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Influence of the background estimation on the superresolution properties of non-linear image restoration algorithms

Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 179-189页

作者： van Kempen, G.M.P. van Vliet, L.J. Unilever Research Vlaardingen Netherlands

ISBN: (纸本)0819430757;9780819430755

A study was carried out focusing on the effect of the background estimation on the performance of the Tikhonov-Miller algorithm (ICTM), Carrington, and Richardson-Lucy algorithms. A comparison was also made with the linear Tikhonov-Miller restoration filter. It was found that an underestimation of the background will make the non-negativity constraint ineffective which results in a performance that does not differ much from the performance obtained by the linear restoration filter. A novel method was devised to estimate the background based on the dependency of non-linear restoration algorithms on the background.

关键词： image reconstruction

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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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Single-Channel Stereoscopic Video Imaging Modality Based on a Transparent Rotating Deflector 22

Single-Channel Stereoscopic Video Imaging Modality Based on ...

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

作者： Radfar, Edalat Park, Jihoon Jun, Eunkwon Ha, Myungjin Lee, Sangyeob Yu, Sung Kon Jang, Seul Gi Jung, Byungjo Yonsei Univ Dept Biomed Engn Wonju Gangwon Do South Korea

ISBN: (纸本)9781628414202

This paper introduces a stereoscopic video imaging modality based on a transparent rotating deflector (TRD). Sequential two- dimensional (2D) left and right images were obtained by rotating the TRD on a stepping motor synchronized with a complementary metal-oxide semiconductor camera, and the components of the imaging modality were controlled through general purpose input/output ports using a microcontroller unit. In this research, live stereoscopic videos were visualized on a personal computer by both active shutter 3D and passive polarization 3D methods. The imaging modality was characterized by evaluating the stereoscopic video image generation, rotation characteristics of the TRD. The level of 3D conception was estimated in terms of simplified human stereovision. The results show that single-channel stereoscopic video imaging modality has the potential to become an economical compact stereoscopic device as the system components are amenable to miniaturization;and could be applied in a wide variety of fields.

关键词： three-dimensional (3D) imaging Stereo microscope Single-channel Transparent rotating deflector (TRD) Stereoscopic quality

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Managing multiple image stacks from confocal laser scanning microscopy

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Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 228-235页

作者： Zerbe, Joerg Goetze, Christian Zuschratter, Werner Fraunhofer Inst for Computer Graphics Rostock Germany

ISBN: (纸本)0819430757;9780819430755

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： Optical microscopy

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Temporal focusing-based multiphoton excitation fluorescence images with background noise cancellation via Hilbert-Huang transform 26

Temporal focusing-based multiphoton excitation fluorescence ...

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

作者： Hu, Yvonne Yuling Luo, Yuan-Rong Lin, Chun-Yu Chang, Chia-Yuan Chen, Shean-Jen Natl Cheng Kung Univ Dept Photon Tainan Taiwan Natl Cheng Kung Univ Dept Engn Sci Tainan Taiwan Natl Cheng Kung Univ Dept Mech Engn Tainan Taiwan Natl Chiao Tung Univ Inst Imaging & Biomed Photon Coll Photon Tainan Taiwan

ISBN: (纸本)9781510624092

Temporal focusing multiphoton excitation microscopy has wide field-of-view and optical sectioning. By using digital micromirror device, it provides patterned illumination. However, without filling the back aperture of objective lens, the axial confinement is limited to micron-meters, leading the out-of-focus fluorophores excited and image blurred. In this study, Hilbert-Huang transform is proposed to reduce the background noise. The empirical mode decomposition is first applied to disassemble the image into intrinsic mode functions and then reconstruct by Hilbert transform after diminishing background residues. The axial confinement can be enhanced from 2.79 mu m to 0.73 mu m with structure frequency in 1.06 mu m(-1).

关键词： Temporal focusing Digital micromirror device Structured illumination microscopy Hilbert-Huang transform

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Bright field segmentation tomography (BFST) for use as surface identification in stereomicroscopy

Bright field segmentation tomography (BFST) for use as surfa...

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Conference on 3-dimensional and multidimensional microscopy - image acquisition and processing XI

作者： Thiesse, J Namati, E de Ryk, J Hoffman, E McLennan, G Univ Iowa Iowa City IA 52242 USA

ISBN: (纸本)0819452327

Stereomicroscopy is an important method for use in image acquisition because it provides a 3D image of an object when other microscopic techniques can only provide the image in 2D. One challenge with this type of imaging is determining the top surface of a sample that has otherwise indistinguishable surface and planar characteristics. We have developed a system that creates a form of oblique illumination and in conjunction with image processing, the top surface can be viewed. The use of BFST consists of the Leica MZ16A stereomicroscope with a unique attached lighting source, and is part of the Large image Microscope Array (LIMA) [1]. The lighting source consists of eight light emitting diodes (LED's) that are separated by 45-degree angles. Each LED in this system illuminates with a 20-degree viewing angle once per cycle with a shadow over the rest of the sample. Subsequently, eight segmented images are taken per cycle. After the images are captured they are stacked through image addition to achieve the full field of view, and the surface is then easily identified. image processing techniques, such as skeletonization, can be used for further enhancement and measurement. With the use of BFST, advances can be made in detecting surface features from metals to tissue samples, such as in the analytical assessment of pulmonary emphysema using mean linear intercept.

关键词： stereomicroscopy illumination surface identification image processing

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Microlymphatic flow using fast video microscopy

Microlymphatic flow using fast video microscopy

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

作者： Dixon, JB Zawieja, DC Gashev, AA Coté, GL Texas A&M Univ Dept Biomed Engn College Stn TX 77843 USA

ISBN: (纸本)0819456756

Despite advances in the measurement of lymphatic flow, little is known about the actual velocities of flow in microlymphatic (similar to 100 um diameter) vessels. In this paper, video microscopy and particle tracking methods were adapted and integrated with an ultra high-speed imaging camera to obtain measurements of high-speed lymph velocities that previous systems were incapable of measuring. In this study, a mesenteric microlymphatic vessel in a loop of the small intestine of a male Sprague-Dawley rat was exteriorized and imaged at a rate of 500 frames per second (fps) for several contraction sequences. Lymph velocity was shown to fluctuate cyclically with the vessel wall contractions and ranged from -1 to 4 mm/sec through a ten second sequence.

关键词： Lymphatic system Cameras Velocity measurements Video microscopy Microfluidics Imaging systems High speed cameras Microfluidic imaging Video Particles

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Digital aberration correction of fluorescent images with coherent holographic image reconstruction by phase transfer (CHIRPT) 13

Digital aberration correction of fluorescent images with coh...

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

作者： Field, Jeffrey J. Bartels, Randy A. Colorado State Univ Microscopy Core Facil Ft Collins CO 80523 USA Colorado State Univ Dept Elect & Comp Engn Ft Collins CO 80523 USA Colorado State Univ Dept Biochem & Mol Biol Ft Collins CO 80523 USA Colorado State Univ Inst Genome Architecture & Funct Ft Collins CO 80523 USA Colorado State Univ Sch Biomed Engn Ft Collins CO 80523 USA

ISBN: (纸本)9781628419474

Coherent holographic image reconstruction by phase transfer (CHIRPT) is an imaging method that permits digital holographic propagation of fluorescent light. The image formation process in CHIRPT is based on illuminating the specimen with a precisely controlled spatio-temporally varying intensity pattern. This pattern is formed by focusing a spatially coherent illumination beam to a line focus on a spinning modulation mask, and image relaying the mask plane to the focal plane of an objective lens. Deviations from the designed spatio-temporal illumination pattern due to imperfect mounting of the circular modulation mask onto the rotation motor induce aberrations in the recovered image. Here we show that these aberrations can be measured and removed non-iteratively by measuring the disk aberration phase externally. We also demonstrate measurement and correction of systematic optical aberrations in the CHIRPT microscope.

关键词： fluorescence holography aberration correction microscopy

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Design of adaptive objective lens for ultrabroad near infrared imaging 13

Design of adaptive objective lens for ultrabroad near infrar...

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

作者： Lan, Gongpu Li, Guoqiang Ohio State Univ Dept Ophthalmol & Visual Sci Columbus OH 43212 USA Ohio State Univ Dept Elect & Comp Engn 1330 Kinnear Road Columbus OH 43212 USA

ISBN: (纸本)9781628419474

We present a compound adaptive objective lens in which a water-filled membrane lens is inserted into a front group (one lens) and a back group (two lenses). This adaptive objective lens works in the ultrabroad near infrared waveband (760nm similar to 920nm) with the volume scan of > 1mm(3) and the resolution of 2.8 mu m (calculated at the wavelength of 840 nm). The focal range is 19.5mm similar to 20.5mm and the numerical number is 0.196. The size of the adaptive lens is 10mm (diameter) x 17mm (length). This kind of lens can be widely used in three-dimensional (3D) volume biomedical imaging instruments, such as confocal microscope, optical coherence tomography (OCT), two photon microscope, etc.

关键词： Active optics adaptive objective lens extended depth of field three-dimensional microscopy confocal microscopy optical coherence tomography two-photon microscopy optical design

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k-microscopy - resolution beyond the diffraction limit

k-microscopy - resolution beyond the diffraction limit

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

作者： Geissbuehler, Matthias Lasser, Theo Leitgeb, Rainer A. Ecole Polytech Fed Lausanne Lab Opt Biomed CH-1015 Lausanne Switzerland

ISBN: (纸本)9780819470362

We present a novel Fourier domain method for microscopic imaging - so-called k-microscopy- with lateral resolution independent of the detection numerical aperture. The concept is based on sample illumination by a lateral fringe-pattern of varying spatial frequency, which probes the lateral spatial frequency or k- spectrum of the sample structure. The illumination pattern is realized by interference of two collimated coherent beams. Wavelength tuning is employed for modulation of the fringe spacing. The uniqueness of the proposed system is that a single point detector is sufficient to collect the total light corresponding to a particular position in the sample k-space. By shifting the phase of the interference pattern, we get full access to the complex frequencies. An inverse Fourier transformation of the acquired band in the frequency- or k-space will reconstruct the sample. The resulting lateral resolution will be defined by the temporal coherence length associated with the detected light source spectrum as well as by the illumination angle. The feasibility of the concept has been demonstrated in 1D.

关键词： k-microscopy sub-diffraction limit microscopy wavelength tuning biomedical imaging Fourier optics

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