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

作者： Shaked, Natan T. Satterwhite, Lisa L. Telen, Marilyn J. Truskey, George A. Wax, Adam Tel Aviv Univ Dept Biomed Engn Fac Engn IL-69978 Tel Aviv Israel Duke Univ Fitzpatrick Inst Photon Dept Biomed Engn Durham NC 27708 USA Duke Univ Med Ctr Dept Med Div Hematol Durham NC 27708 USA

ISBN: (纸本)9780819488701

We have applied wide-field digital interferometric techniques to quantitatively image sickle red blood cells (RBCs) [1] in a noncontact label-free manner, and measure the nanometer-scale fluctuations in their thickness as an indication of their stiffness. The technique can simultaneously measure the fluctuations for multiple spatial points on the RBC and thus yields a map describing the stiffness of each RBC in the field of view. Using this map, the local rigidity regions of the RBC are evaluated quantitatively. Since wide-field digital interferometry is a quantitative holographic imaging technique rather than one-point measurement, it can be used to simultaneously evaluate cell transverse morphology plus thickness in addition to its stiffness profile. Using this technique, we examine the morphology and dynamics of RBCs from individuals who suffer from sickle cell disease, and find that the sickle RBCs are significantly stiffer than healthy RBCs. Furthermore, we show that the technique is sensitive enough to distinguish various classes of sickle RBCs, including sickle RBCs with visibly-normal morphology, compared to the stiffer crescent-shaped sickle RBCs.

关键词： Interferometric phase microscopy holographic microscopy red blood cells sickle cell disease

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Localization accuracy in single molecule microscopy using electron-multiplying charge-coupled device cameras

Localization accuracy in single molecule microscopy using el...

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

作者： Chao, Jerry Ward, E. Sally Ober, Raimund J. Univ Texas Dallas Dept Elect Engn Richardson TX 75083 USA Univ Texas Southwestern Med Ctr Dallas Dept Immunol Dallas TX USA

ISBN: (纸本)9780819488701

The electron-multiplying charge-coupled device (EMCCD) is a popular technology for imaging under extremely low light conditions. It has become widely used, for example, in single molecule microscopy experiments where few photons can be detected from the individual molecules of interest. Despite its important role in low light microscopy, however, little has been done in the way of determining how accurately parameters of interest (e. g., location of a single molecule) can be estimated from an image that it produces. Here, we develop the theory for calculating the Fisher information matrix, and hence the Cramer-Rao lower bound-based limit of the accuracy, for estimating parameters from an EMCCD image. An EMCCD operates by amplifying a weak signal that would otherwise be drowned out by the detector's readout noise as in the case of a conventional charge-coupled device (CCD). The signal amplification is a stochastic electron multiplication process, and is modeled here as a geometrically multiplied branching process. In developing our theory, we also introduce a "noise coefficient" which enables the comparison of the Fisher information of different data models via a scalar quantity. This coefficient importantly allows the selection of the best detector (e.g., EMCCD or CCD), based on factors such as the signal level, and regardless of the specific estimation problem at hand. We apply our theory to the problem of localizing a single molecule, and compare the calculated limits of the localization accuracy with the standard deviations of maximum likelihood location estimates obtained from simulated images of a single molecule.

关键词： Branching process Cramer-Rao lower bound electron multiplication Fisher information matrix single molecule microscopy

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In vivo multiphoton microscopy associated to 3D image processing for human skin characterization

<i>In vivo</i> multiphoton microscopy associated to 3D image...

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conference on Multiphoton microscopy in the Biomedical Sciences xii

作者： Baldeweck, T. Tancrede, E. Dokladal, P. Koudoro, S. Morard, V. Meyer, F. Decenciere, E. Pena, A. -M. LOreal Res & Innovat Aulnay Sous Bois France Hop St Louis Ctr Rech Bioclin LOreal Res & Innovat Paris France MINES Paris Tech Ctr Morphol Math Math & Syst Fontainebleau France

ISBN: (纸本)9780819488695

Multiphoton microscopy has emerged in the past decade as a promising non-invasive skin imaging technique. The aim of this study was to assess whether multiphoton microscopy coupled to specific 3D image processing tools could provide new insights into the organization of different skin components and their age-related changes. For that purpose, we performed a clinical trial on 15 young and 15 aged human female volunteers on the ventral and dorsal side of the forearm using the DermaInspect(R) medical imaging device. We visualized the skin by taking advantage of intrinsic multiphoton signals from cells, elastic and collagen fibers. We also developed 3D image processing algorithms adapted to in vivo multiphoton images of human skin in order to extract quantitative parameters in each layer of the skin (epidermis and superficial dermis). The results show that in vivo multiphoton microscopy is able to evidence several skin alterations due to skin aging: morphological changes in the epidermis and modifications in the quantity and organization of the collagen and elastic fibers network. In conclusion, the association of multiphoton microscopy with specific image processing allows the three-dimensional organization of skin components to be visualized and quantified thus providing a powerful tool for cosmetic and dermatological investigations.

关键词： second harmonic generation two-photon excited fluorescence fibrillar collagen elastic fibers skin in vivo multiphoton microscopy 3D image processing quantification

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Neuronal cell growth on polymeric scaffolds studied by CARS microscopy

Neuronal cell growth on polymeric scaffolds studied by CARS ...

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conference on Multiphoton microscopy in the Biomedical Sciences xii

作者： Enejder, Annika Fink, Helen Kuhn, Hans-Georg Chalmers Univ Technol Dept Chem & Biol Engn S-41296 Gothenburg Sweden Gothenburg Univ Inst Neurosci & Physiol S-41124 Gothenburg Sweden

ISBN: (纸本)9780819488695

For studies of neuronal cell integration and neurite outgrowth in polymeric scaffold materials as a future alternative for the treatment of damages in the neuronal system, we have developed a protocol employing CARS microscopy for imaging of neuronal networks. The benefits of CARS microscopy come here to their best use;(i) the overall three-dimensional (3D) arrangement of multiple cells and their neurites can be visualized without the need for chemical preparations or physical sectioning, potentially affecting the architecture of the soft, fragile scaffolds and (ii) details on the interaction between single cells and scaffold fibrils can be investigated by close-up images at sub-micron resolution. The establishment of biologically more relevant 3D neuronal networks in a soft hydrogel composed of native Extra Cellular Matrix (ECM) components was compared with conventional two-dimensional networks grown on a stiff substrate. images of cells in the hydrogel scaffold reveal significantly different networking characteristics compared to the 2D networks, raising the question whether the functionality of neurons grown as layers in conventional cultivation dishes represents that of neurons in the central and peripheral nervous systems.

关键词： Neurons microscopy 3D image processing Optical parametric oscillators Polymers Microscopes Signal detection Visualization Imaging systems Neural networks

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The study of muscle remodeling in Drosophila metamorphosis using in vivo microscopy and bioimage informatics

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BMC BIOINFORMATICS 2012年第17期13卷 1-13页

作者： Chinta, Rambabu Tan, Joo Huang Wasser, Martin ASTAR Bioinformat Inst BII Imaging Informat Div Live Cell Imaging & Automat Image Anal Grp Singapore Singapore Natl Univ Singapore Dept Biol Sci Singapore 117548 Singapore

Background: Metamorphosis in insects transforms the larval into an adult body plan and comprises the destruction and remodeling of larval and the generation of adult tissues. The remodeling of larval into adult muscles promises to be a genetic model for human atrophy since it is associated with dramatic alteration in cell size. Furthermore, muscle development is amenable to 3D in vivo microscopy at high cellular resolution. However, multi-dimensional image acquisition leads to sizeable amounts of data that demand novel approaches in image processing and analysis. Results: To handle, visualize and quantify time-lapse datasets recorded in multiple locations, we designed a workflow comprising three major modules. First, the previously introduced TLM-converter concatenates stacks of single time-points. The second module, TLM-2D-Explorer, creates maximum intensity projections for rapid inspection and allows the temporal alignment of multiple datasets. The transition between prepupal and pupal stage serves as reference point to compare datasets of different genotypes or treatments. We demonstrate how the temporal alignment can reveal novel insights into the east gene which is involved in muscle remodeling. The third module, TLM-3D-Segmenter, performs semi-automated segmentation of selected muscle fibers over multiple frames. 3D image segmentation consists of 3 stages. First, the user places a seed into a muscle of a key frame and performs surface detection based on level-set evolution. Second, the surface is propagated to subsequent frames. Third, automated segmentation detects nuclei inside the muscle fiber. The detected surfaces can be used to visualize and quantify the dynamics of cellular remodeling. To estimate the accuracy of our segmentation method, we performed a comparison with a manually created ground truth. Key and predicted frames achieved a performance of 84% and 80%, respectively. Conclusions: We describe an analysis pipeline for the efficient handling and

关键词： Maximum Intensity Projection Pupal Stage Surface Evolution Genetic Perturbation Prepupal Stage

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General Fusion Approaches for the Age Determination of Latent Fingerprint Traces: Results for 2D and 3D Binary Pixel Feature Fusion

General Fusion Approaches for the Age Determination of Laten...

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conference on Three-dimensional image processing (3DIP) and Applications II

作者： Merkel, Ronny Gruhn, Stefan Dittmann, Jana Vielhauer, Claus Braeutigam, Anja Otto von Guericke Univ Res Grp Multimedia & Secur Dept Comp Sci Univ Pl 2 D-39106 Magdeburg Germany Brandenburg Univ Appl Sci Dept Informat & Media D-14770 Brandenburg Germany State Criminal Police Off Saxony Anhalt D-39124 Magdeburg Germany

ISBN: (纸本)9780819489371

Determining the age of latent fingerprint traces found at crime scenes is an unresolved research issue since decades. Solving this issue could provide criminal investigators with the specific time a fingerprint trace was left on a surface, and therefore would enable them to link potential suspects to the time a crime took place as well as to reconstruct the sequence of events or eliminate irrelevant fingerprints to ensure privacy constraints. Transferring imaging techniques from different application areas, such as 3D image acquisition, surface measurement and chemical analysis to the domain of lifting latent biometric fingerprint traces is an upcoming trend in forensics. Such non-destructive sensor devices might help to solve the challenge of determining the age of a latent fingerprint trace, since it provides the opportunity to create time series and process them using pattern recognition techniques and statistical methods on digitized 2D, 3D and chemical data, rather than classical, contact-based capturing techniques, which alter the fingerprint trace and therefore make continuous scans impossible. In prior work, we have suggested to use a feature called binary pixel, which is a novel approach in the working field of fingerprint age determination. The feature uses a Chromatic White Light (CWL) image sensor to continuously scan a fingerprint trace over time and retrieves a characteristic logarithmic aging tendency for 2D-intensity as well as 3D-topographic images from the sensor. In this paper, we propose to combine such two characteristic aging features with other 2D and 3D features from the domains of surface measurement, microscopy, photography and spectroscopy, to achieve an increase in accuracy and reliability of a potential future age determination scheme. Discussing the feasibility of such variety of sensor devices and possible aging features, we propose a general fusion approach, which might combine promising features to a joint age determination scheme in

关键词： latent fingerprint traces crime scene forensics age determination binary pixel non-invasive fingerprint lifting fusion CWL spectroscopy microscopy

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High-speed focal modulation microscopy using acousto-optical modulators for visualization of thick biological specimens

High-speed focal modulation microscopy using acousto-optical...

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

作者： Chong, Shau Poh Gao, Guangjun Sheppard, Colin J. R. Chen, Nanguang Natl Univ Singapore Div Bioengn Singapore 117576 Singapore

ISBN: (纸本)9780819484413

In recent development of fluorescence microscopy, the out-of-focus fluorescence background that arises when imaging deep inside biological tissues is critical in determining the image quality and penetration depth. Focal modulation microscopy (FMM) is an advanced fluorescence technique that can provide high subcellular resolution when imaging thick specimens mainly by preserving the signal-to-background ratio.

关键词： Fluorescence microscopy multiple scattering penetration depth

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Heart Synchronization for SPIM microscopy of Living Zebra Fish

Heart Synchronization for SPIM Microscopy of Living Zebra Fi...

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

作者： Taylor, J. M. Saunter, C. D. Chaudhry, B. Henderson, D. J. Love, G. D. Girkin, J. M. Univ Durham Dept Phys Ctr Adv Instrumentat South Rd Durham DH1 3LE England Newcastle Univ Inst Human Genet Newcastle Upon Tyne NE1 3BZ Tyne & Wear England

We describe work on producing a selective plane illumination microscope for cardiac imaging in zebra fish embryos. The system has a novel synchronization system for imaging oscillating structures (e.g. the heart) and ... 详细信息

ISBN: (纸本)9780819484413

关键词： Optical microscopy Selective Plane Illumination Synchronization

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Three-dimensional data acquisition with aberrations correction for video-rate microscopy

Three-dimensional data acquisition with aberrations correcti...

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

作者： Samim, Masood Cisek, Richard Sandkuijl, Daaf Tretyakov, Ian Siu, Sam Musikhin, Sergey Barzda, Virginijus Univ Toronto Dept Chem & Phys Sci Mississauga ON L5L 1C6 Canada

ISBN: (纸本)9780819484413

We demonstrate a multimodal, multifocal, differential nonlinear optical microscope, which is equipped with a pair of deformable mirrors and a Shack-Hartmann sensor for dynamic wavefront manipulation. The optical wavefronts of a home built Yb:KGW femtosecond (1028 nm) laser-beams are engineered to perform multidepth focusing in differential mode with simultaneous corrections for optical aberrations. The 39-actuator deformable mirrors provide fast reshaping of the wavefront and optical aberrations correction of the diffraction-limited focal volume allowing for fast axial scanning. Combination of similar to 200 frames per second lateral scanning with fast refocusing enables a three-dimensional video rate scanning capability, which is essential for studying rapid dynamics in biological organisms, such as blood flow, cardiac contractions, and motility of microorganisms in a three-dimensional volume.

关键词： Deformable mirrors adaptive optics nonlinear optical microscopy harmonic generation microscopy non-invasive in vivo imaging three-dimensional imaging multi-contrast microscopy

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Autostereoscopic visualization of 3D time-varying complex objects in volumetric image sequences

Autostereoscopic visualization of 3D time-varying complex ob...

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

作者： Benassarou, A. Valette, G. Debons, D. Remion, Y. Lucas, L. CReSTIC SIC Rue CrayeresBP 1035 F-51687 Reims 2 France 3DTV Solut F-51100 Reims France

ISBN: (纸本)9780819484413

This paper presents 4dVizMed, a framework for interactive analysis and autostereoscopic visualization of 3d time-varying objects in volumetric image sequences. It combines a deformable surface model which automatically tracks volumetric features, real-time multi-view stereo volume rendering, and some interactive tools for manipulation and quantization. Our method is based on a topological feature tracking process, using a flow-based paradigm and a deformable surface model. It tracks through time the evolution of the components of an isosurface and their interaction with other components. We focus on the difficulties of visualizing 4d volume data, and we report the results of preliminary experiments designed to evaluate the utility of autostereoscopic displays for this purpose.

关键词： Visualization 3D modeling 3D image processing Autostereoscopic displays image visualization image segmentation 3D displays 3D visualizations image processing Volume rendering

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