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

共 591 条记录，以下是441-450 订阅

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Design and performance evaluation of reflection confocal microscopy using acousto-optical deflector and slit detector

Design and performance evaluation of reflection confocal mic...

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

作者： Lee, SW Kang, DK Yoo, HK Gweon, DG Lee, SW Kim, KS Korea Adv Inst Sci & Technol Dept Mech Engn Taejon South Korea

ISBN: (纸本)0819452327

We describe the design and the implementation of reflection confocal scanning microscopy (CSM) using an acousto-optical deflector (AOD) for the fast horizontal scan and a galvanometer mirror (GM) for the slow vertical scan. In the beam scanning system it is important to maintain the lateral and the axial performance during scanning operation. We propose a simple method to design a scanning system using the finite ray tracing and the diffraction theory. We define a cost function which contains the effect of aberrations on the performance of microscopy. We construct the designed system and evaluate its performance. The OSLO simulation shows that the performances of CSM are not changed with deflection angle. So we conclude that the beam scanning system is properly designed. In addition, we propose an image formation method and show images obtained with the system.

关键词： confocal microscopy beam scanning system acousto-optical deflector slit detector finite ray tracing relay optics image formation

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Optimization of spinning disk confocal microscopy: Synchronization with the ultra-sensitive EMCCD

Optimization of spinning disk confocal microscopy: Synchroni...

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

作者： Chong, FK Coates, CG Denvir, DJ McHale, N Thornbury, K Hollywood, M Andor Technol Ltd Belfast BT12 7AL Antrim North Ireland

ISBN: (纸本)0819452327

The advent of Electron Multiplying Charge Coupled Device (EMCCD) technology and it's ability to overcome previous hurdles in low-light fluorescence microscopy, such as phototoxicity to live cells, photobleaching of fluorophores and exposure time restrictions, has resulted in a significant resurgence of interest in use of confocal spinning disk techniques for live cell microscopy. Here provide an understanding of, and technical solutions to, the issues of synchronization that have previously marred the coupling of fast CCD camera technology to confocal spinning disk arrangements. We examine the challenges arising from both old and new models of the Nipkow spinning disk confocal unit and suggest solutions throughout based on a sound comprehension of both (a) relative scan/exposure times;(b) relative orientation of the coupled devices;(c) optimisation of EMCCD clocking parameters.

关键词： CCD EMCCD Nipkow confocal spinning disk confocal microscopy live cell microscopy electron multiplying CCD single photon sensitivity

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LEDs for fluorescence microscopy

LEDs for fluorescence microscopy

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

作者： Young, IT Garini, Y Dietrich, HRC van Oel, W Lung, GL Delft Univ Technol Fac Sci Appl Dept Image Sci & Technol Quantitat Imaging Grp NL-2628 CJ Delft Netherlands

ISBN: (纸本)0819452327

Traditional light sources for fluorescence microscopy have been mercury lamps, xenon lamps, and lasers. These sources have been essential in the development of fluorescence microscopy but each can have serious disadvantages: lack of near monochromaticity, heat generation, cost, lifetime of the light source, and possible distortions due to coherence effects. We are examining the possibility of using the new high-power light-emitting diode (LED) sources as alternatives to the above mentioned sources. LED sources are near monochromatic, are inexpensive, produce little heat, have no coherence problems, have extended lifetimes, are small, and can easily be modulated. We describe experiments comparing various LEDs to other light sources. We compare, for example, a 530 nm LED to the 546 nm line from a mercury lamp on a fluorophore whose absorption maximum is broad and in the middle between these two wavelengths.

关键词： high-power light-emitting diodes fluorescence microscopy multi-spectral sources quantitative microscopy

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Bright field segmentation tomography (BFST) for use as surface identification in stereomicroscopy

Bright field segmentation tomography (BFST) for use as surfa...

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

作者： Thiesse, J Namati, E de Ryk, J Hoffman, E McLennan, G Univ Iowa Iowa City IA 52242 USA

ISBN: (纸本)0819452327

Stereomicroscopy is an important method for use in image acquisition because it provides a 3D image of an object when other microscopic techniques can only provide the image in 2D. One challenge with this type of imaging is determining the top surface of a sample that has otherwise indistinguishable surface and planar characteristics. We have developed a system that creates a form of oblique illumination and in conjunction with image processing, the top surface can be viewed. The use of BFST consists of the Leica MZ16A stereomicroscope with a unique attached lighting source, and is part of the Large image Microscope Array (LIMA) [1]. The lighting source consists of eight light emitting diodes (LED's) that are separated by 45-degree angles. Each LED in this system illuminates with a 20-degree viewing angle once per cycle with a shadow over the rest of the sample. Subsequently, eight segmented images are taken per cycle. After the images are captured they are stacked through image addition to achieve the full field of view, and the surface is then easily identified. image processing techniques, such as skeletonization, can be used for further enhancement and measurement. With the use of BFST, advances can be made in detecting surface features from metals to tissue samples, such as in the analytical assessment of pulmonary emphysema using mean linear intercept.

关键词： stereomicroscopy illumination surface identification image processing

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Mueller polarimetric microscopy

Mueller polarimetric microscopy

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

作者： Laude-Boulesteix, B De Martino, A Le Naour, G Genestie, C Schwartz, L Garcia-Caurel, E Drévillon, B Ecole Polytech Phys Interfaces & Couches Minces Lab CNRS UMR 7647 F-91128 Palaiseau France

ISBN: (纸本)0819452327

We present a multispectral polarimetric imagine, system well suited for complete Mueller matrix microscopy. The source is a spectrally filtered halogen light bulb, and the image is formed on a fast CCD camera The light polarization is modulated before the sample and analyzed after the sample by using nematic liquid crystal modulators.. The whole Mueller matrix image of the sample is typically measured over 5 seconds for a good signal-to-noise ratio. The instrument design, together with an original and easy-to-operate calibration procedure provides a high polarimetric accuracy over wide ranges of wavelengths and magnifications. Mueller polarimetry provides separate images of scalar and vector retardation and dichroism of the sample, together with its depolarizing power, while all these effects do contribute simultaneously to the contrasts observed in standard polarized microsopy. Polarimetric images of several samples, namely an unstained rabbit cornea, a picrosirius red stained hepatic biopsy, and a rat artery specifically stained for collagen III are shown and discussed.

关键词： imaging systems polarimetry medical optics and biotechnology histology

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Development of micro-optical projection tomography for 3D analysis of single cells

Development of micro-optical projection tomography for 3D an...

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

作者： Fauver, M Seibel, EJ Rahn, JR Patten, FW Nelson, AC Univ Washington Coll Engn Human Interface Technol Lab Seattle WA 98195 USA

ISBN: (纸本)0819452327

The Micro-Optical Projection Tomographic Microscope (muOPTM) is an instrument that is being developed for three-dimensional (3D) imaging of cells and subcellular components. The target application for the muOPTM is the early detection of lung cancer by revealing the complex 3D information about chromatin redistribution in the nucleus. The muOPTM employs a scanning objective lens (100x, N.A.=1.25) to create an extended depth-of-field image, similar to a shadowgram or projection, that we call a pseudo-projection. A large number of pseudoprojections (90+) are acquired, from which a tomographically reconstructed 3D image is computed using a filtered backprojection algorithm. The prototype muOPTM uses a single objective lens, so the object (cell) must be rotated for each new pseudo-projection. A custom microtube stage minimizes the lateral and axial motion of the sample tube during scanning and rotation so that registration between successive pseudo-projections is maintained. image processing methods are used to correct any remaining registration errors. The media inside and outside the tube are refractive index-matched to each other and to the tube (Deltan(avg) < 0.02). The index-matched materials are pressed between two flat parallel windows, providing a nearly distortion-free image. Custom phantoms using microspheres have been constructed and images of these 3D test targets acquired. The minimum resolvable feature size is better than 3 microns. The first 3D image of a cell using muOPTM is also shown.

关键词： optical tomography optical projection tomography three-dimensional imaging computed tomography microscopy depth of field cancer screening

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Optical computed-tomography microscope using digital spatial light modulation

Optical computed-tomography microscope using digital spatial...

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

作者： Chamgoulov, RO Lane, PM MacAulay, CE British Columbia Canc Res Ctr Canc Imaging Dept Vancouver BC V5Z 1L3 Canada

ISBN: (纸本)0819452327

A computed-tomography microscope using a digital spatial light modulator has been developed and has demonstrated cytological and histological three-dimensional (3-D) image acquisition. The system consists of two high NA objective lenses, a Digital Micromirror Device (DMD) placed conjugate to the back pupil plane of the illumination objective, a sample stage, a light source, and a CCD detector. Each DMD micromirror can control the illumination of a specific angle. A single 3-D reconstruction is obtained from parallel ray projections acquired by changing the polar angle of illumination -phi (max) less than or equal to phi less than or equal to phi (max) while holding the azimuthal angle theta constant. The polar angle is limited by the NA of the objective. To compensate for an incompleteness of information due to the limited polar angles, several reconstructions acquired at multiple azimuthal angles are combined to create a final reconstruction. A reconstruction algorithm was developed using simulation software based on the 3-D Radon transformation and 3-D synthetic objects. Microscopic 3-D volume reconstructions of quantitatively absorption-stained cells have been demonstrated. 3-D reconstructed images enables the analysis of cell morphology and tissue architecture, as well as virtual two-dimensional slices with the distance between slices of 0.3 mum.

关键词： computed-tomography optical microscopy 3-D imaging Radon transformation digital spatial light modulator

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Object segmentation, quantification, counting, and tracking

Object segmentation, quantification, counting, and tracking

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

作者： Abu-Tarif, A Khiznichenko, V Northan, B Holmes, T Yoon, P Girroir, J Brathwaite, P AutoQuant Imaging Inc Watervliet NY 12189 USA

ISBN: (纸本)0819452327

A complete system for object segmentation, counting, quantification, and tracking from microscopic images was implemented. We found that image deconvolution and reconstruction operations are essential to the success of any general-purpose segmentation algorithm and hence are of paramount importance for a counting and tracking software system. Wavelet-based image enhancement, background equalization, and noise suppression routines are the components in our novel general-purpose segmentation algorithm. Simple object recognition based on averages and preset tolerances suffices for most applications. As expected, boundary smoothing is important if watershed-based blob separation is to be used. One of the challenges of a general-purpose counting and tracking system is the need for a large number of object quantification components (features). In tracking we found that incorporating weighted features into an error function improves the accuracy over just the path coherence criterion and that evaluating correspondences over multiple time frames improves the accuracy over using only two consecutive time frames.

关键词： deconvolution image reconstruction segmentation object counting tracking

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High-speed confocal laser scanning microscopy using acousto-optic deflectors and a digital micromirror device

High-speed confocal laser scanning microscopy using acousto-...

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

作者： Bansal, V Patel, S Saggau, P Rice Univ Dept Bioengn Houston TX 77251 USA

ISBN: (纸本)0819452327

Neurons are known to possess active computational properties. To investigate these properties, it is desirable to study the electrical and chemical properties not only at a living neuron's cell body, but also at many sites within its dendritic arborization. However, currently available recording techniques force a tradeoff between spatial and temporal resolution. To overcome these limitations, we have developed a confocal microscope that can make multisite optical recordings at an effective frame rate that is sufficient to measure fast neuronal events, such as action potentials, that occur on a timescale of milliseconds. We accomplished this by combining acousto-optic deflectors for addressable point illumination with a digital micromirror device for addressable point detection. After developing a registration algorithm to ensure synchronicity between point illumination and point detection, we used light-scattering test preparations to demonstrate that our system is capable of optical sectioning and therefore capable of imaging in living brain tissue. Furthermore, we have shown that fluorescence changes can be monitored at an effective frame rate of 25 kHz.

关键词： confocal acousto-optic deflector AOD digital micromirror device DMD neuron

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Reconstruction of objects with a limited number of non-zero components in fluorescence microscopy

Reconstruction of objects with a limited number of non-zero ...

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

作者： Davis, BJ Karl, WC Swan, AK Goldberg, BB Ünlü, MS Goldberg, MB Boston Univ Dept Elect & Comp Engn Boston MA 02215 USA

ISBN: (纸本)0819452327

A reconstruction algorithm is developed that uses specific a-priori knowledge to produce higher resolution images than standard approaches. Deconvolution is an important image reconstruction tool in fluorescence microscopy. This is especially true for modern interferometric instruments (such as I-5 M and 4Pi systems), as they may have complicated oscillatory point spread functions. Current methods are designed to work on an arbitrary object - i.e. it is assumed that there is no available a-priori knowledge of the object (with the possible exception of a non-negative condition on the fluorophore-emission intensities). In situations where there is a-priori knowledge of the object, it may be possible to use this information to produce a higher quality reconstruction of the object. A useful a-priori condition is investigated here. It is assumed that the object can be represented by the sum of not more than L basis functions. The simplest example of this is when the basis functions are impulses - this leads to an object of L or less non-zero points on a background of zeros. This a-priori condition can be applied directly;applied to a limited region of the object;applied in one dimension (for an object with a layered structure such as lipid bilayers);or applied in two dimensions (for an object with a filamentary structure such as actin fibers.) A reconstruction algorithm is described and applied to some illustrative simulated examples. The results are found for several fluorescence microscopy methodologies and compared to the results produced by standard deconvolution methods.

关键词： fluorescence microscopy deconvolution prior knowledge image reconstruction super-resolution

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