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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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Automated 3D dendritic spine detection and analysis from two-photon microscopy

Automated 3D dendritic spine detection and analysis from two...

引用

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

作者： Koh, IYY Lindquist, WB SUNY Stony Brook Dept Appl Math & Stat Stony Brook NY 11794 USA

ISBN: (纸本)0819439398

The functional significance of dendritic spines and their plasticity to a wide spectrum of developmental and pathological conditions has led to extensive studies based on spine morphology. The advances in image acquisition techniques and the associated generation of large 3D data sets of optical micrographs have not been accompanied by comparable advances in data analysis techniques. We present an automated 3D spine detection and quantification procedure suitable for images obtained by laser scanning microscopy. The image is first processed by deconvolution and the dendritic phase consisting of the neuronal cytoplasm is extracted by segmentation. Spines are detected as geometrical protrusions relative to the dendritic backbone, ks very thin necks may not be imaged, some spine 'heads' may be detached from the dendrite and are detected as detached components. These detected heads are merged with spine 'bases' where appropriate. Morphological characterizations on spine length, volume, density and shape classifications are obtained. For time-lapse data, images are registered and individual spines are traced through the image sequence. Successful comparison results on spine lengths and densities with manual analysis are obtained. This method is highly automatic and allows detailed and objective quantification of the structure and dynamics of dendritic spines, which can be important predictors for the function of neural networks.

关键词： image processing dendrite spine two-photon medial axis laser scanning microscopy

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Programmable array microsope employing two ferroelectric liquid crystal spatial light modulators

Programmable array microsope employing two ferroelectric liq...

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

作者： Taylor, CM McCabe, EM Univ Dublin Trinity Coll Dept Phys Dublin 2 Ireland

ISBN: (纸本)0819439398

We present a programmable array microscope (PAM) which uses a pair of ferroelectric liquid crystal spatial light modulators (FLC SLMs). The system is similar to a confocal imaging system called a direct-view microscope (DVM), in which scanned aperture arrays are used to obtain real-time confocal images. The PAM, unlike the DVM, allows arrays to be scanned electronically rather than mechanically. In our system, one SLM is placed in the source plane of a conventional microscope system;the other is placed in the detector plane. Confocal aperture arrays are displayed and scanned synchronously on the SLMs and confocal imaging results. The resolution is improved when compared to a similar previously presented system which employed a single SLR I. We present axial resolution measurements for a variety of array dimensions and investigate the use of aperture correlation techniques to improve the light throughput in the devices.

关键词： programmable array microscopy ferroelectric liquid crystal spatial light modulator confocal microscope direct-view microscopy

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Focused Ion Beam techniques for the analysis of biological samples: a revolution in ultramicroscopy?

Focused Ion Beam techniques for the analysis of biological s...

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

作者： Ballerini, M Milani, M Batani, D Squadrini, F Univ Milan Dipartimento Sci Mat I-20126 Milan Italy

ISBN: (纸本)0819439398

Focused ion Beam (FIB) is a novel technique that allows easy target cell selection, fast operation, high resolution, 3D imaging and sample manipulation during imaging. The FIB technique of microscopy and nanomachining, that is widely spread in semiconductor technology, is reshuffled to open new horizons in the field of life sciences at cellular and subcellular level. FIB is a source of ions which can be precisely oriented and focused on the sample, supported also by an electron source. The resolution can be as low as 15 nm. Two different operation modes are available: "cutting" and "etching" operations of very high precision and at the same time the resulting secondary ions and electrons provide follow up sample imaging. FIB tomography ability has been tested to provide information on cell characterisation, cell division time sequence, view of inner structures and investigation of membrane structural properties. We compare the performances and the advantages of different high-resolution microscopy techniques: Soft X-ray Contact microscopy (SXCM), Focused Ion Beam (FIB) and Transmission Electron microscopy (TEM). These have been used to image Saccharomyces cerevisiae yeast cells, a rather well known sample of great biological interest. TEM is an established technique used as a benchmark for comparison.

关键词： microscopy cell imaging Focused Ion Beam( FIB) cell ultrastructure

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Proceeding of SPIE - The International Society for Optical Engineering

Proceeding of SPIE - The International Society for Optical E...

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three-dimensional and multidimensional microscopy: image Asquisition and Processin VIII

The proceedings contains 18 papers from SPIE 2001 Conference on three dimensional and multidimensional microscopy: image acquisition and processing VIII. The topics discussed include: inverse scattering;fiber optic co... 详细信息

关键词： Optical microscopy

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One-step calibration of specimen induced focal shifts and spherical aberration for quantitative 3D microscopy:: Approach and first results

One-step calibration of specimen induced focal shifts and sp...

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

作者： Thomann, D Stüssi, E Danuser, G Swiss Fed Inst Technol Biomech Lab CH-8952 Schlieren Switzerland

ISBN: (纸本)0819439398

With the advent of laser scanning confocal and multiphoton microscopy, 3D life tissue characterization has been rendered possible. This involves the restoration of thick section images (in the depth range of 100 microns) of biological samples. In contrast to thin samples new effects become important when imaging thick samples: Because of changes of the refractive index across the specimen or the embedding medium, strong (> 1 micron) focal shifts and spherical aberration occur and scattering effects get more prominent. For tissue mapping it is essential to correct for such aberrations and distortion effects. In this paper. Re propose a calibration framework, which allows us to determine thick section focal shifts and spherical aberrations in a one-step-procedure, Gradients in the focal shift induce a scaling in the Z-direction of the observed sample. A second effect arises with depth-dependent spherical aberration. We model our microscope as a linear-shift-NON-invariant system (LSNI) where multiple depth classes are assigned distinct point spread functions (PSFs). We measure these two effects in a 3D sample of randomly distributed fluorescent focal check beads. The beads are embedded in gelatine, a medium with high resemblance to real biological tissue. The PSF is approximated by a mathematical parametric model. While estimating the parameters of the PSF with object-constrained deconvolution, we track depth dependent changes of the observed bead diameter. This allows us to determine the gradient of the focal shift across a thick section. By numerical integration along the optical axis we obtain the focal shift values as a function of sample depth. In the end, these values will be applied as a correction to compensate depth distortions in the tissue images.

关键词： confocal microscopy calibration spherical aberration focal shifts deconvolution

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4D image processing in microscopy by combined methods

4D image processing in microscopy by combined methods

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

作者： Kriete, A Image Proc Lab Inst Anat & Cell Biol D-35385 Giessen Germany

ISBN: (纸本)081943535X

In time-lapse microscopy image quality is limited by instrumental and specimen characteristics. Moreover, an enormous amount of data may be accumulated. This challenges not only the development of new tools for image processing, but typically forces methods to be executed in combination. To remedy essential problems, an image processing pipeline is employed which features image restoration, image compression, visualization and analysis. To control the flow of information within this pipeline and to optimize the parameter settings like compression rates or number of iterations, the information content is evaluated on the basis of the ratio of signal and noise.

关键词： time-lapse microscopy 4D image processing compression image restoration visualization and analysis

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Inhomogeneous polarization in scanning optical microscopy

Inhomogeneous polarization in scanning optical microscopy

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

作者： Youngworth, KS Brown, TG Univ of Rochester Rochester United States

ISBN: (纸本)081943535X

Polarization control and analysis have proven to be a powerful tool both in conventional and confocal microscopies, but most of these approaches have used homogeneously polarized pupils. Other groups have had varying levels of success producing beams with spatially inhomogeneous polarization. In this paper, we describe the generation and focusing properties of such beams and their application to scanning laser microscopy. We specifically consider microscopy using the lowest-order azimuthally polarized beam.

关键词： spatially inhomogeneous polarization scanning optical microscopy azimuthal polarization radial polarization cylindrical-vector beam polarization converter

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three-dimensional image of sugar content visualization in a melon by spectral information

Three-dimensional image of sugar content visualization in a ...

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

作者： Sugiyama, J Ogawa, Y Natl Food Res Inst Tsukuba Ibaraki 3058642 Japan

ISBN: (纸本)081943535X

In order to visualize sugar content of a melon, the relationship between sugar content and absorption spectra was investigated using a near-infrared (NIR) spectrometer. The absorbance at 676nm, which is close to the chlorophyll absorption band, had a high inverse correlation with sugar content. A high-resolution cooled CCD imaging camera with the band-pass filter of 676nm was used to capture the spectral absorption image. The calibration method for converting the absorbance on the image into Brix sugar content was developed in accordance with MR techniques. Applying this method to each pixel of the absorption image, a color distribution map of the sugar content was constructed. In addition, a special slicing device that can cut a melon in each 5mm thickness was developed in order to create a 3-D image of sugar content distribution.

关键词： three-dimensional near infrared spectral image sugar content melon

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Near-field optical and atomic force constraints for super-resolution 3D deconvolution in far-field optical microscopy

Near-field optical and atomic force constraints for super-re...

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

作者： Axelrod, N Radko, A Ben-Yosef, N Khatchatouriants, A Treinin, M Lewis, A Hebrew Univ Jerusalem Dept Appl Phys IL-91904 Jerusalem Israel

ISBN: (纸本)081943535X

We demonstrate that near-field optical and atomic force microscopy data can be used for super-resolution three-dimensional (3-D) image restoration in optical sectioning fluorescence microscopy. A crucial feature in this approach is the full integration of such data sets with digital far-field images. The scanned probe data is used to provide modalities for boundary constraints which define surface optical information and spatial domains of optical alterations in a sample with a spatial precision that has been unachievable in the past. A restoration algorithm that can use such a complex of data for 3-D image deconvolution is presented. It uses a Tikhonov-Miller regularization scheme and allows for the imposition of different types of constraints to obtain super-resolution deconvolved images. Performance was tested by using simulated 3-D imaging. An example is given of the restoration of a 3-D wide field optical image of a biological specimen in the presence of atomic force constraints.

关键词： image restoration deconvolution algorithm constraints probe microscopy NSOM super-resolution

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Variable-focus microlenses as a potential technology for endoscopy

Variable-focus microlenses as a potential technology for end...

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

作者： Smith, PJ McCabe, EM Taylor, CM Selviah, DR Day, SE Commander, LG Univ Dublin Trinity Coll Dept Phys Dublin 2 Ireland

ISBN: (纸本)081943535X

Using a new device which contains an array of microlenses whose focal lengths can be electrically varied, we have been able to control the input from one microlens to a single mode fibre using an applied voltage. For such a microlens array many closely-spaced focal spots can be generated in parallel, and electrically switched to address, potentially, an array of receiver fibres. We show how the particular switching characteristics of the device, whereby the lenses switch from diverging to converging, serves in turn to disperse light and to focus it into the fibre.

关键词： microlenses endoscopy direct view microscopy

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