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检索条件"任意字段=Conference on 3-Dimensional and Multidimensional Microscopy - Image Acquisition and Processing XII"

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Tracking on tree-like structures in 3-D confocal images

引用

conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Herzog, A Krell, G Michaelis, B Zuschratter, W Univ Magdeburg Inst Measurement Technol & Elect D-39016 Magdeburg Germany

ISBN: (纸本)0819427004

Confocal microscopy is qualified to perform volume scans of nerve cells with dendrites and spines. Length and diameter of the dendrite branches and the spines should be determined to analyze the influence of learning processes. A prerequisite for that is the recognition of the dendritic structure with branching off and spines. Because the microscope operates at the resolution limit the images are blurry, noisy and only poorly sampled. In contrast to other methods which are based on binary images and thinning algorithms, our method tracks the dendritic tree faster and in the gray-level domain using simple geometric models. An explicit segmentation is unnecessary and knowledge about shape and structure of the dendrite is included as a-priori information (nearly circular cross-section, approximate diameter and branching off angle). For large trees, first a low resolution scan is captured to create a rough model. The algorithm allows to refine this model using higher resolution scans for interesting regions along the dendrite. The large unimportant areas between the dendrite branches are not scanned at high resolution to save time and disc space. In a second step, the parameters of the model are adapted to the microscope image by minimizing the deviation of the microscope image from the model image convolved by the microscope point spread function. Features like number, diameter, length and position of the dendrite branches and spines can be easily calculated from the model. An interactive user intervention is possible at the model domain.

关键词： tracking confocal microscopy modeling

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Computational model of image formation process in DIC microscopy

引用

conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Kagalwala, F Kanade, T Carnegie Mellon Univ Pittsburgh PA 15213 USA

ISBN: (纸本)0819427004

A computational model of the image formation process has been developed for the Nomarski differential interference contrast (DIC) microscope. The DIC microscope images variations of the phase of the light wave transmitted through the specimen. In the study of biological phenomena, the DIC microscope is used to visualize live cells which are highly transparent in the visible spectra but distort the phase of the impinging light wave. Within the microscope, a birefringent prism splits the transmitted light wave into two laterally sheared wavefronts. An interference pattern is imaged when the wavefronts recombine. The computational model we developed uses polarization ray-tracing techniques. Rays propagating through different microscope components and the specimen are traced. A specimen is represented by a 3-D grid of voxels, each containing a complex refractive index. At the image plane, a coherent superposition of the diffracted field due to each ray contributes to the image intensity. Partial coherence at the image plane is also simulated by wavefronts with different propagation directions. By computing the image intensity at different positions along the axial direction, we can obtain optically sectioned images. In order to evaluate our model, we compared simulated images to the images taken under a real DIC microscope. We constructed test specimens of known shape and properties, using polystyrene beads in optical cement and an etched glass wafer. As the next step, we plan to use this computational model for the reverse problem, i.e. to reconstruct the 3D refractive index distribution of an imaged specimen.

关键词： differential interference contrast microscope simulation ray-tracing

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Parametric blind deconvolution of microscopic images: Further results

引用

conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Markham, J Conchello, JA Washington Univ Inst Biomed Comp St Louis MO 63110 USA

ISBN: (纸本)0819427004

Blind deconvolution microscopy, the simultaneous estimation of the specimen function and the point spread function (PSF) of the microscope is an under-determined problem with non-unique solutions. The non-uniqueness is commonly avoided by enforcing constraints on both the specimen function and the PSF, such as non-negativity and band limitation. These constraints are some times enforced in ad hoc ways. In addition, many of the existing methods for blind deconvolution estimate the PSF pixel by pixel thus greatly increasing the number of parameters to estimate and slowing the convergence of the algorithm. We derived a maximum-likelihood-based method for blind deconvolution in which we assume that the PSF follows a mathematical expression that depends on a small number of parameters (e.,g. less than 20). The algorithm then estimates the unknown parameters together with the specimen function. The mathematical model ensures that all the constraints of the PSF are satisfied and the maximum likelihood approach ensures that the specimen is non-negative. This parametric blind deconvolution method successfully removes out-of-focus blur but its degree of success depends on the features of the specimen. Specimen features that fall in mostly the null space of the PSF are more difficult to recover and make PSF estimation more difficult.

关键词： blind deconvolution 3D microscopy maximum likelihood

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Confocal fluorescence microscopy using intensity-modulated multiple-wavelength scanning (IMS): Evaluation of results from spectral and lifetime imaging

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Carlsson, K Liljeborg, A Royal Inst Technol Dept Biomed Phys SE-10044 Stockholm Sweden

ISBN: (纸本)0819427004

In this paper we summarize the imaging possibilities and performance offered by the Intensity-modulated Multiple-wavelength Scanning (IMS) technique. The IMS technique is based on intensity-modulated laser illumination of the specimen in combination with lock-in detection of the fluorescent light. We have used this technique with multiple illumination wavelengths, spectral separation of the fluorescent light, and multiple detectors. It is then possible to substantially extend the imaging possibilities of confocal fluorescence microscopy. Among the new possibilities are: 1) Improved spectral separation when recording the light intensities of multiple fluorophores, 2) Separation of intensity signals from multiple fluorophores based on lifetime differences, 3) Simultaneous lifetime recording of multiple fluorophores. We have investigated how different system parameters influence the channel separation and signal-to-noise ratio (SNR). In order to maximize the SNR, factors such as fluorophore lifetimes and modulation frequency of the laser light are important. In our investigations we have obtained good agreement between theoretical and experimental results. In summary, our results show that when using the IMS technique to scan multiple-fluorophore specimens, very high channel separation can be obtained for both lifetime and intensity images. For lifetime images, however, the SNR is worse by a factor of approximately four compared with intensity images.

关键词： confocal microscopy fluorophore separation lifetime imaging signal-to-noise ratio

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Raman chemical imaging using high speed curve resolution chemometrics

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Hancewicz, TM Andrew, JJ Unilever Res US Edgewater Lab Edgewater NJ 07020 USA

ISBN: (纸本)0819427004

The relationship between chemical composition and microstructure is becoming increasingly more important in industry as a way of identifying key performance related properties of fully formulated products. Raman micro-spectroscopic imaging has become the preferred method of obtaining this type of information due to the wealth of detail inherent in Raman spectra pertaining to both microstructure and chemical composition. Concurrently, rapid chemometric data elucidation methodology is required to be able to simultaneously extract pertinent chemical and concentration information in a self-modeling manner from these typically huge and complex data sets. Examples of Raman micro-spectroscopic imaging will be presented as a demonstration of how the principles of Raman imaging can be applied to complex, multi-component, multi-phase systems of inherently low contrast. The use of two-way multivariate curve resolution (MCR) methodology is described as a means of rapidly processing the extremely large, three-way Raman chemical images, dramatically simplifying data analysis. The results from MCR analysis provide the number of chemical species present in the sample, the spectrum of each species for identification, and the concentration image for each species. The additional benefit of image noise reduction is also described for the MCR approach. A comparison of results is given describing the two-way PFA methods currently in use and how they compare with MCR methodology.

关键词： multivariate curve resolution multivariate image analysis principal factor analysis PFA orthogonal projection approach OPA Raman imaging spectral imaging

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Determination of direction-independent optical path-length distribution of cells using rotational-diversity transmitted-light differential interference contrast (DIC) images

引用

conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Preza, C van Munster, EB Aten, JA Snyder, DL Rosenberger, FU Washington Univ Inst Biomed Comp St Louis MO 63130 USA

ISBN: (纸本)0819427004

Two different phase-estimation methods that have been developed for the computation of the optical path-length (OPL) distribution of a specimen from DIC images are compared in this paper. The first method is based on the Wiener filter approach and uses only a single DIC image for the determination of the OPL distribution. The second phase-estimation method is based on the conjugate-gradient optimization method and estimates the OPL distribution using rotational-diversity DIC images;i.e. multiple DIC images obtained by rotating the specimen. For this study, 24 different DIC images of a single bovine spermatozoa head were acquired by rotating the cell by approximately 15 degrees between images. The images were registered and aligned using fiducial marks, and then processed with both methods. Results obtained with the filtering method were found to be dependent on the orientation of the cell with respect to the shear direction. Comparison of the integrated optical path length (IOPL) computed with the the filtering method and the rotational-diversity method using two, four and eight DIC images at different rotation angles showed that the IOPL estimated with the rotational-diversity method is less dependent on the rotation angle, even when only two images separated by 90-degree cell rotation are used for the phase estimation. Our results show that the use of rotational-diversity images in the determination of the OPL distribution is very beneficial because it overcomes the directional dependence of DIC imaging.

关键词： computational Nomarski DIC microscopy phase estimation optical path length

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Three-dimensional imaging of interphase cell nuclei with laser scanning microscopy.

Three-dimensional imaging of interphase cell nuclei with las...

引用

conference on Applications of Digital image processing XXI

作者： Bocker, W Radtke, T Streffer, C Univ Klinikum Essen Inst Med Strahlenbiol D-45122 Essen Germany

ISBN: (纸本)0819429155

During the past decade 3-dimensional image processing has become an important key component in biological research mainly due to two different developments. The first is based on an optical instrument, the so-called confocal laser scanning microscope, allowing optical sectioning of the biological specimen. The second is a biological preparatory method, the so-called FISH-technique (Fluorescence-In-Situ-Hybridization), allowing labeling of certain cellular and sub-cellular compartments with highly specific fluorescent dyes. Both methods make it possible to investigate the 3-dimensional biological framework within cells and nuclei. image acquisition with confocal laser scanning microscopy must deal with different limits of resolution along and across the optical axis. Although lateral resolution is about 0.7 times better than in non-confocal arrangements, axial resolution is more than 3- 4 times poorer than that of the lateral (depending on the pinhole size). For 3D reconstruction it is desirable to improve axial resolution in order to provide nearly identical image information across the 3 dimensional specimen space. This presentation will give an overview of some of the most popular restoration and deblurring algorithms used in 3D image microscopy. After 3D image restoration, segmentation of certain details of the cell structure is usually the next step in image processing. We compared two different kinds of algorithms for segmentation of chromosome territories in interphase cell nuclei. One is based on Mathematical Morphology, the other on Split & Merge methods. The segmented image regions provided the basis for chromosome domain reconstruction as well as for regional localization for subsequent quantitative measurements. As a result the chromatin density within certain chromosome domains as well as some terminal DNA sequences (telomere signals) could be measured.

关键词： 3-D image processing confocal laser scanning microscopy image restoration mathematical morphology automatic segmentation chromosome reconstruction

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An aperture correlation approach to confocal microscopy

An aperture correlation approach to confocal microscopy

引用

conference on Three-dimensional microscopy - image acquisition and processing IV

作者： Wilson, T Juskaitis, R Neil, MAA Kozubek, M Department of Engineering Science University of Oxford Oxford OX1 3PJ Parks Road United Kingdom

We describe a new form of light-efficient real time confocal microscope which requires neither a laser nor scanning in order to obtain a real time confocal image. The approach is based on aperture correlation techniques.

ISBN: (纸本)0819423955

关键词： Optical engineering

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Principles for modeling and functional simulation of biological microstructures

Principles for modeling and functional simulation of biologi...

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conference on Three-dimensional microscopy - image acquisition and processing IV

作者： Kriete, A Univ. of Giessen (Germany)

ISBN: (纸本)0819423955

This paper discusses some aspects of computer based modeling of biological microstructures. The workflow to model and simulate a biological structure is described as a feedback - loop. Beside the system definition by structural and dynamical properties, the simulation is discussed as a mathematical representation coupled with a computer visualization. As an example, the investigation of the functional behavior of lung structures is described with special emphasis to the modeling of respiratory units.

关键词： Mathematical modeling Computer simulations Lung 3D modeling Visualization 3D acquisition Data modeling Process modeling Systems modeling Convection

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image reconstruction for three-dimensional transmitted-light DIC microscopy

Image reconstruction for three-dimensional transmitted-light...

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conference on Three-dimensional microscopy - image acquisition and processing IV

作者： Preza, C Snyder, DL Conchello, JA Washington Univ. (United States)

ISBN: (纸本)0819423955

The derivation of an iterative method for phase reconstruction from Differential-Interference-Contrast (DIG) images is presented here. Because DIC imaging is direction sensitive, in our approach we estimate a specimen's phase function using multiple DIC images obtained by rotating the specimen, Results obtained from testing the method via two-dimensional simulations demonstrate that the use of multiple DIC images at different specimen rotations yield phase reconstructions that more closely resemble the phase function of phantoms than the unprocessed DIC images. Improvement in resolution was also achieved: two points separated by half the Rayleigh resolution limit for coherent illumination not resolved in the unprocessed DIC image, were successfully resolved in the reconstructed phase images. Our results show that phase reconstructions are quantitatively bet-ter and resolution is improved when two or more DIC images are used in the reconstruction.

关键词： computational Nomarski DIC microscopy image reconstruction 3D microscopy

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