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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201-210 订阅

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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...

引用

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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Time-resolved wide-field optically sectioned fluorescence microscopy

Time-resolved wide-field optically sectioned fluorescence mi...

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

作者： Dupuis, Guillaume Benabdallah, Nadia Chopinaud, Aurelien Mayet, Celine Leveque-Fort, Sandrine Univ Paris 11 CPBM Federat LUMAT FR 2764 Orsay France CNRS Orsay France Univ Paris 11 Inst Sci Mol Orsay UMR 8214 Orsay France

ISBN: (纸本)9780819493583

We present the implementation of a fast wide-field optical sectioning technique called HiLo microscopy on a fluorescence lifetime imaging microscope. HiLo microscopy is based on the fusion of two images, one with structured illumination and another with uniform illumination. Optically sectioned images are then digitally generated thanks to a fusion algorithm. HiLo images are comparable in quality with confocal images but they can be acquired faster over larger fields of view. We obtain 4-D imaging by combining HiLo optical sectioning, time-gated detection, and z-displacement. We characterize the performances of this set-up in terms of 3-D spatial resolution and time-resolved capabilities in both fixed- and live-cell imaging modes.

关键词： fluorescence microscopy fluorescence lifetime imaging microscopy (FLIM) structured illumination microscopy (HiLo) wide-field optical sectioning

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three-dimensional quantitative phase imaging: Current and future perspectives

Three-dimensional quantitative phase imaging: Current and fu...

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

作者： Dragomir, Nicoleta M. Goh, Xiao Ming Roberts, Ann Univ Melbourne Sch Phys Melbourne Vic Australia

ISBN: (纸本)9780819470362

Great effort has been made in the recent past to develop new non-destructive imaging modalities for both two and three dimensional objects, based on the phase properties of a specimen. Quantitative phase tomography (QPT) is a hybrid technique that has been proposed to provide three-dimensional (3D) refractive index (RI) profiling of irregular phase objects by combining transverse phase measurements with traditional tomographic reconstruction techniques. This profiling is accomplished through measurements of sets of projections which are ultimately related to the RI values of the object's transverse cross-section. This is particularly useful for 3D refractive index determination of specimens where staining is not appropriate or for materials that cannot be stained and is essential to many applications in photonics and biotechnology. This article reviews recent developments in quantitative phase tomography as they are presently available and suggests future applications based on current research on the 3D RI. The enabling elements for 3D QPT in the context of four key areas are discussed: the effect of the refractive index of the surrounding matching fluid, spatial resolution, phase accuracy and optimal defocus. Recent progress and future perspectives related to each of these areas is presented with regard to various test objects of known optical properties.

关键词： phase imaging refractive index tomography three-dimensional microscopy

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Dual detection confocal fluorescence microscopy: depth imaging without depth scanning

Dual detection confocal fluorescence microscopy: depth imagi...

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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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Reducing noise in extended depth of field microscope images by optical manipulation of the point spread function

Reducing noise in extended depth of field microscope images ...

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

作者： Zahreddine, Ramzi N. Cormack, Robert H. Cogswell, Carol J. Univ Colorado Dept Elect Comp & Energy Engn Boulder CO 80309 USA

ISBN: (纸本)9780819484413

This work describes improved methods and algorithms for implementing extended depth of field (EDF) microscopy through point spread function (PSF) engineering. It utilizes adaptive optics to create a test bed on which to evaluate new phase shapes for EDF. Being able to quickly and cheaply design novel PSFs is essential to overcome limitations of EDF that have prevented the technology from reaching mainstream use. Further improvement is made by reducing the noise normally seen in EDF images. Computational optics principles are used to first encode the noise with an identifiable pattern and a specially-tailored non-linear algorithm then removes the noise. This approach improves a microscope's imaging capabilities in photon-starved applications such as live-cell fluorescence and object tracking.

关键词： Extended Depth of Field PSF Engineering 3D microscopy Adaptive Optics Neural Networks Noise Removal

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LABEL-FREE CLASSIFICATION OF CELL TYPES BY IMAGING OF CELL MEMBRANE FLUCTUATIONS USING LOW-COHERENT FULL-FIELD QUANTITATIVE PHASE microscopy

LABEL-FREE CLASSIFICATION OF CELL TYPES BY IMAGING OF CELL M...

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

作者： Yamauchi, Toyohiko Sugiyama, Norikazu Sakurai, Takashi Iwai, Hidenao Yamashita, Yutaka Hamamatsu Photon KK Hamakita Ku 5000 Hirakuchi Hamamatsu Shizuoka 4348601 Japan MIT Spectroscopy Lab Cambridge MA 02139 USA Hamamatsu Univ Sch Med Photon Med Res Ctr Hamamatsu Shizuoka 4313192 Japan

ISBN: (纸本)9780819479662

Cell membrane motions of living cells are quantitatively measured in nanometer resolution by low-coherent full-field quantitative phase microscopy. Our setup is based on a full-field phase shifting interference microscope with a very low-coherent light source. The reflection mode configuration and the low-coherent illumination make it possible to differentiate the weak reflection light from the cell membrane from the strong reflection from the glass substrate. Two cell populations are quantitatively assessed by the power spectral density of the cell surface motion and show different trends.

关键词： quantitative phase microscope cell imaging plasma membrane

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Supercritical self-interference fluorescence microscopy for full-field membrane imaging

Supercritical self-interference fluorescence microscopy for ...

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

作者： Barroca, Thomas Bon, Pierre Leveque-Fort, Sandrine Fort, Emmanuel ESPCI ParisTech Inst Langevin Ctr Imageries Plasmon Appl CNRSUMR 7587 1 Rue Jussieu F-75231 Paris 05 France Univ Paris Sud CNRS CLUPS UMR 8214 F-91405 Orsay France

ISBN: (纸本)9780819493583

We present a new technique based on the self-interference of Supercritical Angle Fluorescence (SAF) emission in order to perform full-field cell membrane imaging. We show that our Point Spread Function (PSF) engineering technique allows us to obtain a 100 nm axial sectioning while conserving the original lateral resolution of the microscope. The images are acquired using an optical module that can be connected to any fluorescent microscope to simultaneously monitor in real time both the cell membrane and in-depth phenomena.

关键词： Cell membrane imaging Supercritical Angle Fluorescence (SAF) Fluorescence microscopy Dictyostelium discoideum

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Performance evaluation of point-spread function engineering to reduce the impact of depth-induced aberrations on extended depth-of-field microscopy

Performance evaluation of point-spread function engineering ...

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

作者： Yuan, Shuai Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9780819488701

In this study, we evaluated a point-spread function (PSF) engineered using wave front encoding (WFE) and a generalized cubic phase mask (GCPM) design selected to reduce the impact of depth-induced spherical aberration (SA) on extended depth-of-field (EDOF) microscopy with high NA lenses. Mean-square-error based metrics computed from three-dimensional (3D) depth-variant WFE-PSFs with increasing amounts of SA were used to quantify the engineered PSF's sensitivity to SA and to compare it to the sensitivity of the cubic-phase mask (CPM) PSF traditionally used for EDOF microscopy. The potential performance of the engineered PSF with resilience to SA was further evaluated with simulations in which EDOF images of a 3D object were obtained by processing WFE images with and without SA. A qualitative and quantitative comparison of the EDOF images with the true object show that the WFE-PSF engineering with the selected GCPM design provides better performance in reducing the impact of SA. In addition, the GCPM-based EDOF images do not suffer from the known lateral shift of object features located away from the plane of focus encountered in traditional CPM-based EDOF images.

关键词： Extended depth-of-field microscopy point-spread function engineering wavefront encoding spherical aberration

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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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