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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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Absolute and relative quantification and calibration for sectioning fluorescence microscopy using standardized uniform fluorescent layers and SIPchart based correction procedures

Absolute and relative quantification and calibration for sec...

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

作者： Zwier, J. M. Oomen, L. Brocks, L. Jalink, K. Brakenhoff, G. J. Univ Amsterdam Swammerdam Inst Life Sci Kruislaan 316 NL-1098 SM Amsterdam Netherlands Netherlands Canc Inst NL-1066 CX Amsterdam Netherlands

ISBN: (纸本)9780819465566

The total or integrated fluorescence intensity of a through-focus series of a thin standardized uniform fluorescent or calibration layer is shown to be suitable for image intensity correction and calibration in sectioning microscopy. This integrated intensity can be derived from the earlier introduced Sectioned Imaging Property or SIPcharts,(1) derived from the 3D layer datasets. By correcting the 3D image of an object with the 3D image of the standardized uniform fluorescent layer obtained under identical conditions one is able to express the object fluorescence in units fluorescence of the calibration layer. With object fluorescence intensities in 'fluorescence layer unit's or FLU's the object image intensities becomes independent of microscope system and imaging conditions. A direct result is that the often-appreciable lateral intensity variations present in confocal microscopy are eliminated (shading correction). Of more general value is that images obtained with different objectives, magnifications or from different microscope systems can be quantitatively related to each other. The effectiveness of shading correction and relating images obtained under various microscope conditions is demonstrated on images of standard fluorocent beads. Expressing the object fluorescence in FLU units seems to be a promising approach for general quantification of sectioning imaging enabling cross-correlation of imaging results over time and between imaging systems.

关键词： fluorescence microscopy shading correction sectioning microscopy confocal microscopy sectioned imaging 3D microscopy image correction SIPcharts

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Evaluation of the use of wavefront encoding to reduce depth-induced aberration in structured illumination microscopy 25

Evaluation of the use of wavefront encoding to reduce depth-...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Patwary, Nurmohammed Sola-Pikabea, Jorge Doblas, Ana Saavedra, Genaro Martinez-Corral, Manuel Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt Burjassot Spain

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Three-dimensional imaging is affected by depth-induced spherical aberration (SA) when imaging deep into an optically thick sample. In this work, we evaluate the impact of SA on the performance of incoherent grating-projection structured illumination microscopy (SIM). In particular, we analyze the reduction of the contrast in the structured pattern and compare the reconstructed SIM images for different amounts of SA. In order to mitigate the impact of SA, we implement and evaluate in SIM a wavefront encoded imaging system using a square cubic (SQUBIC) phase mask, an approach shown previously to be successful in conventional microscopy.

关键词： Spherical aberration wavefront encoding fluorescence microscopy structured illumination microscopy

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Super-resolution stereo light microscopy for real-time tasks using normal flow and geometric constraints

Super-resolution stereo light microscopy for real-time tasks...

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

作者： Danuser, G Swiss Fed Inst Technol ETH BioMicroMetr Grp Lab Biomech CH-8952 Schlieren Switzerland

ISBN: (纸本)081943535X

Prior knowledge about the observed scene represents the key to recovering frequencies beyond the passband of an imaging system (super-resolution). With regard to microscopy mainly two super-resolution mechanisms have been reported: 1.) analytic continuation of the frequency spectrum and 2.) constrained image deconvolution. This paper describes an alternative super-resolution mechanism. Prior knowledge is introduced on a higher, more symbolic level of visual inference. We exemplify our concept based on the 3D reconstruction of a micro-pipette moving in close proximity to immobile target objects. Information about the shape and the mobility of the pipette is incorporated in order to localize the pipette tip at sub-Rayleigh distances to the target. The algorithm was tested in a micro-robot environment. A machine vision module using stereo light microscopy automatically controlled the manipulation of microscopic objects, e.g. latex beads or diamond mono-crystals. In the theoretical part of the paper we prove that knowledge of the form "the pipette has moved between two consecutive frames of the movie" must result in a twofold increase of resolution. We used the normal flow of an image to decode positional measures from motion. In practice, super-resolution factors between 3 to 5 were obtained. The additional gain originates from the geometric constraints which were included to reconstruct the pipette position.

关键词： light microscopy stereo super-resolution machine vision normal flow geometric constraints

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Investigation of image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy

Investigation of image properties in super-resolution micros...

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

作者： Iketaki, Yoshinori Watanabe, Takeshi Bokor, Nandor Fujii, Masaaki Olympus Corp Adv Core Technol Dept 2-3 Kuboyama Cho Hachioji Tokyo 1928512 Japan Budapest Univ Technol Econ Dept Phys H-1111 Budapest Hungary Tokyo Inst Technol Chem Resources Lab Midori Ku Yokohama Kanagawa 2268503 Japan

ISBN: (纸本)9780819465566

We quantitatively investigated image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy. To evaluate the properties, the point spread function (PSF) and contrast transfer fuNction (CTF) were measured using a fluorescent scale together with a fluorescent bead. From the CTF, it has been found that visible light can resolve a 100 mn line-and-space pattern by microcopy, and provide a contrast of 10%. The CTF corresponds to a PSF with a FWHM of 130 mn. The value is two times finer than the diffraction limit size. An evaluation using a 100 run 0 fluorescent bead consistently supports the result given by the CTF for super-resolution microscopy. The measured CTF shows that super-resolution microscopy can indeed improve the optical properties of fluorescent images and enable us to observe a structure with a spatial resolution overcoming the diffraction limit.

关键词： two-color dip spectroscopy contrast transfer function point spread function fluorescence-depletion process super-resolution microscopy Rhodamine 6G

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An integrated approach to determine prior information for improved wide-field imaging models from computational interference microscopy 22

An integrated approach to determine prior information for im...

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

作者： Hossain, Md Shohag King, Sharon V. Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA

ISBN: (纸本)9781628414202

Knowledge of prior sample information, such as a refractive index (RI) map, can be used to improve image formation models enabling more accurate three-dimensional (3D) restoration in fluorescence microscopy. RI is an indicator of cell composition and structure that allows a more comprehensive representation of the 3D structure of a specimen than fluorescence alone. Due to the integral nature of sample phase, the challenge to compute the RI map is to decouple RI and thickness. Our work investigates the feasibility of determining RI of a specimen from differential interference contrast (DIC) microscopy data acquired by using different wavelengths in illumination. This spectral diversity in the data is exploited to determine sample thickness and RI. Results from simulated and experimental data of polystyrene bead samples are presented to analyze this approach. Phase images were estimated from the DIC data using an alternating minimization algorithm. This study shows that the maximum estimated phase delay is accurate within approximately 7 percent error relative to the 2D phase model. The sensitivity of this integrated approach allows RI to be computed within approximately 0.4 percent error relative to values from the literature.

关键词： Quantitative phase differential interference contrast microscopy alternating minimization algorithm rotational diversity

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Multi-wavelength phase imaging interference microscopy

Multi-wavelength phase imaging interference microscopy

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

作者： Warnasooriya, Nilanthi Kim, Myung K. Univ S Florida Dept Phys Tampa FL 33620 USA

ISBN: (纸本)0819461326

Multi-wavelength phase imaging interferography is a technique that combines phase-shifting interferometry with multiwavelength phase unwrapping. It can be used to obtain phase profile of an object without 2 pi ambiguities inherent to single wavelength phase images. In this technique, a Michelson-type interferometer is illuminated by an LED and the reference mirror is dithered for obtaining interference images at four phase quadratures, which are combined to calculate the phase of the object surface. The 2 pi discontinuities are removed by repeating the experiment using two or three LEDs at different wavelengths, which yields phase images of effective wavelength much longer than the original. The resulting image is a profile of the object surface with a height range of several microns and noise levels of 10's of nm. The technique is applied to imaging of phase profile of microscopic objects. The interferographic images using broadband source are significantly less affected by coherent noise, and there are a number of other advantages, such as lower cost and ease of operation.

关键词： interference microscopy phase shift phase measurement

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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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Restoration of high-resolution AFM images captured with broken probes

Restoration of high-resolution AFM images captured with brok...

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

作者： Wang, Y. F. Corrigan, D. Forman, C. Jarvis, S. Kokaram, A. Univ Dublin Trinity Coll Dept Elect Engn Dublin 2 Ireland Univ Coll Dublin Conway Grp Nanoscale Funct Grp Dublin Ireland

ISBN: (纸本)9780819488701

A type of artefact is induced by damage of the scanning probe when the Atomic Force Microscope (AFM) captures a material surface structure with nanoscale resolution. This artefact has a dramatic form of distortion rather than the traditional blurring artefacts. Practically, it is not easy to prevent the damage of the scanning probe. However, by using natural image deblurring techniques in image processing domain, a comparatively reliable estimation of the real sample surface structure can be generated. This paper introduces a novel Hough Transform technique as well as a Bayesian deblurring algorithm to remove this type of artefact. The deblurring result is successful at removing blur artefacts in the AFM artefact images. And the details of the fibril surface topography are well preserved.

关键词： Atomic force microscopy Distortion Hough transforms Deconvolution image analysis image processing Convolution Gaussian filters image resolution Atomic force microscope

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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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The spectral topography of histopathology samples

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

作者： Lerner, JM Lu, TW Vari, S LightForm Inc Belle Mead NJ 08502 USA

ISBN: (纸本)0819427004

The goal of imaging spectroscopy is to obtain independent spectra from individual objects in a field-of-view. In the case of biological materials, such as histopathology samples, it has been well established that spectral characteristic can be indicative of specific diseases including cancer. Diagnosis can be enhanced by the use of probes and stains to indicate the presence of individual genome or other biologically active cell components or substances. To assess a specimen through a microscope is directly analogous to observing the Earth from space to assess natural features. This paper describes a simple and inexpensive Imaging Spectrometer, with an origin in remote sensing, that demonstrates that it is possible to rapidly identify evidence of disease in histopathology samples using spatially resolved spectral data. The PARISS imaging spectrometer enables a researcher to acquire multispectral images that yield functional maps, showing what and where biological molecules are located within a structure. It is the powerful combination of imaging and spectroscopy that provides the tools not readily available to the Life Sciences. The PARISS system incorporates a powerful hybrid Neural Network (HNN) analysis to break the data logjam that is often associated with the acquisition and processing of multiple spectra.

关键词： imaging spectroscopy spectral topography histopathology spectroscopy micro-imaging microscopy

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