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

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A whole organ serial sectioning and imaging system for correlation of pathology to computer tomography

A whole organ serial sectioning and imaging system for corre...

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

作者： de Ryk, J Namati, E Reinhardt, JM Piker, C Xu, Y Liu, L Hoffman, EA McLennan, G Univ Iowa Iowa City IA 52242 USA

ISBN: (纸本)0819452327

Randomly selected pathology sections of lung tissue are used to correlate lung pathology with Computer Tomography (CT) images. The randomly selected pathology sections provide physicians with little freedom to thoroughly investigate specific areas of interest as identified via CT images. A Large image Microscope Array (LIMA) was designed to serially section and image entire organs for direct correlation between lung pathology and CT. The LIMA consists of a novel vibratome, capable of sectioning tissue down to a thickness of 40mum at specimen dimensions of 20cm by 30cm to a total depth of 30cm. A camera. and a stereomicroscope, mounted on a XYZ gantry above the vibratome is moved through an automated raster scan to capture the entire surface area of the tissue via many high magnification images. A custom software program was developed to automate all hardware components. The alignment and stitching of the images is achieved though custom C++ code in conjunction with the Insight Segmentation and Registration Toolkit (ITK). The resulting high magnification, high-resolution pathology images are registered with corresponding CT images. Through point-to-point correlation between the two imaging techniques a pathological and CT ground truth may be established.

关键词： microscopy pathology serial sectioning computer tomography ITK registration image montage

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Detection, imaging and kinetics of sub micron organelles of chondrocytes by multiple beam interference microscopy

Detection, imaging and kinetics of sub micron organelles of ...

引用

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

作者： Joshi, NV Medina, H Barboza, JM Colantuoni, G Quintero, M Hosp Univ Los Andes Dept Physiol Fac Med Merida Venezuela

ISBN: (纸本)0819452327

Chondrocytes, obtained from testosterone treated human articular cartilage, were examined by a recently developed Multiple Beam Interference microscopy (MBIM) attached to a confocal set up, Video-enhanced differential interference microphotography and also by cinematography. In the MBIM, the intensity of the transmitted pattern is given by the Airy function which increases the contrast dramatically as the coefficient of the reflectance of the parallel plates increases. Moreover, in this configuration, the beam passes several times through a specific organelle and increases its optical path difference both because of the increase in the trajectory and refractive index (high density) of the organelle. The improved contrast enhances the resolving power of the system and makes visible several structural details of sub micron dimensions like nucleolus, retraction fibers, podia, etc. which are not possible to reveal with such a clarity by conventional techniques such as bright field, phase contrast or DIC. This technique permits to detect the oscillatory and rotational motions of unstained cilia for the first time. The frequency of oscillations was found to be 0.8 HZ. Keywords: Multiple Beam Interference Microscope, Chondrocytes Nucleolus and microfilaments, cilium and oscillatory & rotatory motion.

关键词： multiple beam interference microscope chondrocytes nucleolus and microfilaments cilium and oscillatory & rotatory motion

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Enhanced visualization of inner ear structures

Enhanced visualization of inner ear structures

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

作者： Niemczyk, K Kucharski, T Kujawinska, M Bruzgielewicz, A Med Univ Warsaw Dept Otolaryngol PL-02097 Warsaw Poland

ISBN: (纸本)0819452327

Recently surgery requires extensive support from imaging technologies in order to increase effectiveness and safety of operations. One of important tasks is to enhance visualisation of quasi-phase (transparent) 3D structures. Those structures are characterized by very low contrast. It makes differentiation of tissues in field of view very hard. For that reason the surgeon may be extremely uncertain during operation. This problem is connected with supporting operations of inner ear during which physician has to perform cuts at specific places of quasi-transparent vellums. Conventionally during such operations medical doctor views the operating field through stereoscopic microscope. In the paper we propose 3D visualization system based on Helmet Mounted Display. Two CCD cameras placed at the output of stereoscopic microscope perform acquisition of stereo pairs of images. The images are processed in real-time with the goal of enhancement of quasi-phase structures. The main task is to create an algorithm that is not sensitive to changes in intensity distribution. The disadvantages of existing algorithms are their lack of adaptation to occurring reflexes and shadows in field of view. The processed images from both left and right channels are overlaid on the actual images exported and displayed at LCD's of Helmet Mounted Display. A physician observes by EMD a stereoscopic image of operating field with indication of the regions of special interest. The authors present the hardware, procedures applied and initial results of inner ear structure visualization. Several problems connected with processing of stereo-pair images are discussed.

关键词： inner ear structures steroscopic visualisation image enhancement helmet mounted display

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Performance of dual axes confocal microscope for in vivo molecular and cellular imaging

Performance of dual axes confocal microscope for <i>in vivo<...

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

作者： Kino, G Wang, T Contag, C Mandella, M Chan, N Stanford Univ Stanford CA 94305 USA

ISBN: (纸本)0819452327

The application of conventional confocal microscopes with high numerical aperture (NA) to in vivo imaging is limited by the objectiveis large physical dimensions and short working distance. We are developing a confocal microscope that uses simple low NA lenses oriented in a dual axes configuration for miniaturization and in vivo imaging. This architecture achieves a long working distance, micron level axial resolution, and reduced noise from scattered light outside the focal volume. Combined with the novel method of post-objective scanning, this design can be scaled down to millimeter dimensions. We derive the dual axes response from diffraction theory, and construct two tabletop prototypes to demonstrate the performance of this approach. We collect images from freshly excised biopsy specimens of human esophagus and transgenic mouse cerebellum expressing GFP. With horizontal cross-sectional images, we achieve 1 to 2 mum resolution and collect reflectance and fluorescence images. With vertical cross-sectional images, we achieve 4 to 5 mum resolution, dynamic range of 70 dB, and tissue penetration over 1 mm. An instrument miniaturized with this configuration could be used for in vivo cellular and molecular imaging.

关键词： confocal microscopy imaging fluorescence reflectance in vivo NMMS

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Dissecting key components of the Ca²⁺ homeostasis game by multi-functional fluorescence imaging

Dissecting key components of the Ca<SUP>2+</SUP> homeostasis...

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

作者： Bastianello, S Ciubotaru, C Beltramello, M Mammano, F VIMM I-35129 Padua Italy

ISBN: (纸本)0819452327

Different sub-cellular compartments and 2 organelles, such as cytosol, endoplasmic reticulum and mitochondria, are known to be differentially involved in Ca2+ homeostasis. It is thus of primary concern to develop imaging paradigms that permit to make out these diverse components. To this end, we have constructed a complete system that performs multi-functional imaging under software control. The main hardware components of this system are a piezoelectric actuator, used to set objective lens position, a fast-switching monochromator, used to select excitation wavelength, a beam splitter, used to separate emission wavelengths, and a I/O interface to control the hardware. For these demonstrative experiments, cultured HeLa cells were transfected with a Ca2+ sensitive fluorescent biosensor (cameleon) targeted to the mitochondria (mtCam), and also loaded with cytosolic Fura2. The main system clock was provided by the frame-valid signal (FVAL) of a cooled CCD camera that captured wide-field fluorescence images of the two probes. Excitation wavelength and objective lens position were rapidly set during silent periods between successive exposures, with a minimum inter-frame interval of 2 ms. Triplets of images were acquired at 340, 380 and 430 nm excitation wavelengths at each one of three adjacent focal planes, separated by 250 nm. Optical sectioning was enhanced off-line by applying a nearest-neighbor deconvolution algorithm based on a directly estimated point-spread function (PSF). To measure the PSF, image stacks of sub-resolution fluorescent beads, incorporated in the cell cytoplasm by electroporation, were acquired under identical imaging conditions. The different dynamics of cytosolic and mitochondrial Ca2+ signals evoked by histamine could be distinguished clearly, with sub-micron resolution. Other FRET-based probes capable of sensing different chemical modifications of the cellular environment can be integrated in this approach, which is intrinsically suitable for th

关键词： fluorescence microscopy deconvolution multi-spectral analysis piezoelectric actuator CCD camera mitochondria camelcon FRET Fura2

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Proceedings of SPIE: three- dimensional and multidimensional microscopy - image acquisition and processing

Proceedings of SPIE: Three- Dimensional and Multidimensional...

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three-dimensional and multidimensional microscopy:image acquisition and processing X

The proceedings contains 20 papers from the conference on three-dimensional and multidimensional microscopy: image acquisition and processing X. The topics discussed include: high-resolution imaging of biological samples;benifits and applications of programmable spectral devices in confocal microscopes;logitudinal field imaging and confocal endoscopy.

关键词： Medical imaging

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High-resolution three-dimensional imaging using multiple nanometric probes

High-resolution three-dimensional imaging using multiple nan...

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

作者： Dubois, A Moneron, G Lecaque, R Lequeux, F Boccara, AC Ecole Super Phys & Chim Ind Ville Paris CNRS UPR A0005 Lab Opt Phys F-75231 Paris 5 France

We present a new imaging modality using nanometric beads as multiple local probes. The positions of the beads are determined in three dimensions using white-light interference microscopy, by over-sampling and fitting ... 详细信息

ISBN: (纸本)0819447641

关键词： interference microscopy three-dimensional microscopy scanning probe microscopy

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The three-dimensional imaging of microspheres in confocal and conventional polarization microscopes

The three-dimensional imaging of microspheres in confocal an...

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

作者： Yang, LS Taylor, CM Rakovich, Y McCabe, EM Univ Dublin Trinity Coll Dept Phys Dublin 2 Ireland

ISBN: (纸本)0819447641

images of the microspheres are studied in three-dimensions by using the confocal and conventional scanning polarization microscopes. It is found that the polarization of the detected signals is mainly parallel to the initial polarization which is due to the high extinction coefficient of the confocal system. Arc pairs are observed at the edge of the microspheres with the conventional polarization microscope with a crossed analyzer. Theoretical analysis are given by using the vector field theory and the image formations of the two systems.

关键词： three-dimensional microscopy polarization microsphere

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image estimation accounting for point-spread function depth variation in three-dimensional fluorescence microscopy

Image estimation accounting for point-spread function depth ...

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

作者： Preza, C Conchello, JA Washington Univ Dept Elect Engn St Louis MO 63130 USA

ISBN: (纸本)0819447641

An approximate model for optical-sectioning microscopy describing depth-varying imaging is developed. The model incorporates changes in the point-spread function due to refractive index mismatch between the immersion medium and the specimen. which causes spherical aberration that worsens with increasing depth tinder the coverslip. Comparison of model predictions to measured images from a bead phantom shows that the approximate model captures the main features in the data. The model presented in this paper is the first step towards depth-variant image estimation for optical-sectioning microscopy. An expectation maximization algorithm for maximum-likelihood restoration based on this model is also presented.

关键词： 3D microscopy depth-variant imaging model maximum likelihood estimation

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Tracking fluorescent spots in biological video microscopy

Tracking fluorescent spots in biological video microscopy

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

作者： Genovesio, A Olivo-Marin, JC Inst Pasteur Quantitat Image Anal Grp F-75724 Paris France

ISBN: (纸本)0819447641

The analysis of cell and pathogen movement and motility is a major topic in cell biology for which computerized methods are most needed. This study proposes a method to detect and track multiple moving biological objects in image sequences acquired through fluorescence video microscopy. The method enables the analysis of video microscopy image sequences in order to obtain reliable quantitative data such as number, position, speed and movement phases. The method consists of three stages. A stage of detection is performed through a multi-scale analysis of images using an undecimated wavelet transform. The next stage is the prediction of the state of each detected spot in the next frame using a Kalman filter and an adapted model. Then comes a stage of data association which constructs the tracks and refines the filters. Once all moving objects have been assigned with unique spatio-temporal paths, trajectories are analyzed in terms of different parameters relevant to the motility analysis of biological objects.

关键词： image processing tracking wavelets Kalman filter data associations biological video microscopy fluorescent spots

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