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

共 591 条记录，以下是221-230 订阅

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Dual detection confocal fluorescence microscopy: depth imaging without depth scanning

Dual detection confocal fluorescence microscopy: depth imagi...

引用

Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XX

作者： Lee, Dong-Ryoung Kim, Young-Duk Jeong, Hyoung-Jun Lee, Jong-Min Gweon, Dae-Gab Yoo, Hongki KAIST (Korea Republic of) Hanyang Univ. (Korea Republic of)

ISBN: (纸本)9780819493583

We propose a new method for three-dimensional fluorescence imaging without depth scanning that we refer to as the dual detection confocal fluorescence microscopy (DDCFM). Compared to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically to collect a three-dimensional images, DDCFM is able to obtain depth information without depth scanning. DDCFM utilizes two photo multiplier tubes (PMTs) in the confocal detection system. The emitted fluorescence is divided by the beam splitter and received by the two PMTs through pinholes with different size. Each PMT signal generates different axial response curve according to the pinhole diameter, which decides stiffness of the curve. Since the PMT signal is determined by the intensity of the fluorescent emitter and the distance from the focal point, we can acquire depth position of a fluorescent emitter by comparing two intensity signals from the PMTs. Since the depth information can be obtained by a single excitation without depth scanning, DDCFM has many advantages. The measurement time is dramatically reduced for volume imaging. Also, photo-bleaching and photo-toxicity can be minimized. The system can be easily miniaturized because no mechanical depth scan is needed. Here, we demonstrate the feasibility of the proposed method by phantom study using fluorescent beads.

关键词： Confocal fluorescence microscopy three-dimensional imaging depth scanning

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Cell morphology classification in phase contrast microscopy image reducing halo artifact

Cell morphology classification in phase contrast microscopy ...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing Xix

作者： Kang, Mi-Sun Song, Soo-Min Lee, Hana Kim, Myoung-Hee Ewha Womans Univ Dept Comp Sci & Engn Seoul 120750 South Korea

ISBN: (纸本)9780819488701

Since the morphology of tumor cells is a good indicator of their invasiveness, we used time-lapse phase-contrast microscopy to examine the morphology of tumor cells. This technique enables long-term observation of the activity of live cells without photobleaching and phototoxicity which is common in other fluorescence-labeled microscopy. However, it does have certain drawbacks in terms of imaging. Therefore, we first corrected for non-uniform illumination artifacts and then we use intensity distribution information to detect cell boundary. In phase contrast microscopy image, cell is normally appeared as dark region surrounded by bright halo ring. Due to halo artifact is minimal around the cell body and has non-symmetric diffusion pattern, we calculate cross sectional plane which intersects center of each cell and orthogonal to first principal axis. Then, we extract dark cell region by analyzing intensity profile curve considering local bright peak as halo area. Finally, we examined cell morphology to classify tumor cells as malignant and benign.

关键词： cell morphology phase contrast microscopy image halo artifact cell classification

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New Approaches for the analysis of confluent cell layers with quantitative phase digital holographic microscopy 13

New Approaches for the analysis of confluent cell layers wit...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Pohl, L. Kaiser, M. Ketelhut, S. Pereira, S. Goycoolea, F. Kemper, Bjoern Univ Munster Fac Med Biomed Technol Ctr Mendelstr 17 D-48149 Munster Germany Univ Munster Inst Plant Biol & Biotechnol Schlossgarten 3 D-48149 Munster Germany

ISBN: (纸本)9781628419474

Digital holographic microscopy (DHM) enables high resolution non-destructive inspection of technical surfaces and minimally-invasive label-free live cell imaging. However, the analysis of confluent cell layers represents a challenge as quantitative DHM phase images in this case do not provide sufficient information for image segmentation, determination of the cellular dry mass or calculation of the cell thickness. We present novel strategies for the analysis of confluent cell layers with quantitative DHM phase contrast utilizing a histogram based-evaluation procedure. The applicability of our approach is illustrated by quantification of drug induced cell morphology changes and it is shown that the method is capable to quantify reliable global morphology changes of confluent cell layers.

关键词： digital holographic microscopy quantitative phase imaging live cell imaging minimally-invasive label-free stain-free confluent cell layers

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Microlymphatic flow using fast video microscopy

Microlymphatic flow using fast video microscopy

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

作者： Dixon, JB Zawieja, DC Gashev, AA Coté, GL Texas A&M Univ Dept Biomed Engn College Stn TX 77843 USA

ISBN: (纸本)0819456756

Despite advances in the measurement of lymphatic flow, little is known about the actual velocities of flow in microlymphatic (similar to 100 um diameter) vessels. In this paper, video microscopy and particle tracking methods were adapted and integrated with an ultra high-speed imaging camera to obtain measurements of high-speed lymph velocities that previous systems were incapable of measuring. In this study, a mesenteric microlymphatic vessel in a loop of the small intestine of a male Sprague-Dawley rat was exteriorized and imaged at a rate of 500 frames per second (fps) for several contraction sequences. Lymph velocity was shown to fluctuate cyclically with the vessel wall contractions and ranged from -1 to 4 mm/sec through a ten second sequence.

关键词： Lymphatic system Cameras Velocity measurements Video microscopy Microfluidics Imaging systems High speed cameras Microfluidic imaging Video Particles

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Digital aberration correction of fluorescent images with coherent holographic image reconstruction by phase transfer (CHIRPT) 13

Digital aberration correction of fluorescent images with coh...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Field, Jeffrey J. Bartels, Randy A. Colorado State Univ Microscopy Core Facil Ft Collins CO 80523 USA Colorado State Univ Dept Elect & Comp Engn Ft Collins CO 80523 USA Colorado State Univ Dept Biochem & Mol Biol Ft Collins CO 80523 USA Colorado State Univ Inst Genome Architecture & Funct Ft Collins CO 80523 USA Colorado State Univ Sch Biomed Engn Ft Collins CO 80523 USA

ISBN: (纸本)9781628419474

Coherent holographic image reconstruction by phase transfer (CHIRPT) is an imaging method that permits digital holographic propagation of fluorescent light. The image formation process in CHIRPT is based on illuminating the specimen with a precisely controlled spatio-temporally varying intensity pattern. This pattern is formed by focusing a spatially coherent illumination beam to a line focus on a spinning modulation mask, and image relaying the mask plane to the focal plane of an objective lens. Deviations from the designed spatio-temporal illumination pattern due to imperfect mounting of the circular modulation mask onto the rotation motor induce aberrations in the recovered image. Here we show that these aberrations can be measured and removed non-iteratively by measuring the disk aberration phase externally. We also demonstrate measurement and correction of systematic optical aberrations in the CHIRPT microscope.

关键词： fluorescence holography aberration correction microscopy

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Adaptive Optics for Wide-field microscopy

Adaptive Optics for Wide-field Microscopy

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XVIII

作者： Bourgenot, C. Saunter, C. D. Girkin, J. M. Love, G. D. Univ Durham Dept Phys Ctr Adv Instrumentat Durham DH1 3LE England

ISBN: (纸本)9780819484413

We report on recent developments in the use of adaptive optics (AO) in wide-field microscopy to remove both system and sample induced aberrations. We describe progress on using both a full AO system and image optimization techniques (wavefront sensorless AO). In the latter system the determination of the best mirror shape is found via two routes. In the first an optimization algorithm using a Simplex search pattern is used with an initial random set of mirror shapes. We then explore the use of specific Zernike terms as our starting basis set. In both cases the final optimization performance is not affected by the choice of optimization metric. We then describe an open loop AO system in which the equivalent of a laser guide star is used as the light source for the wavefront sensor.

关键词： Optical microscopy Aberration correction Adaptive Optics

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

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

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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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Confocal Fluorescence lifetime imaging microscopy based on a realtime sampling method

Confocal Fluorescence lifetime imaging microscopy based on a...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Bae, Yoon. S. Lee, Dongsoo Moon, Sucbei Won, Young Jae Kim, Dug Y. Gwangju Inst Sci & Technol Dept Informat & Commun 1 Oryong Dong Kwangju 500712 South Korea

ISBN: (纸本)9780819465566

We present a fluorescence lifetime imaging microscope (FLIM) based on a real-time waveform acquisition method. The fluorophores were excited by a 635-mn gain-switched laser diode,, which produced short pulses with duration similar to 50 ps in a 20-MEz repetition rate. The fluorescence signals were detected by a silicon avalanche photo-diode (APD) in addition to a wide-band electric amplifier. The converted electric pulses were sampled by a high-speed digitizer of which sampling rate was 2 GS/s. In order to reduce the sampling interval for analyzing sub-nanosecond lifetimes, an interleaved data acquisition technique was used. The effective sampling rate was increased to 10 GS/s. In addition, the impulse response was measured simultaneously with the lifetime signals by an interleaving manner and was used in calibration of the system. By using these methods, accurate lifetime information was acquired in a short time less than 8 mu s.

关键词： confocal microscopy fluorescence lifetime imaging microscopy interleaved data acquisition

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k-microscopy - resolution beyond the diffraction limit

k-microscopy - resolution beyond the diffraction limit

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XV

作者： Geissbuehler, Matthias Lasser, Theo Leitgeb, Rainer A. Ecole Polytech Fed Lausanne Lab Opt Biomed CH-1015 Lausanne Switzerland

ISBN: (纸本)9780819470362

We present a novel Fourier domain method for microscopic imaging - so-called k-microscopy- with lateral resolution independent of the detection numerical aperture. The concept is based on sample illumination by a lateral fringe-pattern of varying spatial frequency, which probes the lateral spatial frequency or k- spectrum of the sample structure. The illumination pattern is realized by interference of two collimated coherent beams. Wavelength tuning is employed for modulation of the fringe spacing. The uniqueness of the proposed system is that a single point detector is sufficient to collect the total light corresponding to a particular position in the sample k-space. By shifting the phase of the interference pattern, we get full access to the complex frequencies. An inverse Fourier transformation of the acquired band in the frequency- or k-space will reconstruct the sample. The resulting lateral resolution will be defined by the temporal coherence length associated with the detected light source spectrum as well as by the illumination angle. The feasibility of the concept has been demonstrated in 1D.

关键词： k-microscopy sub-diffraction limit microscopy wavelength tuning biomedical imaging Fourier optics

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Prospective gating for 3D imaging of the beating zebrafish heart in embryonic development studies

Prospective gating for 3D imaging of the beating zebrafish h...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing Xix

作者： Taylor, J. M. Saunter, C. D. Love, G. D. Girkin, J. M. Univ Durham Dept Phys Ctr Adv Instrumentat Durham DH1 3LE England

ISBN: (纸本)9780819488701

We demonstrate the use of prospective gating from continuously acquired brightfield images of zebrafish embryos to trigger the acquisition of fluorescence images with the heart at a precisely selected position in its cycle. The laser exposure of the sample is reduced by an order of magnitude compared to alternative techniques which acquire many separate fluorescence images for each section before selecting the most appropriate ones to build up a consistent 3D image stack. We present results obtained using our SPIM system including 3D reconstructions of the living, beating heart, acquired using optical gating without the need for any pharmacological or electrophysiological intervention, and discuss possible wider applications of our technique.

关键词： Optical microscopy Selective Plane Illumination Synchronization Fluorescence imaging

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