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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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Hybrid of two-photon microscopy and optical multimodality imaging for multi-scale imaging of small animals 25

Hybrid of two-photon microscopy and optical multimodality im...

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

作者： Li, Tianmeng Hui, Hui Ma, He Yang, Xin Tian, Jie Northeastern Univ Sinodutch Biomed & Informat Engn Sch Shenyang Liaoning Peoples R China Chinese Acad Sci Inst Automat Key Lab Mol Imaging Beijing 100190 Peoples R China Beijing Key Lab Mol Imaging Inst Automat Beijing 100190 Peoples R China

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Non-invasive imaging technologies, such as magnetic resonance imaging (MRI) and optical multimodality imaging methods, are commonly used for diagnosing and supervising the development of inflammatory bowel disease (IBD). These in vivo imaging methods can provide morphology changes information of IBD in macro-scale. However, it is difficult to investigate the intestinal wall in molecular and cellular level. State-of-art light-sheet and two-photon microscopy have the ability to acquire the changes for IBD in micro-scale. The aim of this work is to evaluate the size of the enterocoel and the thickness of colon wall using both MRI for in vivo imaging, and light-sheet and two-photon microscope for in vitro imaging. C57BL/6 mice were received 3.5% Dextran sodium sulfate (DSS) in the drinking water for 5 days to build IBD model. Mice were imaged with MRI on days 0, 6 to observe colitis progression. After MRI imaging, the mice were sacrificed to take colons for tissue clearing. Then, light-sheet and two-photon microscopies are used for in vitro imaging of the cleared samples. The experimental group showed symptoms of bloody stools, sluggishness and weight loss. It showed that the colon wall was thicker while the enterocoel was narrower compare to control group. The more details are observed using light-sheet and two-photon microscope. It is demonstrated that hybrid of MRI in macro-scale and light-sheet and two-photon microscopy in micro-scale imaging is feasible for colon inflammation diagnosing and supervising.

关键词： Multi-scale molecular imaging Inflammatory bowel disease Magnetic resonance imaging Two-photon microscopy Light-sheet microscopy

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Validation of Excitation-Scan Hyperspectral Multi-faceted Mirror Array Prototype System Advancements to Hyperspectral Imaging Applications 29

Validation of Excitation-Scan Hyperspectral Multi-faceted Mi...

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

作者： Parker, Marina Mayes, Sam A. Browning, Craig M. Deal, Joshua Gunn-Mayes, Samantha Rich, Thomas C. Leavesley, Silas J. Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Ctr Lung Biol Mobile AL 36688 USA Univ S Alabama Dept Syst Engn Mobile AL 36688 USA

ISBN: (纸本)9781510648043;9781510648036

Hyperspectral imaging technologies (HSI) have undergone rapid development since their beginning stages. While original applications were in remote sensing, other uses include agriculture, food safety and medicine. HSI has shown great utility in fluorescence microscopy for detecting signatures from many fluorescent molecules;however, acquisitions speeds have been slow due to light losses associated with spectral filtering. Therefore, we designed a novel light emitting diode (LED)-based rapid excitation scanning hyperspectral imaging platform allowing users to obtain simultaneous measurements of fluorescent labels without compromising acquisition speeds. Previously, we reported our results of the optical ray trace simulations and the geometrical capability of designing a multifaceted mirror imaging system as an initial approach to combine light at many wavelengths. The design utilized LEDs and a multifaceted mirror array to combine light sources into a liquid light guide. The computational model was constructed using Monte Carlo optical ray software (TracePro, Lambda Research Corp.). Recent prototype validation results show that when compared to a commercial emission scanning spectral confocal microscope (Zeiss-LSM-980), the novel LED-based excitation scanning HSI prototype successfully detected and separated six fluorescent labels from a custom 6-label African green monkey kidney epithelial cells. We report on the prototype's ability to overcome limitations of acquisition speeds, sensitivity, and specificity present in conventional systems. Future work will evaluate prototype's light losses to determine latent design modifications needed to demonstrate the system's feasibility as a promising solution for overcoming HSI acquisition speeds. This work was supported by NSF award MRI1725937.

关键词： Systems Design LED Tissue imaging HSI Spectroscopy microscopy Transmission Spectral Fluorescence Molecules Cells

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Quantification of trabecular bone structure from three-dimensional μMR images

Quantification of trabecular bone structure from three-dimen...

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International conference on image processing (ICIP 2001)

作者： Accardo, A Candido, G Toffanin, R Vittur, F Jellús, V Univ Trieste DEEI Trieste Italy

ISBN: (纸本)0780367251

Magnetic resonance imaging (MRI) has recently been applied to the study of osteoporosis. Accurate acquisition techniques and image analysis protocols for the determination of trabecular bone structure are yet to be defined. In this study two different approaches of three-dimensional acquisition and reconstruction were exploited to derive the structural parameters of trabecular bone explants. In particular, MR microscopy (muMRI) was used to obtain conventional (FT) and projection reconstruction (PR) spin-echo images (41x41x41 mum(3)/voxel). Segmentation of the high resolution images into bone and bone marrow was obtained by using a suitable threshold. The main bone structural parameters were then derived by using a 3D version of the directed secant algorithm applied to a series of contiguous sections. The efficiency of the two methods in the assessment of trabecular bone architecture is discussed.

关键词： Magnetic resonance imaging

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Probing neural tissue with Airy light-sheet microscopy: investigation of imaging performance at depth within turbid media 24

Probing neural tissue with Airy light-sheet microscopy: inve...

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

作者： Nylk, Jonathan McCluskey, Kaley Aggarwal, Sanya Tello, Javier A. Dholakia, Kishan Univ St Andrews Sch Phys & Astron SUPA St Andrews KY16 9SS Fife Scotland Univ St Andrews Sch Med St Andrews KY16 9TF Fife Scotland

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Light-sheet microscopy (LSM) has received great interest for fluorescent imaging applications in biomedicine as it facilitates three-dimensional visualisation of large sample volumes with high spatiotemporal resolution whilst minimising irradiation of, and photo-damage to the specimen. Despite these advantages, LSM can only visualise superficial layers of turbid tissues, such as mammalian neural tissue. Propagation-invariant light modes have played a key role in the development of high-resolution LSM techniques as they overcome the natural divergence of a Gaussian beam, enabling uniform and thin light-sheets over large distances. Most notably, Bessel and Airy beam-based light-sheet imaging modalities have been demonstrated. In the single-photon excitation regime and in lightly scattering specimens, Airy-LSM has given competitive performance with advanced Bessel-LSM techniques. Airy and Bessel beams share the property of self-healing, the ability of the beam to regenerate its transverse beam profile after propagation around an obstacle. Bessel-LSM techniques have been shown to increase the penetration-depth of the illumination into turbid specimens but this effect has been understudied in biologically relevant tissues, particularly for Airy beams. It is expected that Airy-LSM will give a similar enhancement over Gaussian-LSM. In this paper, we report on the comparison of Airy-LSM and GaussianLSM imaging modalities within cleared and non-cleared mouse brain tissue. In particular, we examine image quality versus tissue depth by quantitative spatial Fourier analysis of neural structures in virally transduced fluorescent tissue sections, showing a three-fold enhancement at 50 p.m depth into non-cleared tissue with AiryLSM. Complimentary analysis is performed by resolution measurements in bead-injected tissue sections.

关键词： Light-sheet microscopy LSM Airy beam Tissue imaging turbid media neuroscience

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Effect of apodization on axial resolution with a high-aperture objective

Effect of apodization on axial resolution with a high-apertu...

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conference on Three-dimensional microscopy - image acquisition and processing III

作者： Gu, M Victoria Univ. of Technology (Australia)

ISBN: (纸本)0819420298

We investigate the effects of the apodization functions of a high-aperture objective on axial resolution in confocal bright-field microscopy. It is found that the measured apodization function for a commercial objective deviates significantly from the function predicted under the sine condition, which is usually assumed to be satisfied by a commercial objective. This result may explain the long-time discrepancy of the axial resolution between theoretical prediction and measured results, and also the observation of smoothly oscillating side peaks in the axial response when the objective is operated at an incorrect tube length. This finding is of importance when one tries to compensate for the spherical aberrations, with a certain accuracy, by the alteration of tube length of the objective. The deviation of apodization of a practical objective from the sine condition may be also responsible for the observation of the broadened axial response in confocal fluorescence microscopy including 4Pi confocal systems.

关键词： resolution high-aperture objective scanning microscopy confocal microscopy 3-D imaging

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THREE-dimensional SEGMENTATION OF VESICULAR NETWORKS OF FUNGAL HYPHAE IN MACROSCOPIC microscopy image STACKS 24

THREE-DIMENSIONAL SEGMENTATION OF VESICULAR NETWORKS OF FUNG...

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24th IEEE International conference on image processing (ICIP)

作者： Saponaro, P. Treible, W. Kolagunda, A. Rhein, S. Caplan, J. Kambhamettu, C. Wisser, R. Univ Delaware Newark DE 19716 USA

ISBN: (纸本)9781509021758

Automating the extraction and quantification of features from three-dimensional (3-D) image stacks is a critical task for advancing many applications, such as the analysis of biological structures. The quantification of biological resistance of a plant tissue to fungal infection through the analysis of attributes such as fungal penetration depth, fungal mass, and branching of the fungal network at large scales requires the union of 3D image acquisition and analysis. From an image processing perspective, these tasks reduce to segmentation of vessel-like structures and the extraction of features from their skeletonization. In order to sample multiple infection events for analysis, we have developed an approach we refer to as macroscopic microscopy. However, macroscopic microscopy produces high-resolution image stacks that pose challenges to routine approaches and are difficult for a human to annotate to obtain ground truth data. We present a synthetic hyphal network generator, a comparison of several vessel segmentation methods, and a minimum spanning tree method for connecting small gaps resulting from imperfections in imaging or incomplete skeletonization of hyphal networks. Qualitative results are shown for real microscopic data. We believe the comparison of vessel detectors on macroscopic microscopy data, the synthetic vessel generator, and the gap closing technique are beneficial to the image processing community.

关键词： Segmentation Skeletonization Fungal Hyphae Macroscopic microscopy

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Fluorescence Fluctuation Analysis of Mixed Chromophores from a Line-Scanning Hyperspectral Imaging System

Fluorescence Fluctuation Analysis of Mixed Chromophores from...

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

作者： Davis, Ryan W. Aaron, Jesse S. Rempe, Susan L. Timlin, Jerilyn A. Sandia Natl Labs 7011 East Ave Livermore CA 94550 USA Sandia Natl Labs Albuquerque NM 87123 USA

ISBN: (纸本)9780819479662

Fluorescence fluctuation analysis of dilute biomolecules can provide a powerful method for fast and accurate determination of diffusion dynamics, local concentrations, and aggregation states in complex environments. However, spectral overlap among multiple exogenous and endogenous fluorescent species, photobleaching, and background inhomogeneities can compromise quantitative accuracy and constrain useful biological implementation of this analytical strategy in real systems. In order to better understand these limitations and expand the utility of fluctuation correlation methods, spatiotemporal fluorescence correlation analysis was performed on spectrally resolved line scanned images of modeled and real data from mixed fluorescent nanospheres in a synthetic gel matrix. It was found that collecting images at a pixel sampling regime optimal for spectral imaging provides a method for calibration and subsequent temporal correlation analysis which is insensitive to spectral mixing, spatial inhomogeneity, and photobleaching. In these analyses, preprocessing with multivariate curve resolution (MCR) provided the local concentrations of each spectral component in the images, thus facilitating correlation analysis of each component individually. This approach allowed quantitative removal of background signals and showed dramatically improved quantitative results compared to a hypothetical system employing idealized filters and multi-parameter fitting routines.

关键词： Fluorescence correlation spectroscopy hyperspectral imaging spatiotemporal image correlation spectroscopy fluorescence fluctuation multivariate curve resolution line-scan imaging autofluorescence photobleaching

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Tissomics:: two- and threedimensional distribution of nuclei in brain tissue using Laser Scanning Cytometry (LSC)

Tissomics:: two- and threedimensional distribution of nuclei...

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

作者： Lenz, D Mittag, A Mosch, B Bocsi, J Arendt, T Tárnok, A Univ Leipzig Heart Ctr Leipzig Leipzig Germany

ISBN: (纸本)0819456756

Automated quantitative (i.e. stochiometric) analysis of tissues is of eminent importance in the understanding of all interactions between cells in their natural environment. In tissue cytometry a solid trigger is necessary in order to unequivocally differentiate between cellular and non-cellular events. This can be best performed by nuclear staining. Aim of this study was to analyze a brain tissue section by laser scanning cytometry (LSC) in order to depict the threedimensional distribution of nuclei in the tissue. To this end the section was measured in several foci and different nuclei detected in several depths of the tissue were assigned to the respective layer. Frozen sections of formalin-fixed rat or human brain tissue (120 mu m thickness) were incubated with propidiumiodide (PI) (50 mu g/ml) and covered on slides. For analysis by the LSC propidiumiodide was used as trigger. After a first analysis focussed on the top of the tissue, the focus was adjusted in 30 mu m steps deeper into the tissue. Per analysis data of at least 50,000 cells were acquired. After finishing measurements from all depths of the field were merged, i.e. data were combined into a composite data file. With the special features of the LSC it was possible to develop a method depicting the threedimensional distribution of the nuclei in solid tissue sections. LSC can be useful tool for this relatively new field of solid tissue cytometry termed tissomics. After evaluation of methods like this, so far not available data can be analysed for diagnostic purposes. By these studies we intend to demonstrate the power of the LSC for the routine pathological use. This should add up to the bright versatility of applications for the LSC as a cytometric instrument suitable for high throughput and high content analysis.

关键词： Alzheimer's disease cell cycle DNA-polyploidy laser scanning cytometry cytomics

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image acquisition, processing and reconstruction in holographic and tomographic diffractive microscopy

Image acquisition, processing and reconstruction in holograp...

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Digital Holography and Three-dimensional Imaging, DH 2014

作者： Bailleul, Jonathan Liu, Hui Simon, Bertrand Debailleul, Matthieu Haeberlé, Olivier Laboratoire MIPS EA-2332 Université de Haute-Alsace 61 rue Albert Camus 68093 Mulhouse Cedex France

ISBN: (纸本)9781557523082

Tomographic diffractive microscopy allows for imaging unlabeled specimens, with a better resolution than conventional microscopes, giving access to the index of refraction distribution within the specimen, and possibly at high speed. Principles of image formation and reconstruction are presented, and hint towards real-time, three-dimensional acquisition, image reconstruction and final display, are discussed. © 2014 OSA.

关键词： image reconstruction

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Comparison of 3D microscopy methods by imaging a well characterized test object

Comparison of 3D microscopy methods by imaging a well charac...

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conference on Three-dimensional microscopy - image acquisition and processing IV

作者： McNally, JG Cogswell, CJ Fekete, PW Conchello, JA Washington Univ. (United States) Univ. of Sydney (United States) Univ. of Sydney (Australia)

ISBN: (纸本)0819423955

We used a 10-mu m-diameter fluorescent bead as a test object for 3D microscopy, and independently determined its structure by examining 1-mu m-thick physical sections of the bead. images of the full bead on different 3D microscopes revealed a number of aberrations and distortions. images also showed evidence for absorption and/or scattering on all. confocal and wide-field microscopes tested, but not on a two-photon microscope. The best 3D images of the bead came from deconvolution of either wide-field or certain confocal images. For deconvolution of partially confocal data, a region extending 7 mu m beyond the top and bottom of the bead contained out-of-focus-light information essential for correct restoration. Fully confocal images required 10-fold less computer time for deconvolution, but 1000-fold more excitation light than a wide-field image, even though the resultant restorations were reasonably comparable. Laser imaging of the bead appeared to produce an artifactual image for which we currently have no explanation.

关键词： 3D microscopy comparison confocal wide field two photon

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