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At least 1,135 records · Page 63Linked to original sources

Near-field imaging of ultrathin magnetic films with in-plane magnetization.

A new approach to near-field magneto-optical imaging was developed capable of visualization of in-plane magnetization of ultrathin magnetic structures. The approach relies on the magneto-optical effect specific for thin magnetic layers and employs near-field transmission measurements of longitudinal and/or transverse magneto-optical effect arising from the presence of thin film interfaces. The near-field magneto-optical contrast of in-plane domain structure of ultrathin Co film has been demonstrated in different polarization configurations.

Light↗

Confocal laser scanning microscopy and 3-D reconstructions of neuronal structures in human brain cortex.

Human brain material was studied with Lucifer yellow (LY) microinjections, indirect Texas red immunofluorescence, and confocal laser scanning microscopy (CLSM). The scanned images were transferred to a Silicon Graphics (SG) IRIS computer equipped with software for reconstructing the 3-D architecture of cells. By employing dual channel CLSM (Bio-Rad MRC 600), LY-injected cells and Texas red immunofluorescence could be studied simultaneously. Autopsy material with 2- to 48-h postmortem delays (6 control and 2 Rett's syndrome cases) as well as biopsy material (14 cases with therapy-resistant partial epilepsy--TRPE--undergoing neurosurgery) were used. In each specimen, 100-200 pyramidal and nonpyramidal neurons were visualized by LY microinjection. Single neurons were imaged and 2-D reconstructions of each neuron were made using z-projections of serial optical images; 3-D reconstructions and rotations were computed using the SG workstation, with VoxelView software from Vital Images (UK), and stored in a "neuronal library" on laser or magnetic optical disks. In Ret's syndrome cases and in patients with TRPE various abnormalities in the dendritic geometry of pyramidal and nonpyramidal cells have been found. The combination of LY injections with immunofluorescence allows the investigation of transmitter-related substances around the LY-injected cells. Using antibodies to synaptic vesicle proteins, presynaptic elements docking onto individual spines have been demonstrated. This approach may contribute to the understanding of different neurological and psychiatric disorders and may be useful in the Mapping of the Human Brain project. It may also be integrated with functional imaging by PET scan and with the human genome project.

Biopsy↗

[Three-dimensional imaging of the optic nerve using magnetic resonance angiography--application to anterior communicating artery aneurysm and craniopharingioma].

The purpose of this investigation was to analyze three-dimensional images of the optic nerve obtained by magnetic resonance angiography (MRA) in cases of anterior communicating artery aneurysm and craniopharingioma. Four ruptured anterior communicating artery aneurysms, five non-ruptured anterior communicating artery aneurysms and two craniopharingiomas were examined. The images were taken using MR/i Hispeed Plus 1.5T Infinity version, and analyzed by Advantage Work station AW4.1. The routine MR imaging parameters are shown in Table. The imaging time was about 10 minutes. Analysis was made by reformation of images parallel to the optic nerve obtained from the original MRA images. The optic nerve and brain tumor were traced with paintbrush from one sheet to another of the reformed images after subtraction of the blood vessels around the anterior communicating artery in these reformed images, and then three-dimensional images were constructed. Three-dimensional images of the blood vessels were reconstructed from MIP (maximum intensity projection) images using the threshold method. The optic nerve and anterior communicating arterial aneurysm or brain tumor were both observed in the overlapped 3D-SSD (shaded surface display) images. The analysis time was about 15 minutes. Three-dimensional images of the optic nerve and anterior communicating artery aneurysm or brain tumor were able to be made in all cases. As a preoperative investigation for anterior communicating artery aneurysm or suprasellar brain tumor, we considered that three-dimensional imaging of the optic nerve is useful in the operative approach because the optic nerve acts as a merkmal for the anterior communicating aneurysm or brain tumor.

Adult↗

Nanosecond freezing of water under multiple shock wave compression: optical transmission and imaging measurements.

Water samples were subjected to multiple shock wave compressions, generating peak pressures of 1-5 GPa on nanosecond time scales. This loading process approximates isentropic compression and leads to temperatures where the ice VII phase is more stable than the liquid phase above 2 GPa. Time resolved optical transmission and imaging measurements were performed to determine the solidification rate under such conditions. Freezing occurred faster at higher pressures as water was compressed further into the ice VII phase, in agreement with classical micleation theory. Water consistently froze when in contact with a silica window, whereas no solidification occurred in the presence of sapphire windows. The transition was determined to be a surface initiated process--freezing began via heterogeneous nucleation at the water/window interface and propagated over thicknesses greater than 0.01 mm. The first optical images of freezing on nanosecond time scales were obtained. These images demonstrate heterogeneous nucleation and irregular solid growth over 0.01-0.10 mm lateral length scales and are consistent with latent heat emission during the transformation. The combination of optical transmission and imaging measurements presented here provide the first consistent evidence for freezing on short time scales.

Journal Article↗

Screening unlabeled DNA targets with randomly ordered fiber-optic gene arrays.

We have developed a randomly ordered fiber-optic gene array for rapid, parallel detection of unlabeled DNA targets with surface immobilized molecular beacons (MB) that undergo a conformational change accompanied by a fluorescence change in the presence of a complementary DNA target. Microarrays are prepared by randomly distributing MB-functionalized 3-microm diameter microspheres in an array of wells etched in a 500-microm diameter optical imaging fiber. Using several MBs, each designed to recognize a different target, we demonstrate the selective detection of genomic cystic fibrosis related targets. Positional registration and fluorescence response monitoring of the microspheres was performed using an optical encoding scheme and an imaging fluorescence microscope system.

Base Sequence↗

Experimental realization of an imaging system with an extended depth of field.

We describe the experimental realization of an all-optical imaging system with an extended depth of field (DOF). The core of the system is a phase mask consisting of 16 Fresnel lenses (FLs) that are spatially multiplexed and mutually exclusive. Because each FL, in tandem with the primary lens, is designed to produce a sharp image for a specific object plane location, jointly the FLs achieve a wide DOF. However, the resultant image exhibits reduced resolution. The acquired image, onto which we did not apply any postprocessing, clearly is sharper than that acquired with a clear-aperture imaging system with the same pupil size.

Journal Article↗

Near-infrared fluorescence imaging of HER-2 protein over-expression in tumour cells.

The aim of this study was to evaluate in vitro and in vivo imaging of HER-2-over-expressing tumours using near-infrared optical imaging. A fluorochrome probe was designed by coupling Cy5.5 to anti-HER-2 antibodies. Cells over-expressing (SK-BR-3 cells) or normally expressing (PE/CA-PJ34 cells) the HER-2 protein were incubated with the probe. After removing unbound probe molecules, fluorescence intensities were determined (a.u.: arbitrary units). Cells were additionally investigated using FACS and laser scanning microscopy. The probe was also injected intravenously into tumours bearing SK-BR-3 ( n=3) or PE/CA-PJ34 ( n=3). Whole-body fluorescence images were generated and analysed. The incubation of SK-BR-3 cells with the probe led to higher fluorescence intensities [2,133 (+/-143) a.u.] compared to controls [975 (+/-95) a.u.]. The results from FACS and immunocytochemical analysis were in agreement with these findings. A distinct dependency between the fluorescence intensity and the cell number used in the incubations was detected. In vivo, the relative fluorescence intensities in SK-BR-3 tumours were higher than in PE/CA-PJ34 tumours at 16-24 h after probe application. HER-2-over-expressing tumours were depictable in their original size. Labelling of HER-2 with Cy5.5 is suitable for in vitro and in vivo detection of HER-2-over-expressing tumour cells.

Animals↗

The effect of hypercapnia on the neural and hemodynamic responses to somatosensory stimulation.

Modern non-invasive imaging techniques utilize the coupling between neural activity and changes in blood flow, volume and oxygenation to map the functional architecture of the human brain. An understanding of how the hemodynamic response is influenced by pre-stimulus baseline perfusion is important for the interpretation of imaging data. To address this issue, the present study measured hemodynamics with optical imaging spectroscopy and laser Doppler flowmetry, while multi-channel electrophysiology was used to record local field potentials (LFP) and multi-unit activity (MUA). The response to whisker stimulation in rodent barrel cortex was recorded during baseline (normocapnia) and elevated perfusion rates produced by two levels of hypercapnia (5 and 10%). With the exception of the 'washout' of deoxyhemoglobin, which was attenuated, all aspects of the neural and hemodynamic response to whisker stimulation were similar during 5% hypercapnia to those evoked during normocapnia. In contrast, 10% hypercapnia produced cortical arousal and a reduction in both the current sink and MUA elicited by stimulation. Blood flow and volume responses were reduced by a similar magnitude to that observed in the current sink. The deoxyhemoglobin 'washout', however, was attenuated to a greater degree than could be expected from the neural activity. These data suggest that imaging techniques based on perfusion or blood volume changes may be more robust to shifts in baseline than those based on the dilution of deoxyhemoglobin, such as conventional BOLD fMRI.

Anesthesia↗

Fabrication and characterization of probes for combined scanning electrochemical/optical microscopy experiments.

A technique that combines scanning electrochemical microscopy (SECM) and optical microscopy (OM) was implemented with a new probe tip. The tip for scanning electrochemicaVoptical microscopy (SECM/OM) was constructed by insulating a typical gold-coated near-field scanning optical microscopy tip using electrophoretic anodic paint. Once fabricated, the tip was characterized by steady-state cyclic voltammetry, as well as optical and electrochemical approach experiments. This tip generated a stable steady-state current and well-defined SECM approach curves for both conductive and insulating substrates. Durable tips whose geometry was a ring with < 1 microm as outer ring radius could be consistently fabricated. Simultaneous electrochemical and optical images of an interdigitated array electrode were obtained with a resolution on the micrometer scale, demonstrating good performance of the tip as both an optical and an electrochemical probe for imaging microstructures. The SECM feedback current measurements were successfully employed to determine tip-substrate distances for imaging.

Journal Article↗

[Free-vision refraction determination].

A refraction process is called free vision if the patient's visual field is not restricted at all, or at most by the same amount as when using normal spectacles. Free-vision refraction measurement is performed with the help of virtual lenses, which are real optical images of material lenses. In certain circumstances virtual lenses act as normal optical lenses. The advantage of free-vision refraction measurement lies above all in the fact that remote refraction measurement can also be performed from a distance of less than five meters. Moreover, it permits over-refraction, which enables the duration of the measuring procedure to be shortened considerably.

Eyeglasses↗

Noninvasive quantitative imaging of protein-protein interactions in living subjects.

We are developing methods to image molecular and cellular events in living subjects. In this study, we validate imaging of protein-protein interactions in living mice by using bioluminescent optical imaging. We use the well studied yeast two-hybrid system adapted for mammalian cells and modify it to be inducible. We employ the NF-kappaB promoter to drive expression of two fusion proteins (VP16-MyoD and GAL4-ID). We modulate the NF-kappaB promoter through tumor necrosis factor alpha. Firefly luciferase reporter gene expression is driven by the interaction of MyoD and ID through a transcriptional activation strategy. We demonstrate the ability to detect this induced protein-protein interaction in cell culture and image this induced interaction in living mice by using transiently transfected cells. The current approach will be a valuable and potentially generalizable tool to noninvasively and quantitatively image protein-protein interactions in living subjects. The approaches validated should have important implications for the study of protein-protein interactions in cells maintained in their natural in vivo environment as well as for the in vivo evaluation of new pharmaceuticals targeted to modulate protein-protein interactions.

Animals↗

Minimal stochastic complexity image partitioning with unknown noise model.

We present a generalization of a new statistical technique of image partitioning into homogeneous regions to cases where the family of the probability laws of the gray-level fluctuations is a priori unknown. For that purpose, the probability laws are described with step functions whose parameters are estimated. This approach is based on a polygonal grid which can have an arbitrary topology and whose number of regions and regularity of its boundaries are obtained by minimizing the stochastic complexity of the image. We demonstrate that efficient homogeneous image partitioning can be obtained when no parametric model of the probability laws of the gray levels is used and that this approach leads to a criterion without parameter to be tuned by the user. The efficiency of this technique is compared to a statistical parametric technique on a synthetic image and is compared to a standard unsupervised segmentation method on real optical images.

Algorithms↗

Analyzing focal adhesion structure by atomic force microscopy.

Atomic force microscopy (AFM) can produce high-resolution topographic images of biological samples in physiologically relevant environments and is therefore well suited for the imaging of cellular surfaces. In this work we have investigated focal adhesion complexes by combined fluorescence microscopy and AFM. To generate high-resolution AFM topographs of focal adhesions, REF52 (rat embryo fibroblast) cells expressing YFP-paxillin as a marker for focal adhesions were de-roofed and paxillin-positive focal adhesions subsequently imaged by AFM. The improved resolution of the AFM topographs complemented the optical images and offered ultrastructural insight into the architecture of focal adhesions. Focal adhesions had a corrugated dorsal surface formed by microfilament bundles spaced 127+/-50 nm (mean+/-s.d.) apart and protruding 118+/-26 nm over the substratum. Within focal adhesions microfilaments were sometimes branched and arranged in horizontal layers separated by 10 to 20 nm. From the AFM topographs focal adhesion volumes could be estimated and were found to range from 0.05 to 0.50 microm(3). Furthermore, the AFM topographs show that focal adhesion height increases towards the stress-fiber-associated end at an angle of about 3 degrees . Finally, by correlating AFM height information with fluorescence intensities of YFP-paxillin and F-actin staining, we show that the localization of paxillin is restricted to the ventral half of focal adhesions, whereas F-actin-containing microfilaments reside predominantly in the membrane-distal half.

Actin Cytoskeleton↗

Static point-spread function correction dominating higher-order speckle terms at high adaptive correction.

At high adaptive correction, the randomly shifting speckles familiar in conventional astronomical imaging become organized into patterns with distinct regularities that may permit partial suppression of the image noise they produce. Mathematically, the phase exponential in the Fourier-optical imaging expression may be expanded in a Taylor series in remnant phase phi, which is small at very high correction, leading to a perturbed point-spread function (PSF) that is a sum of algebraic terms, each of distinct spatial symmetry. At sufficiently high correction, one need deal with only a few of the lowest-order terms. A first-order expansion gives an ideal PSF plus two terms, linear and quadratic, describing the two brightest, physically most relevant kinds of speckle. A second-order expansion gives three new terms, the brightest of which is primarily a static correction to the PSF, with a much smaller true speckle component. When the correction is great enough to isolate individual speckle terms, the two terms from the first-order expansion alone determine the essential physics. A general observational strategy is outlined for reducing speckle noise in highly corrected companion searches, dominated by a few speckle terms of definite spatial symmetry.

Journal Article↗

Functional specificity of callosal connections in tree shrew striate cortex.

Although callosal connections have been shown to link extensive regions of primary visual cortex, the distribution of these connections with respect to the map of visual space and the map of orientation preference remains unclear. Here we combine optical imaging of intrinsic signals with injection of fluorescent microspheres to assess the functional specificity of callosal connections in the tree shrew. By imaging both hemispheres simultaneously while presenting a series of spatially restricted stimuli, we find that a substantial region of visual space is represented bilaterally. Each hemisphere includes a representation of the ipsilateral visual field that is highly compressed relative to that of the contralateral visual field and is most extensive in the lower visual field, where approximately 30(o) of central visual space are represented bilaterally. Callosal connections extend throughout the region of bilateral representation but terminate in a spatially restricted manner that links visuotopically corresponding sites in the two hemispheres. In contrast, callosal connections appear to terminate without regard for the map of orientation preference, showing little sign of the orientation-specific modular and axial specificity that is characteristic of long-range horizontal connections. By coordinating the activity in the two hemispheres in a way that preserves nearest neighbor relationships, callosal connections may best be viewed as elements of local circuits that operate within a single bilateral representation of visual space.

Animals↗

[Diagnosis of tuberculous pleural effusion].

The endoscopic-macroscopic images of thoracoscopies performed in the presence of a pleural effusion, are analysed. When comparing the various forms of the disease, the image must be resolved into its elements. It is found that the optical image is characteristic for the disease patterns; the appearance of the image elements also depends on the time at which the examination is performed.

Humans↗

Birefringence of single and bundled microtubules.

We have measured the birefringence of microtubules (MTs) and of MT-based macromolecular assemblies in vitro and in living cells by using the new Pol-Scope. A single microtubule in aqueous suspension and imaged with a numerical aperture of 1.4 had a peak retardance of 0.07 nm. The peak retardance of a small bundle increased linearly with the number of MTs in the bundle. Axonemes (prepared from sea urchin sperm) had a peak retardance 20 times higher than that of single MTs, in accordance with the nine doublets and two singlets arrangement of parallel MTs in the axoneme. Measured filament retardance decreased when the filament was defocused or the numerical aperture of the imaging system was decreased. However, the retardance "area," which we defined as the image retardance integrated along a line perpendicular to the filament axis, proved to be independent of focus and of numerical aperture. These results are in good agreement with a theory that we developed for measuring retardances with imaging optics. Our theoretical concept is based on Wiener's theory of mixed dielectrics, which is well established for nonimaging applications. We extend its use to imaging systems by considering the coherence region defined by the optical set-up. Light scattered from within that region interferes coherently in the image point. The presence of a filament in the coherence region leads to a polarization dependent scattering cross section and to a finite retardance measured in the image point. Similar to resolution measurements, the linear dimension of the coherence region for retardance measurements is on the order lambda/(2 NA), where lambda is the wavelength of light and NA is the numerical aperture of the illumination and imaging lenses.

Animals↗