Direct labeling and visualization of blood vessels with lipophilic carbocyanine dye DiI
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Confocal interference microscopy can be used to obtain phase information in a confocal microscope. Different methods of recovering phase are discussed. A fiber optic confocal interference microscope has been developed. Confocal interference can be used to investigate aberrations in a confocal microscope. Methods of profiling surfaces are reviewed. Depth profiling of stratified media is considered.
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Confocal theta microscopy is a novel method by which the axial resolution in confocal fluorescence microscopy can be substantially improved. The basic idea is to observe the sample with two or more objective lenses and to detect the emission light at an angle theta to the illumination axis. The observation volume is considerably decreased when the detection axis is rotated by 90 degree(s) relative to the illumination axis. This leads to an almost spherical observation volume, which is three times smaller than in a comparable confocal fluorescence microscope. Confocal theta microscopy can be combined with 4Pi-confocal microscopy and is the first viable method proposed for fully exploiting the resolution increase achievable with 4Pi-techniques. The axial side lobes of the 4Pi-point spread function are suppressed, and the observation volume is reduced by a factor of 4. The resolution properties of confocal theta fluorescence microscopies are investigated for single- and two-photon absorption. Evaluations of confocal theta point spread functions are presented and the resolution improvement achieved by theta observation is discussed.
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Precise examination of the corneal endothelium has become increasingly important due to the growing number of intraocular and corneal procedures. The purpose of this study was to compare prospectively the corneal endothelial cell count in normal eyes obtained by confocal and specular microscopy. Central corneal endothelial cell densities of 42 eyes from 42 patients were measured by confocal and contact specular microscopy. Endothelial cells were analyzed with the same software in a manual, an automated and a semi-automated mode. The mean endothelial cell density obtained by confocal microscopy was (in the manual, automated and semi-automated modes) 3,069 ± 285, 2,791 ± 344 and 3,077 ± 286 cells/mm<sup>2</sup>, and obtained by specular microscopy 3,076 ± 298, 2,796 ± 271 and 3,082 ± 282 cells/m<sup>2</sup>, respectively. No statistically significant difference of endothelial cell density between confocal and specular microscopy was found. Endothelial cell count was significantly lower in the automated than in the semi-automated and manual analysis both with confocal and with specular microscopy. In conclusion, endothelial cell count measurements with confocal and contact specular microscopy are comparable.
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Due to shrinkage of design rule, optical in-line defect inspection with white-light source is reaching its detection limit. To overcome the limitation, a defect inspection system using UV confocal microscopy was recently introduced. In this paper, we investigated characteristics of UV confocal microscopy, which is confocal microscopy using UV light source, by analyzing TDI images captured by a defect inspection system with UV confocal microscopy. The results of this study showed that UV confocal microscopy has higher sensitivity and is more efficient for detection of 0.1 /spl mu/m-level small defects rather than white-light source or conventional microscopy.
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Three-dimensional (3D) Dammann confocal microscopy is proposed based on introducing 3D Dammann gratings into traditional confocal microscopy. The conventional confocal microscopy usually has a single focal point. Using threedimensional Dammann gratings, it shows a new confocal microscopy which could obtain three-dimensional information of object, therefore, novel Dammann-based microscopy should be developed for practical applications.
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Fluorescence confocal laser scanning microscopy and reflectance confocal laser scanning microscopy are up-to-date highend study methods. Confocal microscopy is used in cell biology and medicine. By using confocal microscopy, it is possible to study bioplasts and localization of protein molecules and other compounds relative to cell or tissue structures, and to monitor dynamic cell processes. Confocal microscopes enable layer-by-layer scanning of test items to create demonstrable 3D models. As compared to usual fluorescent microscopes, confocal microscopes are characterized by a higher contrast ratio and image definition.
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We present an overview of recent theories for describing specimen‐induced spherical aberration in confocal microscopy. One of these theories is used to compute numerically the role of spherical aberration in general confocal, and especially in biological confocal, microscopy for a variety of three‐layer specimen structures. In particular, we study the effect of specimen‐induced spherical aberration on the maximum value of the overall confocal point spread function, the accompanying focal shift and the size of the optical probe in both fluorescence and brightfield confocal microscopy.
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