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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX 2022"

共 602 条记录，以下是141-150 订阅

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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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Investigating the usage of point spread functions in point source and microsphere localization 13

Investigating the usage of point spread functions in point s...

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

作者： Chao, Jerry Ram, Sripad Ward, E. Sally Ober, Raimund J. Texas A&M Univ Dept Biomed Engn College Stn TX 77843 USA Texas A&M Hlth Sci Ctr Dept Mol & Cellular Med College Stn TX 77843 USA Univ Texas Dallas Dept Elect Engn Richardson TX 75080 USA Texas A&M Hlth Sci Ctr Dept Microbial Pathogenesis & Immunol College Stn TX 77843 USA

ISBN: (纸本)9781628419474

Using a point spread function (PSF) to localize a point-like object, such as a fluorescent molecule or microsphere, represents a common task in single molecule microscopy image data analysis. The localization may differ in purpose depending on the application or experiment, but a unifying theme is the importance of being able to closely recover the true location of the point-like object with high accuracy. We present two simulation studies, both relating to the performance of object localization via the maximum likelihood fitting of a PSF to the object's image. In the first study, we investigate the integration of the PSF over an image pixel, which represents a critical part of the localization algorithm. Specifically, we explore how the fineness of the integration affects how well a point source can be localized, and find the use of too coarse a step size to produce location estimates that are far from the true location, especially when the images are acquired at relatively low magnifications. We also propose a method for selecting an appropriate step size. In the second study, we investigate the suitability of the common practice of using a PSF to localize a microsphere, despite the mismatch between the microsphere's image and the fitted PSF. Using criteria based on the standard errors of the mean and variance, we find the method suitable for microspheres up to 1 mu m and 100 nm in diameter, when the localization is performed, respectively, with and without the simultaneous estimation of the width of the PSF.

关键词： Cramer-Rao lower bound Fisher information localization accuracy maximum likelihood estimation microsphere pixel integration point spread function single molecule microscopy

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Development of a temporal multiplexed 3D beam-scanning Lissajous trajectory microscope for rapid, multimodal volumetric imaging 13

Development of a temporal multiplexed 3D beam-scanning Lissa...

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

作者： Newman, Justin A. Sullivan, Shane Z. Dinh, Janny Sarkar, Sreya Simpson, Garth J. Purdue Univ Dept Chem 560 Oval Dr W Lafayette IN 47907 USA

ISBN: (纸本)9781628419474

A beam-scanning microscope is described based on a temporally multiplexed Lissajous trajectory for achieving 1 kHz frame rate 3D imaging. The microscope utilizes two fast-scan resonant mirrors to direct the optical beam on a circuitous, Lissajous trajectory through the field of view. acquisition of two simultaneous focal planes is achieved by implementation of an optical delay line, producing a second incident beam at a different focal plane relative to the initial incident beam. High frame rates are achieved by separating the full time-domain data into shorter sub-trajectories resulting in undersampling of the field of view. A model-based image reconstruction (MBIR) 3D in-painting algorithm is utilized for interpolating the missing data to recover full images. The MBIR algorithm uses a maximum a posteriori estimation with a generalized Gaussian Markov random field prior model for image interpolation. Because images are acquired using photomultiplier tubes or photodiodes, parallelization for multi-channel imaging is straightforward. Preliminary results obtained using a Lissajous trajectory beam-scanning approach coupled with temporal multiplexing by the implementation of an optical delay line demonstrate the ability to acquire 2 distinct focal planes simultaneously at frame rates >450 Hz for full 512 x 512 images. The use of multi-channel data acquisition cards allows for simultaneous multimodal image acquisition with perfect image registry between all imaging modalities. Also discussed here is the implementation of Lissajous trajectory beam-scanning on commercially available microscope hardware.

关键词： Lissajous multi-channel beam-scanning temporal multiplexing 3D imaging nonlinear optics image interpolation

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Investigating the performance of reconstruction methods used in structured illumination microscopy as a function of the illumination pattern's modulation frequency 13

Investigating the performance of reconstruction methods used...

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

作者： Shabani, H. Sanchez-Ortiga, E. Preza, C. Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt E-46100 Burjassot Spain

ISBN: (纸本)9781628419474

Surpassing the resolution of optical microscopy defined by the Abbe diffraction limit, while simultaneously achieving optical sectioning, is a challenging problem particularly for live cell imaging of thick samples. Among a few developing techniques, structured illumination microscopy (SIM) addresses this challenge by imposing higher frequency information into the observable frequency band confined by the optical transfer function (OTF) of a conventional microscope either doubling the spatial resolution or filling the missing cone based on the spatial frequency of the pattern when the patterned illumination is two-dimensional. Standard reconstruction methods for SIM decompose the low and high frequency components from the recorded low-resolution images and then combine them to reach a high-resolution image. In contrast, model-based approaches rely on iterative optimization approaches to minimize the error between estimated and forward images. In this paper, we study the performance of both groups of methods by simulating fluorescence microscopy images from different type of objects ( ranging from simulated two-point sources to extended objects). These simulations are used to investigate the methods' effectiveness on restoring objects with various types of power spectrum when modulation frequency of the patterned illumination is changing from zero to the incoherent cut-off frequency of the imaging system. Our results show that increasing the amount of imposed information by using a higher modulation frequency of the illumination pattern does not always yield a better restoration performance, which was found to be depended on the underlying object. Results from model-based restoration show performance improvement, quantified by an up to 62% drop in the mean square error compared to standard reconstruction, with increasing modulation frequency. However, we found cases for which results obtained with standard reconstruction methods do not always follow the same trend.

关键词： structured illumination modulation frequency 3D microscopy optical sectioning super-resolution

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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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A multi-dimensional microscopic imaging system and reconstruction methods

A multi-dimensional microscopic imaging system and reconstru...

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International Congress on image and Signal processing, BioMedical Engineering and Informatics (CISP-BMEI)

作者： Yi Huang Mei Zhou Qingli Li Hongying Liu Fangmin Guo Shanghai Key Laboratory of Multidimensional Information Processing East China Normal University Shanghai China

ISBN: (纸本)9781509037117

With the in-depth study of microscopic objects, traditional optical microscopes no longer suffice for scientific researches in some occasions because of its limited depth of the imaging device. In this paper, we propose a new method of four-dimensional image information processing to realize four-dimensional reconstruction for smaller objects. This new method combines the 3D reconstruction method and hyperspectral imaging technology together, which realizes 4D information reconstruction of 3D model and its corresponding spectral data. In order to verify this method, a multidimensional micro imaging system is proposed. In the system, the hardware is expanded, the multidimensional image acquisition and multi-dimensional data processing is realized. The experiment results show that the new method and the proposed system have satisfied performance on four-dimensional reconstruction for smaller objects.

关键词： Hyperspectral imaging three-dimensional displays microscopy image reconstruction Optical microscopy Hardware

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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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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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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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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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