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

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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

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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...

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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

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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...

引用

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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Interferometric computed-microtomography of 3D phase objects

Interferometric computed-microtomography of 3D phase objects

引用

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

作者： Vishnyakov, GN Levin, GG Zakarian, CS Russian Research Institute for Optophysical Measurements (Russia) Moscow State Technical Univ. (Russia)

ISBN: (纸本)0819423955

Measurements of refractive index spatial distribution within optically transparent phase 3-D samples with computerized interferomertric microscopes is proposed. For the reconstruction of the phase information the phase shifting interferometry is used, Interferometric computed-tomography microscope are fully automated integrated systems that incorporate CCD camera, frame grabber, computed operated PZT mirror as well as intelligible software for interferogram processing, tomographic reconstruction and refractive index distribution are displayed in 2-D and 3-D, sections, isolines, etc. Experimental results for interferometric microscopy and tomography of blood cells are presented.

关键词： computed tomography 3D microscopy interferometry phase object phase-shifting method refractive index erythrocyte

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Quantitative DIC microscopy using a geometric phase shifter

Quantitative DIC microscopy using a geometric phase shifter

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

作者： Cogswell, CJ Smith, NI Larkin, KG Hariharan, P Univ. of Sydney (United States) Univ. of Sydney (Japan) Univ. of Sydney (Australia)

ISBN: (纸本)0819423955

In this paper we investigate the use of a geometric phase-shifting (GPS) technique which allows us to convert conventional transmission or reflection differential interference contrast (DIC) microscopy into a quantitative mode. A phase-shifting algorithm is employed to extract the specimen phase gradient from the mixture of phase and amplitude information which is common in DIC. Fourier techniques are then used to recover the exact phase (i.e. optical path length variations) throughout the biological specimen viewed. In addition to this quantitative ''phase map,'' we demonstrate that the GPS process simultaneously yields an ''amplitude-only'' representation in which-various absorption and transmission properties of the specimen are displayed as intensity variations in the image, similar to brightfield microscopy. These two resulting images can then be analysed or further processed in a number of ways that are not possible with conventional DIC and which improve the microscopist's ability to correctly identify, interpret and measure features in the specimen.

关键词： 3D microscopy interference DIC phase-shifting image processing biomedical microscopy

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