velate protoplasmic astrocytes and oligodendrocytes were also identified. The fibrous stratum appeared to be formed by compact bundles of thick and thin myelinated axons, running horizontally beneath the Purkinje cell layer and apparently belonging to ascending climbing fibers and descending Purkinje cell axons. At the Purkinje cell layer a selective removal of Bergmann glial cells was observed allowing the visualization of the pericellular basket and the pinceaux. Climbing fiber stems and their tendril collaterals were seen on their way to the molecular layer ascending parallel to the Purkinje dendritic ramifications. Stellate neuron processes were found passing through the fan-like arborescence of Purkinje cell dendrites.     

Morphological observation and characterization of the Pseudoregma bambucicola with the scanning electron microscope

Both leica microscopic camera system and scanning electron microscopy was used to observe and characterize the feet, back, abdomen, antennae and mouthparts of the Pseudoregma bambucicola from the bamboo, Bambusa multiplex. The possible functions of all the external morphological characteristics of the P. bambucicola were described and discussed in detail, which offers a basis for further enriching the biology, phylogeny and ecological niche of the P. bambucicola. Moreover, the morphological results should contribute to morphological identification and differentiation of the P. bambucicola from other aphids in the same family.     

Implementation of subcellular water mapping by electron energy loss spectroscopy in a medium-voltage scanning transmission electron microscope

The water concentration in biological cells plays a predominant role in cellular life. Using electron energy loss spectroscopy, the feasibility to measure the water content in cells has already been demonstrated. In this paper, we present an upgrade of water measurement in hydrated cryosections by spectrum imaging mode in a medium-voltage scanning transmission electron microscope. The electron energy loss spectra are recorded in spectrum imaging mode in a 2ⁿ×2ⁿ pixels array. Each spectrum is processed in order to determine the water mass content in the corresponding pixel. Then a parametric image is obtained in which grey levels are related to water concentration. In this image, it is possible to recognize the different subcellular compartments. By averaging the water concentration over the relevant pixels, we can determine the water mass content in the concerned subcellular compartment. As an example, we present water mass content measurement at subcellular level in rat hepatocytes.     

Scanning electron microscope study of human cervical mucus

The ultrastructure of human cervical mucus was studied by scanning electron microscopy. In fertile women, naturally occurring midcycle cervical mucus showed an arrangement of parallel fibers oriented along the main axis of the mucus sample. Sperm migration through a column of cervical mucus in vitro was also studied. It was found that sperm present among the longitudinal fibers were oriented parallel to the them and to the main axis of the mucus sample.     

A scanning electron microscope study of a microsporidian with a pansporoblast: Thelohania maenadis

Air drying is not adequate for the preservation of the pansporoblastic membrane of Thelohania maenadis (Protozoa, Microsporidia), a parasite of the crabs Carcinus mediterraneus and Carcinus maenas. Freeze-drying and critical point drying preserve the pansporoblast membrane and reveal that isolated spores of the microsporidian are covered with a thick hairy coat. This coat originates as secretory product within the pansporoblast cavity.     

Cosmos pollen ontogeny: A scanning electron microscope study

Summary Ontogenetic data concerning pollen not only clarifies the mode of deposition of the elaborate walls but has considerable functional and taxonomic relevance. Hitherto such studies have used optical or transmission electron microscopy but here a recently devised preparative technique has enabled pollen development inCosmos bipinnatus to be studied using the scanning electron microscope. The technique involves freeze-fracturing of osmium fixed, cryoprotected anthers, maceration in dilute osmium tetroxide, critical point drying, sputter coating and examination. The processes of pollen wall development can then be observed in three dimensions, an important aid to understanding the spatial relationships involved in the determination of ornamentation and apertures. Details of the pollen and tapetum are described at various stages between meiosis and anthesis. A close conformity is demonstrated between the results obtained and those of earlier transmission electron microscopic studies of the same and related species although very different interpretations are made.     

Attempts to label matrix synthesis of human root cementum in vitro

The present study describes the dynamic process of both acellular extrinsic (AEFC) and acellular/cellular intrinsic fiber cementum (AIFC/CIFC) matrix production on growing human teeth. Selected erupting maxillary and mandibular premolars with roots grown to about 70%–95% of their final length were placed in organ culture immediately following extraction. Twelve teeth for short-time labeling were pulse-incubated for 15 min in medium containing ³H-proline and chased for various times in order to follow the migration and secretion of the tracer. Eight teeth for long-time incubation were labeled continuously for 5 h before being chased for 1–8 days in order to label cementum matrix accumulation. After decalcification in ethylene diaminetetraacetic acid (EDTA), their roots were subdivided into about 20 slices each. Epon-embedded sections were prepared for light- and electron-microsopic as well as autoradiographic examination. During CIFC-formation, cementoblasts revealed high intracytoplasmic silver grain concentrations within the first hour after ³H-proline administration. The release of the tracer occurred between 60 to 120 min after administration. After 2 h, cementoblasts and the cementum matrix appeared to be labeled about equally. After 5 h, most of the labeled proteins appeared to be localized in the cementoid. Silver grains increased in number over the cementum matrix from 5–24 h. Very high intracellular grain concentrations within very large cementoblasts corresponded to regions of rapid cementum formation. Tracer-halos around entrapped cells lend support to a multipolar mode of matrix production during CIFC-initiation. The fate of the tracer during the development of early AEFC-matrix was less clear. However, fibroblasts revealed dense intracytoplasmic grain accumulations within the first hour after ³H-proline administration. Thereafter, the tracer localization was vague. This indistinct grain localization reflected the particular mode of AEFC-matrix production characterized by addition of new fibril segments to pre-existing fibers of a collagenous fringe.     

A combined scanning and transmission electron microscopic study and electron probe microanalysis of human pineal acervuli

Summary Untreated, decalcified and trypsinized acervuli from human pineal bodies were studied with the scanning and transmission electron microscope as well as by electron probe microanalysis. The mulberry-like acervuli are composed of a various number of spherical lobes (135–800 m) between which clustered groups of globuli (4–14 urn in diameter) are observed. The acervular lobes are very probably formed by an aggregation of these globuli. Small round particles 125–500 Å in diameter are observed on the surface of the pineal concretions. These are not influenced by either decalcification or trypsin treatment. The acervular mineral corresponds morphologically to hydroxyapatite. The electron probe microanalysis reveals the existence of calcium and phosphorus as main components of the acervuli. Small quantities of magnesium and strontium were also detected.Dedicated to Professor Berta Scharrer on the occasion of her 70th birthdayWith the technical assistance of Mr. P.A. MilliquetThe author wishes to thank Mr. Bauer and Mr. Fryder (Nestec SA, La Tour de Peilz) for the use of the Cambridge Stereoscan electron microscope and Dr. T. Jalanti (C.M.E., Lausanne) for his help with the use of the X-ray microanalyser     

蘑菇属九个种担孢子的电镜观察

蘑菇属 (Agaricus L.)的分类中 ,担孢子形态和大小是区分种的一个重要指标。虽然现在世界许多研究所都在运用分子生物学技术和相互亲和性来重新评价该属的分类 ,但担孢子的大小仍然是种划分的重要指标。介绍了一种新的供扫描电镜观察担孢子的简易的样品制作技术 ,并对野蘑菇 (A.arvensis) ,大紫蘑菇 (A.augustus) ,双孢蘑菇 (A.bisporus) ,A.essettei,短柄蘑菇 (A.maleolens) ,白杵蘑菇 (A.nivescens) ,双环林地蘑菇(A.placomyces) ,亚绒毛蘑菇 (A.subf loccosus)和白林蘑菇 (A.sylvicola)共 9个种的担孢子进行观察测量 ,发现 :(1 )用扫描电镜测量的结果与用光镜测量的结果具有较一致的线性相关关系 (R2 =0 .985 7) ;(2 )在种间或者种内菌株间的孢子形态和大小存在着较大的差异 ,可以作为分类的一个重要指标 ;(3 )在精确的孢子测量方面扫描电镜是一种快速、方便的方法。     

柑橘大实蝇成虫超微感器的扫描电镜观察

利用扫描电镜观察了柑橘大实蝇Bactrocera minax成虫触角、下颚须、平衡棒、前足跗节和产卵器的感器。共发现5种感受器,包括毛形感受器、微毛感受器、刺形感受器、锥形感受器和腔锥形感受器,其中,柑橘大实蝇的触角分布有毛形感受器、微毛感受器、刺形感受器和锥形感受器,下颚须有微毛感受器、刺形感受器和锥形感受器分布,平衡棒分布有微毛感受器和刺形感受器,足跗节有毛形感受器和刺形感受器分布。产卵管针突缩入产卵器基节中,其上有毛形感受器和腔锥形感受器分布,产卵管基节有较多毛形感受器。     

烟草甲触角感器的扫描电镜观察

利用扫描电镜观察烟草甲Lasioderma serricorne(Fabricius)成虫触角感器的形态和分布。结果表明,烟草甲成虫触角由柄节、梗节和鞭节组成,其中鞭节由9个亚节组成;在触角上共观察到毛形感器、刺形感器、锥形感器Ⅰ、锥形感器Ⅱ、锥形感器Ⅲ、锥形感器Ⅳ、钟形感器、B hm氏鬃毛和球状感器9种感器类型,其中锥形感器Ⅳ、钟形感器、B hm氏鬃毛和球状感器为新发现的烟草甲触角感器。此外,还讨论感器结构与功能的关系。     

A comparative scanning electron microscopic analysis of the human cerebral ventricular system

Summary A scanning electron microscopic analysis of the adult human third ventricular wall revealed ultra-architectural differences between dorsal and ventral portions. In the brains of thirteen and sixteen week old human fetuses regional differences in the surface organization of lining ependyma were more sharply defined than those of the adult. Alterations in the luminal surfaces of ependyma may reflect differences in the functional capacity of various ventricular areas. The potential role of certain ependyma (tanycytes) and their putative participation in neuroendocrine events is discussed.Supported by U.S.P.H.S. Grant NS 08171.U.S.P.H.S. Career Development Awardee K 04 GM 70001.     

沙蒿大粒象触角感器的扫描电镜观察

应用扫描电镜技术,对沙蒿大粒象Adosomus sp.触角感器的类型、分布、数量以及雌雄之间的差异进行了详细研究。结果表明:沙蒿大粒象触角上共有5种感器类型,包括板形感器(Ⅰ、Ⅱ和Ⅲ型)、毛形感器和刺形感器。触角各节的感器类型和分布差异较大,以板形感器的分布最广、数量最多,在触角各节均有分布;刺形感器分布在索节第1亚节和棒节,而毛形感器仅分布在棒节且数量较少。雌雄触角的感器类型及分布基本一致,但分布数量上雄虫明显多于雌虫。     

德国小蠊触角感器的扫描电镜观察

用电子显微镜观察德国小蠊Blattella germanica(L.)雌成虫、雄成虫、老熟若虫和低龄若虫的触角,发现其上分布有大量感器,为刺形感器(Ⅰ、Ⅱ型)、锥形感器、毛形感器、弧形感器、帽形感器、边缘感器和椭圆感器。其中刺形感器(Ⅰ、Ⅱ型)、锥形感器、毛形感器、弧形感器和边缘感器在各种虫态均有分布,帽形感器仅见于成虫触角鞭节上。在各种不同虫态中,雌雄成虫含有所有种类的感器,老熟若虫触角亚节数最多,低龄若虫的各类感器的感毛长度和基部横径普遍较小。     

烟盲蝽成虫触角感器的扫描电镜观察

烟盲蝽Nesidiocoris tenuis(Reuter)作为一种杂食性昆虫,是蔬菜害虫的重要捕食者。利用扫描电镜对烟盲蝽雌雄成虫的触角进行观察,结果表明:烟盲蝽触角由基节、柄节、梗节和鞭节组成。绝大部分触角感器位于触角的腹面和外侧面。触角感器共有9种,分别为毛形感器、刺形感器、锥形感器、钟形感器、腔锥形感器、具弯钩形感器、腔形感器、乳形感器和Bhm氏鬃毛。腔锥形感器仅见于雌性触角,其他8种感器在雌雄两性触角上的类型和分布没有明显的区别。     

Scanning electron microscope study of in vitro prepenetration gamete interactions

Hamster and mouse capacitated spermatozoa were interacted in vitro with hamster and mouse eggs in homologous and heterologous combinations. Also, fertilized and trypsin treated unfertilized hamster eggs, and unfertilized rat eggs were made to interact with capacitated hamster spermatozoa. The surface of the zona pellucida was then examined with the scanning electron microscope. It was found that sperm attachment, followed by sperm binding and penetration through the zona pellucida, was observed only when homologous gamete combinations were used. Binding of the spermatozoa to the zona was evidenced by the lytic effect of the acrosomal enzymes on the zona substance. When fertilized eggs and trypsin-treated unfertilized hamster eggs were mixed with capacitated hamster spermatozoa as well as in the heterologous gamete combinations, we found that the spermatozoa were able to establish attachment but not binding. Under these conditions the outer surface of the zona pellucida was never found to have penetration tracks made by the spermatozoa. Failure of heterologous spermatozoa to cross the foreign zona pellucida is believed to be associated with the inability of the foreign spermatozoa to establish binding and to the inability of their acrosomal enzymes to digest the zona. A similar mechanism is believed to work in zona-reacted and in trypsin-treated hamster eggs inseminated in vitro with homologous spermatozoa.