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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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Multimodal light-sheet microscopy for fluorescence live imaging

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

引用

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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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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Extension of multidimensional microscopy to ultra-sensitive applications with maximum-likelihood analysis

Extension of multidimensional microscopy to ultra-sensitive ...

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

作者： Davis, Lloyd M. Shen, Guoqing Univ Tennessee Inst Space Ctr Laser Applicat Tullahoma TN 37388 USA

ISBN: (纸本)9780819465566

multidimensional fluorescence microscopy is finding service in forefront biological studies that require separation of images from different fluorophores. For example, commercial microscopes are available with multi-band analog detectors and user-friendly software for "linear unmixing" of species with overlapping emission spectra. To extend such techniques to ultrasensitive and single-molecule applications, we have developed a custom-built microscope, which incorporates two tunable-wavelength picosecond dye lasers for pulse-interleaved laser excitation, angle-tuned reflection of the laser beams from narrow-band Raman notch filters to introduce epi-illumination and provide strong rejection of scattered laser wavelengths, diffraction-limited confocal imaging with 3-dimensional piezo-scanning, an adjustable prism spectrometer for high-throughput resolution of collected fluorescence into 4 spectral bands, and a 4-channel high-quantum efficiency avalanche diode for sub-nanosecond-resolved single-photon detection. Custom software enables multi-band fluorescence correlation spectroscopy and identification of photon bursts for single-molecule detection. For umnixing of spectrally-overlapping signatures for ultrasensitive molecular imaging applications, we find that maximum-likelihood analysis can out-perform least-squares-based linear unmixing in the regime of low photon numbers per spectral/temporal channel. Also, the likelihood surface provides the confidence of the parameter estimates and the covariance of the species contributions. Monte Carlo simulations show that bias in the results of the analysis, which stems from the constraint that photon numbers should be positive, becomes more pronounced at low signal levels, for both maximum-likelihood and least-squares based uninixing.

关键词： fluorescence microscopy spectral-FLIM maximum-likelihood multidimensional microscopy photon counting

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Modeling images of phase information for three-dimensional objects

Modeling images of phase information for three-dimensional o...

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three-dimensional and multidimensional microscopy: image acquisition and processing XV

作者： Sierra, Heidy DiMarzio, Charles A. Brooks, Dana H. Electrical and Computer Engineering Department Northeastern University 360 Huntington Avenue Boston MA 02115 United States

ISBN: (纸本)9780819470362

In this work we present a model that describes the electric field in the image plane of a transparent 3D object. The imaged object is modeled as a stack of independent planes along the optical axis and the phase from all the object planes at the image plane is considered as a superposition of the 2D phase contributions from all slices. The object is defined as a phase transmission function and the backscattered field is neglected. We present and discuss examples of simulated data for 3D objects with phase variations along the optical axis.

关键词： Electric fields

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Adaptive compensation of aberrations in ultrafast 3D microscopy using a deformable mirror

Adaptive compensation of aberrations in ultrafast 3D microsc...

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

作者： Sherman, L Albert, O Schmidt, C Vdovin, G Mourou, G Norris, TB Univ Michigan Ctr Ultrafast Opt Sci Ann Arbor MI 48109 USA

ISBN: (纸本)081943535X

3D imaging using a multiphoton scanning confocal microscope is ultimately Limited by aberrations of the system. We describe a system to adaptively compensate the aberrations with a deformable mirror. We have increased the transverse scanning range of the microscope by three with compensation of off-axis aberrations. We have also significantly increased the longitudinal scanning depth with compensation of spherical aberrations from the penetration into the sample. Our correction is based on a genetic algorithm that uses second harmonic or two-photon fluorescence signal excited by femtosecond pulses from the sample as the enhancement parameter. This allows us to globally optimize the wavefront without a wavefront measurement. To improve the speed of the optimization we use Zernike polynomials as the basis for correction. Corrections can be stored in a database for look-up with future samples.

关键词： scanning confocal microscopy aberration correction

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Studying the illumination puzzle towards an isotropic increase of optical resolution - art. no. 686112

Studying the illumination puzzle towards an isotropic increa...

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

作者： Cella, Francesca Ronzitti, Emiliano Vicidomini, Giuseppe Mondal, Parta Pratim Diaspro, Alberto Univ Genoa LAMBS IFOM MicroScoBio Res Ctr Dept Phys DIFI I-16145 Genoa Italy

ISBN: (纸本)9780819470362

The aim of this work is to propose and analyze optical schemes to obtain an improvement of resolution in optical fluorescence microscopy. This goal can be achieved by implementing interfering illumination beams. We start from the insertion, on the illumination arm of the confocal microscope, of appropriately phase plates inducing laterally interfering beams, and then we propose to exploit two-photon excitation, too. We plan to implement solutions for shaping also the axial component of the point spread function by use of phase-only pupil filters and binary filters. In order to implement such schemes we use a computational simulation mainly based on a vectorial approach coupled to experimental procedures utilizing ultra-thin fluorescent layers and thick gels containing immobile fluorescent molecules as 2D and 3D phantoms, respectively. As well, image processing and successive views can be recombined to get a final isotropic improvement of resolution.

关键词： confocal microscopy two-photon excitation microscopy superresolution filters

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Simulating structured-illumination microscopy in the presence of spherical aberrations

Simulating structured-illumination microscopy in the presenc...

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

作者： Preza, Chrysanthe Univ Memphis Computat Imaging Res Lab Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9780819484413

The effect of depth-induced spherical aberrations (SA) on structured illumination microscopy (SIM)(1) is investigated. SIM is a technique used in three-dimensional (3D) fluorescence microscopy to improve resolution in optical sections acquired from 3D specimens. A 3D depth-variant imaging model was developed to predict the intermediate SIM or grid images that are used by the SIM approach to compute improved optical sections. The model incorporates SA due to imaging depth within a sample when there is a refractive index (RI) mismatch between the average RI of the specimen and the RI of the immersion medium of the lens. The model was implemented using a stratum-based model approximation and multiple depth-variant point-spread functions (PSFs)(2). SIM optical sections were computed using the subtraction algorithm(1, 3) and simulated grid images that include SA predicted by our model. Simulations were performed for different imaging conditions by varying the grid frequency, the amount of SA and the level of noise added to the grid images. Simulated results demonstrate that SIM images are less accurate in the presence of SA, and confirm that the SIM approach is very sensitive to system noise resulting in a reduced SNR in the optically sectioned images.

关键词： Structured illumination microscopy optical sectioning microscopy 3D fluorescence microscopy spherical aberrations depth-variant imaging depth-variant point spread functions

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Autostereoscopic visualization of 3D time-varying complex objects in volumetric image sequences

Autostereoscopic visualization of 3D time-varying complex ob...

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

作者： Benassarou, A. Valette, G. Debons, D. Remion, Y. Lucas, L. CReSTIC SIC Rue CrayeresBP 1035 F-51687 Reims 2 France 3DTV Solut F-51100 Reims France

ISBN: (纸本)9780819484413

This paper presents 4dVizMed, a framework for interactive analysis and autostereoscopic visualization of 3d time-varying objects in volumetric image sequences. It combines a deformable surface model which automatically tracks volumetric features, real-time multi-view stereo volume rendering, and some interactive tools for manipulation and quantization. Our method is based on a topological feature tracking process, using a flow-based paradigm and a deformable surface model. It tracks through time the evolution of the components of an isosurface and their interaction with other components. We focus on the difficulties of visualizing 4d volume data, and we report the results of preliminary experiments designed to evaluate the utility of autostereoscopic displays for this purpose.

关键词： Visualization 3D modeling 3D image processing Autostereoscopic displays image visualization image segmentation 3D displays 3D visualizations image processing Volume rendering

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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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Evaluation of a combined two-color phase plate forming three-dimensional dark holes in super resolution microscopy 27

Evaluation of a combined two-color phase plate forming three...

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

作者： Nagai, Koumei Maruyama, Takashi Kodaira, Akira Kumagai, Hiroshi Bokor, Nandor Iketaki, Yoshinori NTT Adv Technol Corp 3-1 Morinosato Wakamiya Atsugi Kanagawa 2430124 Japan Kitasato Univ Sch Allied Hlth Sci Phys 1-15-1 Kitasato Sagamihara Kanagawa 2520373 Japan Budapest Univ Technol & Econ Dept Phys Budafoki Ut 8 H-1111 Budapest Hungary Olympus Corp 2-3 Kuboyama Cho Hachioji Tokyo 1928512 Japan

ISBN: (纸本)9781510632547

In this study, a combined two-color phase plate (CTPP) was designed for super-resolution microscopy based on up conversion fluorescence depletion (FD), fabricated, and evaluated. It is composed of two types of phase plates, a spiral phase plate and an annular phase plate. A two-color phase plate modulates the phase of the erase beam while maintaining the phase of the pump beam. SRM performed using the proposed CTPP is expected to enable super resolution in both the focal plane and in the optical axis direction. Despite its complex structure, a highly accurate CTPP was obtained by using the exposure and etching processes used in semiconductor manufacturing.

关键词： Super-resolution microscopy Phase modulation Fluorescence depletion Optical-axis resolution

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