共查询到20条相似文献,搜索用时 0 毫秒
1.
We investigate both theoretically and experimentally wavelength division multiplexed confocal imaging by using white light supercontinuum. We show that with the optimized pinhole diameter an axial resolution of 0.75 μm and detection efficiency of 80% can be achieved. In addition, we applied the axial WDM confocal system to 3D surface measurement and the result agreed well with that measured by commercially available surface profilometer. The measured sensitivity of the system is 3.25 nm. Finally, we demonstrated lateral confocal imaging by using supercontinuum. An effective lateral scanning range of 130 μm was obtained. 相似文献
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Reported in this paper are the experimental results on fibre-optical confocal scanning microscopy (FOCSM). We have performed various measurements in the FOCSM involving axial responses, edge responses, signal levels and interference axial responses in order to demonstrate the differences in confocal scanning imaging between using optical fibres and using pinholes. Experimentally measured results are compared with our theoretical predictions, revealing good agreements. It is shown convincingly that the FOCSM is a purely coherent imaging system as predicted in our previous theory. 相似文献
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Common path noise and disturbance in light source and ambient lighting affect detection accuracy of edge contours greatly in optical microscopy. In order to solve this problem, a lateral differential confocal microscopy is proposed based on principle of lateral difference and confocal microscopy. The approach proposed uses confocal dual-receiving light path arrangement and real-time heterodyne subtraction of two signals with lateral offsets by a differential detector to improve the system′s accuracy and detection sensitivity. Theoretical analyses have been presented. In addition, a simple prototype system has been built based on theoretical analyses. Related experiments have been performed under laboratory conditions. Different from former image processing methods, common path noise and additive disturbance in light source and ambient lighting are eliminated before they are recorded. Theoretical analyses and experimental results indicate a more accurate and sensitive result. 相似文献
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Confocal microscopy with serrated divided apertures is presented. Compared with using two smooth D-shaped apertures in confocal microscopy, the serrated divided apertures can suppress the out-of-focus central bright spot, thus increasing the efficiency of rejection of scattered light. Diffraction analysis shows that the serrated apertures maintain the optical sectioning strength while attenuating the background coming from far from the focal plane. In addition, the signal to background ratio (S/B) is also improved. 相似文献
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Confocal microscopy is widely used to measure the surface topography of specimen with a precision in the micrometer range. The measurement uncertainty and quality of the acquired data of confocal microscopy depends on various effects, such as optical aberrations, vibrations of the measurement setup and variations in the surface reflectivity. In this article, the influence of steep edges and undercuts on measurement results is examined. Steep edges on the specimen's surface lead to a reduced detector signal which influences the measurement accuracy and undercuts cause surface regions, which cannot be captured in a measurement. The article describes a method to overcome the negative effects of steep edges and undercuts by capturing several measurements of the surface with different angles between the surface and the optical axis of the objective. An algorithm is introduced which stitches different angle measurements together without knowledge of the exact position and orientation of the rotation axis. Thus, the measurement uncertainty due to steep edges and undercuts can be avoided without expensive high-precision rotation stages and time consuming adjustment of the measurement setup. 相似文献
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We discover that the slight transverse offset of a point detector results in a shift of the axial intensity response curve in a dual-axes confocal microscopy (DCM). Based on this, we propose a new dual-axes differential confocal microscopy (DDCM) with high axial resolution and long working distance, in which two point detectors are placed symmetrically about the collection axis. And a signal is obtained through the differential subtraction of two signals received simultaneously by the two point detectors. Theoretical analyses and preliminary experiments indicate that DDCM is feasible and suitable for the high precision tracing measurement of microstructures and surface contours. 相似文献
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Photobleaching was studied during recording of confocal scanning laser microscopy. Studies on fluorescent gels of FITC-labeled
dextran were used to evaluate differential bleaching along thez-axis. Differential bleaching along the z-axis was observed and it was seen that this was related to the numerical aperture
of the objective in use. This points to the conclusion that photon energy flux density is an important parameter in photobleaching.
To check if photon energy flux density heterogeneity is affected by local variation in the refractive index of the sample,
photobleaching rates were calculated for different fluorescent objects (sections of seeds, animal cells stained with nuclear
stains, immunocytochemistry preparations) and a pronounced similarity was found between photobleaching rates and DIC images. 相似文献
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In order to achieve a higher lateral resolution required for ultraprecision measurement of microstructural workpieces, phase-only pupil filtering differential confocal microscopy (PFDCM), a new approach is proposed based on the differential confocal microscopy (DCM), which uses a three-zone phase-only pupil filter with lateral super-resolution capability obtained through optimized design to change the distribution of DCM three-dimensional point spread function, so that the DCM lateral resolution is therefore significantly improved while its axial resolution is slightly improved. Preliminary experimental comparison and analyses indicate that, the lateral and axial resolutions of PFDCM are better than 0.2 μm and 2 nm, respectively, when wavelength of incidence laser beam , numerical aperture of measuring lens NA=0.85, and lateral spot size with a three-zone phase-only pupil filter GT=0.65. It is therefore concluded that PFDCM is a new approach to further improvement of lateral resolution in laser probe measurement systems. 相似文献
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With the growing demand for microoptics in different areas the importance of the characterization increases. Methods for a fast defect detection in microlens arrays are developed. We present a technique where the confocal principle is applied for determining the variation and the absolute value of the focal length. Additionally, using a self–filtering method the deviation of the periodic structure of microlens arrays is investigated theoretically and experimentally. Point-like defects as well as aberrations have been detected. The introduced methods allow the fast, parallel characterization of microlens arrays. 相似文献
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H.J. Tiziani R. Achi R.N. Krämer T. Hessler M.T. Gale M. Rossi R.E. Kunz 《Optics & Laser Technology》1997,29(2):85-91
A new; high resolution measuring system based on confocal microscopy has been developed for the evaluation of microlens arrays; in particular for applications in confocal microscopy itself. Lenslet arrays for parallel scanning and processing in confocal microscopy were designed as phase-matched Fresnel lenslets and fabricated by direct laser writing. Replica arrays were produced by ultraviolet embossing and hot embossing techniques. Fabricated arrays with a numerical aperture of 0.28 exhibited near diffraction limited performance and a focal length standard deviation of 120 nm in a nominal value of 250 μm. The technique developed represents a convenient and powerful technique for the characterization of lenslet arrays in general. 相似文献
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We propose a comprehensive treatment of theta microscopy based on dipole emission, which better describes fluorescence emission than the isotropic emission model, as fluorescence emission is often polarized. Formulas describing the point spread function for polarized confocal fluorescence theta microscopy are given. Examples are given and some advantages of polarized theta fluorescence microscopy are presented. To cite this article: O. Haeberlé et al., C. R. Physique 3 (2002) 1445–1450. 相似文献
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Fading or bleaching of fluorescence intensity during continuous illumination of stained objects is a serious problem in fluorescence microscopy. Fluorescence intensity as well as bleaching characteristics of dyes are dependent primarily upon physical parameters such as molecular constants (absorption rate and quantum efficiency), excitation energy and brightness (causes photon saturation), and environmental parameters (pH, ions, binding to proteins, etc.) that can strongly influence the properties of fluorochrome molecules. We have studied the effect of various antifading reagents on the behavior of the common dyes fluorescein isothiocyanate (FITC) and phycoerythrin (PE) using immunofluorescent-stained living cells in suspension or membrane-permeabilized dried cells as test systems. As expected, fading cannot be completely eliminated but may be reduced to varying degrees. In our hands, the most efficient antifading reagent for FITC isn-propyl gallate (NPG) dissolved in glycerol. No additive was found to retard fading, but complete dehydration of the cell suspension reduces this effect. 相似文献
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A sub-diffraction limit fluorescence localization microscope was constructed using a standard cooled 1.4 mega-pixel fluorescence charge-coupled device (CCD) camera to simultaneously resolve closely adjacent paired quantum dots on a flat surface with emissions of 540 and 630 nm. The images of the overlapping Airy discs were analyzed to determine the center of the point spread function after noise reduction using Fourier transformation analysis. The Cartesian coordinates of the centers of the point spread functions were compared in serial images. Histograms constructed from serial images fit well to Gaussian functions for resolving two quantum dots separated by as little as 10 nm in the x–y coordinates. Statistical analysis of multiple pairs validated discrimination of inter-fluorophore distances that vary by 10 nm. The method is simple and developed for x–y resolution of dilute fluorophores on a flat surface, not serial z sectioning. 相似文献
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A shaped annular beam superresolution approach is proposed to improve a lateral resolution of a small probe laser confocal measurement system (LCMS). The approach proposed enables lateral superresolution measurement of LCMS to be achieved by using a binary optical diffractive element to shape a He–Ne Gaussian laser beam into an annular beam with an inner diameter of 0.87 mm and an outer diameter of 1.8 mm required for superresolution measurement, and shift the beam spatial frequency from low to high. And a binary optical element (BOE) with 16 phase levels is designed and fabricated to shape a Gaussian laser beam into an annular beam. Preliminary experimental results indicate that an intensity distribution of a shaped annular beam is in agreement with simulation results, the diffractive efficiency is 87.2%; LCMS lateral and axial resolutions of 0.2 μm and 3 nm are achieved, respectively, and its measurement range is expanded nearly to double, when BOE is used in LCMS and , NA=0.85. 相似文献
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Neil Everall 《Journal of Raman spectroscopy : JRS》2014,45(1):133-138
This article compares the quality of Raman images obtained using metallurgical and oil immersion objectives to map complex structures in two and three dimensions. While the performance of these objectives for depth profiling planar structures has been discussed at length in the literature, the same comparison has not yet been made for 3D mapping of complex objects, where additional complications are introduced by non‐planar geometries. Studying samples with increasing complexity shows that the oil immersion objective is strongly preferred because it yields brighter images with better contrast, and eliminates some new and confusing artefacts that do not arise with simple planar objects. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
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Although confocal fluorescence microscopes are widely used in biology and have been proven to be promising diagnostic tools in dermatologic diagnostics, they are at present uncommon in medical practice. This is mainly due to high costs of acquisition and their large and complex outline. With the integration of a MEMS scanner we present a demonstration system of a confocal fluorescence laser scanning microscope which is affordable and portable. It has a field of view of 500 μm × 500 μm and is mainly composed of off-the-shelf components. 相似文献
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A three-dimensional (3D) resolution measure for the conventional optical microscope is introduced which overcomes the drawbacks of the classical 3D (axial) resolution limit. Formulated within the context of a parameter estimation problem and based on the Cramer-Rao lower bound, this 3D resolution measure indicates the accuracy with which a given distance between two objects in 3D space can be determined from the acquired image. It predicts that, given enough photons from the objects of interest, arbitrarily small distances of separation can be estimated with prespecified accuracy. Using simulated images of point source pairs, we show that the maximum likelihood estimator is capable of attaining the accuracy predicted by the resolution measure. We also demonstrate how different factors, such as extraneous noise sources and the spatial orientation of the imaged object pair, can affect the accuracy with which a given distance of separation can be determined. 相似文献