Ultrastructure of cells cultured on polycarbonate membranes.

A method is described for preparing undisturbed cell cultures for both scanning and transmission electron microscopy. Cells were propagated on polycarbonate membranes with pores of 0.2 micrometer or less. Cultured cells together with their supports were prepared for both scanning electron microscopy and transmission electron microscopy using routine methods. For transmission electron microscopy a rapid schedule of infiltration and polymerization was used. The method described in this report yielded good results and it allowed the fine structure of cultured cells to be viewed in situ by both scanning electron microscopy and transmission electron microscopy.     

Ultrastructure of Cells Cultured on Polycarbonate Membranes

A method is described for preparing undisturbed cell cultures for both scanning and transmission electron microscopy. Cells were propagated on polycarbonate membranes with pores of 0.2 pm or less. Cultured cells together with their supports were prepared for both scanning electron microscopy and transmission electron microscopy using routine methods. For transmission electron microscopy a rapid schedule of infiltration and polymerization was used. The method described in this report yielded good results and it allowed the fine structure of cultured cells to be viewed in situ by both scanning electron microscopy and transmission electron microscopy.     

Structure of the cell surface of the yeast Candida tropicalis and its relation to hydrocarbon transport

The surface structure of the hypdrocarbon-utilizing yeast Candida tropicalis was investigated by scanning and transmission electron microscopy (SEM and TEM respectively). The sample preparation technique was based on a rapid cryofixation without any addition of cryoprotectants. In subsequently freeze-dried samples the surface structure was analysed by scanning electron microscopy. Thin sections were prepared from freeze substituted samples. Both techniques revealed hair-like structures at the surface of hydrocarbon-grown cells. The hairy surface structure of the cells was less expressed in glucose-grown cells and it was absent completely after proteolytic digestion of the cells. When cells were incubated with hexadecane prior to cyryofixation a contrast-rich region occured in the hair fringe of thin sections as revealed by TEM. Since these structures were characteristic for hexadecane-grown cells and could not be detected in glucose-grown or proteasetreated cells it was concluded that they originate from hexadecane adhering to the cell surface and are functionally related to hexadecane transport. The structure of the surface and its relation to hydrocarbon transport are discussed in view of earlier results on the chemical composition of the surface layer of the cell wall.Abbreviations SEM Scanning electron microscopy - TEM transmission electron microscopy     

Ultrastructure of the Secondary Egg Envelope of Cyprinodontidae of the Genus Epiplatys Gill, 1862 (Pisces,Teleostei)

Abstract The structure of the secondary egg envelope of seven species of Cyprinodontidae belonging to the genus Epiplatys was observed by transmission and scanning electron microscopy. The transverse structure of the secondary envelope examined by transmission electron microscopy is a constant feature in all the species studied and represents a good criterion for identification. In contrast, the surface structure of the secondary envelope when observed by scanning electron microscopy exhibits a great diversity of form. It is possible to distinguish each of the seven different species by the pattern of ornamentation, the width of the polygons and the size of the granules comprising them on the secondary egg envelope.     

Elastofibroma dorsi. Cytologic, histologic, immunohistochemical and ultrastructural studies.

Cytologic, light and electron microscopic, and immunohistochemical studies were conducted on a case of elastofibroma. Aspiration cytology showed a characteristic "braidlike" or "fern leaf-like" structure. Immunohistochemically the accumulate was shown to be elastin. Transmission electron microscopy indicated electron-dense, granular aggregates surrounded by microfilaments and collagen, while scanning electron microscopy revealed balls with a ball-of-yarn-like structure consisting of small fibrils, probably of elastin. These structures are unique to this disease and useful for diagnosis.     

Cellular and body scale morphology of Heterocapsa huensis sp. nov. (Peridiniales, Dinophyceae) found in Hue, Vietnam

Cellular and body scale structure of a new armored dinoflagellate Heterocapsa huensis , collected from Hue, Vietnam were investigated. Morphology of motile cell was observed by light, fluorescent and scanning electron microscopy, and body scale structure was examined by whole mounts of transmission electron microscopy. Cells of H. huensis were ellipsoid with a spherical nucleus located in the posterior and multiple pyrenoids located above the nucleus; this arrangement was similar to that of Heterocapsa pygmaea . Transmission electron microscopy revealed ultrastructure of the body scales consisted of a rounded triangular basal plate and three-dimensional ornaments. Structure of the basal plate resembles that of Heterocapsa illdefina ; however, the number of the peripheral spine is different from that of H. illdefina and this structure has never been reported from Heterocapsa species. A new Heterocapsa species, H. huensis Iwataki et Matsuoka sp. nov., is described based on positions of organelles and body scale ultrastructure.     

兰属春剑叶艺突变体叶片结构的研究

以兰属春剑(Cymbidium tortisepalum Fukuy.var.longibracteatum)叶色突变体‘隆昌素’及其亲本为材料,采用石蜡切片、扫描电镜及透射电镜对叶片的组织结构、扫描结构和超微结构进行了观察。结果显示,春剑叶色突变体叶片的表皮厚度和叶肉细胞面积均大于亲本,但导管数量少于亲本;叶片上、下表皮纹饰与亲本具有明显差异;气孔密度和面积均小于亲本,且气孔闭合度较大,占总气孔数的56%;突变体的叶绿体呈高密度的囊泡状结构,内部严重解体,体积较小的嗜锇滴聚集分布,无基粒片层,无淀粉粒。本研究表明经组培诱变的春剑叶色突变体的叶片结构与亲本存在明显差异。     

Capsulation of in vitro and in vivo grown Bacteroides species

By centrifugation on a four step Percoll density gradient cells of Bacteroides species could be separated according to the size of extracellular structure. The difference in size was visible by both light and electron microscopy. Two structures were observed on Bacteroides fragilis by electron microscopy, namely a fibrous network and an electron dense layer. An electron dense layer was visible on Bacteroides ovatus only when stained with ruthenium red. B. fragilis cells grown in the mouse peritoneal cavity did not produce a large fibrous network. An electron dense layer was observed on some cells in the presence of ruthenium red stain and cells possessing this layer were phagocytosed in vivo.     

Electron microscope studies of fish lymphocytes and thrombocytes

No observable differences were noted in electron microscopic studies between brown trout, Salmo trutta L., and carp, Cyprinus carpio L., lymphocytes and thrombocytes separated by the Ficoll-Paque technique. Transmission electron microscopy showed a similar ultra-structure of fish lymphocytes to those of other reported species. Scanning electron microscopy showed that these lymphocytes studied all had a villus structure and the thrombocytes clearly showed their spindle-shaped structure.     

小鼠海马锥体细胞树突棘形态的电镜三维重建

大多数神经元的复杂三维结构是很难直接观察的。激光扫描共聚焦显微镜技术结合染料标记技术可以重建神经元的三维形态,但精细结构的识别需要电子显微镜。利用透射电子显微镜技术,可以得到连续超薄组织切片的高分辨率图像,结合计算机支持的三维重建技术就可进一步获得神经细胞精细结构的三维信息。通过电镜三维重建技术对未成熟和成熟小鼠海马锥体细胞树突棘的形态进行了观察和分析,并对其关键步骤的操作技巧进行了重点说明。实验结果为进一步利用成像技术研究树突棘的结构、功能和可塑性提供了重要信息。     

Depositing electron microscopy maps

A meeting was held at the European Bioinformatics Institute (EBI) in Hinxton, United Kingdom to discuss recent progress in the development of EMD, a database for maps determined by electron microscopy that is now integrated with MSD, the macromolecular structure database at EBI. This meeting of representatives of many of the major image processing groups in electron microscopy also discussed possible software developments that would ease the documentation and deposition of such datasets. The meeting concluded with a strong endorsement of map deposition in electron microscopy and its linkage with the family of archival databases in biomedical research.     

Characterization of linear-oblong pyrenoids with cp-DNA along their sides in Nitzschia sigmoidea (Bacillariophyceae)

In the past 10 years Nitzschia sigmoidea (Nitzsch) W. Sm. has begun to occur in Japanese rivers in various areas. It is a common diatom in Europe but was previously absent in Japan. Each chloroplast of N. sigmoidea contains many unusual linear‐oblong structures. The internal structure of the chloroplast in this species was observed using epifluorescence and electron microscopy with immunolocalization techniques. The linear‐oblong structures in the chloroplasts could hardly be observed by conventional light microscopy of living cells, but were obvious in cells stained with propionocarmine. Transmission electron microscopy showed that the cross sections of this structure were lanceolate to fusiform with penetration by a single thylakoid. In cells stained with DAPI, chloroplast DNA was detected along both sides of the linear‐oblong structures, and DNA fibrils were detected by electron microscopy. Immunofluorescence microscopy of sectioned cells and also immunoelectron microscopy revealed specific localization of Rubisco between these DNA‐containing areas, which divided at the same time as the chloroplast. Our observations confirmed that the linear‐oblong structures are pyrenoids. The diversity of localization patterns of chloroplast DNA in diatoms is discussed.     

Ultrastructural changes in murine lymphocytes induced by aflatoxin B1

This investigation sought to determine whether splenic lymphocytes obtained from Balb/C mice exposed to aflatoxin B1 (AFB1) showed any ultrastructural changes which could account for the immunodysfunction attributable to aflatoxins. Lymphocytes obtained from Balb/C mice administered aflatoxin B1 in olive oil daily for three weeks were studied using both transmission and scanning electron microscopy. The lymphocytes demonstrated ultrastructural changes primarily in the mitochondria where marked internal dissociation of the cristae was revealed by transmission electron microscopy. All other cellular organelles were unaffected. No significant alterations in external structure were observed under scanning electron microscopy. The findings of this study indicate that AFB1 administration does not affect the surface topography of lymphocytes, but AFB1, by causing extensive mitochondrial damage, may affect the way in which these cells function. This could be a possible explanation for the immunodysfunction associated with AFB1.Abbreviations AFB1 Aflatoxin B1 - SEM scanning electron microscopy - TEM transmission electron microscopy     

Distribution and movement of membrane-associated platelet glycoproteins: use of colloidal gold with correlative video-enhanced light microscopy, low-voltage high-resolution scanning electron microscopy, and high-voltage transmission electron microscopy

Scanning electron microscopy (SEM), especially low-voltage (1 KeV) high-resolution SEM, can be used in conjunction with stereo pair high-voltage (1 MeV) transmission electron microscopy (HVEM) of whole spread cells or thick sections effectively to correlate surface structure with internal structure. Surface features such as microvilli, pits, pseudopodia, ruffles, attached virus, and other surface-related morphologic characteristics can be identified using SEM, while underlying cytoskeletal structure and organelle organization can be viewed by HVEM of the same preparation. However, the need to "prepare" cells for electron microscopy precludes observation in the living state. The use of several types of video-enhanced light microscopy (VLM) permits observation of living cells such that certain surface and internal features can be observed at a relatively high level of resolution or detection. Thus, changes in living cells can be followed, and at appropriate times the cells may be chemically fixed or rapidly frozen and prepared for ultrastructural examination by electron microscopy. We have utilized VLM in conjunction with SEM and HVEM to correlate changes in shape and surface structure with changes in the internal structure of platelets. In addition, we have found it advantageous to use colloidal gold-labeling procedures, because these markers are detectable by all three forms of microscopy. Using this approach we have labeled platelet membrane GPIIb/IIIa, a receptor for RGD-containing adhesive proteins, with gold-fibrinogen or gold-anti-IIb/IIIa. The initial binding and subsequent movement of gold-fibrinogen-IIb/IIIa complexes in living platelets was followed by VLM. The movement of individual labels could be mapped. Subsequent observation by low-voltage (1 KeV) high-resolution SEM and HVEM permits visualization of the same individual receptors tracked by LM. The final position on the membrane or the position-in-transit when fixative was added was determined relative to surface ultrastructure (SEM) and internal, particularly cytoskeletal, ultrastructure (HVEM).     

New electron microscopy database and deposition system

To manage, organize and disseminate data on the structure of biological macromolecules solved by 3D electron microscopy, an electron microscopy database has been set up at the European Bioinformatics Institute.     

3D electron microscopy of biological nanomachines: principles and applications

Transmission electron microscopy is a powerful technique for studying the three-dimensional (3D) structure of a wide range of biological specimens. Knowledge of this structure is crucial for fully understanding complex relationships among macromolecular complexes and organelles in living cells. In this paper, we present the principles and main application domains of 3D transmission electron microscopy in structural biology. Moreover, we survey current developments needed in this field, and discuss the close relationship of 3D transmission electron microscopy with other experimental techniques aimed at obtaining structural and dynamical information from the scale of whole living cells to atomic structure of macromolecular complexes. Presented at the joint biannual meeting of the SFB-GEIMM-GRIP, Anglet France, 14–19 October, 2006.     

百合细胞中内生菌的发现

在健康的百合鳞茎、地上茎、叶和幼胚等组织的细胞中发现了“内生菌”。通过光学显微镜、电镜和组培等观察,“内生菌”不影响百合细胞的正常分裂和分化,再生植株健壮。这些细菌还能随着宿主细胞的分裂转移到子细胞。“内生菌”椭圆形或近圆球形。电镜切片菌壁厚,单层,均质,为典型革兰氏阳性菌壁结构。电镜观察结果与光学显微镜观察内生菌的革兰氏染色反应是一致的。     

生物胆汁液晶态的研究

本文报导了用偏光显微镜、电子显微镜研究人体胆汁中的液晶态.结果表明:胆汁中存在有双射折的微粒,它们的图象指出是具有片层结构的液晶态.胆汁中还存在有类脂球滴液晶.     

Structural damages in adsorbed vaccines affected by freezing

This study was planned to evaluate structural damages in adsorbed vaccines affected by freezing using scanning electron microscopy and X-ray analysis of the elements. Randomly selected 42 vials of eight different types of WHO pre-qualified adsorbed freeze-sensitive vaccines from 10 manufacturers were included in the study. Vaccines were kept at 5 °C. Selected numbers of vials from each type were then exposed to ?25 °C for 24 h periods. All samples were evaluated for their structure using scanning electron microscopy, X-ray analysis of the elements and precipitation time. Scanning electron microscopy of vaccines affected by freezing showed either smooth or rough surfaced conglomerates associated with phosphate content of the precipitate. These vaccines precipitated 2–15 times faster compared to non-frozen samples. Non-frozen samples showed uniform flocculent structure either dense or dispersed. X-ray analysis of precipitates in frozen samples confirmed that the precipitate is mainly aluminium clutters. Scanning electron microscopy confirmed that the lattice structure of bonds between adsorbent and the antigen is broken and aluminium forms conglomerates that grow in size and weight. The precipitation time of vaccines affected by freezing is 4.5 times faster on average compared to non-frozen samples. These facts form the basis of the "shake test".     

Protein-lipid particles of medicinal leech salivary gland secretion; Their size and morphology

The relative location of proteins and lipids in particles of medicinal leech salivary gland secretion (SGS) is revealed for the first time. Their sizes and morphology are described. Using scanning electron microscopy and transmission electron microscopy, it was determined that SGS consists of particles of different sizes and form. This picture is supported by confocal laser scanning microscopy of SGS preparations treated with fluorescein isothiocyanate. After incubation with nonionic detergents (Brij 35 and Tween 20), transmission electron microscopy revealed the dissociation of fragments composing protein-lipid particles (PLP), and in this case an increase in free protein concentration determined by a modification of the Lowry method was observed. Perylene probing of lipids in SGS preparations showed that they are concentrated mainly inside PLP and are almost absent on the surface. Cholesterol was detected during SGS probing using the cholesteryl-Bodipy (hydrophobic fluorescent analog of cholesterol) on surface sections during confocal analysis of electron microphotographs of SGS. This analysis detected PLP structures in SGS resembling caveoles full of cholesterol. SGS, preliminary frozen at −70°C, transformed into a multitude of similar small particles visualized by transmission electron microscopy, whose fixed distribution resembled water crystal structure.