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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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3D Microscope Imaging Robust to Restoration Artifacts Introduced by Optically Thick Specimens 22

3D Microscope Imaging Robust to Restoration Artifacts Introd...

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

作者： Patwary, Nurmohammed King, Sharon V. Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9781628414202

We demonstrate 3D microscope imaging using computational optical sectioning microscopy (COSM) with an engineered point-spread function (PSF) robust to depth-induced spherical aberration (SA). Earlier we demonstrated that wavefront encoding (WFE) using a squared cubic (SQUBIC) phase mask reduces the PSF depth-variance in the presence of SA and that space-invariant (SI) restoration of simulated images using a single WFE-PSF does not lead to artifacts as in the conventional case. In this study, we show experimental verification of our WFE COSM approach. The WFE system used is a commercial microscope with a modified side port imaging path, where a spatial light modulator projects the SQUBIC phase mask on the back focal plane of the imaging lens. High resolution images of a test sample with 6 mu m in diameter microspheres embedded in UV-cured optical cement (RI = 1.47) were captured using both the engineered and the conventional imaging paths of the system. The acquired images were restored using a regularized SI expectation maximization algorithm based on Tikhonov-Miller regularization with a roughness penalty. A comparative study quantified in terms of the correlation coefficients between the XZ medial sections of the restored images, from experimental data, shows an 11% reduction in depth sensitivity in the SQUBIC system compared to the conventional system.

关键词： Three-dimensional microscopy wavefront encoding point-spread function engineering three-dimensional image restoration spherical aberration

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Wave-optical calibration of a light-field fluorescence microscopy 26

Wave-optical calibration of a light-field fluorescence micro...

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

作者： Pribosek, J. Steinbrener, J. Baumgart, M. Bereczki, T. Harms, K. Tortschanoff, A. Kenda, A. Carinthian Tech Res Europastr 12 A-9524 Villach Austria Mol Devicess Lagerhausstr 45 A-5071 Wals Austria

ISBN: (纸本)9781510624092

In light-field microscopy, a single point emitter gives rise to a complex diffraction pattern, which varies with the position of the emitter in object space. In order to use deconvolution-based wave-optical reconstruction schemes for light-field imaging systems, established methods rely on theoretical estimation of such diffraction patterns. In this paper we propose a novel method for direct experimental estimation of the light-field point spread function. Our approach relies on a modified reversed micro-Hartmann test to acquire a composite light-field point spread function of several thousand point emitters in the object plane simultaneously. By using fiducial markers and a custom image processing algorithm we separate the contributions of individual point emitters directly in raw light-field images and allow the construction of the forward imaging process without any prior assumption about the optical system required. The constructed forward imaging model can finally be applied in the 3D-deconvolution based wave-optical reconstruction scheme.

关键词： light-field microscopy light-field point spread function wave-optical calibration fluorescence imaging

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Amplitude, phase, and polarization control with a single spatial light modulator

Amplitude, phase, and polarization control with a single spa...

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

作者： Theisen, Michael J. Head, Stephen T. Brown, Thomas G. Gillmer, Steven R. Ellis, Jonathan D. Univ Rochester Inst Opt Rochester NY 14627 USA Univ Rochester Dept Mech Engn Rochester NY 14627 USA

ISBN: (纸本)9780819498625

Point spread function engineering is usually accomplished by controlling the amplitude, phase and/or polarization of the pupil fields. We analyze and test an optical design for full amplitude, phase, and polarization control of the pupil fields using a single spatial light modulator. In our scheme, the beam is spatially split into four components whose relative phases provide the four degrees of freedom necessary for amplitude, phase, and polarization control.

关键词： Spatial Light Modulators Polarization Control microscopy Pupil Engineering

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Tomographic incoherent phase imaging, a diffraction tomography alternative for any white-light microscope

Tomographic incoherent phase imaging, a diffraction tomograp...

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

作者： Bon, Pierre Aknoun, Sherazade Savatier, Julien Wattellier, Benoit Monneret, Serge Aix Marseille Univ Inst Fresnel Ecole Cent Marseille CNRSUMR7249 Domaine Univ St Jerome F-13397 Marseille France CNRS ESPCI UMR 7587 F-75005 Paris France PHSICS SA Campus Ecole Polytech F-91128 Palaiseau France

ISBN: (纸本)9780819493583

In this paper, we discuss the possibility of making tomographic reconstruction of the refractive index of a microscopic sample using a quadriwave lateral shearing interferometer, under incoherent illumination. A Z-stack is performed and the acquired incoherent elecromagnetic fields are deconvoluted before to retrieve in a quantitative manner the refractive index. The results are presented on polystyrene beads and can easily be expanded to biological samples. This technique is suitable to any white-light microscope equipped with nanometric Z-stack module.

关键词： Wavefront sensor Quantitative phase microscopy Diffraction Tomography Unlabeled biological samples

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Investigation of the SQUBIC phase mask design for depth-invariant widefield microscopy point-spread function engineering

Investigation of the SQUBIC phase mask design for depth-inva...

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

作者： Doblas, Ana King, Sharon V. Patwary, Nurmohammed Saavedra, Genaro Martinez-Corral, Manuel Preza, Chrysanthe Univ Valencia Dept Opt E-46100 Burjassot Spain Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9780819498625

Point-spread function engineering (PSF), achieved by placing a phase mask at the pupil plane of the imaging lens to encode the wavefront emerging from an imaging system, can be employed to reduce the impact of spherical aberration (SA) in 3D microscopy. In a previous study, the effect of SA on a confocal scanning microscope using a squared cubic phase mask (SQUBIC) was investigated using computer simulations. Here the effect of the SQUBIC design parameter on the insensitivity of the engineered PSF to SA is investigated using a metric based on the second-order moment of the axial variability of the PSF. We show that it is possible to find the optimum SQUBIC for the insensitization to SA.

关键词： Point-spread function engineering wavefront coding spherical aberration

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Managing multiple image stacks from confocal laser scanning microscopy

Managing multiple image stacks from confocal laser scanning ...

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing VI

作者： Zerbe, J Götze, C Zuschratter, W Fraunhofer Inst Comp Graph D-18059 Rostock Germany

ISBN: (纸本)0819430757

A major goal in neuroanatomy is to obtain precise information about the functional organization of neuronal assemblies and their interconnections. Therefore, the analysis of histological sections frequently requires high resolution images in combination with an overview about the structure. To overcome this conflict we have previously introduced a software for the automatic acquisition of multiple image stacks (3D-MISA) in confocal laser scanning microscopy. Here, we describe a Windows NT based software for fast and easy navigation through the multiple images stacks (MIS-browser), the visualization of individual channels and layers and the selection of user defined subregions. In addition, the MIS browser provides useful tools for the visualization and evaluation of the datavolume, as for instance brightness and contrast corrections of individual layers and channels. Moreover, it includes a maximum intensity projection, panning and zoom in/out functions within selected channels or focal planes (x/y) and tracking along the z-axis. The import module accepts any tiff-format and reconstructs the original image arrangement after the user has defined the sequence of images in x/y and z and the number of channels. The implemented export module allows storage of user defined subregions (new single image stacks) for further 3D-reconstruction and evaluation.

关键词： multiple-image-stacks confocal laserscanning microscopy image handling alignment 3D-image acquisition

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Membrane-substrate separation distance assessed by normalized total internal reflection fluorescence microscopy

Membrane-substrate separation distance assessed by normalize...

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

作者： Dos Santos, Marcelina Cardoso Vezy, Cyrille Jaffiol, Rodolphe Univ Technol Troyes CNRS UMR 6281 Lab Nanotechnol & Instrumentat OptInst Charles D F-10004 Troyes France

ISBN: (纸本)9780819498625

As a consequence of the recent progress in nanoscale technology, more and more sensitive methods are developed to characterize and understand the dynamic of cell membrane adhesion process. In this paper we present a new quantitative method to measure the separation distances between the membrane and the substrate. This technique is based on a normalization of Total Internal Reflection Fluorescence (TIRF) images by usual epi-illumination images. This simple method allows to achieve a nanometric axial resolution, typically 10 nm. We demonstrate the potential of our technique through the study of phospholipids membranes such as Giant Unilamellar Vesicles (GUVs), which are usual biomimetic systems to investigate membrane-substrate interactions.

关键词： Total Internal Fluorescence (TIRF) nanoscale fluorescence imaging cell adhesion Giant Unilamellar Vesicles (GUV)

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Resolving power in direct oblique plane imaging 22

Resolving power in direct oblique plane imaging

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

作者： Kim, Jeongmin Li, Tongcang Wang, Yuan Zhang, Xiang NSF Nanoscale Sci & Engn Ctr Berkeley CA 94720 USA Purdue Univ Dept Phys & Astron W Lafayette IN 47907 USA Univ Calif Berkeley Lawrence Berkeley Natl Lab Div Mat Sci Berkeley CA 94720 USA

ISBN: (纸本)9781628414202

Direct oblique plane imaging is a high-speed microscopy technique that observes a sample's plane that is inclined to the focal plane of the microscope objective lens. This wide-field microscopy is suitable for a study of fast dynamics of living samples where the principle plane of interest is tilted to the focal plane. A way to implement this imaging technique is to use remote focusing together with a tilted mirror, which involves asymmetrical pupil function of the imaging system. We rigorously study the anisotropic resolving power of the oblique plane imaging using a vectorial diffraction theory. From the derived effective pupil function, we calculate vectorial point spread function (PSF) and optical transfer function (OTF). We show that the two-dimensional (2D) PSF of the direct oblique plane imaging is not merely an oblique cross-section of the 3D PSF of circular aperture system. Similarly, 2D OTF of the oblique plane imaging is different from 2D oblique projection of conventional 3D OTF in circular aperture system.

关键词： Oblique plane imaging oblique plane microscopy vectorial diffraction theory asymmetric pupil function point spread function optical transfer function optical resolution remote focusing

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Self-reference extended depth-of-field quantitative phase microscopy

Self-reference extended depth-of-field quantitative phase mi...

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

作者： Jang, Jaeduck Bae, Chae Yun Park, Je-Kyun Ye, Jong Chul Korea Adv Inst Sci & Technol Dept Bio & Brain Engn Bio Imaging & Signal Proc Lab Gwahangno 335 Daejeon South Korea Korea Adv Inst Sci & Technol Dept Bio & Brain Engn Nano Biotech Lab Daejeon South Korea

ISBN: (纸本)9780819479662

This paper describes a novel quantitative phase microscopy based on a simple self-referencing scheme using Michelson interferometry. In order to achieve the homogeneous reference field for accurate phase measurement, the imaging field-of-view (FOV) is split onto the sample and homogenous background areas. The reference field can be generated by rotating the relative position of the sample and homogenous background in the object arm. Furthermore, our system is realized using an extended depth-of-field (eDOF) optics, which allows quantitative phase measurement for an increase of the depth-of-field without moving objective lens or specimen. The proposed method is confirmed by experimental results using various samples such as polystyrene beads and red blood cells (RBCs).

关键词： cell analysis microscopy phase imaging

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An algorithm for image stitching and blending

An algorithm for image stitching and blending

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing XII

作者： Rankov, V Locke, RJ Edens, RJ Barber, PR Vojnovic, B Mt Vernon Hosp Gray Canc Inst Adv Technol Dev Grp Northwood HA6 2JR Middx England

ISBN: (纸本)0819456756

In many clinical studies, including those of cancer, it is highly desirable to acquire images of whole tumour sections whilst retaining a microscopic resolution. A usual approach to this is to create a composite image by appropriately overlapping individual images acquired at high magnification under a microscope. A mosaic of these images can be accurately formed by applying image registration, overlap removal and blending techniques. We describe an optimised, automated, fast and reliable method for both image joining and blending. These algorithms can be applied to most types of light microscopy imaging. Examples from histology, from in vivo vascular imaging and from fluorescence applications are shown, both in 2D and 3D. The algorithms are robust to the varying image overlap of a manually moved stage, though examples of composite images acquired both with manually-driven and computer-controlled stages are presented. The overlap-removal algorithm is based on the cross-correlation method;this is used to determine and select the best correlation point between any new image and the previous composite image. A complementary image blending algorithm, based on a gradient method, is used to eliminate sharp intensity changes at the image joins, thus gradually blending one image onto the adjacent 'composite'. The details of the algorithm to overcome both intensity discrepancies and geometric misalignments between the stitched images will be presented and illustrated with several examples.

关键词： image stitching blending mosaic images

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