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

共 590 条记录，以下是311-320 订阅

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3D microscopy with a double-helix point spread function

3D microscopy with a double-helix point spread function

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

作者： Pavani, Sri Rama Prasanna Piestun, Rafael Univ Colorado Dept Elect & Comp Engn Boulder CO 80309 USA

ISBN: (纸本)9780819474308

Double-helix point spread function (DH-PSF) is an engineered three-dimensional (3D) PSF specifically designed for 3D position estimation and imaging. It exhibits two lobes that rotate continuously around the optical axis with propagation. An information theoretical analysis shows that the DH-PSF carries higher and more uniform Fisher Information than the standard PSF through the 3D volume of interest. Experiments with DH-PSF demonstrate nanometer scale position localization accuracies in all three dimensions. Further, a variety of microscopy techniques such as bright-field, dark-field, and fluorescence can be directly transformed into their DH-PSF counterparts by placing an appropriate phase mask in the imaging path. The flexibility and performance make the DH-PSF attractive for 3D position localization in both photon-limited and photon-unlimited regimes.

关键词： three-dimensional microscopy Point spread function engineering Bright-field Dark-field Fluorescence Computational Optical sensing and imaging

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3D Effects in DIC images of Extended Objects

3D Effects in DIC Images of Extended Objects

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

作者： Sierra, Heidy DiMarzio, Charles A. Brooks, Dana H. Northeastern Univ Dept Elect & Comp Engn Boston MA 02115 USA

ISBN: (纸本)9780819474308

Phase microscopy has been widely used to acquire information about unstained transparent biological objects. These objects are well visualized by techniques such as Differential Interference Contrast (DIC) microscopy, but standard DIC systems don't provide quantitative phase information. Quantitative analysis is limited by the nonlinear relation between the intensity image and the object phase. In this work we combine nonlinearity concepts of the image formation process to model 3D DIC images, specifically the point spread function (PSF) formulation;capturing behavior of measured images along the optical axis that are not obtained when using other models. We verify our model by comparing simulated images to real measured images. This model is designed so that the reconstruction of the 3D properties of imaged specimens can be done in a simple form.

关键词： Differential Interference Contrast (DIC) thick objects coherent microscopy image formation theory

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Coherence measurements applied to critical and Kohler vortex illumination

Coherence measurements applied to critical and Kohler vortex...

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

作者： Brown, Dean P. Brown, Thomas G. Univ Rochester Inst Opt Rochester NY 14627 USA

ISBN: (纸本)9780819474308

Both coherence and polarization play an important role in microscopy. It was long ago established that, in the context of scalar treatments, Kohler and critical illumination produce equivalent coherence functions in an object plane. This paper examines this equivalence in the context of polarization vortex illumination. Using a reversed-wavefront Young interferometer (RWYI), we show measurements of the coherence and correlation properties of the output illumination plane of various illumination systems. We compare the coherence properties of critical and Kohler vortex illumination schemes and look at the effects these properties have in an imaging system.

关键词： Coherence Polarization Vortex Partial Coherent Imaging Critical Illumination Kohler Illumination Polarization microscopy

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Fluorescence microscopy with a coded aperture snapshot spectral imager

Fluorescence microscopy with a coded aperture snapshot spect...

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

作者： Fernandez, Christy A. Wagadarikar, Ashwin Brady, David J. McCain, Scott C. Oliver, Tim Duke Univ Fitzpatrick Inst Photon Durham NC 27708 USA Applied Quantum Technol Durham NC 27707 USA Duke Univ Dept Cell Biol Durham NC 27710 USA

ISBN: (纸本)9780819474308

This paper describes the application of a dual-dispersive (DD) coded aperture snapshot spectral imager (CASSI) to fluorescence microscopy. CASSI records an interleaved spatially varying, spectrally filtered map of an object onto a two-dimensional (2D) focal plane. Using a compressive sensing framework, the spectrally encoded 2D scene is reconstructed into a three-dimensional (3D) data cube. CASSI also records a 3D dataset at video rate making it suitable for dynamic cellular imaging. A convex optimization technique combining least squares QR factorization with a total variance constraint is used to reconstruct the image. The system records 32 spectral channels that span the spectral range between 450nm and 750nm with 10nm spectral resolution for any pixel in a 2D image. This paper illustrates the application of CASSI imaging to fluorescence microscopy applications. We report on the reconstruction of fluorescent microspheres used in fluorescence microscopy as calibration standards. images are compared with a multi-spectral confocal system.

关键词： spectral imaging microscopy fluorescence optical systems

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Two-dimensional resolution improvement in standing wave microscopy using fast acousto-optic laser scanning

Two-dimensional resolution improvement in standing wave micr...

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

作者： Gliko, Olga Brownell, William E. Saggau, Peter Baylor Coll Med Dept Otorhinolaryngol Houston TX 77030 USA Baylor Coll Med Dept Neurosci Houston TX 77030 USA Rice Univ Dept Bioengn Houston TX USA

ISBN: (纸本)9780819474308

We present a versatile scheme for two-dimensional (2D) resolution enhancement in standing wave fluorescence microscopy (SWFM). This SWFM scheme consists of an interferometer, where both beams are focused at the back focal plane of the objective lens. Their position is controlled by a pair of acousto-optic deflectors (AODs). This results in two collimated beams that interfere in the focal plane, creating a lateral periodic excitation pattern with controlled fringe spacing and orientation. The phase of the standing wave (SW) pattern is controlled by the phase delay between two RF sinusoidal signals driving the AODs. An enlarged fluorescence image formed using the same objective lens is captured by a cooled CCD camera. Data collection involves acquiring images with excitation pattern of three equi-polar orientations (0 degrees, 60 degrees and 120 degrees) and three different phases (0 degrees, 120 degrees, 240 degrees) for each orientation. The SWFM image is algebraically reconstructed from these 9 acquired images. The SWFM image has enhanced 2D lateral resolution of about 100 nm with nearly isotropic effective point-spread function (PSF). As a result of the acousto-optic scanning, the total acquisition time can be as short as 100 mu s and is only further limited by the fluorescence intensity, as well as sensitivity and speed of the CCD camera. Utilizing acousto-optic laser scanning for advanced SWFM provides the exceptional precision and speed necessary for real-time imaging of subresolution processes in living biological systems.

关键词： super resolution fluorescence microscopy interference phase shift

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Arbitrary Two-dimensional Multiphoton Excitation Patterns with Temporally Focused Digital Holograms

Arbitrary Two-Dimensional Multiphoton Excitation Patterns wi...

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

作者： Oron, Dan Papagiakomou, Eirini de-Sars, Vincent Emiliani, Valentina Weizmann Inst Sci Dept Phys Complex Syst IL-76100 Rehovot Israel Univ Paris 05 Neurophysiol & New Microscopies Lab Paris France

ISBN: (纸本)9780819474308

Multiphoton excitation has recently found application in the fields of bioimaging, uncaging and lithography. In order to fully exploit the advantages of nonlinear excitation, in particular the axial resolution due to nonlinearity, most systems to date operate with point or multipoint excitation, while scanning either the laser beam or the sample to generate the illumination pattern. Here we combine the recently introduced technique of scanningless multiphoton excitation by temporal focusing with recent advances in digital holography to generate arbitrarily shaped, depth resolved, two-dimensional excitation patterns completely without scanning. This is of particular importance in applications requiring uniform excitation of large areas over short time scales, such as neuronal activation by multiphoton uncaging of neurotransmitters. We present an experimental and theoretical analysis of the effect of spatial patterning on the depth resolution achieved in temporal focusing microscopy. It is shown that the depth resolution for holographic excitation is somewhat worse than that achieved for uniform illumination. This is also accompanied by the appearance of a speckle pattern at the temporal focal plane. The origin of the two effects, as well as means to overcome them, are discussed.

关键词： Temporal focusing Digital holography Multiphoton microscopy

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Live TIRF fluorescence imaging at 100nm resolution through structured illumination

Live TIRF fluorescence imaging at 100nm resolution through s...

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

作者： Kner, P. Chhun, B. Griffis, E. Winoto, L. Shao, L. Gustafsson, M. G. L. Univ Georgia Fac Engn Driftmier Engn Ctr Athens GA 30602 USA Univ Calif San Francisco Dept Biochem & Biophys San Francisco CA 94158 USA

ISBN: (纸本)9780819474308

Linear Structured Illumination is a powerful technique for increasing the resolution of a fluorescence microscope by a factor of two beyond the diffraction limit. Previously this technique has only been used to image fixed samples because the implementation, using a mechanically rotated fused silica grating, was too slow. Here we describe a microscope design, using a ferroelectric spatial light modulator to structure the illumination light, capable of linear structured illumination at frame rates up to 11Hz. We show live imaging of GFP labeled Tubulin and Kinesin in Drosophila S2 cells.

关键词： Structured illumination Super-resolution microscopy Bio-imaging

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Testing for nonrandom shape similarity between sister cells using automated shape comparison

Testing for nonrandom shape similarity between sister cells ...

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

作者： Guo, Monica Marshall, Wallace F. Univ Calif San Francisco Dept Biochem & Biophys Integrat Program Quantitat Biol San Francisco CA 94143 USA

ISBN: (纸本)9780819474308

Several reports in the biological literature have indicated that when a living cell divides, the two daughter cells have a tendency to be mirror images of each other in terms of their overall cell shape. This phenomenon would be consistent with inheritance of spatial organization from mother cell to daughters. However the published data rely on a small number of examples that were visually chosen, raising potential concerns about inadvertent selection bias. We propose to revisit this issue using automated quantitative shape comparison methods which would have no contribution from the observer and which would allow statistical testing of similarity in large numbers of cells. In this report we describe a first order approach to the problem using rigid curve matching. Using test images, we compare a pointwise correspondence based distance metric with a chamfer matching strategy and find that the latter provides better correspondence and smaller distances between aligned curves, especially when we allow nonrigid deformation of the outlines in addition to rotation.

关键词： Morphological image processing curve matching microscopy cytoskeleton actin cytokinesis cell shape

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Thickness and index measurements of a transparent specimen by full-field optical coherence microscopy

Thickness and index measurements of a transparent specimen b...

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

作者： Na, Jihoon Choi, Woo June Choi, Hae Young Ryu, Seon Young Choi, Eun Seo Lee, Byeong Ha Gwangju Inst Sci & Technol Dept Informat & Commun 261 Cheomdan Gwagiro Gwangju 500712 South Korea Chosun Univ Dept Phys Gwangju 501759 South Korea

ISBN: (纸本)9780819474308

We present the method measuring the thickness and the refractive index of a transparent specimen at a same time based on full-field optical coherence tomography. As a sample a small drop of epoxy was placed on a flat plate and the high-resolution depth resolved en-face images of the epoxy drop were taken. With adopting the plate surface as a reference plane, the physical thickness and the refractive index distribution could be obtained. Owing to the full-field imaging capability, we could obtain the transverse distributions of the thickness and the refractive index without any transverse scanning. The measured thickness at the center of the sample was 24 mu m and the average index was 1.4055 with the standard deviation of 0.0002.

关键词： Refractive index measurement Full-field optical coherence microscopy Medical imaging equipment

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Biological imaging beyond the diffraction limit by saturated excitation (SAX) microscopy

Biological imaging beyond the diffraction limit by saturated...

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

作者： Yamanaka, M. Kawano, S. Fujita, K. Smith, N. I. Kawata, S. Osaka Univ Dept Frontier Biosci 2-1 Yamadaoka Suita Osaka 5650871 Japan Osaka Univ Dept Appl Phys Suita Osaka 5650871 Japan RIKEN Nanophoton Lab Wako Saitama 3510198 Japan

ISBN: (纸本)9780819474308

We present an alternative high-resolution fluorescence imaging technique, saturated excitation (SAX) microscopy, for observations of biological samples. In the technique, we saturate the population of fluorescence molecules at the excited state with high excitation intensity. Under this condition, the fluorescence intensity is no longer proportional to the excitation intensity and the relation of the fluorescence and excitation intensity shows strong nonlinearity. In the centre of laser focus, the nonlinear responses induced by the saturation appear notably because of higher excitation intensity. By detecting fluorescence signals from the saturated area, we can push the spatial resolution beyond the diffraction barrier in three dimensions. SAX microscopy can be realized with a simple optics, where a laser intensity modulation sisytem and a lock-in amplifier are simply added to a conventional confocal microscope system. Using the SAX microscope, we demonstrated high-resolution imaging of a biological sample by observing mitochondria in HeLa cells. We also examined the nonlinear response of commercially available dyes under saturated excitation conditions.

关键词： High resolution saturated excitation fluorescence microscopy confocal microscopy biological imaging

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