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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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Improved optical method for measuring of AFM cantilever deflection

引用

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

作者： Baibyrin, VB Konnov, NP Volkov, UP Saratov Tech Univ Saratov 410016 Russia

ISBN: (纸本)0819427004

In our lab have been developed an atomic force microscope (AFIM) for biological and technical applications with improved optical method for measuring of cantilever deflection. We use conventional technique of measuring the change in angle of light reflected off the cantilever with an additional lens placed between cantilever and four-segment photodiode, The lens forms an image of the cantilever in the photodiode plane. Small sizes of the cantilever image improve the resolution of AFM and reduce the requirements to the optical scheme of the microscope. The lens non-linear transmission of the cantilever deflection is compensated by means of computer program. With the A FM were imaged the DNA of plague microbes (Y. pestis EV) and phages of plague and V. Cholera.

关键词： atomic force microscopy cantilever deflection methods phage DNA biology

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Three-dimensional limited-angle microtomography of blood cells: experimental results

引用

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

作者： Levin, GG Vishnyakov, GN Zakarian, CS Likhachov, AV Pickalov, VV Kozinets, GI Novoderzhkina, JK Streletskaya, EA Russian Res Inst Optophys Measurements Moscow 119361 Russia

ISBN: (纸本)0819427004

Tomographic measurements of the 3D refractive index spatial distribution within optically transparent phase samples with computerized interferometric microscopes are proposed. Phase shifting interferometric microtomography applications for the 3D image reconstruction of the blood cells (lymphocytes) are represented. The immersion 100x, N.A.=1.25 objective was used to increase the spatial resolution and observation angle range to 90 degree. ART, combined ART and iterative Gerchberg-Papoulis 3D algorithm were used for the tomogram reconstruction. To determine the accuracy and spatial resolution of the blood cells image reconstruction by means of the interferometric microtomographic method the numerical simulations were implemented.

关键词： microtomography interferometric microscope refractive index living cells and lymphocyte

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Comparing methods for 3D microscopy

引用

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

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

ISBN: (纸本)0819427004

In previous studies, we examined three-dimensional images of a latex bead containing a surface layer of fluorescence whose thickness was determined by physical sectioning. Confocal images of the bead from six different microscopes all exhibited a significantly thicker fluorescent shell than actually present. In contrast, deconvolved wide-field images of the bead produced an image with the correct shell thickness. We have now repeated some of these studies using a new latex bead containing a much thinner layer of surface fluorescence. In contrast to earlier studies, confocal images of this bead appear to show an accurate thickness for the fluorescent shell. These particular confocal microscopes (a Biorad 1024 and a Zeiss 510) also yielded essentially aberration free images, which was not the case for some of the earlier microscopes tested. In these recent studies however, deconvolution of the bead appeared less robust than earlier. Of three independent wide-held images of the bead, only one yielded a substantially artifact-free image upon deconvolution.

关键词： 3D microscopy comparison confocal wide held deconvolution

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On-line multi-spectral image analysis using unsupervised neural network

引用

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

作者： Lu, T Tan, SZ Lu, RQ Lerner, JM In Harmony Technol Corp Petaluma CA 94952 USA

ISBN: (纸本)0819427004

The overwhelming size of a hyperspectral image creates serious problem for users to understand and utilize the data. A novel unsupervised neural network (UNN) model is presented. The UNN is designed to analyze the spectral contents of the multi-spectral images. The UNN automatically grows its layers and neurons by scanning the training images and by learning spectral features. The learning strategy is optimized to ensure fast convergence. At the end of learning, the UNN provides a table of the spectral content of the images. The image contents are categorized based on spectral similarities. The resulting spectral classes are then mapped onto the image, thus the UNN effectively compresses a hyperspectral image cube into a single image. The UNN is also able to automatically recognize objects by their spectral features. The ability of the UNN in identifying subtle spectral differences is shown.

关键词： hyperspectral multispectral image analysis unsupervised neural networks automated object recognition

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Data correction methods for 3D light microscopy

引用

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

作者： Wang, CL O'Connor, NJ Wang, M Turner, JN Holmes, TJ AutoQuant Imaging Inc Watervliet NY 12189 USA

ISBN: (纸本)0819427004

A cooled or video-rate CCD camera is often used to collect optical slices from many modalities of microscopy including widefield-fluorescence (WF) and transmitted-light brightfield (TLB). Raw optical slices collected from both of these types of cameras are contaminated by imperfect performance of the CCD camera. Optical slice image data must be corrected for the bias level and nonuniformity in the photometric response of CCD elements (flatfield). Fluctuation of the exposure time in a cooled CCD camera needs to be calibrated as well. Bad pixels due to the severely low sensitivity of some of the CCD elements often occur and otherwise impair the conventional schemes of correction and calibration. An adaptive median filter has been introduced by us previously to treat these bad pixels. In this study a Wiener-type regularization scheme is proposed as robust, fast, and practical alternative treatment of these bad pixels. One advantage of using this scheme is that doing so provides a nicely modularized design of the computer algorithm. As such, the correction scheme is software readily integrated as a software component in a three-dimensional (3D) microscopy system and may operate independently without affecting the design of other software components in the system.

关键词： data correction 3D light microscopy adaptive median filter Wiener-type regularization

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Proceedings of Three-dimensional and multidimensional microscopy: image acquisition and processing V

Proceedings of Three-Dimensional and Multidimensional Micros...

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Proceedings of Three-dimensional and multidimensional microscopy: image acquisition and processing V

This Volume 3261 of the conference proceedings contains 36 papers. Topics discussed include optical instrumentation, microscopies for challenging biological imaging and measurement, improved methods and instruments fo... 详细信息

关键词： Imaging techniques

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The spectral topography of histopathology samples

引用

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

作者： Lerner, JM Lu, TW Vari, S LightForm Inc Belle Mead NJ 08502 USA

ISBN: (纸本)0819427004

The goal of imaging spectroscopy is to obtain independent spectra from individual objects in a field-of-view. In the case of biological materials, such as histopathology samples, it has been well established that spectral characteristic can be indicative of specific diseases including cancer. Diagnosis can be enhanced by the use of probes and stains to indicate the presence of individual genome or other biologically active cell components or substances. To assess a specimen through a microscope is directly analogous to observing the Earth from space to assess natural features. This paper describes a simple and inexpensive Imaging Spectrometer, with an origin in remote sensing, that demonstrates that it is possible to rapidly identify evidence of disease in histopathology samples using spatially resolved spectral data. The PARISS imaging spectrometer enables a researcher to acquire multispectral images that yield functional maps, showing what and where biological molecules are located within a structure. It is the powerful combination of imaging and spectroscopy that provides the tools not readily available to the Life Sciences. The PARISS system incorporates a powerful hybrid Neural Network (HNN) analysis to break the data logjam that is often associated with the acquisition and processing of multiple spectra.

关键词： imaging spectroscopy spectral topography histopathology spectroscopy micro-imaging microscopy

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Two-dimensional defocusing correction using artificial neural nets

引用

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

作者： de Solorzano, GO Gonzalez, V Santos, A del Pozo, F Univ Calif Berkeley Lawrence Berkeley Lab Berkeley CA 94720 USA

ISBN: (纸本)0819427004

The aim of this work is to show that properly trained Artificial Neural Nets (ANN) can be used as an image restoration tool to correct the effect of defocusing on 2D optical microscopy images. The proposed method can be applied to correct the results of inaccurate range focusing algorithms on fully automated imaging and analysis systems and to compensate the effect of the limited of the depth of focusing on high numerical aperture system. One type of ANN was used : Feedforward Multilayer Perceptron, with supervised backpropagation training. Its performance has been tested with both synthetic images (artificially defocused) and real images (in-focus and out-of-focus) from latex microspheres. The network was trained with sets of pairs of images : each pair consisted of a defocused image and its corresponding in-focus version. Different levels of defocusing were used. The criteria used to select the algorithm parameters to tune the networks and to train them will be presented. The results of the experiments performed to test their ability to 'learn' to correct the defocusing and to generalize the results will also be shown. The results show that, when trained with images with some levels of defocusing, the network was able to learn and accurately correct these defocusing levels, but it can also generalize the results and correct other levels of defocusing.

关键词： image restoration focus correction neural networks

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

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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

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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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