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At least 181 records · Page 10Linked to original sources
PubMed1997

Correlation of vascular smooth muscle cell morphology observed by scanning electron microscopy with transmission electron microscopy.

Smooth muscle cell (SMC) morphology was studied in rat large mesenteric artery using scanning electron microscopy (SEM), after removal of connective tissue elements with the OsO4/HCl method. Transmission electron microscopy (TEM) was used to correlate the findings of SEM and to assess the effects of digestion on cell ultrastructure. When viewed with SEM, relaxed SMC possessed a smooth, regular surface morphology with longitudinal striations or grooves. TEM examination of cross-sectional profiles of SMC suggested that these striations corresponded to the arrangement of membrane dense bodies within the SMC. The presence of cell contacts among SMC as observed with SEM were confirmed using TEM. Two types of SMC contacts were found: simple appositional contacts and interdigitations. Gap junctions were not observed between SMC in the media. Although structures suggesting nerve networks were observed in the medial-adventitial border with SEM, this could not be confirmed using TEM as a result of alteration in cell ultrastructure with the digestive method. We conclude that the OsO4/HCl method, although useful for SEM, is not suitable for TEM study, because it caused significant alterations of SMC ultrastructure such that organelles and other structures were no longer evident when viewed with TEM.

Animals↗
PubMedDate not supplied

Analysis of immunolabeled cells by atomic force microscopy, optical microscopy, and flow cytometry.

In this study we investigated the applicability of the (silver-enhanced) immunogold labeling method for atomic force microscopy. Human lymphocytes were labeled with anti-CD3 conjugated to fluorescein isothiocyanate and a secondary antibody (goat anti-mouse) linked with 1- or 30-nm colloidal gold particles. Silver enhancement was applied on these labeled cells to increase the size of the labels. In a setup combining an inverted optical microscope and a stand-alone atomic force microscope, a direct correlation was made between the force and the fluorescent images. Additionally, we performed flow cytometric analysis. From the results we conclude that immunogold labeling using small labels (1 nm) in combination with silver enhancement (30 min) proves to be a reliable method for high-resolution cell surface antigen detection in atomic force microscopy.

Animals↗
PubMed2000

Complementary visualization of mitotic barley chromatin by field-emission scanning electron microscopy and scanning force microscopy.

The surface structure of mitotic barley chromatin was studied by field-emission scanning electron microscopy (FESEM) and scanning force microscopy (SFM). Different stages of the cell cycle were accessible after a cell suspension was dropped onto a glass surface, chemical fixed, and critically point dried. Imaging was carried out with metal-coated specimen or uncoated specimen (only for SFM). The spatial contour of the chromatin could be resolved by SFM correlating to FESEM data. The experimentally determined volume of the residue chromatin during mitosis was within the range of 65-85 microm(3). A comparison with the theoretically calculated volume indicated a contribution of about 40% of internal cavities. Decondensation of chromosomes by proteinase K led to a drastic decrease in the chromosome volume, and a 3-D netlike architecture of the residue nucleoprotein material, similar to that in the intact chromosome, was obvious. Incubation of metaphase chromosomes in citrate buffer permitted access to different levels of chromatin packing. We imaged intact chromosomes in liquid by SFM without any intermediate drying step. A granular surface was obvious but with an appreciably lower resolution. Under similar imaging conditions proteinase K-treated chromosomes exhibited low topographic contrast but were susceptible to plastic deformations.

Chromatin↗
PubMed1987

Cell surface characteristics and DNA content of macrophages in murine bone marrow cultures. A study using simultaneous scanning electron microscopy and fluorescence microscopy.

An instrument combining scanning electron microscopy (SEM) and light microscopy (LM) was used to study the cell surface characteristics and DNA content of macrophages in murine bone marrow cultures. After a quantitative Feulgen DNA staining, the DNA content of the individual macrophages was measured and their cell surface morphology was studied immediately thereafter with the SEM part of the instrument. The cells were divided into six groups according to the number of microvilli and/or microridges present on their surface. A proportion of macrophages showed a DNA content more than occurs in diploid cells, which could indicate a future division. No special surface morphology could be detected in this cell type.

Animals↗
PubMed2006

An endolithic microbial community in dolomite rock in central Switzerland: characterization by reflection spectroscopy, pigment analyses, scanning electron microscopy, and laser scanning microscopy.

A community of endolithic microorganisms dominated by phototrophs was found as a distinct band a few millimeters below the surface of bare exposed dolomite rocks in the Piora Valley in the Alps. Using in situ reflectance spectroscopy, we detected chlorophyll a (Chl a), phycobilins, carotenoids, and an unknown type of bacteriochlorophyll-like pigment absorbing in vivo at about 720 nm. In cross sections, the data indicated a defined distribution of different groups of organisms perpendicular to the rock surface. High-performance liquid chromatography analyses of pigments extracted with organic solvents confirmed the presence of two types of bacteriochlorophylls besides chlorophylls and various carotenoids. Spherical organisms of varying sizes and small filaments were observed in situ with scanning electron microscopy and confocal laser scanning microscopy (one- and two-photon technique). The latter allowed visualization of the distribution of phototrophic microorganisms by the autofluorescence of their pigments within the rock. Coccoid cyanobacteria of various sizes predominated over filamentous ones. Application of fluorescence-labeled lectins demonstrated that most cyanobacteria were embedded in an exopolymeric matrix. Nucleic acid stains revealed a wide distribution of small heterotrophs. Some biological structures emitting a green autofluorescence remain to be identified.

Bacteria↗
PubMed2005

Characterization of a peg-like terminal NOR structure with light microscopy and high-resolution scanning electron microscopy.

An atypical peg-like terminal constriction ("peg") on metaphase chromosomes of the plant genus Oziroë could be identified as a nucleolus organizing region (NOR) by detecting 45S rDNA with correlative light microscopy (LM) and scanning electron microscopy (SEM) in situ hybridization (ISH). Using high-resolution 3D analytical SEM, the architecture and DNA distribution of the peg-like NOR were characterized as typical for chromosomes, albeit with significantly smaller chromomeres. ISH procedure was improved for SEM concerning signal localization, labeling efficiency, and structural preservation, allowing 3D SEM analysis of the peg-like NOR structure and rDNA distribution for the first time. It could be shown that implementation of FluoroNanogold markers is an attractive tool that allows efficient immunodection in both LM and SEM. A model is proposed for the peg structure and its mode of condensation.

Cell Cycle↗
PubMed2004

Surface morphology and chemistry of Prunus laurocerasus L. leaves: a study using X-ray photoelectron spectroscopy, time-of-flight secondary-ion mass spectrometry, atomic-force microscopy and scanning-electron microscopy.

The surface properties of the plant cuticle play a crucial role in plant-pathogen interactions and the retention and penetration of agriculturally important chemicals. This paper describes the use of X-ray photoelectron spectroscopy (XPS), time-of-flight secondary-ion mass spectrometry (ToF-SIMS), tapping-mode atomic force microscopy (TM-AFM) and scanning electron microscopy (SEM) to determine surface-specific chemical and material properties of the adaxial surface of Prunus laurocerasus L. leaves. XPS data, derived from the uppermost few nanometres (< 10 nm) of the leaf surface, were consistent with the wax components and functionality known to be present within the waxes. ToF-SIMS provided molecular speciation from the outermost monolayer of the leaf surface, indicating the importance of a family of acetates with chain lengths ranging from C20 to C34. The presence of alkanes with C29 and C31 chain lengths was also confirmed. SEM and TM-AFM topography images revealed a textured granular surface, while simultaneously recorded AFM phase images revealed heterogeneous material properties at the nanoscale. The relevance of these data to plant cuticle development, allelochemistry and agrochemical delivery is discussed.

Microscopy, Atomic Force↗
PubMed2005

Atomic force microscopy and laser confocal scanning microscopy analysis of callose fibers developed from protoplasts of embryogenic cells of a conifer.

Efficiency of novel fiber formation was much improved in protoplast culture of embryogenic cells (ECs) of a conifer, Larix leptolepis (Sieb. et Zucc.) Gord., by pre-culturing ECs in a medium containing a high concentration of glutamine (13.7 mM). The fibrillar substructures of large and elongated fibers of protoplasts isolated from Larix ECs were investigated by laser confocal scanning microscopy (LCSM) after Aniline Blue staining and atomic force microscopy (AFM) using a micromanipulator without any pre-treatment. Fibers were composed of bundles of fibrils and subfibrils, whose diameters were defined as 0.7 and 0.17 mum, respectively, by image analysis after LCSM and AFM. These fibers were proven to be composed of callose by using specific degrading enzymes for beta-1,4-glucan and beta-1,3-glucan.

Carbohydrate Conformation↗
PubMed1997

Expression of intercellular adhesion molecule-1 and lymphocytefunction-associated antigen-1 in experimental Schistosoma mansoni infection and in synchronous periparticular hepatic granulomas in mice: immunohistochemistry, confocal laser scanning microscopy, and immunoelectron microscopy.

Adhesion molecules play an important role in inflammatory and immunological responses. We assessed the expression pattern of intercellular adhesion molecule-1 (ICAM-1) and lymphocytefunction-associated antigen-1 (LFA-1) in the livers of mice experimentally infected with Schistosoma mansoni and in synchronous hepatic granulomas induced by injection of soluble egg antigen (SEA)-coupled Sepharose beads in a mesenteric vein of mice. By immunohistochemistry, confocal laser scanning microscopy, and immunoelectron microscopy, ICAM-1 was localized on endothelial cells, sinusoidal-lining cells (Kupffer cells and sinusoidal endothelium), the hepatocyte cell membrane facing Disse's space, and inflammatory cells in the granuloma. LFA-1 was visualized on the inflammatory cells of the granuloma and on phagocytic sinusoidal-lining cells, most likely Kupffer cells. ICAM-1- and LFA-1-immunoreactive cells were present in the granuloma as early as at 3 days after injection of SEA-coupled beads and persisted with time. As granulomas became older, nonimmunoreactive granuloma cells appeared. We conclude that adhesion molecules play an important role in the genesis of the schistosomal granuloma.

Animals↗
PubMed2003

Macrophages and dendritic cells in the rat meninges and choroid plexus: three-dimensional localisation by environmental scanning electron microscopy and confocal microscopy.

The present investigation provides novel information on the topographical distribution of macrophages and dendritic cells (DCs) in normal meninges and choroid plexus of the rat central nervous system (CNS). Whole-mounts of meninges and choroid plexus of Lewis rats were incubated with various anti-leucocyte monoclonal antibodies and either visualised with gold-conjugated secondary antibody followed by silver enhancement and subsequent examination by environmental scanning electron microscopy or by the use of fluorochromes and confocal microscopy. Large numbers of MHC class II(+) putative DCs were identified on the internal or subarachnoid aspect of dural whole-mounts, on the surface of the cortex (pia/arachnoid) and on the surface of the choroid plexus. Occupation of these sites would allow DCs access to cerebrospinal fluid (CSF) and therefore allow antigens into the subarachnoid space and ventricles. By contrast, macrophages were less evident at sites exposed to CSF and were more frequently located within the connective tissue of the dura/arachnoid and choroid plexus stroma and also in a sub-pial location. The present data suggest that DC may be strategically located within the CNS to sample CSF-borne antigens. Furthermore, the data suggest that CNS tissue samples collected without careful removal of the meninges may inadvertently be contaminated by DCs and meningeal macrophages.

Animals↗
PubMed1998

Sutural mineralization of rat calvaria characterized by atomic-force microscopy and transmission electron microscopy.

The application of transmission electron microscopy (TEM) and atomic-force microscopy (AFM) aid the acquisition of detailed structural information on the process of hard tissue formation. The sutural mineralization of rat calvaria is taken as a model for a collagen-related mineralization system. After cryofixation or chemical fixation an anhydrous tissue preparation technique with no staining procedures is used. The atomic-force microscope and the transmission electron microscope are used for structural analysis of the mineralizing region of the sutural tissue. With the application of AFM the collagen macroperiod is shown to be well represented in the unmineralized sutural tissue. At the mineralization front the collagen fibrils are found to be thickened and to change to a characteristic stacked platelet structure. Using TEM the macroperiod is faintly visible before mineral crystallites have formed and is more prominent after the apatite crystallization has started in the fibrils. In this step a needle-like structure of the newly formed apatitic crystals is visible.

Animals↗
PubMed1989

Study of the vaginal mucous membrane following tampon utilisation; aspect on colposcopy, scanning electron microscopy and transmission electron microscopy.

The effect of periodic tampons was studied in 17 young women during the menstrual and inter-menstrual cycle. Biopsies (n = 19) were analysed on scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The effect of medium absorbant tampons was compared with that of super-absorbant tampons. On colposcopy, dryness was noted in 89% cases, peeling in 47% cases. Microulcerations were observed only once (5%). Dryness and peeling was most often encountered with the super-absorbant tampons and during the inter-menstrual phase. TEM revealed cellular destruction affecting all layers of the epithelium: lysis of superficial and intermediate cells, destruction of desmosomes accompanied by the creation of inter-cellular spaces, lipidic vacuoles in the lysed cells. SEM revealed epithelial peeling with cleavage and severe cellular desquamation. Cellular anomalies were common at the level of the cytoplasmic membrane, which presented either defects, a porous aspect, or appeared swollen by air-bubbles.

Adult↗
PubMedDate not supplied

Fluorescence microscopy used in conjunction with horseradish peroxidase localization and electron microscopy for studying sympathetic nuclei of the rat spinal cord.

After application of horseradish peroxidase (HRP) to the transected cervical sympathetic trunk, labeled sympathetic neurons were seen in the intermediolateral nucleus (IML), central autonomic nucleus (CAN) and intercalated nucleus of rat spinal cords. As studied by glyoxylic acid-induced histofluorescence microscopy (FM), catecholaminergic (CA) terminals were most densely packed in the IML. There were 60 +/- 2 CA varicosities per 2,200 mu2 area in 20 mu thick sections through the IML. A combined FM/HRP method confirmed that preganglionic sympathetic neurons are heavily innervated by CA terminals. CA boutons were tagged with 5-hydroxydopamine for EM identification, and in the IML, 56% of CA boutons were seen to make synaptic contacts as compared to 60% in the CAN. It seems likely that virtually all CA boutons may form synapses but serial sections through boutons were not studied. It is inferred that preganglionic sympathetic neurons of both IML and CAN are served by CA inputs, and electron microscopy has revealed that most or all of these CA terminals transmit their signals via synapses; which may permit more precise structural sorting than non-synaptic transmission.

Adrenergic Fibers↗
PubMed2006

Nanostructure of the neurocentral growth plate: Insight from scanning small angle X-ray scattering, atomic force microscopy and scanning electron microscopy.

In this study, the experimental techniques scanning electron microscopy (SEM) including energy-dispersive X-ray analysis, atomic force microscopy (AFM) and scanning small angle X-ray scattering (SAXS) have been exploited to characterize the organization of large molecules and nanocrystallites in and around the neurocentral growth plate (NGP) of a pig vertebrae L4. The techniques offer unique complementary information on the nano- to micrometer length scale and provide new insight in the changes in the matrix structure during endochondral bone formation. AFM and SEM imaging of the NGP reveal a fibrous network likely to consist of collagen type II and proteoglycans. High-resolution AFM imaging shows that the fibers have a diameter of approximately 100 nm and periodic features along the fibers with a periodicity of 50-70 nm. This is consistent with the SAXS analysis that yields a cross-sectional diameter of the fibers in the range of 90 to 112 nm and a predominant orientation in the longitudinal direction of the NGP. Furthermore, we find inhomogeneities around 7 nm in the NGP by SAXS analysis. Moving towards the bone in the direction perpendicular to the growth plate, a systematic change in apparent thickness is observed, while the large-scale structural features remain constant. In the region of bone, the apparent thickness equals the mean mineral thickness and increases from 2 nm to approximately 3.5 nm as a function distance from the NGP. The mineral particles are organized as plates in a rather compact network structure. We have demonstrated that SEM, AFM and SAXS are valuable tools for the investigation of the organization of large molecules and nanocrystallites in the NGP and adjacent trabecular bone. Our findings will be an important basis for future work into identifying the defects on nanometer length scale responsible for idiopathic scoliosis and other growth-plate-related diseases.

Animals↗
PubMed2005

Assessing transferrin modification of liposomes by atomic force microscopy and transmission electron microscopy.

Site-specific delivery of drugs and therapeutics can significantly reduce drug toxicity and increase the therapeutic effect. Transferrin (Tf) is one suitable ligand to be conjugated to drug delivery systems to achieve site-specific targeting, due to its specific binding to transferrin receptors (TfR), expressed on several cell types of therapeutic interest. TfRs have been reported to be highly expressed on the surfaces of tumour cells and the well-characterised and efficient mechanism of internalisation of Tf has been exploited for the delivery of anticancer drugs, proteins, and therapeutic genes into primarily proliferating malignant cells. Liposomes are effective vehicles for drugs, genes and vaccines and can be easily modified with proteins, antibodies, and other appropriate ligands, resulting in attractive formulations for targeted drug delivery. In this study, we used atomic force microscopy (AFM) and transmission electron microscopy (TEM) to confirm the conjugation of Tf to liposomes by three different coupling methods. In addition, the conventional assays for quantification of protein amount (BCA) and phospholipid content (according to Steward) were performed. AFM and TEM were able to display Tf-molecules on the liposomal surfaces and can be routinely used to obtain additional visual information on the protein-drug carrier conjugation in a fast and reliable manner.

Drug Delivery Systems↗
PubMed2006

In vivo and in vitro study of porcine oviductal epithelial cells, cumulus oocyte complexes and granulosa cells: a scanning electron microscopy and inverted microscopy study.

The morphology and structure of porcine oviductal epithelial cells (POEC), cumulus-oocyte complexes (COCs) and granulosa cells (GC) were investigated in vivo and in vitro conditions using scanning electron microscopy (SEM) and inverted microscopy. The POEC contained columnar ciliated cells and spherical shaped non-ciliated cells. Both non- and ciliated cells appeared either in groups or distributing among each other. However, the isolation of cells was observed after culture for 48 h. A total of 921 oocytes from 20 ovaries was isolated resulting in an average of 46 oocytes per ovary. They were round in shape, surrounded by zona pellucida with layers of cumulus cells ranging between 89.16 and 144.68 microm in size. As for COCs, they were classified into 4 types; intact-, multi-, partial-cumulus cell layers and completely denuded oocyte. Interestingly, changes in morphology of COCs with intact and multi-cumulus cell layers were observed in the in vitro study. The GCs in the follicular fluid were also round in shape and found as clusters. After culturing in in vitro for 48 h, no change in morphology was observed. The GC appeared in smaller clusters or were present as single cells and their sizes ranged from 6 to 8 microm. The results obtained from this study allow us to have a better understanding of the morphology and nature of cells under both in vivo and in vitro conditions. This information is also important for the study of their secretions and biochemical compositions, which is of great importance to the use of cells as feeder cells in in vitro fertilization in current studies.

Animals↗
PubMed2000

Atomic force microscopy and electron microscopy analysis of retrovirus Gag proteins assembled in vitro on lipid bilayers.

We have used an in vitro system that mimics the assembly of immature Moloney murine leukemia virus (M-MuLV) particles to examine how viral structural (Gag) proteins oligomerize at membrane interfaces. Ordered arrays of histidine-tagged Moloney capsid protein (his-MoCA) were obtained on membrane bilayers composed of phosphatidylcholine (PC) and the nickel-chelating lipid 1, 2-di-O-hexadecyl-sn-glycero-3-(1'-2"-R-hydroxy-3'N-(5-amino-1-carboxy pentyl)iminodiacetic acid)propyl ether (DHGN). The membrane-bound arrays were analyzed by electron microscopy (EM) and atomic force microscopy (AFM). Two-dimensional projection images obtained by EM showed that bilayer-bound his-MoCA proteins formed cages surrounding different types of protein-free cage holes with similar cage holes spaced at 81.5-A distances and distances between dissimilar cage holes of 45.5 A. AFM images, showing topological features viewed near the membrane-proximal domain of the his-MoCA protein, revealed a cage network of only symmetrical hexamers spaced at 79-A distances. These results are consistent with a model in which dimers constitute structural building blocks and where membrane-proximal and distal his-MoCA regions interact with different partners in membrane-bound arrays.

Capsid↗
PubMed1997

Starch granule surface imaging using low-voltage scanning electron microscopy and atomic force microscopy.

High resolution imaging of wheat and potato starch granule surfaces has been performed using low-voltage scanning electron microscopy and atomic force microscopy. The complimentary images of uncoated granules demonstrate that the two starch types possess substantially different surface topologies; potato starch has many protrusions (100-300 nm in diameter), above a flatter surface containing 20-50 nm structures, whilst wheat starch possesses far fewer protrusions and generally has a smoother surface composed of approximately 20 nm structures. The protrusions are believed to be carbohydrate in nature and thus represent the ends of amylopectin side-chain clusters at the granule surface.

Microscopy, Atomic Force↗