Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Microscopy, Interference”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,045 records · Page 58Linked to original sources
PubMed1997

Occurrence of birefringent retinal inclusions in cynomolgus monkeys after high doses of canthaxanthin.

PURPOSE: To reproduce and investigate in a primate animal model the phenomenon of the red carotenoid canthaxanthin (beta, beta-carotene-4'4'-dione) to induce crystal-like retinal deposits as they have been observed in the ocular fundus of humans after high canthaxanthin intake (i.e., more than 30 mg/day). METHODS: Groups of four cynomolgus monkeys (Macaca fascicularis) per gender and dose were administered 5.4, 16.2, or 48.6 mg canthaxanthin/kg body weight daily by oral gavage for 2.5 years. Eight control animals received placebo. In vivo ophthalmoscopy was performed at intervals of 3 months along with electroretinography after 12 and 24 months and retinal biomicroscopy just before the monkeys were killed. Retinal wholemounts or frozen sections were investigated postmortem by polarization, bright field, and differential interference contrast microscopy. Retinal and preterminal plasma canthaxanthin concentrations were determined by high-performance liquid chromatography (HPLC). RESULTS: By ophthalmoscopy and retinal biomicroscopy in vivo, no crystals or other light-reflecting particles were observed in the central paramacular retina. However, postmortem polarization microscopy of all 24 canthaxanthin-treated animals showed a circular zone in the peripheral retina containing birefringent, polymorphous red, orange, or white inclusions. The density of these inclusions was diminished within 1 to 8 mm posterior to the ora serrata. These inclusions were located mainly in the inner retinal layers, that is the nerve fiber layer and ganglion cell layer, inner plexiform layer, and inner nuclear layer. Twelve of the 24 canthaxanthin-treated animals showed yellow, golden birefringent inclusions in the macula. Retinas of placebo-treated monkeys were free of birefringent, crystal-like inclusions. The HPLC confirmed the presence of all-trans canthaxanthin, and 4-OH-echinenone and isozeaxanthin as well, in the retinas of all canthaxanthin-treated animals. Neither electroretinography nor histopathology indicated any adverse effects of the canthaxanthin-induced retinal inclusions seen in this study. CONCLUSIONS: A high intake of canthaxanthin for 2.5 years led to the deposition of crystal-like birefringent inclusions in the inner layers of the peripheral retina and, to some extent, the central retina of cynomolgus monkeys. The presence of these deposits did not interfere with morphology nor with retinal function.

Animals↗
PubMed1979

Sectioning of Epon blocks in the 20 m to 60 micromicron range for histological studies.

Thick sections of tissue, (20 micron--60 micron), are useful in studying the relationship between individual large cells and cell layers in organized neural structures. The ability of the Nomarski Differential Interference-Contrast Microscope to bring a single thin layer into sharp focus makes the examination of such sections feasible. Although celloidin is the classical embedding medium for large, thick sections of neural tissue, the time necessary for this preparation is most inconvenient. Epon is an excellent embedding medium; however, it is extremely hard and brittle. By heating the Epon block face, thick sections can be cut. To avoid the cumbersome, often detrimental use of heat, a modification of this technique was found. Epon blocks, trimmed to a 1 millimeter square face, may be sectioned at room temperature on the sliding microtome at 20 micron to 60 micron with ease. The simple method of preparing such sections is described.

Animals↗
PubMed2002

Epitope-mapping of transglutaminase with parallel label-free optical detection.

The gastrointestinal disorder coeliac disease (CD) is induced by the ingestion of wheat gluten and is characterized by damage of the typical structure of the intestinal mucosa. The enzyme tissue transglutaminase (tTGase) was identified as the major target of disease-specific antibodies in-patients. We performed an epitope fine-mapping with a series of pentadecapeptides synthesized using parallel multiple peptide synthesis. For the detection of biomolecular interactions a label-free parallel method, reflectometric interference spectroscopy (RIfS), was used. This is the first optical label-free method adapted to a high throughput screening (HTS) format and the experimental results demonstrate its applicability as a biological screening device. A high titer of anti-tTGase antibodies is found in the serum of coeliac patients. We have taken the first step towards a fast non-surgical test for the detection of these antibodies. In order to identify and characterize a continuous epitope with high affinity against the anti-tTGase antibody a screening of 21 pentadecapeptides has been accomplished with the parallel RIfS system. A single channel RIfS-system with high resolution was used to determine binding constants of identified peptides with high affinity.

Animals↗
PubMed1979

A scanning microinterferometer with correction of errors caused by overlapping 'ghost' images.

An experimental setup has been built to correct the errors caused by overlapping 'ghost' images when scanning and integrating measurements are carried out with the Jamin-Libedef interference microscope. First a reference field with the object, producing the 'ghost' image, is scanned and the values of the optical path difference (OPD) of each point in the reference field are stored in a computer. Subsequently, the OPD of the object to be measured is calculated for each point by adding the measuring result to the average OPD of five points, located around the corresponding point in the reference field. The applicability of the setup has been tested by measurements with Sepharose beads, Hela cells and the Zeiss microinterference refractometer. The difference between measurements on the same object with and without overlapping 'ghost' image was about 10%. The reproducibility was tested by repeated measurements on the same cell whereby a standard deviation of 1.1% was found.

Computers↗
PubMed1976

The M. omohyoideus of the mouse as a convenient mammalian muscle preparation. A study of junctional and extrajunctional acetylcholine receptors by noise analysis and cooperativity.

Muscles from cats, rats, guinea pigs and mice have been investigated as preparations for visualizing mammalian neuromuscular junctions with the aid of Nomarski interference optics. The M. omohyoideus of the mouse was found to be most convenient. Electrophysiological investigations showed that an endplate is normally surrounded by a population of perijunctional receptors. For junctional receptors in the endplate, a Hill coefficient of nH = 2.6 for acetylcholine was determined at 38 degrees C, decreasing to a value of 2.3 at room temperature. For both perijunctional and extrajunctional receptors (the latter occurring after denervation), the coefficient nH was 1.9. Noise analysis revealed a channel conductance gamma which changed abruptly from 22.4 +/- 1.0 pS (10--23 degrees C) to 45.6 +/- 3.9 pS (34--39 degrees C) in a very small temperature range around 25.5 degrees C. The mean channel lifetime tau was 0.3 ms at 39 degrees C and 1.0 ms at 23 degrees C.

Animals↗
PubMed1993

Degeneration of human respiratory cell ciliary beat in monolayer cell cultures.

The degeneration of ciliary beat of human respiratory cells was studied in monolayer cell cultures by using a differential interference microscope equipped with a high speed video system. This method for studying ciliary beat in cell cultures on collagen-coated cover glasses is quite advantageous, because it allows for detailed study of all parts of ciliary function and not just ciliary beat frequency (CBF). In the present study both CBF and ciliary beat amplitude (CBA) were found to decrease continuously from the 1st day after plating but the wave form of ciliary beat did not change. Cultures with high cell density provided better preservation of normal ciliary beat for a longer period. In contrast, ciliary beat degenerated quickly in cultures with low cell density. CBF and CBA in cell cultures less than 5 days after plating were always high, supporting use of these cultures for studies of normal ciliary motility.

Cell Movement↗
PubMed2004

Ultrahigh-resolution full-field optical coherence tomography.

We have developed a white-light interference microscope for ultrahigh-resolution full-field optical coherence tomography of biological media. The experimental setup is based on a Linnik-type interferometer illuminated by a tungsten halogen lamp. En face tomographic images are calculated by a combination of interferometric images recorded by a high-speed CCD camera. Spatial resolution of 1.8 microm x 0.9 microm (transverse x axial) is achieved owing to the extremely short coherence length of the source, the compensation of dispersion mismatch in the interferometer arms, and the use of relatively high-numerical-aperture microscope objectives. A shot-noise-limited detection sensitivity of 90 dB is obtained in an acquisition time per image of 4 s. Subcellular-level images of plant, animal, and human tissues are presented.

Animals↗
PubMed2006

Laser-scanning lithography (LSL) for the soft lithographic patterning of cell-adhesive self-assembled monolayers.

We report the development of laser-scanning lithography (LSL), which employs a laser-scanning confocal microscope to pattern photoresists that can be utilized, for example, in the fabrication of masters for use in soft lithography. This convenient technique provides even exposure across the entire view field and facilitates accurate alignment of successive photoresist exposures. Features on the scale of 3 microm have been achieved to date with a 10x objective (NA 0.45). Virtual masks, instructions for laser irradiation, were drawn using the Region of Interest (ROI) function of a Zeiss LSM 510 microscope. These regions were then exposed to a 458 nm argon laser for 32 micros (0.9 mW/microm(2)). Differential interference contrast (DIC) imaging was utilized with a non-destructive 514 nm argon laser as an immediate quality check of each exposure, to align successive exposures, and to reduce chromatic aberration between imaging and exposure. Developed masters were replica-molded with poly(dimethylsiloxane) (PDMS); these masters were then utilized for microcontact printing of cell-adhesive self-assembled monolayers (SAMs) to demonstrate the utility of this process. Initial studies confirmed that human dermal fibroblast adhesion and spreading were limited to cell-adhesive SAM areas. LSL is a rapid, flexible, and readily available technique that will accelerate master design and preparation; moreover, it can be applied to additional forms of photolithography and photopolymerization for studies in cell biology, biomaterials design and evaluation, materials science, and surface chemistry.

Alkanes↗
PubMed2003

Cell membrane alignment along adhesive surfaces: contribution of active and passive cell processes.

Cell adhesion requires nanometer scale membrane alignment to allow contact between adhesion receptors. Little quantitative information is presently available on this important biological process. Here we present an interference reflection microscopic study of the initial interaction between monocytic THP-1 cells and adhesive surfaces, with concomitant determination of cell deformability, using micropipette aspiration, and adhesiveness, using a laminar flow assay. We report that 1), during the first few minutes after contact, cells form irregular-shaped interaction zones reaching approximately 100 micro m(2) with a margin extension velocity of 0.01-0.02 micro m/s. This happens before the overall cell deformations usually defined as spreading. 2), These interference reflection microscopic-detected zones represent bona fide adhesion inasmuch as cells are not released by hydrodynamic forces. 3), Alignment is markedly decreased but not abolished by microfilament blockade with cytochalasin or even cell fixation with paraformaldehyde. 4), In contrast, exposing cells to hypotonic medium increased the rate of contact extension. 5), Contacts formed in presence of cytochalasin, after paraformaldehyde fixation or in hypotonic medium, were much more regular-shaped than controls and their extension matched cell deformability. 6), None of the aforementioned treatments altered adhesiveness to the surface. It is concluded that adhesive forces and passive membrane deformations are sufficient to generate initial cell alignment to adhesive surfaces, and this process is accelerated by spontaneous cytoskeletally-driven membrane motion.

Cell Adhesion↗
PubMed1970

The optics of the compound eye of the honeybee (Apis mellifera).

The optical system of the compound eye of the worker honeybee, as a representative of the closed-rhabdom type of eye, was investigated and its function analyzed. Measurements of refractive indices of the elements of the optical system were made with an interference microscope. With the use of the resulting measurements, the optical system was analyzed by means of a ray-tracing procedure implemented for the IBM 7094 digital computer, and by means of the Gaussian thick lens formulae. The more detailed results of the ray-tracing technique were used for further analyses. Direct visual confirmation of the focal point was obtained. The rhabdom and the surrounding zone of lower refractive index act together as a wave guide, as demonstrated by the presence of several wave guide modes in the rhabdom. An admittance function was defined as the percentage of the rays reaching the rhabdom with respect to those entering the ommatidium. Good agreement with experimental results was found. The characterization of the visual field of an ommatidium by means of an admittance function permits the analysis of the influence of different stimuli on the eye.

Animals↗
PubMed1993

Effect of exogenous ATP on ciliary beat of human ciliated cells studied with differential interference microscope equipped with high speed video.

To study the detailed effects of exogenous adenosine triphosphate (ATP) on ciliary function, we used the differential interference microscope equipped with high speed video and evaluated ciliated cells from the human sinus mucosa in monolayer culture. With this system it was possible to evaluate all parts of ciliary motility with a minimum of interference from the mucous membrane, secretory cells and the autonomic nervous system. The best direct ciliostimulative effect of exogenous ATP on ciliary beat frequency (CBF) and ciliary beat amplitude (CBA) was observed at concentrations of ATP ranging from 10(-5) M to 10(-3) M. Exogenous ATP appeared to normalize the slightly damaged ciliary motility in groups with an initial CBF of 14 Hz or higher. When the CBF was less than 14 Hz, ATP produced an increase in CBF greater than 40%, an increase in CBA greater than 30%, but these did not reach the normal level in 5 min. The biggest increases: 59.9% in CBF and 40.7% in CBA were seen in the group with an initial CBF less than 9 Hz. In the cells with a low initial CBF and unsynchronized motility exogenous ATP increased the synchrony of ciliary movement together with an increase in CBF and CBA.

Adenosine Triphosphate↗
PubMed1998

Two-dimensional tracking of ncd motility by back focal plane interferometry.

A technique for detecting the displacement of micron-sized optically trapped probes using far-field interference is introduced, theoretically explained, and used to study the motility of the ncd motor protein. Bead motions in the focal plane relative to the optical trap were detected by measuring laser intensity shifts in the back-focal plane of the microscope condenser by projection on a quadrant diode. This detection method is two-dimensional, largely independent of the position of the trap in the field of view and has approximately 10-micros time resolution. The high resolution makes it possible to apply spectral analysis to measure dynamic parameters such as local viscosity and attachment compliance. A simple quantitative theory for back-focal-plane detection was derived that shows that the laser intensity shifts are caused primarily by a far-field interference effect. The theory predicts the detector response to bead displacement, without adjustable parameters, with good accuracy. To demonstrate the potential of the method, the ATP-dependent motility of ncd, a kinesin-related motor protein, was observed with an in vitro bead assay. A fusion protein consisting of truncated ncd (amino acids 195-685) fused with glutathione-S-transferase was adsorbed to silica beads, and the axial and lateral motions of the beads along the microtubule surface were observed with high spatial and temporal resolution. The average axial velocity of the ncd-coated beads was 230 +/- 30 nm/s (average +/- SD). Spectral analysis of bead motion showed the increase in viscous drag near the surface; we also found that any elastic constraints of the moving motors are much smaller than the constraints due to binding in the presence of the nonhydrolyzable nucleotide adenylylimidodiphosphate.

Calibration↗
PubMed2004

Effects of mannitol on ischemia-induced degeneration in rat hippocampus.

Although mannitol has been used as an osmotherapeutic drug on brain injury, the clinical efficiency of the drug are still controversial. In the present study, we examined the effects of mannitol on the edema in a hippocampal slice due to brief ischemia. To evaluate the effects, we employed an image analysis system that consists of an infrared-differential interference contrast (IR-DIC) microscope, an infrared CCD camera, and a computer with custom-made software. By this system, severity of the edema can be quantified as the coefficient of variation (CV) of digitalized slice images. The dose-dependent improvement on the deteriorated hippocampal slices could be obtained by administration of mannitol (10, 50, and 100 mM) after 10-min ischemia. However, field excitatory postsynaptic potentials (fEPSP) in CA1 stratum radiatum, which disappeared during 10-min ischemia, were never recovered by mannitol after more than 20-min treatment. fEPSP were blocked by the effective dose of mannitol for morphological recovery, but the effects found to be reversible. Although we failed to find positive rescuing effects of mannitol on the synaptic activities after ischemia, the protective effects of the drug on ischemic edema may rescue the secondary damages around the infarct area.

Animals↗
PubMedDate not supplied

Differential interference contrast and confocal reflectance imaging of collagen organization in three-dimensional matrices.

The remodeling of extracellular matrices by cells plays a defining role in developmental morphogenesis and wound healing as well as in tissue engineering. Three-dimensional (3-D) type I collagen matrices have been used extensively as an in vitro model for studying cell-induced matrix reorganization at the macroscopic level. However, few studies have directly assessed the process of 3-D extracellular matrix (ECM) remodeling at the cellular and subcellular level. In this study, we directly compare two imaging modalities for both quantitative and qualitative imaging of 3-D collagen organization in vitro: differential interference contrast (DIC) and confocal reflectance imaging. The results demonstrate that two-dimensional (2-D) DIC images allow visualization of the same population of collagen fibrils as observed in 2-D confocal reflectance images. Thus, DIC can be used for qualitative assessment of fibril organization, as well as tracking of fibril movement in sequential time-lapse 2-D images. However, we also found that quantitative techniques that can be applied to confocal reflectance images, such as Fourier transform analysis, give different results when applied to DIC images. Furthermore, common techniques used for 3-D visualization and reconstruction of confocal reflectance datasets are not generally applicable to DIC. Overall, obtaining a complete understanding of cell-matrix mechanical interactions will likely require a combination of both wide-field DIC imaging to study rapid changes in ECM deformation which can occur within minutes, and confocal reflectance imaging to assess more gradual changes in cell-induced compaction and alignment of ECM which occur over a longer time course.

Cell Line↗
PubMed1999

Extrinsic Fabry-Perot interferometer for measuring the stiffness of ciliary bundles on hair cells.

We have developed an extrinsic Fabry-Perot interferometer (EFPI) to measure displacements of microscopic, living organelles in the inner ear. The EFPI is an optical phase-shifted instrument that can be used to measure nanometer displacements. The instrument transmits a coherent light signal to the end of a single glass optical fiber where the measurement is made. As the coherent light reaches the end of the fiber, part of this incident signal is reflected off the internal face of the fiber end (reference reflection) and part is transmitted through the end of the fiber. This transmitted light travels a short distance and is reflected off the surface whose displacement is to be measured (the target). This sensing reflection then reenters the fiber where it interferes with the reference reflection. The resulting interference signal then travels up the same optical fiber to a detector, where it is converted into a voltage that can be read from an oscilloscope. When the target moves, the phase relation between reference and sensing reflections changes, and the detector receives a modulated signal proportional to the target movement. Reflections of as little as 1% at both the sensor tip and target surfaces produce good results with this system. We use the EFPI in conjunction with fine glass whiskers to measure the stiffness (force per unit deflection) of stereociliary bundles on hair cells of the inner ear. The forces generated are in the tenths of picoNewton range and the displacements are tens of nanometers. Here we describe the EFPI and its development as a method for measuring displacements of microscopic organelles in a fluid medium. We also report experiments to validate the accuracy of the EFPI output and preliminary measurements of ciliary bundle stiffness in the posterior semicircular canal.

Animals↗
PubMed2002

Simultaneous three-dimensional step-height measurement and high-resolution tomographic imaging with a spectral interferometric microscope.

A noncontact, nonmechanical scanning, wide-field spectral interference microscope is developed for simultaneous measurement of three-dimensional step-height of discontinuous objects and tomographic imaging. A superluminescent diode (SLD) is used as a broadband light source and a liquid-crystal Fabry-Perot interferometer (LC-FPI) as a frequency-scanning device. By means of changing the injection current to the SLD, the spectral profile of the SLD is equalized, and a constant light input to the interferometer is achieved over the entire frequency-scan range. The Fourier-transform technique is used to determine both the amplitude and the phase of spectral fringe signals. Three-dimensional height distribution of a discontinuous object is obtained from the phase information, whereas optically sectioned images of the object are obtained either from the amplitude information alone or from the combination of both the amplitude and phase information. Experimental results with submicrometer resolution are presented for both step-height measurement and tomographic sectioning.

Imaging, Three-Dimensional↗
PubMed1988

Differential response to compressive loads of zones of canine hyaline articular cartilage: micromechanical, light and electron microscopic studies.

Thin (100 micron) perpendicular slices of canine femoral condylar cartilage were placed horizontally on the stage of a Nachet microscope and viewed by transmitted light in the differential interference contrast mode. Each slice was held on the microscope stage by a loading rig and tested mechanically in compression. Measured loads to a maximum of approximately 2-3 MN/m2 were applied to the end of the slice corresponding to the articular surface. Photographs were taken of the cartilage before and during loading, and the distance by which selected chondrocytes were displaced was used as an index of mechanical strain, i.e., of change in length/original length. Maximum strains were observed in the superficial cartilage zone. Minimum strains were recorded in the mid-zone, at a depth corresponding to approximately 75% of the total cartilage thickness. The relative concentrations of cartilage collagen (COL) and proteoglycan (PG) were assessed by the light and electron microscopic histochemical study of cartilage sections taken from contiguous blocks. Superficial cartilage, which deformed most, had high concentrations of oriented COL fibres, low concentrations of PG. Mid-zone cartilage, which deformed least, had lower concentrations of randomly arrayed COL fibres but relatively high concentrations of PG.

Animals↗
PubMed1975

Computer analysis of organelle translocation in primary neuronal cultures and continuous cell lines.

Organelle translocation in a number of cell types in tissue culture as seen by high-resolution Zeiss-Nomarski differential interference contrast optics was filmed and analyzed by computer. Principal cell types studied included primary chick spinal cord, chick dorsal root ganglion, ratbrain, and various clones of continuous cell lines. Organelle translocations in all cell types studied exhibited frequent, large changes in velocity during any one translocation. The appearance of particles as seen with Nomarski optics was correlated with their fine structures in one dorsal root ganglion neurite by fixing the cell as it was being filmed and obtaining electron micrographs of the region filmed. This revealed the identity of several organelles as well as the presence of abundant neurotubules but no neurofilaments. Primary cell cultures exhibited more high-velocity organelle movements than continuous cell lines. The net progress of an organelle in a given direction was greater in primary neuronal cells than in fibroblasts or continuous cell lines. These findings are correlated with the literature on organelle translocation and axoplasmic transport.

Animals↗