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Effect of double-helix point-spread functions on 3D imaging in the presence of spherical aberrations

Effect of double-helix point-spread functions on 3D imaging ...

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

作者： Ghosh, Sreya Grover, Ginni Piestun, Rafael Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Colorado Dept Elect Comp & Energy Engn Boulder CO 80309 USA

ISBN: (纸本)9780819484413

Double Helix point-spread functions (DH-PSFs), the result of PSF engineering, are used for super resolution microscopy. The DH-PSF design features two dominant lobes in the image plane which rotate with the change in axial (z) position of the light point source. The center of the DH-PSF gives the precise XY location of the point source, while the orientation of the lobes gives the axial location. In this paper we investigate the effect of spherical aberrations on the DH-PSF. Physical parameters such as the lens used, the size of the particle, refractive index of medium, and depth i.e., location within the underlying object, contribute to the amount of spherical aberration. DH-PSFs with spherical aberrations are computed for different imaging conditions. Three-dimensional images were generated of computer-generated objects using both space-invariant and depth-variant approach. Different approaches to estimate intensity and location of points from these images were investigated. Our results show that the DH-PSFs are susceptible to spherical aberration leading to an apparent shift in the location of the point source with increasing spherical aberrations which is comparable to the conventional PSF. Estimation algorithms like the depth variant expectation maximization (DVEM) can be used to obtain estimates of the true underlying object from the image obtained with DH-PSFs.

关键词： Computational Imaging Double Helix Point Spread Functions Spherical Aberration 3D microscopy Fluorescence microscopy Point spread function engineering

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3D fluorescence microscopy imaging accounting for depth-varying point-spread functions predicted by a strata interpolation method and a principal component analysis method

3D fluorescence microscopy imaging accounting for depth-vary...

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

作者： Yuan, Shuai Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9780819484413

In three-dimensional (3D) computational imaging for wide-field microscopy, estimation methods that solve the inverse imaging problem play an important role. The accuracy of the forward model has a significant impact on the complexity of the estimation method and consequently on the accuracy of the estimated intensity. Previous studies have shown that a forward model based on a depth-varying point-spread function (DV-PSF) leads to a substantial improvement in the resulting images because it accounts for depth-induced aberrations present in the imaging system. In this depth-varying (DV) model, the depth-dependent imaging effects are handled using a stratum-based interpolation method defined on discrete, non-overlapping layers or strata along the Z axis. Recently, a new approximation method based on a principle component analysis (PCA) was developed to predict DV-PSFs(1) with improved accuracy over the DV-PSFs predicted by the strata interpolation method of Ref. [11]. In this study, we implemented the PCA-based forward model for DV imaging to further compare the two approaches. DV-PSFs and forward models were computed using both the strata-based and the new PCA-based approximation schemes. Differences are quantified as a function of the approximation, i.e. the number of bases or strata used in each case respectively. A new PCA-based image estimation method was also developed based on the DV expectation maximization (DV-EM) algorithm of Ref. [11]. Preliminary evaluation of the performance of the PCA-based estimation shows promising results and consistency with previous results obtained in previous studies(1).

关键词： computational microscopy principle component analysis depth-variant image estimation strata interpolation method depth-variant point-spread function spherical aberrations 3D fluorescence microscopy computational optical sectioning microscopy

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Sparse OCT: Optimizing compressed sensing in spectral domain optical coherence tomography

Sparse OCT: Optimizing compressed sensing in spectral domain...

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

作者： Liu, Xuan Kang, Jin U. Johns Hopkins Univ Dept Elect & Comp Engn Baltimore MD 21218 USA

ISBN: (纸本)9780819484413

We applied compressed sensing (CS) to spectral domain optical coherence tomography (SD-OCT). Namely, CS was applied to the spectral data in reconstructing A-mode images. This would eliminate the need for a large amount of spectral data for image reconstruction and processing. We tested the CS method by randomly undersampling k-space SD-OCT signal. OCT images are reconstructed by solving an optimization problem that minimizes the l1 norm to enforce sparsity, subject to data consistency constraints. Variable density random sampling and uniform density random sampling were studied and compared, which shows the former undersampling scheme can achieve accurate signal recovery using less data.

关键词： Optical coherence tomography image reconstruction techniques information theoretical analysis

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Comparison of resolution in tomographic diffractive microscopy using combinations of sample rotation and illumination rotation

Comparison of resolution in tomographic diffractive microsco...

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

作者： Vertu, Stanislas Maeda, Etsuo Fluegge, Jens Delaunay, Jean-Jacques Haeberle, Olivier PTB Bundesallee 100 D-38116 Braunschweig Germany Univ Tokyo Sch Engn Dept Engn Mech Tokyo 1138656 Japan Univ Haute Alsace Lab MIPS EA2332 F-68093 Mulhouse France

ISBN: (纸本)9780819484413

Tomographic Diffractive microscopy (TDM) is a technique, which permits to image transparent living specimens without staining. For weakly diffractive samples, the three-dimensional distribution of the complex Refractive Index (RI) can be reconstructed from the knowledge of the measured scattered fields sampled under various viewing and illumination angles, according to the diffraction tomography theorem. TDM is commonly implemented in two ways, by either rotating the sample illumination keeping the specimen fixed, or by rotating the sample using fixed illumination. Both methods present limitations. Under the first-order Born approximation, the varying illumination direction method presents a strong anisotropic resolution along the optical axis due to the so-called "missing cone" of non captured frequencies. The sample rotation method presents a better isotropic resolution, but with a reduced extension of the captured frequencies. In view of overcoming the limitations of each method, we have studied various techniques for expanding the Optical Transfer Function with a tomographic microscope by combining different configurations of the sample rotation method with the varying illumination direction method, in order to obtain a high and isotropic resolution. Using simulations, we investigate the performances of the different configurations we propose.

关键词： Holography Fourier Optics image Reconstruction Tomographic Diffractive microscopy

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Quadriwave lateral shearing interferometry for quantitative phase microscopy: coupling phase imaging and fluorescence imaging

Quadriwave lateral shearing interferometry for quantitative ...

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

作者： Bon, Pierre Savatier, Julien Wattellier, Benoit Marguet, Didier Monneret, Serge Aix Marseille Univ Inst Fresnel Ecole Cent Marseille CNRSUMR6133 Domaine Univ St Jerome F-13397 Marseille France PHASICS SA Campus Ecole Polytech F-91128 Palaiseau France Univ Aix Marseille 1 CNRS INSERM Ctr Immunol Marseille Luminy F-13288 Marseille France

ISBN: (纸本)9780819484413

Phase imaging with a high-resolution wavefront sensor is a useful setup for biological imaging. Our setup is based on a quadriwave lateral shearing interferometer mounted on a commercial non-modified transmission white-light microscope. That allows us to make simultaneous measurement in both quantitative transmission phase and fluorescence imaging. We propose here to study co-localization between phase and fluorescence on african green monkey kidney COS7 cells. Phase permits an enhanced visualization of the whole cell and intracellular components while the fluorescence allows a complete identification of each component. Post treatments on phase-shift images are proposed and become very interesting for enhanced visualization of small details such as vesicles or mitochondrias.

关键词： Wavefront sensor Quantitative phase measurements Fluorescence co-localization

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Beyond the lateral resolution limit by phase imaging

Beyond the lateral resolution limit by phase imaging

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

作者： Cotte, Y. Toy, M. Fatih Depeursinge, C. Ecole Polytech Fed Lausanne Microvis & Microdiagnost Grp CH-1015 Lausanne Switzerland

ISBN: (纸本)9780819484413

We present a theory stating how to overcome the classical Rayleigh-resolution limit. It is based upon a new resolution criterion in phase of coherent imaging process and its spatial resolution is thought to be only SNR limited. Recently, the experimental observation of systematically occurring phase singularities in coherent imaging of sub-Rayleigh distanced objects has been reported.(1) The phase resolution criterion relies on the unique occurrence of phase singularities. A priori, coherent imaging system's resolution can be extended to Abbe's limit.(2) However, by introducing a known phase difference, the lateral as well as the longitudinal resolution can be tremendously enlarged. The experimental setup is based on Digital Holographic microscopy (DHM), an interferometric method providing access to the complex wave front. In off-axis transmission configuration, sub-wavelength nano-metric holes on a metallic film acts as the customized high-resolution test target. The nano-metric apertures are drilled with focused ion beam (FIB) and controlled by scanning electron microscopy (SEM). In this manner, Rayleighs classical two-point resolution condition can be rebuilt by interfering complex fields emanated from multiple single circular apertures on an opaque metallic film. By introducing different offset phases, enhanced resolution is demonstrated. Furthermore, the measurements can be exploited analytically or within the post processing of sampling a synthetic complex transfer function (CTF).

关键词： Point spread function microscopy super-resolution digital holography phase imaging phase vortices

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Whole cell imaging based on wide-field interferometric phase microscopy and its application to cardiomyocytes

Whole cell imaging based on wide-field interferometric phase...

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

作者： Shaked, Natan T. Satterwhite, Lisa L. Bursac, Nenad Wax, Adam Duke Univ Dept Biomed Engn Fitzpatrick Inst Photon Durham NC 27708 USA

ISBN: (纸本)9780819484413

Whole cell imaging is a novel technique using which the time-dependent quantitative phase profiles of live unstained biological cells are analyzed numerically to learn on the cell functionally. Dynamic phase profiles of the sample are first acquired by wide-field digital interferometry (WFDI), a quantitative holographic approach, without the need for scanning or using exogenous contrast agents. The resulting phase profiles are proportional to the multiplication between the cell thickness profile and its integral refractive index profile. However, many morphological parameters, including cell volume and cell force distribution, are based on the cell thickness profile, rather than on its WFDI phase profile. For cells with heterogeneous refractive index structure, more than a single exposure is typically needed to decouple thickness from integral refractive index using the phase profile, with the risk of losing transient acquisition. The presented whole-cell-imaging approach show that the WFDI phase profiles are useful for numerically analyzing cells even in cases where decoupling of thickness and integral refractive index is not possible or desired. We thus define new numerical parameters that directly utilize the WFDI phase profile and demonstrate their usefulness for characterizing contracting cardiomyocytes, cells with complex and highly-dynamic refractive-index structure.

关键词： Cell analysis interference microscopy phase holography cardiomyocytes

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Design and analysis of confocal-spectral microscopy using wavelength scanning scheme

Design and analysis of confocal-spectral microscopy using wa...

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

作者： Do, Dukho Chun, Wanhee Jeong, Hyeongjun Gweon, Dae-Gab Korea Adv Inst Sci & Technol Dept Mech Engn Nano Opto Mechatron Lab Taejon 305701 South Korea

ISBN: (纸本)9780819484413

The multi-color, or spectral fluorescence microscopy has ability to detect fluorescence spectral signals which are useful in case of studying interactions and phenomena between biological samples. Recently, commercial devices are combining with confocal microscope so to enhance lateral resolution and to have axial direction discernment. Also Acousto-Optic Tunable Filter(AOTF) is used instead of dichroic mirror to divide excitation and emission signals with mininum light efficiency. In addition, AOTF is used in spectral fluorescence microscopy have many advantages, these are very fast switching speed and high resolution in wavelength selection. However it uses acousto-optic interactions in birefringence material, Tellurium Dioxide(TeO(2)), the excitation light interacts with appropriate acoustic signal so that it is diffracted to 1 or -1 order path. But the fluorescence signals from a sample propagate in 0 order path with small different angle according to the polarization state. In this paper, a confocal-spectral microscopy is proposed with the new kind of spectral detector design having wavelength scanning galvano mirror. It makes possible to detect broad wavelength fluorescence signal by single PMT with simply rotating the galvano mirror. Also a new birefringent material, calcite(CaCO(3)) is used to compensate polarization effect. The proposed spectral confocal microscopy with unique spectrometer body has many advantages in comparison with commercial devices. In terms of detection method, it can be easily applied to other imaging modalities. Hence this system will be adapted in many applications.

关键词： wavelength scanning confocal-spectral birefringence compensation spectral detector

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Dynamic phase imaging utilizing a 4-dimensional microscope system

Dynamic phase imaging utilizing a 4-dimensional microscope s...

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

作者： Creath, Katherine 4D Technol Corp Tucson AZ 85706 USA

ISBN: (纸本)9780819484413

This paper describes a new, novel interference Linnik microscope system and presents images and data of live biological samples. The specially designed optical system enables instantaneous 4-dimensional video measurements of dynamic motions within and among live cells without the need for contrast agents. This "label-free", vibration insensitive imaging system enables measurement of biological objects in reflection using harmless light levels with a variety of magnifications and wavelengths with fields of view from several hundred microns up to a millimeter. At the core of the instrument is a phase measurement camera (PMC) enabling simultaneous measurement of multiple interference patterns utilizing a pixelated phase mask taking advantage of the polarization properties of light. Utilizing this technology enables the creation of phase image movies in real time at video rates so that dynamic motions and volumetric changes can be tracked. Objects are placed on a reflective surface in liquid under a coverslip. Phase values are converted to optical thickness data enabling volumetric, motion and morphological studies. Data from a number of different organisms such as flagellates and rotifers will be presented, as will measurements of human breast cancer cells with the addition of various agents that break down the cells. These data highlight examples of monitoring different biological processes and motions.

关键词： phase imaging interference microscopy polarization interferometry cellular imaging cell dynamics optical thickness measurement

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Refractive index reconstruction of biological samples from multimodal phase microscopy

Refractive index reconstruction of biological samples from m...

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

作者： Sierra, Heidy DiMarzio, Charles A. Brooks, Dana H. Rutgers State Univ Dept Biomed Engn 599 Taylor Rd Piscataway NJ 08854 USA Northeastern Univ Dept Elect & Comp Engn Boston MA 02115 USA

ISBN: (纸本)9780819484413

The extraction of quantitative information is important to better understand cellular activity in biological processes. In particular the optical refractive index can be used to analyze the results of cellular processes such as the average dry mass of biological samples. Phase microscopy modalities are widely used to image unstained biological samples because of their ability to obtain high-contrast images without introducing exogenous agents. The most common phase modalities are predominantly qualitative. However quantitative phase microscopy can provide more specific information about optical thickness and refractive index. In biological samples with several internal inhomogeneities and thickness variations, refractive index calculation becomes challenging to achieve by direct analysis of the images. Here we present a multimodal iterative method to reconstruct the spatial distribution of refractive index, combining information from two phase microscopy techniques. We use a constrained boundary iterative method under the assumption that the index of refraction inside the object can be approximated as piecewise constant. The boundary locations of all inhomogeneities are obtained by leveraging measurements from DIC and quantitative phase imaging modalities, and then the index of refraction is estimated based on those boundaries and a quantitative forward model for one modality. Simulations have confirmed the reliability of the proposed method. Experiments with measurement from mouse embryos at several development stages show that the proposed approach can reconstruct the distribution of the refractive index of these samples.

关键词： Differential Interference Contrast (DIC) Multimodal Phase Imaging Inversion Techniques Three-dimensional Reconstruction Index of Refraction Reconstruction Optical Quadrature microscopy (OQM)

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