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At least 145 records · Page 8Linked to original sources
PubMed1981

Topographic examination of sister chromatid differential staining by Nomarski interference microscopy and scanning electron microscopy.

BrdU-substituted Chinese hamster chromosomes were treated with a hog Na2HPO4 solution and stained with Giemsa to produce sister chromatid differential staining (SCD). The process of SCD was examined with the Nomarski differential interference microscope and the scanning electron microscope. After the Na2HPO4 treatment alone, unifilarly BrdU-substituted (TB) chromatids appeared somewhat more severely collapsed than the bifilarly substituted (BB) chromatids. Subsequent Giemsa staining, however, brought about pronounced piling up of the Giemsa dye on the TB-chromatids but not on the BB-ones, causing highly distinct differential Giemsa staining as well as a marked differentiation in surface topography between the sister chromatids. Removal of the Giemsa dye from the differentially Giemsa stained chromosomes resulted in a disappearance of such a pronounced topographic differentiation.

Animals↗
PubMed1983

The effect of primycin on the intracellular monovalent ion and water contents of rat hepatocytes as revealed by energy dispersive X-ray microanalysis and interference microscopy.

Using energy-dispersive X-ray microanalytic and interference microscopic techniques, the intracellular concentration of the monovalent ions (Na+, K+, Cl+) as well as the intracytoplasmic and intracellular water contents were studied in normal and adrenalectomized rat hepatocytes with and without primycin treatment. Although primycin influenced significantly only the intracellular potassium content of the adrenalectomized group, it exerted a marked influence on the intranuclear water content in both the normal and adrenalectomized rats. The intranuclear water content increased significantly in the primycin-treated animals. The conclusion is drawn that the increased level of hydration of the nuclear substances reflects a 'decondensation' of the chromatin which on the other hand, may represent the basis for the various effects of primycin on the induction of certain hepatic enzymes.

Adrenalectomy↗
PubMed2006

Hydrodynamic damping of membrane thermal fluctuations near surfaces imaged by fluorescence interference microscopy.

Hydrodynamic coupling between closely apposed lipid bilayer membranes can substantially slow the time scale of thermal fluctuations. These effects could dominate the kinetics of macromolecular interactions at membrane interfaces, and may exert important influences over the interactions between living cells. Here, we study a reconstituted membrane system consisting of two planar bilayers with average separation distances of 30-60 nm. This spacing range is comparable to that of natural intercellular junctions. Using real-time fluorescence interference contrast imaging, spatial and temporal characteristics of the thermal fluctuation spectra are monitored simultaneously. Strong hydrodynamic damping of the fluctuation time scale is observed whereas fluctuation spatial dimensions are unaltered by proximity to another surface. These observations provide experimental confirmation of recent theoretical models.

Lipid Bilayers↗
PubMed2001

Integration of Raman microscopy, differential interference contrast microscopy, and attenuated total reflection Fourier transform infrared spectroscopy to investigate chlorhexidine spatial and temporal distribution in Candida albicans biofilms.

Two spectroscopic techniques, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and Raman microscopy (RM), were used to characterize transport of chlorhexidine digluconate (CHG) in Candida albicans (CA) biofilms. Different (volumetric) regions of the biofilm are sampled by these two vibrational spectroscopies making them complementary techniques. Simple mathematical models were developed to analyze ATR-FTIR and RM data to obtain an effective diffusion coefficient describing transport through CA biofilms. CA biofilms were composed primarily of yeast and hyphal forms, with some pseudohyphae. Upper regions of biofilms that had become confluent, (i.e., biofilms that completely covered the germanium (Ge) substratum) were composed primarily of a tangled mass of hyphae with openings between germtubes about 10 to 50 microm across. Quantitative analysis of ATR-FTIR kinetic data curves indicated that the effective diffusion coefficient for transport of CHG through confluent biofilms about 200-microm thick was reduced 0.1 to 0.3 times compared to the diffusion coefficient for CHG in water. Effective diffusion coefficients obtained from analysis of RM data were consistently higher than those indicated by ATR-FTIR data suggesting that transport is more hindered in regions near the base of the biofilm than in the outer layers. Analysis of both ATR-FTIR and RM data obtained from thicker films indicated that adsorption of CHG to biofilm components was responsible for a substantial portion of the transport limitation imposed by the biofilm. Comparison of ATR-FTIR and RM data for both types of biofilms indicated that sites of CHG adsorption were more concentrated in the interfacial region than in the bulk biofilm. Comparison of results for ATR-FTIR and RM measurements suggests that these relatively thick CA biofilms can be modeled, for purposes of predicting transport, approximately as a homogeneous thin planar sheet. Thus, these biofilms offer a relatively tractable model system for initial investigations of the relation between antimicrobial transport and kinetics of antimicrobial action.

Anti-Bacterial Agents↗
PubMed2002

Regular intergrowth in the AFI-type crystals: influence on the intracrystalline adsorbate distribution as observed by interference and FTIR-microscopy.

Interference microscopy and FTIR microscopy are applied to study intracrystalline concentration profiles of methanol in CrAPO-5 zeolite crystals. By using both techniques, the high spatial resolution of interference microscopy is complemented by the ability of FTIR spectroscopy to pinpoint adsorbates by their characteristic IR bands. For the first time two-dimensional concentration profiles of an unprecedented quality are reported which show a nonhomogeneous distribution of adsorbate in zeolite crystal under equilibrium with the adsorbate vapor. These nonhomogeneous profiles are attributed to regular intergrowth effects in CrAPO-5. A possible internal structure of CrAPO-5 crystals is suggested.

Journal Article↗
PubMed2001

A three-dimensional distribution of osteocyte processes revealed by the combination of confocal laser scanning microscopy and differential interference contrast microscopy.

Osteocytes are the most numerous cells in bone, embedded within the mineralized bone matrix. Their slender cytoplasmic processes form a complex intercellular network. In addition, these processes are thought to be important structures in the response to mechanical stress. This study provides an extensive analysis of the three-dimensional structure of the osteocyte and its processes in 16-day-old embryonic chick calvariae, based on nondestructive subsurface histotomography using both confocal laser scanning (CLS) microscopy and differential interference contrast (DIC) microscopy. OB7.3, a chicken osteocyte-specific monoclonal antibody, and Texas Red-X-conjugated phalloidin were used to confirm the osteocyte phenotype and to identify whole cells in the calvariae, respectively. Serial CLS images revealed morphological changes in bone cells up to 20 microm in depth. Osteocytes had widely spread their processes into the osteoblast layer, and we found for the first time that some of these processes had elongated to the vascular-facing surface of the osteoblast layer. Furthermore, stereotype images reconstructed from CLS images could show the three-dimensional distribution of these processes. Using the stereopair image, we could evaluate the frequency of processes between osteocytes and osteoblasts. Complementation of DIC microscopy revealed canaliculi and lacunae with high contrast. The distributional pattern of canaliculi generally coincided with that of the osteocyte processes. We consider that the combination method of CLS microscopy and DIC microscopy using a laser scanning microscope is a very useful new technical approach for investigating osteocytes in bone.

Animals↗
PubMed1977

[Selective contrast enhancement of microscopical specimen by optical procedures for automatized quantitative image analysis (author's transl)].

One of the most important problems in automatic image analysis is the discrimination of features by, a certain range of grey levels. To obtain as many as possible different grey levels sufficient contrast is achieved mainly by specific staining. Alternative and supplementary methods are some micrscopical methods, not routinely used. Interference microscopy, interference reflexion microscopy and microfluorimetry are discussed in detail. These optical procedures enhance the contrast of specimen specifically without the necessity for the application of specimen specifically without the necessity for the application of sophisticated staining methods. In interference contrast, tissues can be separately detected by grey level discrimination due to varying concentration of dry mass; this is shown for a cornifying part of fish skin (breeding tubercle of Rutilus rutilus L.). Furthermore, very small amounts of dry matter can be determined with high precision, as demonstrated for a single tissue culture cell (XTH-cell). Automatisation of image analysis provides a unique opportunity for routine application of interferometric measurements. The principles of the procedures are outlined. By interference reflexion microscopy cellular attachment areas to a glass surface are visualized, providing a powerful tool in cellular diagnosis based on grey level discrimination (darkest parts correspond to zones of closest contact to the substratum). A fast migrating lymphocyte and stationary endothelial cells have been chosen for demonstration. Various histochemical problems can be solved elegantly by fluorescence methods, e.g. mitochondria in living cells are specifically stained by a fluorochrome (DASPMI) and the distribution of fluorescence intensity can be followed within the mitochondrial population of a cell. Fluorescence was recorded from fotographic negatives taken with a fluorescence microscope. Additionally a short comment is given on the application of polarisation microscopy for feature detection.

Animals↗
PubMed1995

Cell-substrate interactions and locomotion of Dictyostelium wild-type and mutants defective in three cytoskeletal proteins: a study using quantitative reflection interference contrast microscopy.

Reflection interference contrast microscopy combined with digital image processing was applied to study the motion of Dictyostelium discoideum cells in their pre-aggregative state on substrata of different adhesiveness (glass, albumin-covered glass, and freshly cleaved mica). The temporal variations of the size and shape of the cell/substratum contact area and the time course of advancement of pseudopods protruding in contact with the substratum were analyzed. The major goal was to study differences between the locomotion of wild-type cells and strains of triple mutants deficient in two F-actin cross-linking proteins (alpha-actinin and the 120-kDa gelation factor) and one F-actin fragmenting protein (severin). The size of contact area, AC, of both wild-type and mutant cells fluctuates between minimum and maximum values on the order of minutes, pointing toward an intrinsic switching mechanism associated with the mechanochemical control system. The fluctuation amplitudes are much larger on freshly cleaved mica than on glass. Wild-type and mutant cells exhibit remarkable differences on mica but not on glass. These differences comprise the population median of AC and alterations in pseudopod protrusion. AC is smaller by a factor of two or more for all mutants. Pseudopods protrude slower and shorter in the mutants. It is concluded that cell shape and pseudopods are destabilized by defects in the actin-skeleton, which can be overcompensated by strongly adhesive substrata. Several features of amoeboid cell locomotion on substrata can be understood on the basis of the minimum bending energy concept of soft adhering shells and by assuming that adhesion induces local alterations of the composite membrane consisting of the protein/lipid bilayer on the cell surface and the underlying actin-cortex.

Animals↗
PubMed2005

Probing the structure of supported membranes and tethered oligonucleotides by fluorescence interference contrast microscopy.

Fluorescence interference contrast microscopy (FLIC) is a powerful method to structurally characterize fluorescent objects with nanometer-scale resolution in the z direction. Here we use FLIC to characterize the water layer underlying supported membranes and membrane-tethered double-stranded oligonucleotides. FLIC measurements of supported membranes containing the lipid-anchored fluorescent dye DiI in both leaflets indicate the thickness of the water layer separating the solid support and the lower lipid leaflet is 1.3 +/- 0.2 nm. Addition of cobalt(II) chloride to a DiI-supported membrane quenches the fluorescence in the top leaflet of the supported membrane; FLIC measurements of this system precisely locate the DiI to the bottom leaflet. These experiments confirm the accuracy of the model and parameters used to determine the water layer thickness, demonstrate the ability to differentiate between fluorescent objects whose average position differs by approximately 1.9 nm, and provide a widely applicable method to test the resolution of other high-z-resolution fluorescent microscopies. FLIC measurements of Alexa-labeled double-stranded oligonucleotides tethered to a supported membrane indicate that the DNA double helix is oriented perpendicular to the surface. Complications that arise from uncertainly in the orientation of the fluorophore are discussed. Several improvements in FLIC methodology are described. These stringent tests of the resolution of FLIC and the ability to unambiguously determine fluorescent lipid distribution provide structural insight on assemblies at membrane interfaces and permit the detection of even subtle changes at such interfaces.

Membranes, Artificial↗
PubMed2003

Rapid detection of biofilms and adherent pathogens using scanning confocal laser microscopy and episcopic differential interference contrast microscopy.

Knowledge of biofilm structure and function has changed significantly in the last few years due to advances in light microscopy. One pertinent example is the use of scanning confocal laser microscopy (SCLM) to visualise corrosion pits caused by the biofilm mosaic footprint on corroding metal surfaces. Nevertheless, SCLM has some limitations as to its widespread use, including cost, inability to observe motile bacteria and eukaryotic grazers within biofilms, and difficulty to scan a curved surface. By contrast, episcopic differential interference contrast (EDIC) microscopy has provided a rapid, real time analysis of biofilms on opaque, curved, natural or man-made surfaces without the need for cover slips and oil. EDIC, coupled with epi-fluorescence (EDIC/EF), microscopy has been used successfully to visualise the 3-D biofilm structure, physiological niches, protozoal grazing and iron biomineralization, and the location of specific pathogens such as Legionella pneumophila, Campylobacter jejuni and Cryptosporidium parvum. These species were identified using gold nanoparticles or fluorophores coupled to monoclonal antibodies or 16S rRNA probes, respectively. Among its many potential uses, the EDIC technique will provide a rapid procedure to facilitate the calibration of the modern generation of biofilm-sensing electrodes.

Animals↗
PubMed1995

Red blood cells, phase contrast, interference contrast microscopy and microspectrophotometry.

Following Teorell's (1) observation that the ghosts of hypotonically hemolysed erythrocytes reseal, it was shown that during the time they are permeable to hemoglobin, foreign macromolecules (dextran) can enter and that the hemolysed cell can achieve a final colloid-osmotic equilibrial state containing dextran and some residual Hb. In this way dextran reduces the hemoglobin loss in hypotonic hemolysis. Some hemoglobin loss is, however, inevitable, as it begins with a non-diffusive bulk outflow, sometimes observable as a jet, during which time a diffusive influx of the colloid-osmotic "balancer", dextran, is not possible. Finally, as expected from a process which is for the most part diffusive, transmembrane macromolecular transport is bidirectional; during hemolysis smaller molecules escape to a greater extent than larger ones.

Erythrocytes↗