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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX"

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Near-field optical and atomic force constraints for super-resolution 3D deconvolution in far-field optical microscopy

Near-field optical and atomic force constraints for super-re...

引用

Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Axelrod, N Radko, A Ben-Yosef, N Khatchatouriants, A Treinin, M Lewis, A Hebrew Univ Jerusalem Dept Appl Phys IL-91904 Jerusalem Israel

ISBN: (纸本)081943535X

We demonstrate that near-field optical and atomic force microscopy data can be used for super-resolution three-dimensional (3-D) image restoration in optical sectioning fluorescence microscopy. A crucial feature in this approach is the full integration of such data sets with digital far-field images. The scanned probe data is used to provide modalities for boundary constraints which define surface optical information and spatial domains of optical alterations in a sample with a spatial precision that has been unachievable in the past. A restoration algorithm that can use such a complex of data for 3-D image deconvolution is presented. It uses a Tikhonov-Miller regularization scheme and allows for the imposition of different types of constraints to obtain super-resolution deconvolved images. Performance was tested by using simulated 3-D imaging. An example is given of the restoration of a 3-D wide field optical image of a biological specimen in the presence of atomic force constraints.

关键词： image restoration deconvolution algorithm constraints probe microscopy NSOM super-resolution

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Variable-focus microlenses as a potential technology for endoscopy

Variable-focus microlenses as a potential technology for end...

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Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Smith, PJ McCabe, EM Taylor, CM Selviah, DR Day, SE Commander, LG Univ Dublin Trinity Coll Dept Phys Dublin 2 Ireland

ISBN: (纸本)081943535X

Using a new device which contains an array of microlenses whose focal lengths can be electrically varied, we have been able to control the input from one microlens to a single mode fibre using an applied voltage. For such a microlens array many closely-spaced focal spots can be generated in parallel, and electrically switched to address, potentially, an array of receiver fibres. We show how the particular switching characteristics of the device, whereby the lenses switch from diverging to converging, serves in turn to disperse light and to focus it into the fibre.

关键词： microlenses endoscopy direct view microscopy

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Generation and imaging with the third harmonic

Generation and imaging with the third harmonic

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Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Brakenhoff, GJ Squier, J Univ Amsterdam BioCentrum Dept Mol Cyt NL-1098 SM Amsterdam Netherlands

ISBN: (纸本)081943535X

In this paper we study the spatial distribution and wavefront characteristics of third harmonic generation in relation to some material and interface conditions over the focal region of the fundamental beam. We investigate, mostly from an experimental point of view, the implications the physics of the THG generation process has in situations where THG may be employed for -3D- imaging. Due to the non-linear character of the THG generation process it is inherently suitable for this application. For the first time images of the distribution of the THG radiation, as the interface is moved through focus, are shown. Experiments on closely spaced interfaces or bilayers confirm unambiguously the correctness of the vector model for THG generation (Ward et al. 1969) in uniform media. In view of these and other data the image formation, especially for biological objects, with THG radiation will be discussed.

关键词： THG third harmonic microscopy imaging

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Adaptive compensation of aberrations in ultrafast 3D microscopy using a deformable mirror

Adaptive compensation of aberrations in ultrafast 3D microsc...

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Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Sherman, L Albert, O Schmidt, C Vdovin, G Mourou, G Norris, TB Univ Michigan Ctr Ultrafast Opt Sci Ann Arbor MI 48109 USA

ISBN: (纸本)081943535X

3D imaging using a multiphoton scanning confocal microscope is ultimately Limited by aberrations of the system. We describe a system to adaptively compensate the aberrations with a deformable mirror. We have increased the transverse scanning range of the microscope by three with compensation of off-axis aberrations. We have also significantly increased the longitudinal scanning depth with compensation of spherical aberrations from the penetration into the sample. Our correction is based on a genetic algorithm that uses second harmonic or two-photon fluorescence signal excited by femtosecond pulses from the sample as the enhancement parameter. This allows us to globally optimize the wavefront without a wavefront measurement. To improve the speed of the optimization we use Zernike polynomials as the basis for correction. Corrections can be stored in a database for look-up with future samples.

关键词： scanning confocal microscopy aberration correction

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Multiple beam interference confocal microscopy - a tool for morphological investigation of living cells and tissues

Multiple beam interference confocal microscopy - a tool for ...

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Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Joshi, NV Medina, H Univ Los Andes Fac Med Dept Physiol Merida 5101 Venezuela

ISBN: (纸本)081943535X

Multiple beam interference system is used in conjunction with a conventional scanning confocal microscope to examine the morphology and construction of 3-D images of Histolytic Ameba and parasite Candida Albicans. The present combination permits to adjoin advantages of both systems, namely the vertical high contrast and optical sectioning. The interference pattern obtained from a multiple internal reflection of a sample, sandwiched between the glass plate and the cover plate, was focussed on an objective of a scanning confocal microscope. According to optical path differences, morphological details were revealed. The combined features, namely improved resolution in z axis, originated from the interference pattern and the optical sectioning of the confocal scanning system, enhance the resolution and contrast dramatically. These features permitted to obtain unprecedented images of Histolytic Ameba and parasite Candida Albicans. Because of the improved contrast, several details like double wall structure of candida, internal structure of ameba are clearly visible.

关键词： Confocal microscopy Microscopes Digital image correlation Glasses Tissues Refractive index 3D image processing microscopy Optical scanning systems Phase contrast

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Optical characterization of microfabricated solid immersion lenses

Optical characterization of microfabricated solid immersion ...

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Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Fletcher, DA Crozier, KB Goodson, KE Quate, CF Kino, GS Stanford Univ Ginzton Lab Stanford CA 94305 USA

ISBN: (纸本)081943535X

Circumventing the optical diffraction limit is important for data storage, photolithography, and imaging. The Solid Immersion Lens (SIL), in which light is focussed through a high refractive-index lens held close to a sample, offers a method for improving resolution. Microfabrication of SILs enables the use of lenses as small as a few wavelengths in diameter. When these are integrated onto cantilevers, the SIL can be scanned over topography with control of lens-sample separation. We have extended Mie theory to investigate aberration in small lenses and the effect of lens size on a converging beam. We also report progress on the fabrication and first results from micromachined silicon SILs.

关键词： microlens microfabrication solid immersion lens scanning probe microscopy

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Super-resolution stereo light microscopy for real-time tasks using normal flow and geometric constraints

Super-resolution stereo light microscopy for real-time tasks...

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Conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Danuser, G Swiss Fed Inst Technol ETH BioMicroMetr Grp Lab Biomech CH-8952 Schlieren Switzerland

ISBN: (纸本)081943535X

Prior knowledge about the observed scene represents the key to recovering frequencies beyond the passband of an imaging system (super-resolution). With regard to microscopy mainly two super-resolution mechanisms have been reported: 1.) analytic continuation of the frequency spectrum and 2.) constrained image deconvolution. This paper describes an alternative super-resolution mechanism. Prior knowledge is introduced on a higher, more symbolic level of visual inference. We exemplify our concept based on the 3D reconstruction of a micro-pipette moving in close proximity to immobile target objects. Information about the shape and the mobility of the pipette is incorporated in order to localize the pipette tip at sub-Rayleigh distances to the target. The algorithm was tested in a micro-robot environment. A machine vision module using stereo light microscopy automatically controlled the manipulation of microscopic objects, e.g. latex beads or diamond mono-crystals. In the theoretical part of the paper we prove that knowledge of the form "the pipette has moved between two consecutive frames of the movie" must result in a twofold increase of resolution. We used the normal flow of an image to decode positional measures from motion. In practice, super-resolution factors between 3 to 5 were obtained. The additional gain originates from the geometric constraints which were included to reconstruct the pipette position.

关键词： light microscopy stereo super-resolution machine vision normal flow geometric constraints

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Modal wavefront sensor for adaptive confocal microscopy

Modal wavefront sensor for adaptive confocal microscopy

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three-dimensional and multidimensional microscopy: image acquisition processing VII

作者： Booth, Martin J. Neil, Mark A.A. Wilson, Tony Univ of Oxford Oxford United Kingdom

A confocal microscope permits 3-D imaging of volume objects by the inclusion of a pinhole in the detector path which eliminates out of focus light. This configuration is however very sensitive to aberrations induced by the specimen or the optical system and would therefore benefit from an adaptive optics approach. We present a wavefront sensor capable of measuring directly the Zernike components of an aberrated wavefront and show that it is particularly applicable to the confocal microscope since only those wavefronts originating in the focal region contribute to the measured aberration.

关键词： Medical imaging

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Doubling the lateral resolution of wide-field fluorescence microscopy using structured illumination

Doubling the lateral resolution of wide-field fluorescence m...

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three-dimensional and multidimensional microscopy: image acquisition processing VII

作者： Gustafsson, Mats G.L. Agard, David A. Sedat, John W. Univ of California San Francisco San Francisco United States

Spatial information that exceeds the classical resolution limit by a factor of two can be made visible in the widefield fluorescence microscope by illuminating the sample with spatially structured (patterned) excitation light. By computationally restoring this information to its proper location in reciprocal space, an image with twice the normal lateral resolution can be produced. The method can be applied in three dimensions, and yields an axial sectioning power equal to that of confocal microscopes. Unlike the case in confocal microscopy, however, both the lateral and axial resolution enhancements are achieved without any loss of emission light, resulting in uncompromised sensitivity. The method has been experimentally verified on both test objects and complex biological structures and performs in complete agreement with theoretical predictions. The resulting images possess a visual clarity that dramatically exceeds that of both conventional and confocal microscopes.

关键词： Medical imaging

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3-D image of protein visualization in a whole rice grain using an automatic precision microtome system

3-D image of protein visualization in a whole rice grain usi...

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three-dimensional and multidimensional microscopy: image acquisition processing VII

作者： Ogawa, Yukiharu Ohtani, Toshio Sugiyama, Junichi Hagiwara, Shoji Tanaka, Kunisuke Kudoh, Ken-ichi Higuchi, Toshiro Natl Food Research Inst Tsukuba Japan

The three-dimensional (3-D) image formation technique using confocal microscopy has allowed visualization of the 3-D chemical structure in small parts of the bio-body. However, the large-scale 3-D structure such as the distribution of chemical components throughout the whole body has not been shown. To allow such large scale visualization of the 3-D internal analysis technique for bio-body has been developed. The 3-D internal analysis method has three stages. In the first stage, sequential sections of a whole bio-body were made using the automatic precision microtome system. Although this system is that used for microscopy, the obtained sections were positioned on an adhesion tape for 3-D reconstruction. In the next stage, all sections were stained using a chemical or immunofluorescence technique for visualization of the chemical components. Each stained section was digitally imaged using a spectroscopic stereomicroscope with a CCD camera. The obtained 2-D digital images contained the spectroscopic information representing the chemical components. In the last stage, the 2-D digital images were reconstructed to produce a 3-D image. As a result, the distribution of chemical components in the whole bio-body could be visualized in 3-D. In this study, the distribution of protein in a whole rice grain was visualized using a 3-D image.

关键词： Medical imaging

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