银纹夜蛾幼虫气门形态特征的扫描电镜观察

采用扫描电子显微镜对银纹夜蛾Argyrogramma agnata Staudinger 5龄幼虫气门的细微结构进行观察。结果表明,银纹夜蛾的幼虫气门的外面观呈二唇状,二唇瓣的内侧向外微凸,唇瓣由若干密生的簇生分枝结构(筛板)组成,在2000倍镜下,筛板呈较规则的刺猬状,在17000倍镜下,簇生分枝圆柱状,柱体光滑,顶端钝圆;筛板的内面观呈平行排列的树形结构,分枝长刺状,单生,分枝数量明显减少。气门外围体壁具有尖塔形纹饰。银纹夜蛾气门的超微结构特征是其长期适应和进化的结果,除具有其本身的生理学意义外,也可作为系统分类的参考依据。     

Phosphorylation-dependent translocation of glycogen synthase to a novel structure during glycogen resynthesis

Glycogen metabolism has been the subject of extensive research, but the mechanisms by which it is regulated are still not fully understood. It is well accepted that the rate-limiting enzymes in glycogenesis and glycogenolysis are glycogen synthase (GS) and glycogen phosphorylase (GPh), respectively. Both enzymes are regulated by reversible phosphorylation and by allosteric effectors. However, evidence in the literature indicates that changes in muscle GS and GPh intracellular distribution may constitute a new regulatory mechanism of glycogen metabolism. Already in the 1960s, it was proposed that glycogen was present in dynamic cellular organelles that were termed glycosomas but no such cellular entities have ever been demonstrated. The aim of this study was to characterize muscle GS and GPh intracellular distribution and to identify possible translocation processes of both enzymes. Using in situ stimulation of rabbit tibialis anterior muscle, we show GS and GPh intracellular redistribution at the beginning of glycogen resynthesis after contraction-induced glycogen depletion. We identify a new "player," a new intracellular compartment involved in skeletal muscle glycogen metabolism. They are spherical structures that were not present in basal muscle, and we present evidence that indicate that they are products of actin cytoskeleton remodeling. Furthermore, for the first time, we show a phosphorylation-dependent intracellular distribution of GS. Here, we present evidence of a new regulatory mechanism of skeletal muscle glycogen metabolism based on glycogen enzyme intracellular compartmentalization.     

Scanning electron microscopic evidence for a budding mode of chloroplast multiplication in higher plants

Scanning electron microscopic observations of leaf sections of certain plants show two distinct types of chloroplast multiplication. In the normal mode, chloroplast division is achieved by binary fission. In Bryophyllum pinnatum Kurz. the chloroplast multiplication occurs by a process similar to the budding of yeast. The advantages of using scanning electron microscopy to follow closely the different stages of chloroplast division are demonstrated.     

Scanning electron microscopic study on the structure of the lingual papillae of the Saanen goat

The morphology of lingual papillae of the ten male mature Saanen goats (11 months old, approximately 42 kg in weight and of a known pedigree) was examined by scanning electron microscopy. Tissues were taken from the dorsal and ventral surfaces of the apex, body and root of the tongue, and were prepared accordingly and observed under the scanning electron microscope. On the dorsal and ventro-lateral surfaces of the lingual mucosa, three types of mechanical papillae (filiform, lenticular, and conical) and two types of gustatory papillae (vallate and fungiform) were observed. The structure and density of the filiform papillae differentiated on the anterior, posterior and ventro-lateral aspects of the tongue. Two types of lenticular papillae, both possessing a prominent surrounding papillary groove, were determined. The pyramidal-shaped type I lenticular papilla had a pointed apex while the round-shaped type II lenticular papilla possessed a blunt apex. Certain number of the type I lenticular papillae had double apices. The larger conical papillae were hollow structures, differing structurally from the filiform papillae with their larger size, a tip without projections and lack of the secondary papillae. The vallate papillae were present on both rims of the torus linguae, were encircled by a prominent gustatory furrow which was also surrounded by a thick annular fold. The fungiform papillae were scattered among the filiform papillae in the anterior two-thirds of the dorsal and lateral surfaces, and each of them was highly protected by surrounding filiform papillae, yet encircled by a papillary groove. Our findings indicate that Saanen goat have profuse distribution of papillae on the tongue displaying morphological features characteristic of mechanical function.     

Scanning electron microscopic study of uropathogen adherence to a plastic surface

We used a polyethylene surface to study the adherence of various urinary pathogens to a representative inert surface. The bacteria were suspended in filter-sterilized urine during this adhesion study, and differential adhesion was clearly demonstrated. Pseudomonas aeruginosa adhered most avidly and formed large microcolonies that were surrounded by an extensive amorphous matrix. Staphylococcus saprophyticus also formed microcolonies on the surface of the plastic droppers. In general, piliated strains of Escherichia coli adhered less avidly than the other organisms, but more avidly than nonpiliated strains; however, one piliated strain of E. coli adhered very poorly and behaved like a nonpiliated strain.     

Scanning electron microscopic study of uropathogen adherence to a plastic surface.

We used a polyethylene surface to study the adherence of various urinary pathogens to a representative inert surface. The bacteria were suspended in filter-sterilized urine during this adhesion study, and differential adhesion was clearly demonstrated. Pseudomonas aeruginosa adhered most avidly and formed large microcolonies that were surrounded by an extensive amorphous matrix. Staphylococcus saprophyticus also formed microcolonies on the surface of the plastic droppers. In general, piliated strains of Escherichia coli adhered less avidly than the other organisms, but more avidly than nonpiliated strains; however, one piliated strain of E. coli adhered very poorly and behaved like a nonpiliated strain.     

Local interactions as a structure determinant for protein molecules: II

Van der Waals interactions between sidechains are indicated to be important in determining the native state of the proteins of known structure by the following observations:

• 1.1.|the average radial distribution of polarity increases continuously from the center of the molecule to its periphery.
• 2.2.|nonpolar sidechains tend to occur in clusters.
• 3.3.|the frequencies of long-range nearest-neighbor pairs are markedly non-random; each type of sidechain seeks nearest-neighbors of similar polarity. To investigate how these interactions affect the overall structure of the protein molecule, three simpified models are treated: a sheath-core model composed of independent residues, a modification accounting approximately for the connected nature of the chain, and a model consisting of three concentric spherical phases.

     

An ImageJ-based tool for three-dimensional registration between different types of microscopic images

Three-dimensional (3D) registration (i.e., alignment) between two microscopic images is very helpful to study tissues that do not adhere to substrates, such as mouse embryos and organoids, which are often 3D rotated during imaging. However, there is no 3D registration tool easily accessible for experimental biologists. Here we developed an ImageJ-based tool which allows for 3D registration accompanied with both quantitative evaluation of the accuracy and reconstruction of 3D rotated images. In this tool, several landmarks are manually provided in two images to be aligned, and 3D rotation is computed so that the distances between the paired landmarks from the two images are minimized. By simultaneously providing multiple points (e.g., all nuclei in the regions of interest) other than the landmarks in the two images, the correspondence of each point between the two images, i.e., to which nucleus in one image a certain nucleus in another image corresponds, is quantitatively explored. Furthermore, 3D rotation is applied to one of the two images, resulting in reconstruction of 3D rotated images. We demonstrated that this tool successfully achieved 3D registration and reconstruction of images in mouse pre- and post-implantation embryos, where one image was obtained during live imaging and another image was obtained from fixed embryos after live imaging. This approach provides a versatile tool applicable for various tissues and species.     

An improved approach for the segmentation of starch granules in microscopic images

Background

Starches are the main storage polysaccharides in plants and are distributed widely throughout plants including seeds, roots, tubers, leaves, stems and so on. Currently, microscopic observation is one of the most important ways to investigate and analyze the structure of starches. The position, shape, and size of the starch granules are the main measurements for quantitative analysis. In order to obtain these measurements, segmentation of starch granules from the background is very important. However, automatic segmentation of starch granules is still a challenging task because of the limitation of imaging condition and the complex scenarios of overlapping granules.

Results

We propose a novel method to segment starch granules in microscopic images. In the proposed method, we first separate starch granules from background using automatic thresholding and then roughly segment the image using watershed algorithm. In order to reduce the oversegmentation in watershed algorithm, we use the roundness of each segment, and analyze the gradient vector field to find the critical points so as to identify oversegments. After oversegments are found, we extract the features, such as the position and intensity of the oversegments, and use fuzzy c-means clustering to merge the oversegments to the objects with similar features. Experimental results demonstrate that the proposed method can alleviate oversegmentation of watershed segmentation algorithm successfully.

Conclusions

We present a new scheme for starch granules segmentation. The proposed scheme aims to alleviate the oversegmentation in watershed algorithm. We use the shape information and critical points of gradient vector flow (GVF) of starch granules to identify oversegments, and use fuzzy c-mean clustering based on prior knowledge to merge these oversegments to the objects. Experimental results on twenty microscopic starch images demonstrate the effectiveness of the proposed scheme.

     

The structure of the hypothalamic inferior lobes of the blacktip reef shark: Scanning and transmission electron microscopic observations

The inferior lobes of the shark hypothalamus were examined with light, transmission and scanning electron microscopy. The cells bordering the floor of the lateral recess appear to be typical liquor-contacting neurons. With scanning electron microscopy (SEM) the apical ends of these cells are seen to bulge into the ventricular lumen. In contrast, the roof is lined by a more typical ependymal cell characterized by numerous cilia and microvilli. In addition, SEM reveals several kinds of supraependymal cells with processes that appear to penetrate the ventricular lining. A periventricular nucleus underlies the ependymal cells. Neurons of the periventricular nucleus contain numerous lipofuchsin granules. The rest of the inferior lobe consists of many neuronal fibers. The morphology of the hypothalamic inferior lobe is discussed in relation to its possible role in feeding and aggressive behavior in both elasmobranchs and teleosts.     

Extended morphological processing: a practical method for automatic spot detection of biological markers from microscopic images

Background  

A reliable extraction technique for resolving multiple spots in light or electron microscopic images is essential in investigations of the spatial distribution and dynamics of specific proteins inside cells and tissues. Currently, automatic spot extraction and characterization in complex microscopic images poses many challenges to conventional image processing methods.     

Scanning electron microscopic visualization of a microexudate prepared by the release of cells by urea

The scanning electron microscope (SEM) was used to examine the residue remaining after human fibroblasts, permitted to attach to plastic dishes in the absence of serum, were removed with buffered 1 M urea. The ‘urea carpet’ is entirely different from the “substrate-attached material” (SAM) of Culp [4, 5, 6] in that it contains no “footpad” material. Furthermore, the SEM pictures clearly indicate that urea carpet stimulates the adhesion and spreading of newly added fibroblasts.     

A laminated, flex structure for electronic transport and hybridization of DNA

We have developed the first prototypes of a three-dimensional, electrophoretically driven microlaboratory for the analysis of proteins and DNA. By selecting the appropriate spacing and geometrical configuration, oligonucleotides were transported, in a controlled, rapid fashion, by electrophoresis in free-space. Transport efficiencies over 2 mm distances exceeded 70%. Electrodes of similar design were combined with an electronically addressed DNA hybridization chip to form a fully electrophoretic microlaboratory. In this instance, gold-plated copper electrodes were patterned on a 2 mil thick polyimide substrate. This polyimide layer was stiffened with 20 mil of polyimide to provide support for flip-chip bonding of our standard 100-site Nanochip. This composite structure illustrated three-dimensional transport of target oligonucleotides, through a via in the polyimide, along a series of electrodes and onto the diagnostic chip. Upon reaching the diagnostic chip, electronic hybridization was performed for a competitive reverse dot blot assay. The electronic assay showed a specific to nonspecific ratio in excess of 20:1. These results suggested that this type of structure might be of practical consequence with the development of a microlaboratory for biowarfare applications.     

Design and realization of an on-line database for multidimensional microscopic images of biological specimens

The BioImage database is a new scientific database for multidimensional microscopic images of biological specimens, which is available through the World Wide Web (WWW). The development of this database has followed an iterative approach, in which requirements and functionality have been revised and extended. The complexity and innovative use of the data meant that technical and biological expertise has been crucial in the initial design of the data model. A controlled vocabulary was introduced to ensure data consistency. Pointers are used to reference information stored in other databases. The data model was built using InfoModeler as a database design tool. The database management system is the Informix Dynamic Server with Universal Data Option. This object-relational system allows the handling of complex data using features such as collection types, inheritance, and user-defined data types. Informix datablades are used to provide additional functionality: the Web Integration Option enables WWW access to the database; the Video Foundation Blade provides functionality for video handling.     

Scanning electron microscopic observation of the parasitic forms of Fonsecaea pedrosoi in a human skin lesion

The parasitic form of Fonsecaea pedrosoi in the superficial hyperkeratotic layer of the skin, in a patient with chromoblastomycosis, was observed by a scanning electron microscope using some technical improvements. The fungus elements were comprised of branched septate hyphae with spherical cells and sclerotic cells. It was observed that the sclerotic cells, divided into two or three parts, formed a germ tube.     

Integral equation theories for predicting water structure around molecules

Water plays a crucial role in the structure and function of proteins and other biological macromolecules; thus, theories of aqueous solvation for these molecules are of great importance. However, water is a complex solvent whose properties are still not completely understood. Statistical mechanical integral equation theories predict the density distribution of water molecules around a solute so that all particles are fully represented and thus potentially both molecular and macroscopic properties are included. Here we discuss how several theoretical tools we have developed have been integrated into an integral equation theory designed for globular macromolecular solutes such as proteins. Our approach predicts the three-dimensional spatial and orientational distribution of water molecules around a solute. Beginning with a three-dimensional Ornstein-Zernike equation, a separation is made between a reference part dependent only on the spatial distribution of solvent and a perturbation part dependent also on the orientational distribution of solvent. The spatial part is treated at a molecular level by a modified hypernetted chain closure whereas the orientational part is treated as a Boltzmann prefactor using a quasi-continuum theory we developed for solvation of simple ions. A potential energy function for water molecules is also needed and the sticky dipole models of water, such as our recently developed soft-sticky dipole (SSD) model, are ideal for the proposed separation. Moreover, SSD water is as good as or better than three point models typically used for simulations of biological macromolecules in structural, dielectric and dynamics properties and yet is seven times faster in Monte Carlo and four times faster in molecular dynamics simulations. Since our integral equation theory accurately predicts results from Monte Carlo simulations for solvation of a variety of test cases from a single water or ion to ice-like clusters and ion pairs, the application of this theory to biological macromolecules is promising.