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检索条件"任意字段=Conference on 3-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX"

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Time-resolved confocal microscopy of cryogenic processes in biological tissues

Time-resolved confocal microscopy of cryogenic processes in ...

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

作者： Schellenberg, M. Bergmann, T. Peev, E. Kloster, M. Napier, J. Neu, W. Univ Appl Sci Emden Leer Dept Nat Sci Constantiapl 4 D-26723 Emden Germany Univ Appl Sci Giessen Friedberg Inst Medizin Phys & Strahlenschutz D-35390 Giessen Germany

ISBN: (纸本)9780819484413

Cryogenic procedures are fundamental tools in modern biology, e. g. for conservation or purification of biological materials. The processes occurring in biological cells and tissues during freezing and thawing are subject to ongoing research. Optimization of cell survival rates demands the development and evaluation of exactly defined temperature profiles. 4D-DMD-microscopy is capable of imaging these highly dynamic processes with high spatial and temporal resolution, utilizing well established staining procedures for differentiating structures of interest.

关键词： two-photon DMD confocal microscopy cryogenic process 4D-microscopy cryomicroscopy

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Reducing noise in extended depth of field microscope images by optical manipulation of the point spread function

Reducing noise in extended depth of field microscope images ...

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

作者： Zahreddine, Ramzi N. Cormack, Robert H. Cogswell, Carol J. Univ Colorado Dept Elect Comp & Energy Engn Boulder CO 80309 USA

ISBN: (纸本)9780819484413

This work describes improved methods and algorithms for implementing extended depth of field (EDF) microscopy through point spread function (PSF) engineering. It utilizes adaptive optics to create a test bed on which to evaluate new phase shapes for EDF. Being able to quickly and cheaply design novel PSFs is essential to overcome limitations of EDF that have prevented the technology from reaching mainstream use. Further improvement is made by reducing the noise normally seen in EDF images. Computational optics principles are used to first encode the noise with an identifiable pattern and a specially-tailored non-linear algorithm then removes the noise. This approach improves a microscope's imaging capabilities in photon-starved applications such as live-cell fluorescence and object tracking.

关键词： Extended Depth of Field PSF Engineering 3D microscopy Adaptive Optics Neural Networks Noise Removal

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Simulating structured-illumination microscopy in the presence of spherical aberrations

Simulating structured-illumination microscopy in the presenc...

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

作者： Preza, Chrysanthe Univ Memphis Computat Imaging Res Lab Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9780819484413

The effect of depth-induced spherical aberrations (SA) on structured illumination microscopy (SIM)(1) is investigated. SIM is a technique used in three-dimensional (3D) fluorescence microscopy to improve resolution in optical sections acquired from 3D specimens. A 3D depth-variant imaging model was developed to predict the intermediate SIM or grid images that are used by the SIM approach to compute improved optical sections. The model incorporates SA due to imaging depth within a sample when there is a refractive index (RI) mismatch between the average RI of the specimen and the RI of the immersion medium of the lens. The model was implemented using a stratum-based model approximation and multiple depth-variant point-spread functions (PSFs)(2). SIM optical sections were computed using the subtraction algorithm(1, 3) and simulated grid images that include SA predicted by our model. Simulations were performed for different imaging conditions by varying the grid frequency, the amount of SA and the level of noise added to the grid images. Simulated results demonstrate that SIM images are less accurate in the presence of SA, and confirm that the SIM approach is very sensitive to system noise resulting in a reduced SNR in the optically sectioned images.

关键词： Structured illumination microscopy optical sectioning microscopy 3D fluorescence microscopy spherical aberrations depth-variant imaging depth-variant point spread functions

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High resolution optical projection tomographic microscopy for 3D tissue imaging

High resolution optical projection tomographic microscopy fo...

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

作者： Miao, Qin Hayenga, Jon Meyer, Michael G. Neumann, Thomas Patten, Florence Nelson, Alan C. Seibel, Eric J. Univ Washington Dept Bioengn Seattle WA 98195 USA VisionGate Inc Gig Harbor WA 98335 USA Univ Washington Dept Mech Engn Seattle WA 98195 USA

ISBN: (纸本)9780819484413

Optical projection tomography (OPT) requires a large depth of field (DOF) of a low numerical aperture (NA) lens resulting in low resolution. However, DOF of a high NA objective can be extended by scanning the focal plane through the sample. This extended DOF image is called pseudoprojection, which is used by optical projection tomographic microscope (OPTM) for tomographic reconstruction. The advantage of OPTM is the acquisition of relatively high resolution and large depth of field concurrently. This method requires the working distance of the lens to be larger than the size of the sample, so proper lens should be chosen for samples of different sizes. In this paper, we imaged hematoxylin stained muntjac cells inside capillary tube with two different sizes. Two objective lenses with different NA are used for these two tubes. Experimental results show that resolution improves over 10 times in OPTM compared to conventional OPT, which make it possible for OPTM technique to resolve sub-cellular features for large samples. Therefore, OPTM can be used for 3D histological analysis of hematoxylin & eosin (H&E) stained biopsy specimen with sub-cellular resolution in the future.

关键词： Three-dimensional microscopy Tomographic imaging Tissue imaging Needle biopsy

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Simplified setup for imaging with digital holographic microscopy and enhanced quantitative phase contrast by osmotic stimulation of living cells

Simplified setup for imaging with digital holographic micros...

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

作者： Kemper, Bjoern Przibilla, Sabine Rommel, Christina E. Vollmer, Angelika Ketelhut, Steffi Schnekenburger, Juergen von Bally, Gert Univ Munster Ctr Biomed Opt & Photon Robert Koch Str 45 D-48149 Munster Germany Univ Munster Gastroenterol Mol Cell Biol Dept Med B D-48149 Munster Germany

ISBN: (纸本)9780819484413

Many interferometry-based quantitative phase contrast imaging techniques require the generation of a coherent reference wave, which results in a phase stability decrease and the demand for a precise adjustment of the intensity ratio between object and reference wave. Thus, investigations on a simplified digital holographic microscopy approach that avoids a separate reference wave were performed. Results from live cell investigations demonstrate the capability of the method for quantitative phase contrast imaging. In further experiments the modification of the intracellular refractive index distribution by osmotic stimulation was analyzed. Data from human pancreas tumor cells show that by choice of suitable buffer solutions live cell imaging with enhanced quantitative phase contrast is achieved.

关键词： quantitative phase imaging digital holographic microscopy label-free live cell imaging

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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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