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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101-110 订阅

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A state space based approach to localizing single molecules from multi-emitter images 24

A state space based approach to localizing single molecules ...

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

作者： Vahid, Milad R. Chao, Jerry Ward, E. Sally Ober, Raimund J. Texas A&M Univ Dept Biomed Engn College Stn TX 77843 USA Texas A&M Univ Hlth Sci Ctr Dept Mol & Cellular Med College Stn TX 77843 USA Texas A&M Univ Hlth Sci Ctr Dept Microbial Pathogenesis & Immunol College Stn TX 77843 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Single molecule super-resolution microscopy is a powerful tool that enables imaging at sub-diffraction-limit resolution. In this technique, subsets of stochastically photoactivated fluorophores are imaged over a sequence of frames and accurately localized, and the estimated locations are used to construct a high-resolution image of the cellular structures labeled by the fluorophores. Available localization methods typically first determine the regions of the image that contain emitting fluorophores through a process referred to as detection. Then, the locations of the fluorophores are estimated accurately in an estimation step. We propose a novel localization method which combines the detection and estimation steps. The method models the given image as the frequency response of a multi-order system obtained with a balanced state space realization algorithm based on the singular value decomposition of a Hankel matrix, and determines the locations of intensity peaks in the image as the pole locations of the resulting system. The locations of the most significant peaks correspond to the locations of single molecules in the original image. Although the accuracy of the location estimates is reasonably good, we demonstrate that, by using the estimates as the initial conditions for a maximum likelihood estimator, refined estimates can be obtained that have a standard deviation close to the Cram ' er-Rao lower bound-based limit of accuracy. We validate our method using both simulated and experimental multi-emitter images.

关键词： Fluorescence microscopy frequency response multi-emitter localization single molecule localization single molecule microscopy singular value decomposition state space realization super-resolution microscopy

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Multi-beam optical coherence tomography for microvascular imaging of human skin in vivo 24

Multi-beam optical coherence tomography for microvascular im...

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

作者： Chen, Chaoliang Cheng, Kyle H. Y. Jakubovic, Raphael Jivraj, Jamil Ramjist, Joel Deorajh, Ryan Gao, Wanrong Yang, Victor X. D. Nanjing Univ Sci & Technol Dept Opt Engn Nanjing Jiangsu Peoples R China Ryerson Univ Dept Elect & Comp Engn Biophoton & Bioengn Lab Toronto ON Canada Sunnybrook Hlth Sci Ctr Div Neurosurg Toronto ON Canada Ryerson Univ Dept Biomed Phys Toronto ON Canada Univ Toronto Fac Med Div Neurosurg Toronto ON Canada

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

In this paper, a multi-beam optical coherence tomography (OCT) was used to reconstruct the microvascular image of human skin in vivo with phase resolved Doppler OCT (PRDOCT), phase resolved Doppler variance (PRDV) and speckle variance OCT (svOCT), in which the blood flow image was calculated by averaging the four blood flow images obtained by the four beams. In PRDOCT method, it is difficult to detect the blood flow perpendicular to optical axis of the probe beam for single beam OCT, but the multi-beam scanning method can solve this because the input angles of the four probe beams are slightly different from each other. The proposed method can further improve the signal-to-noise ratio (SNR) of the blood flow signals extracted by the three methods mentioned above.

关键词： Optical coherence tomography medical and biological imaging tomographic image processing

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Simple and fast spectral domain algorithm for quantitative phase imaging of living cells with digital holographic microscopy 24

Simple and fast spectral domain algorithm for quantitative p...

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

作者： Min, Junwei Yao, Baoli Ketelhut, Steffi Kemper, Bjoern Univ Munster Biomed Technol Ctr Mendelstr 17 D-48149 Munster Germany Chinese Acad Sci Xian Inst Opt & Precis Mech State Key Lab Transient Opt & Photon Xian 710119 Shaanxi Peoples R China

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

The modular combination of optical microscopes with digital holographic microscopy (DHM) has been proven to be a powerful tool for quantitative live cell imaging. The introduction of condenser and different microscope objectives (MO) simplifies the usage of the technique and makes it easier to measure different kinds of specimens with different magnifications. However, the high flexibility of illumination and imaging also causes variable phase aberrations that need to be eliminated for high resolution quantitative phase imaging. The existent phase aberrations compensation methods either require add additional elements into the reference arm or need specimen free reference areas or separate reference holograms to build up suitable digital phase masks. These inherent requirements make them unpractical for usage with highly variable illumination and imaging systems and prevent on-line monitoring of living cells. In this paper, we present a simple numerical method for phase aberration compensation based on the analysis of holograms in spatial frequency domain with capabilities for on-line quantitative phase imaging. From a single shot off-axis hologram, the whole phase aberration can be eliminated automatically without numerical fitting or pre-knowledge of the setup. The capabilities and robustness for quantitative phase imaging of living cancer cells are demonstrated.

关键词： Digital holographic microscopy aberration compensation frequency spectrum analysis quantitative phase imaging quantitative live cell analysis

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Accurate Time-Domain Angular Jitter Measurements for a High-Speed Polygon Scanner 24

Accurate Time-Domain Angular Jitter Measurements for a High-...

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

作者： Ji, Youn Young So, Byung Hwy Hwang, Won Sang Kim, Don Geun Kim, Jun Woo Kim, Dug Young Yonsei Univ Dept Phys 50 Yonsei Ro Seoul 120749 South Korea

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Angles of polygon scanners have been measured by using rotary encoders, autocollimators or indexing tables. These methods produce precise angle values but require removal of polygon mirror from its motor. For resolving this inconvenience, we introduce a simple angle measurement method by measuring timing jitters of a scanned beam in the time-domain with a high-speed detector and a digitizer while a polygon scanner is rotating at its full speed. Our setup includes a 635 nm wavelength semiconductor laser, a high-speed photodiode, two lenses, and a high-speed digitizer. A polygon scanner with 12 facets were tested with a rotating frequency of near 350 Hz. To detect the signal of the photodiode, we used a high speed digitizer which has a sampling rate of 2Gs/s with 256MB on-board memory. We obtained repeated pulsed sequential photodiode signals for 12 mirror facets of the scanner. Angle variations and their jitters for 12 scanner mirror facets were successfully calculated from measured data. We have repeated same experiments with a photomultiplier tube and compared results with those measured by a photodiode.

关键词： Polygon mirror Polygon Scanner Angle measurement Laser printer Confocal microscopy Laser scanner

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Comparison of two structured illumination techniques based on different 3D illumination patterns 24

Comparison of two structured illumination techniques based o...

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

作者： Shahani, H. Patwary, N. Doblas, A. Saavedra, G. Preza, C. Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt E-46100 Burjassot Spain

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Manipulating the excitation pattern in optical microscopy has led to several super-resolution techniques. Among different patterns, the lateral sinusoidal excitation was used for the first demonstration of structured illumination microscopy (SIM), which provides the fastest SIM acquisition system (based on the number of raw images required) compared to the multi-spot illumination approach. Moreover, 3D patterns that include lateral and axial variations in the illumination have attracted more attention recently as they address resolution enhancement in three dimensions. A threewave (3W) interference technique based on coherent illumination has already been shown to provide super-resolution and optical sectioning in 3D-SIM. In this paper, we investigate a novel tunable technique that creates a 3D pattern from a set of multiple incoherently illuminated parallel slits that act as light sources for a Fresnel biprism. This setup is able to modulate the illumination pattern in the object space both axially and laterally with adjustable modulation frequencies. The 3D forward model for the new system is developed here to consider the effect of the axial modulation due to the 3D patterned illumination. The performance of 3D-SIM based on 3W interference and the tunable system are investigated in simulation and compared based on two different criteria. First, restored images obtained for both 3D-SIM systems using a generalized Wiener filter are compared to determine the effect of the illumination pattern on the reconstruction. Second, the effective frequency response of both systems is studied to determine the axial and lateral resolution enhancement that is obtained in each case.

关键词： 3D structured illumination microscopy 3D sinusoidal pattern axial and lateral modulation frequency

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Probing neural tissue with Airy light-sheet microscopy: investigation of imaging performance at depth within turbid media 24

Probing neural tissue with Airy light-sheet microscopy: inve...

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

作者： Nylk, Jonathan McCluskey, Kaley Aggarwal, Sanya Tello, Javier A. Dholakia, Kishan Univ St Andrews Sch Phys & Astron SUPA St Andrews KY16 9SS Fife Scotland Univ St Andrews Sch Med St Andrews KY16 9TF Fife Scotland

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Light-sheet microscopy (LSM) has received great interest for fluorescent imaging applications in biomedicine as it facilitates three-dimensional visualisation of large sample volumes with high spatiotemporal resolution whilst minimising irradiation of, and photo-damage to the specimen. Despite these advantages, LSM can only visualise superficial layers of turbid tissues, such as mammalian neural tissue. Propagation-invariant light modes have played a key role in the development of high-resolution LSM techniques as they overcome the natural divergence of a Gaussian beam, enabling uniform and thin light-sheets over large distances. Most notably, Bessel and Airy beam-based light-sheet imaging modalities have been demonstrated. In the single-photon excitation regime and in lightly scattering specimens, Airy-LSM has given competitive performance with advanced Bessel-LSM techniques. Airy and Bessel beams share the property of self-healing, the ability of the beam to regenerate its transverse beam profile after propagation around an obstacle. Bessel-LSM techniques have been shown to increase the penetration-depth of the illumination into turbid specimens but this effect has been understudied in biologically relevant tissues, particularly for Airy beams. It is expected that Airy-LSM will give a similar enhancement over Gaussian-LSM. In this paper, we report on the comparison of Airy-LSM and GaussianLSM imaging modalities within cleared and non-cleared mouse brain tissue. In particular, we examine image quality versus tissue depth by quantitative spatial Fourier analysis of neural structures in virally transduced fluorescent tissue sections, showing a three-fold enhancement at 50 p.m depth into non-cleared tissue with AiryLSM. Complimentary analysis is performed by resolution measurements in bead-injected tissue sections.

关键词： Light-sheet microscopy LSM Airy beam Tissue imaging turbid media neuroscience

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White matter segmentation by estimating tissue optical attenuation from volumetric OCT massive histology of whole rodent brains 24

White matter segmentation by estimating tissue optical atten...

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

作者： Lefebvre, Joel Castonguay, Alexandre Lesage, Frederic Ecole Polytech Montreal CP 6079Succ Ctr Ville Montreal PQ H3C 3A7 Canada Inst Cardiol Montreal 5000 Rue Belanger Est Montreal PQ H1T 1C8 Canada

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

A whole rodent brain was imaged using an automated massive histology setup and an Optical Coherence Tomography (OCT) microscope. Thousands of OCT volumetric tiles were acquired, each covering a size of about 2.5x2.5x0.8 mm3 with a sampling resolution of 4.9x4.9x6.5 microns. This paper shows the techniques for reconstruction, attenuation compensation and segmentation of the sliced brains. The tile positions within the mosaic were evaluated using a displacement model of the motorized stage and pairwise coregistration. Volume blending was then performed by solving the 3D Laplace equation, and consecutive slices were assembled using the cross-correlation of their 2D image gradient. This reconstruction algorithm resulted in a 3D map of optical reflectivity for the whole brain at micrometric resolution. OCT tissue slices were then used to estimate the local attenuation coefficient based on a single scattering photon model. The attenuation map obtained exhibits a high contrast for all white matter fibres, regardless of their orientation. The tissue optical attenuation from the intrinsic OCT reflectivity contributes to better white matter tissue segmentation. The combined 3D maps of reflectivity and attenuation is a step toward the study of white matter at a microscopic scale for the whole brain in small animals.

关键词： Massive histology Volume reconstruction image processing Optical Coherence Tomography Attenuation Coefficient White matter Segmentation

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5D imaging approaches reveal the formation of distinct intracellular cAMP spatial gradients 24

5D imaging approaches reveal the formation of distinct intra...

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

作者： Rich, Thomas C. Annamdevula, Naga Trinh, Kenny Britain, Andrea L. Mayes, Samuel A. Griswold, John R. Deal, Joshua Hoffman, Chase West, Savannah Leavesley, Silas J. Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Ctr Lung Biol Mobile AL 36688 USA Univ S Alabama Basic Med Sci Grad Program Mobile AL 36688 USA Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ S Alabama Biomed Sci Mobile AL 36688 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Cyclic AMP (cAMP) is a ubiquitous second messenger known to differentially regulate many cellular functions. Several lines of evidence suggest that the distribution of cAMP within cells is not uniform. However, to date, no studies have measured the kinetics of 3D cAMP distributions within cells. This is largely due to the low signal-tonoise ratio of FRET-based probes. We previously reported that hyperspectral imaging improves the signal-to-noise ratio of FRET measurements. Here we utilized hyperspectral imaging approaches to measure FRET signals in five dimensions (5D) -three spatial (x, y, z), wavelength (lambda), and time (t) -allowing us to visualize cAMP gradients in pulmonary endothelial cells. cAMP levels were measured using a FRET-based sensor (H188) comprised of a cAMP binding domain sandwiched between FRET donor and acceptor - Turquoise and Venus fluorescent proteins. We observed cAMP gradients in response to 0.1 or 1 mu M isoproterenol, 0.1 or 1 mu M PGE1, or 50 mu M forskolin. Forskolin-and isoproterenol-induced cAMP gradients formed from the apical (high cAMP) to basolateral (low cAMP) face of cells. In contrast, PGE1-induced cAMP gradients originated from both the basolateral and apical faces of cells. Data suggest that 2D (x, y) studies of cAMP compartmentalization may lead to erroneous conclusions about the existence of cAMP gradients, and that 3D (x, y, z) studies are required to assess mechanisms of signaling specificity. Results demonstrate that 5D imaging technologies are powerful tools for measuring biochemical processes in discrete subcellular domains. This work was supported by NIH P01HL066299, R01HL058506, S10RR027535, AHA 16PRE27130004 and the Abraham Mitchell Cancer Research Fund.

关键词： Hyperspectral imaging spectral imaging microscopy FRET Forster resonance energy transfer cAMP second messenger signaling pulmonary endothelium

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Fluorescence microscopy with isotropic resolution using three objectives 13

Fluorescence microscopy with isotropic resolution using thre...

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

作者： Huelsnitz, Thomas Kner, Peter Univ Georgia Coll Engn Athens GA 30602 USA

ISBN: (纸本)9781628419474

Widefield and confocal fluorescence microscopy using a single objective suffer from poor resolution and a strong anisotropy between the lateral and axial resolution. Coherently combining the excitation and emission from two coaxial objectives improves the axial resolution up to sevenfold, but leaves the lateral resolution unchanged. Here we investigate the coherent combination of three objectives to create a point spread function (PSF) that is isotropic with higher resolution in the plane of the objectives. We develop a theoretical framework for simulating the performance of interferometric imaging with three objectives. Using three identical objectives with a large working distance and 0.9 numerical aperture (NA), the full-width half maximum of the confocal PSF is 135 nm compared to the lateral FWHM of 274 nm for imaging with a single objective at a wavelength of 515 nm.

关键词： microscopy fluorescence microscopy interferometric imaging PSF Engineering

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Deformable mirror based remote focusing for fast three-dimensional microscopy 13

Deformable mirror based remote focusing for fast three-dimen...

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

作者： Zurauskas, Mantas Frade, Maria Booth, Martin Univ Oxford Ctr Neural Circuits & Behav Mansfield Rd Oxford OX1 3SR England Univ Oxford Dept Engn Sci Parks Rd Oxford OX1 3PJ England

ISBN: (纸本)9781628419474

We present a deformable mirror based remote focusing method for three-dimensional imaging in high-resolution microscopy systems. The method relies on predefined mirror mode arrays that are obtained during initial mirror training step with a low complexity wavefront-sensing module. The imaging plane can be refocused over distances over a hundred times greater than the original depth of field of the objective lens along the optical axis at millisecond rates. We will demonstrate the combination of the remote focusing method with spatiotemporally focused two-photon excitation applied to three-dimensional imaging of biological samples.

关键词： Remote focusing adaptive optics three dimensional microscopy two photon microscopy

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