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A novel phase shifting structured illumination microscopy 13

A novel phase shifting structured illumination microscopy

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Singh, Veena Dubey, Vishesh Ahmad, Azeem Singh, Gyanendra Mehta, D. S. Indian Inst Technol Delhi Dept Phys New Delhi 110016 India Indian Inst Technol Delhi Instrument Design & Dev Ctr New Delhi 110016 India

ISBN: (纸本)9781628419474

This paper describes a new and novel phase shifting technique for qualitative as well as quantitative measurement in microscopy. We have developed a phase shifting device which is robust, inexpensive and involves no mechanical movement. In this method, phase shifting is implemented using LED array, beam splitters and defocused projection of Ronchi grating. The light from the LEDs are made incident on the beam splitters at spatially different locations. Due to variation in the geometrical distances of LEDs from the Ronchi grating and by sequentially illuminating the grating by switching on one LED at a time the phase shifted grating patterns are generated. The phase shifted structured patterns are projected onto the sample using microscopic objective lens. The phase shifted deformed patterns are recorded by a CCD camera. The initial alignment of the setup involves a simple procedure for the calibration for equal fringe width and intensity such that the phase shifted fringes are at equal phase difference. Three frame phase shifting algorithm is employed for the reconstruction of the phase map. The method described here is fully automated so that the phase shifted images are recorded just by switching of LEDs and has been used for the shape measurement of microscopic industrial objects. The analysis of the phase shifted images provides qualitative as well as quantitative information about the sample. Thus, the method is simple, robust and low cost compared to PZT devices commonly employed for phase shifting.

关键词： Ronchi grating LEDs phase shifting three step phase shifting algorithm quantitative phase imaging

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Volumetric retinal fluorescence microscopic imaging with extended depth of field 13

Volumetric retinal fluorescence microscopic imaging with ext...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Li, Zengzhuo Fischer, Andrew Li, Wei Li, Guoqiang Ohio State Univ Dept Ophthalmol & Visual Sci Columbus OH 43212 USA Ohio State Univ Dept Elect & Comp Engn Columbus OH 43212 USA Ohio State Univ Dept Neurosci Columbus OH 43212 USA NEI Bethesda MD 20892 USA

ISBN: (纸本)9781628419474

Wavefront-engineered microscope with greatly extended depth of field (EDoF) is designed and demonstrated for volumetric imaging with near-diffraction limited optical performance. A bright field infinity-corrected transmissive/reflective light microscope is built with Kohler illumination. A home-made phase mask is placed in between the objective lens and the tube lens for ease of use. General polynomial function is adopted in the design of the phase plate for robustness and custom merit function is used in Zemax for optimization. The resulting EDoF system achieves an engineered point spread function (PSF) that is much less sensitive to object depth variation than conventional systems and therefore 3D volumetric information can be acquired in a single frame with expanded tolerance of defocus. In Zemax simulation for a setup using 32X objective (NA = 0.6), the EDoF is 20 mu m whereas a conventional one has a DoF of 1.5 mu m, indicating a 13 times increase. In experiment, a 20X objective lens with NA = 0.4 was used and the corresponding phase plate was designed and fabricated. Retinal fluorescence images of the EDoF microscope using passive adaptive optical phase element illustrate a DoF around 100 mu m and it is able to recover the volumetric fluorescence images that are almost identical to in-focus images after post processing. The image obtained from the EDoF microscope is also better in resolution and contrast, and the retinal structure is better defined. Hence, due to its high tolerance of defocus and fine restored image quality, EDoF optical systems have promising potential in consumer portable medical imaging devices where user's ability to achieve focus is not optimal, and other medical imaging equipment where achieving best focus is not a necessary.

关键词： microscopy 3D imaging extended depth of field point spread function wavefront engineering passive adaptive optics eye imaging fluorescence imaging

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GPU accelerated Monte-Carlo simulation of SEM images for metrology 30

GPU accelerated Monte-Carlo simulation of SEM images for met...

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30th conference on Metrology, Inspection, and Process Control for Microlithography

作者： Verduin, T. Lokhorst, S. R. Hagen, C. W. Delft Univ Technol Fac Sci Appl Dept Imaging Phys Lorentzweg 1 NL-2628 CJ Delft Netherlands

ISBN: (纸本)9781510600133

In this work we address the computation times of numerical studies in dimensional metrology. In particular, full Monte-Carlo simulation programs for scanning electron microscopy (SEM) image acquisition are known to be notoriously slow. Our quest in reducing the computation time of SEM image simulation has led us to investigate the use of graphics processing units (GPUs) for metrology. We have succeeded in creating a full Monte-Carlo simulation program for SEM images, which runs entirely on a GPU. The physical scattering models of this GPU simulator are identical to a previous CPU-based simulator, which includes the dielectric function model for inelastic scattering and also refinements for low-voltage SEM applications. As a case study for the performance, we considered the simulated exposure of a complex feature: an isolated silicon line with rough sidewalls located on a flat silicon substrate. The surface of the rough feature is decomposed into 408 012 triangles. We have used an exposure dose of 6 mC/cm(2), which corresponds to 6 553 600 primary electrons on average (Poisson distributed) per image. We repeat the simulation for various primary electron energies, 300 eV, 500 eV, 800 eV, 1 keV, 3 keV and 5 keV. At first we run the simulation on a GeForce GTX480 from NVIDIA. The very same simulation is duplicated on our CPU-based program, for which we have used an Intel Xeon X5650. Apart from statistics in the simulation, no difference is found between the CPU and GPU simulated results. The GTX480 generates the images (depending on the primary electron energy) 387 to 894 times faster than a single threaded Intel X5650 CPU. Although this is a tremendous speedup, we actually have not reached the maximum throughput because of the limited amount of available memory on the GTX480. Nevertheless, the speedup enables the fast acquisition of simulated SEM images for metrology. We now have the potential to investigate case studies in CD-SEM metrology, which otherwise would take

关键词： Simulation Monte Carlo methods CUDA dimensional metrology Scanning electron microscopy

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Two-dimensional compressive sensing in spectral domain optical coherence tomography 22

Two-dimensional compressive sensing in spectral domain optic...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Xu, Daguang Huang, Yong Kang, Jin U. Johns Hopkins Univ Dept Elect & Comp Engn Baltimore MD 21218 USA

ISBN: (纸本)9781628414202

In this paper, we proposed a novel compressive sensing (CS) method in spectral domain optical coherence tomography (SD OCT), which reconstructs B-scan image using a subset of the spectral data that is under-sampled in both axial and lateral dimensions. Thus a fraction of the A-scans for a B-scan are acquired;the spectral data of each acquired A-scan is under-sampled. Compared with the previous studies, our method further reduces the overall size of the spectral measurements. Experimental results show that our approach can obtain high quality B-scan image using 25% spectral data, which takes 50% number of A-scans and acquires 50% spectral data for each selected A-scan.

关键词： Compressive sensing optical coherence tomography reconstruction algorithm image processing

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Simultaneous multiplane imaging for 3D confocal microscopy using high-speed z-scanning multiplexing 22

Simultaneous multiplane imaging for 3D confocal microscopy u...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Duocastella, Marti Vicidomini, Giuseppe Diaspro, Alberto Ist Italiano Tecnol Nanophys Via Morego 30 I-16163 Genoa Italy

ISBN: (纸本)9781628414202

One of the key frontiers in optical imaging is to maximize the spatial information retrieved from a sample while minimizing acquisition time. Confocal laser scanning microscopy is a powerful imaging modality that allows real-time and high-resolution acquisition of two-dimensional (2D) sections. However, in order to obtain information from three-dimensional (3D) volumes it is currently limited by a stepwise process that consists of acquiring multiple 2D sections from different focal planes by slow z-focus translation. Here, we present a novel method that enables the capture of an entire 3D sample in a single step. Our approach is based on an acoustically-driven varifocal lens integrated in a commercial confocal system that enables axial focus scanning at speeds of 140 kHz or above. Such high-speed allows for one or multiple focus sweeps on a pixel by pixel basis. By using a fast acquisition card, we can assign the photons detected at each pixel to their corresponding focal plane allowing simultaneous multiplane imaging. We exemplify this novel 3D confocal microscopy technique by imaging different biological fluorescent samples and comparing them with those obtained using traditional z-scanners. Based on these results, we find that image quality in this novel approach is similar to that obtained with traditional confocal methods, while speed is only limited by signal-to-noise-ratio. As the sensitivity of photodetectors increases and more efficient fluorescent labeling is developed, this novel 3D method can result in significant reduction in acquisition time allowing the study of new fundamental processes in science.

关键词： confocal microscopy optofluidics acousto-optics 3D imaging liquid lenses

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Graphics processing unit-accelerated real-time compressive sensing spectral domain optical coherence tomography 22

Graphics processing unit-accelerated real-time compressive s...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Xu, Daguang Huang, Yong Kang, Jin U. Johns Hopkins Univ Dept Elect & Comp Engn Baltimore MD 21218 USA

ISBN: (纸本)9781628414202

In this paper, we systematically demonstrate two real-time CS SD OCT systems based on a conventional desktop having three GPUs. The first one takes fast Fourier transform (FFT) as the sensing technique and under-sampled linear wavenumber spectral sampling as input data, while the second one uses non-uniform fast Fourier transform (NUFFT) and under-sampled nonlinear wavenumber spectral sampling, respectively. The maximum reconstruction speed of 72k and 33.5k A-line/s were achieved for these two systems, respectively, with A-scan size 2048. It is > 100 times faster than the C++ implementation and > 400 times faster than the MATLAB implementation. Finally, we present real-time dispersion compensated image reconstruction for both systems.

关键词： Compressive sensing optical coherence tomography graphics processing unit reconstruction algorithm image processing

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Single-Channel Stereoscopic Video Imaging Modality Based on a Transparent Rotating Deflector 22

Single-Channel Stereoscopic Video Imaging Modality Based on ...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Radfar, Edalat Park, Jihoon Jun, Eunkwon Ha, Myungjin Lee, Sangyeob Yu, Sung Kon Jang, Seul Gi Jung, Byungjo Yonsei Univ Dept Biomed Engn Wonju Gangwon Do South Korea

ISBN: (纸本)9781628414202

This paper introduces a stereoscopic video imaging modality based on a transparent rotating deflector (TRD). Sequential two- dimensional (2D) left and right images were obtained by rotating the TRD on a stepping motor synchronized with a complementary metal-oxide semiconductor camera, and the components of the imaging modality were controlled through general purpose input/output ports using a microcontroller unit. In this research, live stereoscopic videos were visualized on a personal computer by both active shutter 3D and passive polarization 3D methods. The imaging modality was characterized by evaluating the stereoscopic video image generation, rotation characteristics of the TRD. The level of 3D conception was estimated in terms of simplified human stereovision. The results show that single-channel stereoscopic video imaging modality has the potential to become an economical compact stereoscopic device as the system components are amenable to miniaturization;and could be applied in a wide variety of fields.

关键词： Three-dimensional (3D) imaging Stereo microscope Single-channel Transparent rotating deflector (TRD) Stereoscopic quality

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An integrated approach to determine prior information for improved wide-field imaging models from computational interference microscopy 22

An integrated approach to determine prior information for im...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Hossain, Md Shohag King, Sharon V. Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9781628414202

Knowledge of prior sample information, such as a refractive index (RI) map, can be used to improve image formation models enabling more accurate three-dimensional (3D) restoration in fluorescence microscopy. RI is an indicator of cell composition and structure that allows a more comprehensive representation of the 3D structure of a specimen than fluorescence alone. Due to the integral nature of sample phase, the challenge to compute the RI map is to decouple RI and thickness. Our work investigates the feasibility of determining RI of a specimen from differential interference contrast (DIC) microscopy data acquired by using different wavelengths in illumination. This spectral diversity in the data is exploited to determine sample thickness and RI. Results from simulated and experimental data of polystyrene bead samples are presented to analyze this approach. Phase images were estimated from the DIC data using an alternating minimization algorithm. This study shows that the maximum estimated phase delay is accurate within approximately 7 percent error relative to the 2D phase model. The sensitivity of this integrated approach allows RI to be computed within approximately 0.4 percent error relative to values from the literature.

关键词： Quantitative phase differential interference contrast microscopy alternating minimization algorithm rotational diversity

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Resolution analysis in compressive multidimensional microscopy 22

Resolution analysis in compressive multidimensional microsco...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Rodriguez, A. D. Clemente, P. Irles, E. Soldevila, F. Salvador, E. Tajahuerce, E. Lancis, J. Univ Jaume 1 Inst Noves Tecnol Imatge INIT Castellon de La Plana 12071 Spain Univ Jaume 1 CSIC Serv Cent Instrumentacio Cient Castellon de La Plana 12071 Spain

ISBN: (纸本)9781628414202

Despite imaging systems that scan a single-element benefit from mature technology, they suffer from acquisition times linearly proportional to the spatial resolution. A promising option is to use a single-pixel system that benefits from data collection strategies based on compressive sampling. Single-pixel systems also offer the possibility to use dedicated sensors such as a fiber spectrometer for multispectral imaging or a distribution of photodiodes for 3D imaging. The image is obtained by lighting the scene with microstructured masks implemented onto a programmable spatial light modulator. The masks are used as generalized measurement modes where the object information is expressed and the image is recovered through algebraic optimization. The fundamental reason why the bucket detection strategy can outperform conventional optical array detection is the use of a single channel detector that simultaneously integrates all the photons transmitted through the patterned scene. Spatial frequencies that are not transmitted through this low-quality optics are demonstrated to be present in the retrieved image. Our work makes two specific contributions within the field of single-pixel imaging through patterned illumination. First, we demonstrate that single-pixel imaging improves the resolution of conventional imaging systems overcoming the Rayleigh criterion. An analysis of resolution using a low NA microscope objective for imaging at a CCD camera shows that single-pixel cameras are not limited at all by the optical quality of the collection optics. Second, we experimentally demonstrate the capability of our technique to properly recover an image even when an optical diffuser is located in between the sample and the bucket detector.

关键词： Optical microscopy multidimensional Imaging Computational Imaging Aberration Compensation Resolution Compressive Sensing

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Dark-field illuminated fiber bundle endoscopy with iterative l₁-min image reconstruction for honeycomb pattern removal 22

Dark-field illuminated fiber bundle endoscopy with iterative...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Liu, Xuan Zhang, Lijun Kirby, Mitchell Raj, Divyaansh Qi, Shaohai Zhao, Feng New Jersey Inst Technol Dept Elect & Comp Engn Newark NJ 07102 USA Michigan Technol Univ Dept Biomed Engn Houghton MI 49931 USA Sun Yat Sen Univ Affiliated Hosp 1 Dept Burns Guangzhou 510080 Guangdong Peoples R China Livingston High Sch Livingston NJ 07039 USA

ISBN: (纸本)9781628414202

In this study, we developed a dark-field illuminated reflectance fiber-optic microscope (DRFM) along with an algorithm for l(1)-norm minimization of fiber bundle image to provide intrinsic endoscopic imaging with cellular resolution. To suppress specular reflection from fiber bundle facets, we adopted a dark-field configuration. To remove the honeycomb pattern of fiber bundle while preserve image resolution and contrast, we chose to minimize the image l(1) norm using iterative shrinkage thresholding (IST) algorithm.

关键词： Fiber optics imaging image reconstruction-restoration microscopy

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