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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231-240 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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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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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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Time-Resolved Single Molecule microscopy coupled with Atomic Force microscopy

Time-Resolved Single Molecule Microscopy coupled with Atomic...

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

作者： Koenig, M. Koberling, F. Schulz, O. Ros, R. Fore, S. Sackrow, M. Trautmann, S. Erdmann, R. PicoQuant GmbH Rudower Chaussee 29 D-12489 Berlin Germany Arizona State Univ Dept Phys Tempe AZ 85287 USA PicoQuant Photon North Amer Inc Westfield MA 01085 USA

ISBN: (纸本)9780819488701

Time-resolved confocal microscopy is well established to image spectral and spatial properties of samples in biology and material science. Atomic Force microscopy (AFM) in addition enables to investigate properties which are not optically addressable or are hidden by the diffraction limited optical resolution. We present a straight forward combination of single molecule sensitive time-resolved confocal microscopy with different commercially available AFMs. Besides an extra of information about for example a cell surface, the AFM tip can also be used to manipulate the sample on a nanometer scale down to the single molecule level.

关键词： Confocal microscopy Atomic Force microscopy AFM quenching time-resolved fluorescence sub diffraction single molecule

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Performance evaluation of an image estimation method based on principal component analysis (PCA) developed for quantitative depth-variant fluorescence microscopy imaging

Performance evaluation of an image estimation method based o...

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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 3D wide-field computational microscopy, the image formation process is depth variant due to the refractive index mismatch between the imaging layers. In a previous study, an image estimation method based on a principle component analysis (PCA) model for the representation of the depth varying point spread function (DV-PSF) was presented and demonstrated with noiseless simulations(1). In this study, the performance of the PCA-based DV expectation maximization algorithm (PCA-DVEM) was further evaluated with noisy simulations. Different levels of Poisson noise were used in simulated forward images of a synthetic object computed using theoretically-determined DV-PSFs approximated by the PCA approach. The noise influence on the reconstructed images obtained with PCA-DVEM was evaluated. We found that without regularization, the algorithm performs well when the signal-to-noise ratio (SNR) is 14 dB or higher. The relationship of the number of PCA components, B, to the image reconstruction performance was also investigated on both noiseless and noisy simulated data. In both cases, we found that the number of PCA components has limited effect on the image reconstruction performance for B > 1. To reduce computational complexity while maintaining image estimation performance, B = 2 is suggested for processing experimental data.

关键词： computational microscopy principle component analysis spherical aberration 3D image estimation noise analysis

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Background and speckle suppression with a divided pupil and Nomarski prism for reflectance line-scanning confocal microscopy of human tissues

Background and speckle suppression with a divided pupil and ...

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

作者： Glazowski, Christopher Zavislan, James Rajadhyaksha, Milind Mem Sloan Kettering Canc Ctr Dermatol Serv New York NY 10022 USA Univ Rochester Inst Opt Rochester NY 14623 USA

ISBN: (纸本)9780819488701

We present a design of a line-scanning confocal microscope for reduced speckle and improved sectioning performance by the use of two pupil-modification techniques. The first is a divided-pupil configuration in which the illumination and detection paths are separate in object space except in the focal (optical sectioning) plane. The second technique is a novel implementation of a Nomarski prism and quarter-wave retarder, termed NRDIC, which has shown good results in point-scanning confocal microscopy and we will present its translation to line-scanning confocal microscopy. A stable turbid phantom that simulates the background-driven speckle was used for quantitative characterization. Compared to standard full pupil line-scanning, we show improvements in signal to background of 1.8 and 9 for NRDIC and divided pupil, respectively. Preliminary imaging in human skin in-vivo demonstrates the improvements in contrast and reduction of speckle for both the NRDIC and divided pupil modes.

关键词： microscopy turbid speckle confocal line-scanning divided pupil NRDIC

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Multimodal light-sheet microscopy for fluorescence live imaging

Multimodal light-sheet microscopy for fluorescence live imag...

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

作者： Oshima, Y. Kajiura-Kobayashi, H. Nonaka, S. Ehime Univ Hosp Shitukawa Toon City Ehime 7910295 Japan Natl Inst Basic Biol Okazaki Aichi 4448585 Japan

ISBN: (纸本)9780819488701

Light-sheet microscopy, it is known as single plane illumination microscope (SPIM), is a fluorescence imaging technique which can avoid phototoxic effects to living cells and gives high contrast and high spatial resolution by optical sectioning with light-sheet illumination in developmental biology. We have been developed a multifunctional light-sheet fluorescence microscopy system with a near infrared femto-second fiber laser, a high sensitive image sensor and a high throughput spectrometer. We performed that multiphoton fluorescence images of a transgenic fish and a mouse embryo were observed on the light-sheet microscope. As the results, two photon images with high contrast and high spatial resolution were successfully obtained in the microscopy system. The system has multimodality, not only mutiphoton fluorescence imaging, but also hyperspectral imaging, which can be applicable to fluorescence unmixing analysis and Raman imaging. It enables to obtain high specific and high throughput molecular imaging in vivo and in vitro.

关键词： Light-sheet microscopy fluorescence microscopy SPIM DSLM Multiphoton microscopy hyperspectral imaging Raman imaging in vivo imaging

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Calibration of an adaptive microscope using phase diversity

Calibration of an adaptive microscope using phase diversity

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

作者： Debarre, Delphine Vieille, Thibault Facomprez, Aurelie Mahou, Pierre Beaurepaire, Emmanuel Ecole Polytech ParisTech Lab Opt & Biosci CNRS F-91128 Palaiseau France INSERM F-91128 Palaiseau France

ISBN: (纸本)9780819488701

Accurate control over the phase and amplitude modulation in an adaptive microscope is essential to the quality of aberration correction that can be achieved. In this paper we present a robust and compact method for characterising such amplitude and phase modulation in the pupil plane of the focussing objective. This method, based on phase diversity, permits calibrating the microscope as a whole and thus avoids errors in the alignment of the wavefront shaping device after calibration and the resulting imprecision in the induced modulation: by acquiring three 2D images of the point spread function at different distances from the focal plane, we show that the electric field distribution at the pupil plane can be retrieved using an iterative algorithm. We have applied this technique to the characterisation of the phase modulation induced by a deformable mirror when conjugated with the entrance pupil of different objectives, which permits accurate evaluation of the performance of the mirror for subsequent aberration correction.

关键词： Adaptive optics aberration correction deformable mirror phase diversity microscopy

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Analysis of the Effects of Different Resampling Techniques for Optical Coherence Tomography

Analysis of the Effects of Different Resampling Techniques f...

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

作者： Copeland, Christopher N. Ellerbee, Audrey K. Stanford Univ EL Ginzton Lab Stanford CA 94305 USA Stanford Univ Dept Elect Engn Stanford CA 94305 USA

ISBN: (纸本)9780819488701

Fourier domain optical coherence tomography (FD-OCT) uses interferometry and spatially coherent, polychromatic light to acquire cross-sectional images of scattering media such as biological tissue. Phase-sensitive derivatives of FD-OCT, such as spectral domain phase microscopy (SDPM), can perform quantitative phase imaging of cellular dynamics with sub-Angstrom sensitivity. FD-OCT and SDPM images are generated by taking the Fourier Transform of the raw spectral interferogram;the accuracy of the reconstruction depends on the choice of processing algorithms used, system noise, calibration and quantization errors. To decrease the processing time, the Fast Fourier transform (FFT) algorithm can be applied when the data are evenly sampled in wavenumber. Unfortunately, most OCT systems are designed for constant wavelength sampling, not constant wavenumber sampling. While there is general agreement on the qualitative superiority of some resampling methods over others, to the best of our knowledge there has been no study that compares these methods for OCT phase data. We examine the effects of various resampling techniques on simulated phase and amplitude OCT data. Given the trend towards high-speed imaging with OCT, the choice of resampling methods is critical, as one need balance processing time and image accuracy - not merely quality - when analyzing medical image data.

关键词： optical coherence tomography low coherence interferometry phase retrieval microscopy

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Quantifying fluorescence signals in confocal image stacks deep in turbid media

Quantifying fluorescence signals in confocal image stacks de...

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

作者： Beer, S. Maeder, U. Bergmann, T. Burg, J. M. Fiebich, M. Runkel, F. Univ Appl Sci Giessen Tech Hsch Mittelhessen Inst Med Phys & Radiat Protect Wiesenstr 14 D-35390 Giessen Germany Univ Appl Sci Giessen Tech Hsch Mittelhessen Inst Biopharmaceut Technol D-35390 Giessen Germany

ISBN: (纸本)9780819488701

When confocal depth stacks are taken, the collected signal (normally the fluorescence signal), decays dependent of the depth of the confocal slice in the turbid medium. This decay is caused by scattering and absorption of the exciting light and of the fluorescence light. As the attenuation parameters, i.e. scattering and absorption coefficients, are normally unknown when observing a new sample, a method is proposed to compensate for the attenuation of the involved light by correcting the fluorescence signal using the attenuation behavior of the sample measured directly on the spot where the fluorescence stack is taken. The method works without any a priori knowledge about the optical properties of the sample. Using this self-reference technique, a confocal fluorescence depth stack can be created where the signal intensity is not dependent on the scattering and absorption caused intensity decay. The proposed method is tested on fluorescent beads embedded in scattering and absorbing hydrogel phantoms.

关键词： confocal microscopy image processing signal correction

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