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

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Assessment of robust reconstruction algorithms for compressive sensing spectral-domain optical coherence tomography

Assessment of robust reconstruction algorithms for compressi...

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

作者： Xu, Daguang Huang, Yong Kang, Jin U. Johns Hopkins Univ Dept Elect & Comp Engn Baltimore MD 21218 USA

ISBN: (纸本)9780819493583

In this paper, we performed an in-depth assessment of current state-of-the-art compressive sensing (CS) reconstruction algorithms, including YALL1, CSALSA, NESTA, SPGL1, TwIST and SpaRSA for use in spectral domain optical coherence tomography (SD-OCT). A brief description of mentioned algorithms and criterion in assessing performance between constraint and unconstraint algorithms are presented. The performance of all algorithms is initially assessed using a set of artificial noiseless A-scan signals with different spatial-domain dynamic range. Reconstruction error, computation time, noise tolerance and reliability of each algorithm are used as key metrics. A fair speed comparison is then implemented. Finally, computation time, SNR and local contrast of the algorithms are evaluated on real OCT B-scan data. Our results show that SPGL1 and YALL1 have moderately better performance.

关键词： Compressive sensing Optical coherence tomography Reconstruction algorithm image processing

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Light sheet adaptive optics microscope for 3D live imaging

Light sheet adaptive optics microscope for 3D live imaging

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

作者： Bourgenot, C. Taylor, J. M. Saunter, C. D. Girkin, J. M. Love, G. D. Univ Durham Dept Phys Ctr Adv Instrumentat Durham DH1 3LE England

ISBN: (纸本)9780819493583

We report on the incorporation of adaptive optics (AO) into the imaging arm of a selective plane illumination microscope (SPIM). SPIM has recently emerged as an important tool for life science research due to its ability to deliver high-speed, optically sectioned, time-lapse microscope images from deep within in vivo selected samples. SPIM provides a very interesting system for the incorporation of AO as the illumination and imaging paths are decoupled and AO may be useful in both paths. In this paper, we will report the use of AO applied to the imaging path of a SPIM, demonstrating significant improvement in image quality of a live GFP-labeled transgenic zebrafish embryo heart using a modal, wavefront sensorless approach and a heart synchronization method. These experimental results are linked to a computational model showing that significant aberrations are produced by the tube holding the sample in addition to the aberration from the biological sample itself.

关键词： Optical microscopy Aberration correction Adaptive Optics Light sheet microscopy zebra fish

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Increasing the resolution of light sheet microscopy in the presence of aberrations

Increasing the resolution of light sheet microscopy in the p...

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

作者： Vettenburg, T. Dalgarno, H. I. C. Cizmar, T. Gunn-Moore, F. J. Dholakia, K. Univ St Andrews SUPA Sch Phys & Astron St Andrews KY16 9SS Fife Scotland Univ St Andrews Sch Med St Andrews KY16 9SS Fife Scotland Univ St Andrews Sch Biol St Andrews KY16 9SS Fife Scotland

ISBN: (纸本)9780819493583

Single plane illumination microscopy (SPIM) allows rapid imaging of large, three-dimensional, samples of living tissue. The thin light sheet ensures high contrast whilst photo-bleaching and damage are kept to a minimum. However, many specimen of interest have a significant thickness. To date, high axial resolution in such specimen has only been achieved by compromising these key advantages and adding considerable technical complexity. Although the light sheet can propagate several hundreds of micrometers into the tissue, its width can be several orders of magnitude larger than it would be in a homogeneous sample. In this paper we explore the use of pupil-phase modulation to overcome such sample-induced aberrations and produce diffraction-limited deep inside turbid samples.

关键词： selective plane illumination microscopy (SPIM) light sheet fluorescence microscopy (LSFM) digitally scanned light sheet microscopy (DSLM) high resolution

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Photothermal imaging of melanin

Photothermal imaging of melanin

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

作者： Kerimo, Josef DiMarzio, Charles A. Northeastern Univ Dept Elect & Comp Engn Boston MA 02115 USA

ISBN: (纸本)9780819493583

We present photothermal images of melanin using modulation with two laser beams. Strong melanin absorption followed by efficient nonradiative relaxation caused heating and an increase in temperature. This temperature effect was used as an imaging contrast to detect melanin. Melanin from several samples including Sepia officinalis, black human hair, and live zebra fish, were imaged with a high signal-to-noise ratio. For the imaging, we focused two near infrared laser beams (pump and probe) collinearly with different wavelengths and the pump was modulated in amplitude. The thermally induced variations in the refractive index, at the modulation frequency, were detected by the scattering of the probe beam. The Photothermal method brings several imaging benefits including the lack of background interference and the possibility of imaging for an extended period of time without photodamage to the melanin. The dependence of the photothermal signal on the laser power, modulation frequency, and spatial offset of the probe is discussed. The new photothermal imaging method is promising and provides background-free and label-free imaging of melanin and can be implemented with low-cost CW lasers.

关键词： melanin photothermal imaging modulation near-infrared microscopy

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Portable low-coherence interferometer for quantitative phase microscopyof live cells

Portable low-coherence interferometer for quantitative phase...

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

作者： Shaked, Natan T. Girshovitz, Pinhas Tel Aviv Univ Fac Engn Dept Biomed Engn IL-69978 Ramat Aviv Israel

ISBN: (纸本)9780819493583

We present a compact, highly portable and inexpensive interferometric module for obtaining spatial interferograms of microscopic biological samples, without the strict stability and the highly-coherent illumination that are usually required for interferometric microscopy setups. The module is built using off-the-shelf optical elements and can easily operate with low-coherence illumination, while being positioned in the output of a conventional transmission microscope. The interferograms are processed into the quantitative amplitude and phase profiles of the sample. Based on the phase profile, the optical thickness or optical path delay profile of the sample is obtained with temporal and spatial stabilities, at the order of 0.2-0.3 nm. We show several configurations of this interferometer that are suitable for both on-axis and off-axis holographic geometries, and present various experimental results, including imaging live cells in a non-contact label-free manner and transparent elements with nano-scale thickness. Since the interferometer can be connected to the output of a transmission microscope and operate in a simple way, without involvement of an expert user with a knowledge in optics and without complicated alignment prior to every experiment, and still obtain a remarkably high accuracy, we believe that this new setup will make interferometric phase microscopy more accessible and affordable for biologists and clinicians, while significantly broadening its range of applications.

关键词： Holographic microscopy Interferometric microscopy Live cell quantitative imaging

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Using liquid crystal variable retarders for fast modulation of bias and shear direction in quantitative differential interference contrast (DIC) microscope

Using liquid crystal variable retarders for fast modulation ...

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

作者： Shribak, Michael Marine Biol Lab Woods Hole MA 02543 USA

ISBN: (纸本)9780819493583

Conventional DIC microscope shows the two-dimensional distribution of optical path length gradient encountered along the shear direction between two interfering beams. It is therefore necessary to rotate unknown objects in order to examine them at several orientations. We built new DIC beam shearing assembly, which allows the bias to be modulated and shear directions to be switched rapidly without any mechanically rotating the specimen or the prisms. The assembly consists of two standard DIC prisms with liquid crystal cell in between. Another liquid crystal cell is employed for modulating a bias. All components do not require a special design and are available on the market. We describe techniques for measuring parameters of DIC prisms and calibrating liquid crystal cells. One beam-shearing assembly is added to the illumination path and another one to the imaging path of standard microscope. Two sets of raw DIC images at the orthogonal shear directions and two or three different biases are captured and processed within a second. Then the quantitative image of optical path gradient distribution within a thin optical section is displayed on a computer screen. The obtained data are also used to compute the quantitative distribution of optical phase, which represents refractive index gradient or height distribution. It is possible to generate back the enhanced regular DIC images with any desired shear direction. New DIC microscope can be combined with other techniques, such as fluorescence and polarization microscopy.

关键词： Differential interference contrast (DIC) microscopy quantitative phase imaging optical path gradient dry mass liquid crystal devices live cell imaging

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Spatial-spectral coupling in hyperspectral CARS microscopy image formation

Spatial-spectral coupling in hyperspectral CARS microscopy i...

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three-dimensional and multidimensional microscopy: image acquisition and processing XX

作者： Barlow, Aaron M. Popov, Konstantin Andreana, Marco Moffatt, Douglas J. Ridsdale, Andrew Slepkov, Aaron D. Ramunno, Lora Stolow, Albert SDT Emerging Technologies Division National Research Council of Canada 100 Sussex Dr. Ottawa ON Canada Department of Physics University of Ottawa Ottawa ON Canada Department of Physics and Astronomy Trent University Peterborough ON Canada Department of Physics Queen's University Kingston ON Canada

ISBN: (纸本)9780819493583

Hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy has provided an imaging tool for extraction of 3-dimensional volumetric information, as well as chemically-sensitive spectral information. These techniques have been used in a variety of different domains including biophysics, geology, and material science. The measured CARS spectrum results from interference between the Raman response of the sample and a non-resonant background. We have collected four dimensional data sets (three spatial dimensions, plus spectra) and extracted Raman response from the CARS spectrum using a Kramers-Kronig transformation. However, the three dimensional images formed by a CARS microscope are distorted by interference, some of which arises because of the Gouy phase shift. This type of interference comes from the axial position of the Raman resonant object in the laser focus. We studied how the Gouy phase manifests itself in the spectral domain by investigating microscopic diamonds and nitrobenzene droplets in a CARS microscope. Through experimental results and numerical calculation using finite-diference time-domain (FDTD) methods, we were able to demonstrate the relationship between the spatial configuration of the sample and the CARS spectral response in three dimensional space. © 2013 Copyright SPIE.

关键词： Raman spectroscopy

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An optimized two-photon method for in vivo lung imaging reveals intimate cell collaborations during infection

An optimized two-photon method for in vivo lung imaging reve...

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

作者： Fiole, Daniel Deman, Pierre Trescos, Yannick Douady, Julien Tournier, Jean-Nicolas Inst Rech Biomed Armees Dept Microbiol Unite Interact Hote Agents Pathogenes La Tronche France Univ Grenoble 1 CNRS UMR 5588 Lab Interdisciplinaire Phys Grenoble France Ecole Val Grace Paris France

ISBN: (纸本)9780819493583

Lung tissue motion arising from breathing and heart beating has been described as the largest annoyance of in vivo imaging. Consequently, infected lung tissue has never been imaged in vivo thus far, and little is known concerning the kinetics of the mucosal immune system at the cellular level. We have developed an optimized post-processing strategy to overcome tissue motion, based upon two-photon and second harmonic generation (SHG) microscopy. In contrast to previously published data, we have freed the lung parenchyma from any strain and depression in order to maintain the lungs under optimal physiological parameters. Excitation beams swept the sample throughout normal breathing and heart movements, allowing the collection of many images. Given that tissue motion is unpredictably, it was essential to sort images of interest. This step was enhanced by using SHG signal from collagen as a reference for sampling and realignment phases. A normalized cross-correlation criterion was used between a manually chosen reference image and rigid transformations of all others. Using CX3CR1(+/gfp) mice this process allowed the collection of high resolution images of pulmonary dendritic cells (DCs) interacting with Bacillus anthracis spores, a Gram-positive bacteria responsible for anthrax disease. We imaged lung tissue for up to one hour, without interrupting normal lung physiology. Interestingly, our data revealed unexpected interactions between DCs and macrophages, two specialized phagocytes. These contacts may participate in a better coordinate immune response. Our results not only demonstrate the phagocytizing task of lung DCs but also infer a cooperative role of alveolar macrophages and DCs.

关键词： two-photon microscopy in vivo lung imaging host-pathogen interactions infectious disease Bacillus anthracis

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Tunable thin-film optical filters for hyperspectral microscopy

Tunable thin-film optical filters for hyperspectral microsco...

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

作者： Favreau, Peter F. Rich, Thomas C. Prabhat, Prashant Leavesley, Silas J. Univ S Alabama Biomol Chem engn Mobile AL 36688 USA Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Ctr Lung Biol Mobile AL 36688 USA Semrock Inc A Unit Index Mobile AL 36688 USA

ISBN: (纸本)9780819493583

Hyperspectral imaging was originally developed for use in remote sensing applications. More recently, it has been applied to biological imaging systems, such as fluorescence microscopes. The ability to distinguish molecules based on spectral differences has been especially advantageous for identifying fluorophores in highly autofluorescent tissues. A key component of hyperspectral imaging systems is wavelength filtering. Each filtering technology used for hyperspectral imaging has corresponding advantages and disadvantages. Recently, a new optical filtering technology has been developed that uses multi-layered thin-film optical filters that can be rotated, with respect to incident light, to control the center wavelength of the pass-band. Compared to the majority of tunable filter technologies, these filters have superior optical performance including greater than 90% transmission, steep spectral edges and high out-of-band blocking. Hence, tunable thin-film optical filters present optical characteristics that may make them well-suited for many biological spectral imaging applications. An array of tunable thin-film filters was implemented on an inverted fluorescence microscope (TE 2000, Nikon Instruments) to cover the full visible wavelength range. images of a previously published model, GFP-expressing endothelial cells in the lung, were acquired using a charge-coupled device camera (Rolera EM-C2, Q-Imaging). This model sample presents fluorescently-labeled cells in a highly autofluorescent environment. Linear unmixing of hyperspectral images indicates that thin-film tunable filters provide equivalent spectral discrimination to our previous acousto-optic tunable filter-based approach, with increased signal-to-noise characteristics. Hence, tunable multi-layered thin film optical filters may provide greatly improved spectral filtering characteristics and therefore enable wider acceptance of hyperspectral widefield microscopy.

关键词： Hyperspectral imaging spectral imaging microscopy AOTF acousto-optic tunable filter TFTF thin-film tunable filter VersaChrome

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three-dimensional and multidimensional microscopy: image acquisition and processing XIX

Three-Dimensional and Multidimensional Microscopy: Image Acq...

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three-dimensional and multidimensional microscopy: image acquisition and processing XIX

ISBN: (纸本)9780819488701

The proceedings contain 41 papers. The topics discussed include: measuring aberrations in the rat brain by a new coherence-gated wavefront sensor using a linnik interferometer;performance evaluation of point-spread function engineering to reduce the impact of depth-induced aberrations on extended depth-of-field microscopy;simultaneous quantitative depth mapping and extended depth of field for 4D microscopy through PSF engineering;calibration of an adaptive microscope using phase diversity;correction accuracy in image-based adaptive optics for nonlinear microscopy;hyperspectral fluorescence microscopy based on compressed sensing;optimization of the diffractive optical element for snapshot spectral imaging used in fluorescence microscopy;double helix PSF engineering for computational fluorescence microscopy imaging;and three dimensional refractive index imaging with differential interference contrast microscopy.

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