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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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A wide-field microscopy technique using a linear image sensor for obtaining 3D images

A wide-field microscopy technique using a linear image senso...

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

作者： Macedo, MP Barata, AJ Fernandes, AG Correia, CM Inst Engn Coimbra Dept Phys & Math P-3030 Coimbra Portugal

ISBN: (纸本)0819456756

Three-dimensional (3D) microscopy is a huge field that includes several microscopy techniques. Among these techniques the confocal microscopy is widely known and used due to its improved axial resolution or optical sectioning ability. However its high cost and image acquisition time as well as low signal-to-noise ratio are important drawbacks. Other techniques use wide-field structured illumination to get depth information from the sample. The detectors in commercial microscopes are normally of the point detector type (PMTs) in confocal and area sensors (CCDs) in widefield microscopy. Examples of microscopes that take advantage of lower cost and faster readout cycles of linear image sensors are very uncommon. The purpose of this work is to develop a low-cost microscopy technique for obtaining 3D images with lower acquisition time and higher signal-to-noise ratio (SNR) than in classical confocal microcopy. It uses a linear CMOS image sensor and specific reconstruction algorithms will be able to extract 3D information from the distribution of light intensity collected. The sensor readout circuitry has been developed and tests are currently running in a laboratory prototype in reflection mode with an epi-illuminated configuration of scanning-stage type. The light source is a commercial incandescent lamp with regulated intensity. For the development, test and optimization of the reconstruction algorithms the object is mounted on a 3-axis translation stage to scan the object in three perpendicular directions. Contrast and resolution results obtained using a resolution test slide and preliminary images of integrated circuit (IC) bonding are presented.

关键词： 3D microscopy confocal microscopy reconstruction algorithms optical sectioning linear sensor

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Multimodal optical microscope for imaging biological samples

Multimodal optical microscope for imaging biological samples

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

作者： Townsend, DJ DiMarzio, CA Laevsky, G Rajadhvaksha, M Northeastern Univ Ctr Subsurface Sensing & Imaging Syst Boston MA 02115 USA

ISBN: (纸本)0819456756

A multimodal microscope provides a researcher with the ability to capture images with multiple sources of contrast. Previous attempts at multimodal microscopy have been limited to capturing images at different instruments and digitally registering the images empirically using features common in the specimen. Other multimodal microscopes have combined different microscopy techniques, but have been limited in their sources of contrast. We present a unique microscope which contains 5 different imaging modalities, with four different sources of contrast. Quadrature microscopy measures the phase of the electric field of the light transmitted through an optically transparent specimen, by interference with a reference beam, using a polarimetric technique to resolve the ambiguities between phase and amplitude. Differential Interference Contrast (DIC) measures the phase gradient in an optically transparent specimen by combining two spatially separated images interferometrically. Confocal reflectance measures index of refraction changes in a plane or reflection from small scatterers. Confocal fluorescence and 2-photon laser scanning microscopy, measure fluorescent signatures of a specimen. The last three of these are inherently capable of producing three-dimensional images directly, through localized probing.

关键词： multimodal microscopy quadrature confocal

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Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy

Visualization of nanocontractions in cardiomyocytes by simul...

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

作者： Greenhalgh, C Barzda, V Elmore, S van Beeo, JHGM Der Au, JA Squier, J Univ Toronto Dept Phys Mississauga ON L5L 1C6 Canada

ISBN: (纸本)0819456756

Dynamic imaging of cardiomyocytes was performed with a simultaneous detection second harmonic generation (SHG), third harmonic generation (THG) and multiphoton excitation fluorescence (MPF) microscope. The fast scanning system of similar to 12 frames/second synchronized with multichannel detection provided the possibility of imaging three dimensional static and two dimensional dynamic structures of cardiomyocytes. The SHG images highlighted the myofibrils of the cardiomyocytes while THG images revealed the locations of mitochondria. Dynamic data showed that during imaging, chaotic nanocontractions took place inside the cardiomyocytes. The time series of THG images reveled large intensity fluctuations i flickeringi in the regions of mitochondria. The flickering in THG correlated with the flickering in MPF. Addition of the uncoupler FCCP inhibited flickering in THG and MPF, and also inhibited nanocontractions. The simultaneous imaging with SHG, THG and MPF proved to be a very powerful microscopy tool for investigation of interactions of different organelles inside a cell.

关键词： harmonic generation imaging femtosecond laser scanning microscopy dynamic microscopy biological imaging contractility of myocytes mitochondria bioenergetics

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Variable-focal length microlens arrays in confocal microscopy.

Variable-focal length microlens arrays in confocal microscop...

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

作者： Mac Raighne, A Wang, JG Cabe, EM Scharf, T Trinity Coll Dublin Dept Phys Dublin Ireland

ISBN: (纸本)0819456756

Microlenses have been implemented in confocal systems successfully as components of aperture arrays and as arrays of objective lenses. The use of the novel technology of variable focal length (VFL) microlenses in the confocal system is investigated. The use of VFL microlenses as an aperture array in conjunction with an optical fiber as a pinhole array is examined. Axial responses of the system where measured and the Full-Width Half Maximum (FWHM) found to be similar to 16 mu m. VFL microlenses as an array of objective lenses is investigated with a novel method for scanning in the axial direction presented. By variation of the focal length of the lenses the focal plane can be scanned through the sample without the mechanical movement of the sample or the objective lens, we have named this 'focal scanning'. It is shown that the limiting factor with this type of scanning is the low numerical aperture (NA) of the microlenses available. Both focal scanning and conventional scanning are examined for this experimental set-up.

关键词： confocal microscopy variable focal length microlenses axial scanning

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Spectral domain phase microscopy: A new tool for measuring cellular dynamics and cytoplasmic flow

Spectral domain phase microscopy: A new tool for measuring c...

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

作者： McDowell, EJ Choma, MA Ellerbee, AK Izatt, JA Duke Univ Dept Biomed Engn Durham NC 27708 USA

ISBN: (纸本)0819456756

Broadband interferometry is an attractive technique for the detection of cellular motions because it provides depth-resolved interferometric phase information via coherence gating. Here a phase sensitive technique called spectral domain phase microscopy (SDPM) is presented. SDPM is a functional extension of spectral domain optical coherence tomography that allows for the detection of cellular motions and dynamics with nanometer-scale sensitivity. This sensitivity is made possible by the inherent phase stability of spectral domain OCT combined with common-path interferometry. The theory that underlies this technique is presented, the sensitivity of the technique is demonstrated by the measurement of the thermal expansion coefficient of borosilicate glass, and the response of an Amoeba proteus to puncture of its cell membrane is measured. We also exploit the phase stability of SDPM to perform Doppler flow imaging of cytoplasmic streaming in A. proteus. We show reversal of cytoplasmic flow in response to stimuli, and we show that the cytoplasmic flow is laminar (i.e. parabolic) in nature. We are currently investigating the use of SDPM in a variety of different cell types.

关键词： Optical Coherence Tomography interferometry interference microscopy Doppler flow imaging cellular dynamics

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An algorithm for image stitching and blending

An algorithm for image stitching and blending

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

作者： Rankov, V Locke, RJ Edens, RJ Barber, PR Vojnovic, B Mt Vernon Hosp Gray Canc Inst Adv Technol Dev Grp Northwood HA6 2JR Middx England

ISBN: (纸本)0819456756

In many clinical studies, including those of cancer, it is highly desirable to acquire images of whole tumour sections whilst retaining a microscopic resolution. A usual approach to this is to create a composite image by appropriately overlapping individual images acquired at high magnification under a microscope. A mosaic of these images can be accurately formed by applying image registration, overlap removal and blending techniques. We describe an optimised, automated, fast and reliable method for both image joining and blending. These algorithms can be applied to most types of light microscopy imaging. Examples from histology, from in vivo vascular imaging and from fluorescence applications are shown, both in 2D and 3D. The algorithms are robust to the varying image overlap of a manually moved stage, though examples of composite images acquired both with manually-driven and computer-controlled stages are presented. The overlap-removal algorithm is based on the cross-correlation method;this is used to determine and select the best correlation point between any new image and the previous composite image. A complementary image blending algorithm, based on a gradient method, is used to eliminate sharp intensity changes at the image joins, thus gradually blending one image onto the adjacent 'composite'. The details of the algorithm to overcome both intensity discrepancies and geometric misalignments between the stitched images will be presented and illustrated with several examples.

关键词： image stitching blending mosaic images

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En-face optical coherence imaging for three-dimensional microscopy

En-face optical coherence imaging for three-dimensional micr...

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conference on 3-dimensional and multidimensional microscopy: image acquisition and processing IX

作者： Akiba, M Chan, KP Tanno, N Japan Sci & Technol Corp Reg Joint Res Project Yamagata 9902473 Japan

ISBN: (纸本)0819443603

En-face optical coherence tomography (OCT) employing parallel heterodyne detection technique is demonstrated for three-dimensional microscopy. To enable the use of commercially available CCD cameras as two-dimensional heterodyne detector arrays in OCT imaging, a frequency synchronous detection method is Employed. Depth-resolved, full-field images are acquired at 30 Hz video-rate without lateral scanning.

关键词： three-dimensional microscopy optical coherence tomography frequency synchronous detection

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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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Fluorescence lifetime imaging microscopy based on a synchroscan streak camera

Fluorescence lifetime imaging microscopy based on a synchros...

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

作者： Qu, J Li, J Niu, HB Shenzhen Univ Inst Optoelect Shenzhen 518060 Guangdong Prov Peoples R China

ISBN: (纸本)0819452327

A new time-domain two-dimensional fluorescence lifetime detection method and a fluorescence lifetime imaging microscope that is based on a synchroscan streak camera are presented. This system can generate in parallel a complete two-dimensional fluorescence lifetime image without scanning the light. We demonstrate the acquisition of a fluorescence lifetime image of a semiconductor ET material sample at a temporal resolution of 2ps and spatial resolution of 4mum within 40ms.

关键词： fluorescence lifetime fluorescence lifetime imaging fluorescence lifetime imaging microscopy synchroscan scan streak camera temporal resolution spatial resolution

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Three-dimensional optical computed-tomography microscope

Three-dimensional optical computed-tomography microscope

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

作者： Chamgoulov, R Lane, P MacAulay, C British Columbia Canc Res Ctr Canc Imaging Dept Vancouver BC V5Z 1L3 Canada

ISBN: (纸本)0819456756

We present our recent results on the development of three-dimensional (3-D) optical computed-tomography microscope. The instrument is a novel imaging device for the 3-D visualization and quantitative analysis of absorption-stained biological samples. The first instrument developed by our group at the BC Cancer Research Centre used a digital micromirror device (DMD) as a spatial light modulator to control the angles of illumination. This new embodiment employs an optical scanner instead of the DMD. The optical scanner is placed in the illumination path of the microscope system, conjugate to the field plane. The optical system includes also two high numerical aperture objective lenses, a sample stage, a light source, and a CCD camera. Projections are acquired by illuminating a specimen at a number of selected angles within the numerical aperture of the objective (0 <= phi <= 135 degrees). A new reconstruction algorithm that employs both transform-based and iterative methods is developed to address the limited-angle reconstruction problem. A transform-based reconstruction is used as an initial starting point for the following iterative reconstruction. A feedback correction of the reconstructed image is made on each iteration step. The algorithm enables to incorporate previously known information about the object into the reconstruction process, and improves the reconstruction accuracy. Microscopic 3-D volume reconstructions of quantitatively absorption-stained cells have been generated. The system enables one to look at multiple optical levels of a specimen, and at more natural tissue architecture, including intact cells. Axial and lateral resolutions were measured to be better than 6 microns.

关键词： optical microscopy 3-D imaging computed tomography limited-angle reconstruction

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