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

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Dual-mode digital holographic and fluorescence microscopy for the study of morphological changes in cells under simulated microgravity

Dual-mode digital holographic and fluorescence microscopy fo...

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

作者： Toy, M. Fatih Pache, Christophe Parent, Jerome Kuehn, Jonas Egli, Marcel Depeursinge, Christian Ecole Polytech Fed Lausanne Adv Photon Lab CH-1015 Lausanne Switzerland ETH Space Biol Grp CH-8005 Zurich Switzerland

ISBN: (纸本)9780819479662

A dual mode microscope is developed to study morphological evolution of mouse myoblast cells under simulated microgravity in real time. Microscope operates in Digital Holographic microscopy (DHM) and widefield epifluorescence microscopy modes in a time sequential basis. DHM provides information on real time cellular morphology. EGFP transfected actin filaments in mouse myoblast cells function as the reporter for the fluorescence microscopy mode. Experimental setup is fixed in the RPM to observe microgravity induced dynamic changes in live cells. Initial results revealed two different modifications. Disorganized structures become visible in the formed lamellipodias, and proteins accumulate in the perinuclear region.

关键词： microscopy holography digital holographic microscopy fluorescence microscopy microgravity cell morphology cytoskeleton

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Improvement based on FDK reconstruction algorithm of conebeam CT

Improvement based on FDK reconstruction algorithm of conebea...

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

作者： Miao, Hui Wang, Tingting Zhao, Huijuan Gao, Feng Tianjin Univ Coll Precis Instruments & Optoelect Engn Tianjin 300072 Peoples R China

ISBN: (纸本)9780819479662

As a typical 3D image algorithm, Feldkamp-David-Kress reconstruction algorithm is a filtered backprojection very similar to the 2D algorithm. Recent years, it has been widely used for its easy implementation and acceptable reconstruction precision for small cone-beam angle. For big cone-beam angles, however, the reconstruction exactness of horizontal planes will dramatically decrease with the distance from the central plane increases. Therefore, for improving the reconstruction precision in vertical direction, a similar reciprocal Gaussian function as the weighted factor was introduced into FDK algorithm in the article. The validity of the improved FDK algorithm was verified and evaluated through both the computer numerical simulation and the phantom model experiment. The quantitative analysis for the reconstruction results demonstrated that the reconstruction image using the improved FDK algorithm could primely revise the original reconstruction images and restore well closely to the tomography image of the prototype object.

关键词： FDK reconstruction cone-beam CT image reconstruction reconstruction algorithm

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Fully Automated Data acquisition and Fast Data Interpretation in a Customized Multimodal Multiphoton Microscope

Fully Automated Data Acquisition and Fast Data Interpretatio...

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

作者： Rahn, C. D. Meine, H. Gallinat, S. Wenck, H. Wittern, K. P. Fischer, F. Beiersdorf AG R&D Struct Res Lab Unnastr 48 D-20245 Hamburg Germany Univ Hamburg Dept Informat Cognit Syst Lab D-22527 Hamburg Germany

ISBN: (纸本)9780819479662

Examining a volunteer or patient in-vivo usually is a strictly time-constrained process. When complex (custom) multicomponent, multi-modal imaging devices are involved, an examination session might be significantly stressful for the experimenter who is in charge of instrumental control. This is especially true when many parameters, several different devices or several different software applications are to be controlled during a single session. Stress may increase the probability of suboptimal data acquisition and data loss. The experimenter should be focused on the sample, not on instrumental control. In order to ease image acquisition, we present concepts for a new intuitive and interactive software application that integrates control of all hardware components, guides the experimenter through customizable acquisition workflows, presents the recorded data ready for facilitated interpretation using state-of-the-art image processing algorithms and thus supports the experimenter in acquiring optimized and highly valuable image data. Our concept combines into a single application many functions that were previously performed by several different programs, it allows linking together the functionalities, it allows automation, and it adds several convenience and safety features.

关键词： microscopy image acquisition image analysis software automatization

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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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Closed loop adaptive optics for microscopy without a wavefront sensor

Closed loop adaptive optics for microscopy without a wavefro...

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

作者： Kner, Peter Winoto, Lukman Agard, David A. Sedat, John W. Univ Georgia Fac Engn Athens GA 30602 USA Univ Calif San Francisco Dept Biochem & Biophys Keck Adv Microscopy Lab San Francisco CA 94158 USA Univ Calif San Francisco Howard Hughes Med Inst San Francisco CA 94143 USA

ISBN: (纸本)9780819479662

A three-dimensional wide-field image of a small fluorescent bead contains more than enough information to accurately calculate the wavefront in the microscope objective back pupil plane using the phase retrieval technique. The phase-retrieved wavefront can then be used to set a deformable mirror to correct the point-spread function (PSF) of the microscope without the use of a wavefront sensor. This technique will be useful for aligning the deformable mirror in a widefield microscope with adaptive optics and could potentially be used to correct aberrations in samples where small fluorescent beads or other point sources are used as reference beacons. Another advantage is the high resolution of the retrieved wavefont as compared with current Shack-Hartmann wavefront sensors. Here we demonstrate effective correction of the PSF in 3 iterations. Starting from a severely aberrated system, we achieve a Strehl ratio of 0.78 and a greater than 10-fold increase in maximum intensity.

关键词： Adaptive Optics microscopy Biomedical Imaging

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Quadriwave lateral shearing interferometry for quantitative phase microscopy: applications to long-duration imaging

Quadriwave lateral shearing interferometry for quantitative ...

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

作者： Bon, Pierre Wattellier, Benoit Monneret, Serge Marguet, Didier Aix Marseille Univ Ecole Cent Marseille CNRS Inst Fresnel Campus St Jerome F-13013 Marseille France Ecole Polytech PHASICS SA F-91128 Palaiseau France CNRS INSERM Ctr Immuno F-13288 Marseille France

ISBN: (纸本)9780819479662

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 non-modified transmission white-light microscope. We propose here to study long-time duration imaging on different type of adherent cells : green monkey kidney COS7 cells, human breast epithelial MCF10A cells, and human breast cancer derived from MDA-231 cells. This study permits a enhanced visualization of the whole cell life at different levels of confluence. Post treatments on phase-shift images are proposed and become very interesting for enhanced visualization of small details and thresholding.

关键词： Wavefront sensor Quantitative phase measurements Time-lapse imaging

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Hyper-NA imaging with solid-immersion optics and induced polarization imaging

Hyper-NA imaging with solid-immersion optics and induced pol...

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

作者： Zhang, Jun Yang, Seung-Hune Milster, Tom D. Bletscher, Warren Hansen, Del Univ Arizona Coll Opt Sci Tucson AZ 85721 USA Samsung Elect Seoul South Korea

ISBN: (纸本)9780819479662

Solid immersion lens (SIL) is used in microscopic systems for hyper numerical aperture (NA) imaging. The NA of the SIL microscope can be larger than 2 by using the high refractive index SIL. In this paper, examples of hyper-NA (NA>1.4) imaging are illustrated, including a normal SIL microscope for imaging samples like CPU chips, photomasks which have no cover layer and a special SIL microscope for imaging samples like Blu-Ray optical discs which have cover layer to protect the pattern. In both cases, good contrast images can be achieved by minimizing the system aberration. At the end, characteristics of induced polarization imaging (imaging through crossed polarizers) and a two-step solid immersion lens using Gallium Phosphide (GaP) (NA similar to 2) are discussed.

关键词： hyper-NA solid immersion induced polarization microscopy

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Photobleaching Property of Confocal Laser Scanning microscopy with Masked Illumination

Photobleaching Property of Confocal Laser Scanning Microscop...

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

作者： Kim, DongUk Moon, Sucbei Song, Hoseong Yang, Wenzhong Kim, Dug Y. Gwangju Inst Sci & Technol Dept Informat & Commun Kwangju 500712 South Korea

ISBN: (纸本)9780819479662

Confocal laser scanning microscopy (CLSM) has become the tool of choice for high-contrast fluorescence imaging in the study of the three-dimensional and dynamic properties of biological system. However, the high cost and complexity of commercial CLSMs urges many researchers to individually develop low cost and flexible confocal microscopy systems. The high speed scanner is an influential factor in terms of cost and system complexity. Resonant galvo scanners at several kHz have been commonly used in custom-built CLSMs. However, during the repeated illumination for live cell imaging or 3D image formation, photobleaching and image distortion occurred at the edges of the scan field may be more serious than the center due to an inherent property (e. g. sinusoidal angular velocity) of the scan mirror. Usually, no data is acquired at the edges due to large image distortion but the excitation beam is still illuminated. Here, we present the photobleaching property of CLSM with masked illumination, a simple and low cost method, to exclude the unintended excitation illumination at the edges. The mask with a square hole in its center is disposed at the image plane between the scan lens and the tube lens in order to decrease photobleaching and image distortion at the edges. The excluded illumination section is used as the black level of the detected signals for a signal quantizing step. Finally, we demonstrated the reduced photobleaching at the edges on a single layer of fluorescent beads and real-time image acquisition without a standard composite video signal by using a frame grabber.

关键词： Photobleaching confocal laser scanning microscopy video-rate 3D image masked illumination

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Real time focal modulation microscopy

Real time focal modulation microscopy

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

作者： Chen, N. G. Wong, C. H. Chong, S. P. Sheppard, C. J. R. Natl Univ Singapore Dept Elect & Comp Engn Singapore 117548 Singapore Natl Univ Singapore Div Bioengn Singapore Singapore Natl Univ Singapore Dept Radiol Singapore Singapore

ISBN: (纸本)9780819479662

Focal modulation microscopy is a novel microscopy method that can achieve a large penetration depth with single photon excited fluorescence. In this method, excitation intensity within the focal volume is modulated by the use of a time dependent spatial phase modulator. The resultant fluorescence emission is filtered by the spatial gating provided by the pinhole and demodulated with a lock-in amplifier. Previous implementation of the spatial modulator is realised through a mechanical tilt plate phase modulator. This mechanical implementation affords stable modulation and provides satisfactory image quality. However, the slow modulation rate(in kHz range) depreciates the prospect of its employment in real time imaging. A new modulation scheme is proposed in which an acoustic optical modulator(AOM) is utilised. This new arrangement boost the modulation speed and makes real time imaging possible. We report on the development of such a modulation system together with the accompanying electronics. images obtained with this new modulation system will also be presented

关键词： Optical Sectioning microscopy Focal Modulation Synthetic Illumination

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Snapshot Spectral Imaging System

Snapshot Spectral Imaging System

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

作者： Arnold, Thomas De Biasio, Martin McGunnigle, Gerald Leitner, Raimund Ctr Carinthian Tech Res Villach Austria

ISBN: (纸本)9780819479662

Spectral imaging is the combination of spectroscopy and imaging. These fields are well developed and are used intensively in many application fields including industry and the life sciences. The classical approach to acquire hyper-spectral data is to sequentially scan a sample in space or wavelength. These acquisition methods are time consuming because only two spatial dimensions, or one spatial and the spectral dimension, can be acquired simultaneously. With a computed tomography imaging spectrometer (CTIS) it is possible to acquire two spatial dimensions and a spectral dimension during a single integration time, without scanning either spatial or spectral dimensions. This makes it possible to acquire dynamic image scenes without spatial registration of the hyper-spectral data. This is advantageous compared to tunable filter based systems which need sophisticated image registration techniques. While tunable filters provide full spatial and spectral resolution, for CTIS systems there is always a tradeoff between spatial and spectral resolution as the spatial and spectral information corresponding to an image cube is squeezed onto a 2D image. The presented CTIS system uses a spectral-dispersion element to project the spectral and spatial image information onto a 2D CCD camera array. The system presented in this paper is designed for a microscopy application for the analysis of fixed specimens in pathology and cytogenetics, cell imaging and material analysis. However, the CTIS approach is not limited to microscopy applications, thus it would be possible to implement it in a hand-held device for e. g. real-time, intra-surgery tissue classification.

关键词： spectral imaging CTIS computed tomography microscopy snapshot

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