Delivery of Nano-Tethered Therapies to Brain Metastases of Primary Breast Cancer Using a Cellular Trojan Horse

DTIC Science & Technology

2015-10-01

tomography images. The CT image densities in Hounsfield units (HU) of the brain were translated into corresponding optical properties (absorption...derived the Hounsfield units and optical properties of brain tissues such as white/gray matter. 13-15 The segmentation software generated an optical map...treatment protocol. Head CT image densities (in Hounsfield Units /HU) are segmented and translated into optical properties of the brain tissue

Spatial Mapping of Lipids at Cellular Resolution in Embryos of Cotton[W][OA

PubMed Central

Horn, Patrick J.; Korte, Andrew R.; Neogi, Purnima B.; Love, Ebony; Fuchs, Johannes; Strupat, Kerstin; Borisjuk, Ljudmilla; Shulaev, Vladimir; Lee, Young-Jin; Chapman, Kent D.

2012-01-01

Advances in mass spectrometry (MS) have made comprehensive lipidomics analysis of complex tissues relatively commonplace. These compositional analyses, although able to resolve hundreds of molecular species of lipids in single extracts, lose the original cellular context from which these lipids are derived. Recently, high-resolution MS of individual lipid droplets from seed tissues indicated organelle-to-organelle variation in lipid composition, suggesting that heterogeneity of lipid distributions at the cellular level may be prevalent. Here, we employed matrix-assisted laser desorption/ionization–MS imaging (MALDI-MSI) approaches to visualize lipid species directly in seed tissues of upland cotton (Gossypium hirsutum). MS imaging of cryosections of mature cotton embryos revealed a distinct, heterogeneous distribution of molecular species of triacylglycerols and phosphatidylcholines, the major storage and membrane lipid classes in cotton embryos. Other lipids were imaged, including phosphatidylethanolamines, phosphatidic acids, sterols, and gossypol, indicating the broad range of metabolites and applications for this chemical visualization approach. We conclude that comprehensive lipidomics images generated by MALDI-MSI report accurate, relative amounts of lipid species in plant tissues and reveal previously unseen differences in spatial distributions providing for a new level of understanding in cellular biochemistry. PMID:22337917

aMAP is a validated pipeline for registration and segmentation of high-resolution mouse brain data

PubMed Central

Niedworok, Christian J.; Brown, Alexander P. Y.; Jorge Cardoso, M.; Osten, Pavel; Ourselin, Sebastien; Modat, Marc; Margrie, Troy W.

2016-01-01

The validation of automated image registration and segmentation is crucial for accurate and reliable mapping of brain connectivity and function in three-dimensional (3D) data sets. While validation standards are necessarily high and routinely met in the clinical arena, they have to date been lacking for high-resolution microscopy data sets obtained from the rodent brain. Here we present a tool for optimized automated mouse atlas propagation (aMAP) based on clinical registration software (NiftyReg) for anatomical segmentation of high-resolution 3D fluorescence images of the adult mouse brain. We empirically evaluate aMAP as a method for registration and subsequent segmentation by validating it against the performance of expert human raters. This study therefore establishes a benchmark standard for mapping the molecular function and cellular connectivity of the rodent brain. PMID:27384127

Isolating specific cell and tissue compartments from 3D images for quantitative regional distribution analysis using novel computer algorithms.

PubMed

Fenrich, Keith K; Zhao, Ethan Y; Wei, Yuan; Garg, Anirudh; Rose, P Ken

2014-04-15

Isolating specific cellular and tissue compartments from 3D image stacks for quantitative distribution analysis is crucial for understanding cellular and tissue physiology under normal and pathological conditions. Current approaches are limited because they are designed to map the distributions of synapses onto the dendrites of stained neurons and/or require specific proprietary software packages for their implementation. To overcome these obstacles, we developed algorithms to Grow and Shrink Volumes of Interest (GSVI) to isolate specific cellular and tissue compartments from 3D image stacks for quantitative analysis and incorporated these algorithms into a user-friendly computer program that is open source and downloadable at no cost. The GSVI algorithm was used to isolate perivascular regions in the cortex of live animals and cell membrane regions of stained spinal motoneurons in histological sections. We tracked the real-time, intravital biodistribution of injected fluorophores with sub-cellular resolution from the vascular lumen to the perivascular and parenchymal space following a vascular microlesion, and mapped the precise distributions of membrane-associated KCC2 and gephyrin immunolabeling in dendritic and somatic regions of spinal motoneurons. Compared to existing approaches, the GSVI approach is specifically designed for isolating perivascular regions and membrane-associated regions for quantitative analysis, is user-friendly, and free. The GSVI algorithm is useful to quantify regional differences of stained biomarkers (e.g., cell membrane-associated channels) in relation to cell functions, and the effects of therapeutic strategies on the redistributions of biomolecules, drugs, and cells in diseased or injured tissues. Copyright © 2014 Elsevier B.V. All rights reserved.

Iron in Multiple Sclerosis and Its Noninvasive Imaging with Quantitative Susceptibility Mapping

PubMed Central

Stüber, Carsten; Pitt, David; Wang, Yi

2016-01-01

Iron is considered to play a key role in the development and progression of Multiple Sclerosis (MS). In particular, iron that accumulates in myeloid cells after the blood-brain barrier (BBB) seals may contribute to chronic inflammation, oxidative stress and eventually neurodegeneration. Magnetic resonance imaging (MRI) is a well-established tool for the non-invasive study of MS. In recent years, an advanced MRI method, quantitative susceptibility mapping (QSM), has made it possible to study brain iron through in vivo imaging. Moreover, immunohistochemical investigations have helped defining the lesional and cellular distribution of iron in MS brain tissue. Imaging studies in MS patients and of brain tissue combined with histological studies have provided important insights into the role of iron in inflammation and neurodegeneration in MS. PMID:26784172

T2* mapping and delayed gadolinium-enhanced magnetic resonance imaging in cartilage (dGEMRIC) of humeral articular cartilage--a histologically controlled study.

PubMed

Bittersohl, Bernd; Kircher, Jörn; Miese, Falk R; Dekkers, Christin; Habermeyer, Peter; Fröbel, Julia; Antoch, Gerald; Krauspe, Rüdiger; Zilkens, Christoph

2015-10-01

Cartilage biochemical imaging modalities that include the magnetic resonance imaging (MRI) techniques of T2* mapping (sensitive to water content and collagen fiber network) and delayed gadolinium-enhanced MRI of cartilage (dGEMRIC, sensitive to the glycosaminoglycan content) can be effective instruments for early diagnosis and reliable follow-up of cartilage damage. The purpose of this study was to provide T2* mapping and dGEMRIC values in various histologic grades of cartilage degeneration in humeral articular cartilage. A histologically controlled in vitro study was conducted that included human humeral head cartilage specimens with various histologic grades of cartilage degeneration. High-resolution, 3-dimensional (3D) T2* mapping and dGEMRIC were performed that enabled the correlation of MRI and histology data. Cartilage degeneration was graded according to the Mankin score, which evaluates surface morphology, cellularity, toluidine blue staining, and tidemark integrity. SPSS software was used for statistical analyses. Both MRI mapping values decreased significantly (P < .001) with increasing cartilage degeneration. Spearman rank analysis revealed a significant correlation (correlation coefficients ranging from -0.315 to 0.784; P < .001) between the various histologic parameters and the T2* and T1Gd mapping values. This study demonstrates the feasibility of 3D T2* and dGEMRIC to identify various histologic grades of cartilage damage of humeral articular cartilage. With regard to the advantages of these mapping techniques with high image resolution and the ability to accomplish a 3D biochemically sensitive imaging, we consider that these imaging techniques can make a positive contribution to the currently evolving science and practice of cartilage biochemical imaging. Copyright © 2015 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.

Imaging articular cartilage using second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Mansfield, Jessica C.; Winlove, C. Peter; Knapp, Karen; Matcher, Stephen J.

2006-02-01

Sub cellular resolution images of equine articular cartilage have been obtained using both second harmonic generation microscopy (SHGM) and two-photon fluorescence microscopy (TPFM). The SHGM images clearly map the distribution of the collagen II fibers within the extracellular matrix while the TPFM images show the distribution of endogenous two-photon fluorophores in both the cells and the extracellular matrix, highlighting especially the pericellular matrix and bright 2-3μm diameter features within the cells. To investigate the source of TPF in the extracellular matrix experiments have been carried out to see if it may originate from the proteoglycans. Pure solutions of the following proteoglycans hyaluronan, chondroitin sulfate and aggrecan have been imaged, only the aggrecan produced any TPF and here the intensity was not great enough to account for the TPF in the extracellular matrix. Also cartilage samples were subjected to a process to remove proteoglycans and cellular components. After this process the TPF from the samples had decreased by a factor of two, with respect to the SHG intensity.

3D single point imaging with compressed sensing provides high temporal resolution R 2* mapping for in vivo preclinical applications.

PubMed

Rioux, James A; Beyea, Steven D; Bowen, Chris V

2017-02-01

Purely phase-encoded techniques such as single point imaging (SPI) are generally unsuitable for in vivo imaging due to lengthy acquisition times. Reconstruction of highly undersampled data using compressed sensing allows SPI data to be quickly obtained from animal models, enabling applications in preclinical cellular and molecular imaging. TurboSPI is a multi-echo single point technique that acquires hundreds of images with microsecond spacing, enabling high temporal resolution relaxometry of large-R 2 * systems such as iron-loaded cells. TurboSPI acquisitions can be pseudo-randomly undersampled in all three dimensions to increase artifact incoherence, and can provide prior information to improve reconstruction. We evaluated the performance of CS-TurboSPI in phantoms, a rat ex vivo, and a mouse in vivo. An algorithm for iterative reconstruction of TurboSPI relaxometry time courses does not affect image quality or R 2 * mapping in vitro at acceleration factors up to 10. Imaging ex vivo is possible at similar acceleration factors, and in vivo imaging is demonstrated at an acceleration factor of 8, such that acquisition time is under 1 h. Accelerated TurboSPI enables preclinical R 2 * mapping without loss of data quality, and may show increased specificity to iron oxide compared to other sequences.

Real-time hyperspectral fluorescence imaging of pancreatic β-cell dynamics with the image mapping spectrometer

PubMed Central

Elliott, Amicia D.; Gao, Liang; Ustione, Alessandro; Bedard, Noah; Kester, Robert; Piston, David W.; Tkaczyk, Tomasz S.

2012-01-01

Summary The development of multi-colored fluorescent proteins, nanocrystals and organic fluorophores, along with the resulting engineered biosensors, has revolutionized the study of protein localization and dynamics in living cells. Hyperspectral imaging has proven to be a useful approach for such studies, but this technique is often limited by low signal and insufficient temporal resolution. Here, we present an implementation of a snapshot hyperspectral imaging device, the image mapping spectrometer (IMS), which acquires full spectral information simultaneously from each pixel in the field without scanning. The IMS is capable of real-time signal capture from multiple fluorophores with high collection efficiency (∼65%) and image acquisition rate (up to 7.2 fps). To demonstrate the capabilities of the IMS in cellular applications, we have combined fluorescent protein (FP)-FRET and [Ca2+]i biosensors to measure simultaneously intracellular cAMP and [Ca2+]i signaling in pancreatic β-cells. Additionally, we have compared quantitatively the IMS detection efficiency with a laser-scanning confocal microscope. PMID:22854044

Singlet oxygen Triplet Energy Transfer based imaging technology for mapping protein-protein proximity in intact cells

PubMed Central

To, Tsz-Leung; Fadul, Michael J.; Shu, Xiaokun

2014-01-01

Many cellular processes are carried out by large protein complexes that can span several tens of nanometers. Whereas Forster resonance energy transfer has a detection range of <10 nm, here we report the theoretical development and experimental demonstration of a new fluorescence imaging technology with a detection range of up to several tens of nanometers: singlet oxygen triplet energy transfer. We demonstrate that our method confirms the topology of a large protein complex in intact cells, which spans from the endoplasmic reticulum to the outer mitochondrial membrane and the matrix. This new method is thus suited for mapping protein proximity in large protein complexes. PMID:24905026

Harnessing cell-to-cell variations to probe bacterial structure and biophysics

NASA Astrophysics Data System (ADS)

Cass, Julie A.

Advances in microscopy and biotechnology have given us novel insights into cellular biology and physics. While bacteria were long considered to be relatively unstructured, the development of fluorescence microscopy techniques, and spatially and temporally resolved high-throughput quantitative studies, have uncovered that the bacterial cell is highly organized, and its structure rigorously maintained. In this thesis I will describe our gateTool software, designed to harness cell-to-cell variations to probe bacterial structure, and discuss two exciting aspects of structure that we have employed gateTool to investigate: (i) chromosome organization and the cellular mechanisms for controlling DNA dynamics, and (ii) the study of cell wall synthesis, and how the genes in the synthesis pathway impact cellular shape. In the first project, we develop a spatial and temporal mapping of cell-cycle-dependent chromosomal organization, and use this quantitative map to discover that chromosomal loci segregate from midcell with universal dynamics. In the second project, I describe preliminary time- lapse and snapshot imaging analysis suggesting phentoypical coherence across peptidoglycan synthesis pathways.

Increasing the Accuracy of Volume and ADC Delineation for Heterogeneous Tumor on Diffusion-Weighted MRI: Correlation with PET/CT

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gong, Nan-Jie; Wong, Chun-Sing, E-mail: drcswong@gmail.com; Chu, Yiu-Ching

2013-10-01

Purpose: To improve the accuracy of volume and apparent diffusion coefficient (ADC) measurements in diffusion-weighted magnetic resonance imaging (MRI), we proposed a method based on thresholding both the b0 images and the ADC maps. Methods and Materials: In 21 heterogeneous lesions from patients with metastatic gastrointestinal stromal tumors (GIST), gross lesion were manually contoured, and corresponding volumes and ADCs were denoted as gross tumor volume (GTV) and gross ADC (ADC{sub g}), respectively. Using a k-means clustering algorithm, the probable high-cellularity tumor tissues were selected based on b0 images and ADC maps. ADC and volume of the tissues selected using themore » proposed method were denoted as thresholded ADC (ADC{sub thr}) and high-cellularity tumor volume (HCTV), respectively. The metabolic tumor volume (MTV) in positron emission tomography (PET)/computed tomography (CT) was measured using 40% maximum standard uptake value (SUV{sub max}) as the lower threshold, and corresponding mean SUV (SUV{sub mean}) was also measured. Results: HCTV had excellent concordance with MTV according to Pearson's correlation (r=0.984, P<.001) and linear regression (slope = 1.085, intercept = −4.731). In contrast, GTV overestimated the volume and differed significantly from MTV (P=.005). ADC{sub thr} correlated significantly and strongly with SUV{sub mean} (r=−0.807, P<.001) and SUV{sub max} (r=−0.843, P<.001); both were stronger than those of ADC{sub g}. Conclusions: The proposed lesion-adaptive semiautomatic method can help segment high-cellularity tissues that match hypermetabolic tissues in PET/CT and enables more accurate volume and ADC delineation on diffusion-weighted MR images of GIST.« less

Far-field photostable optical nanoscopy (PHOTON) for real-time super-resolution single-molecular imaging of signaling pathways of single live cells

NASA Astrophysics Data System (ADS)

Huang, Tao; Browning, Lauren M.; Xu, Xiao-Hong Nancy

2012-04-01

Cellular signaling pathways play crucial roles in cellular functions and design of effective therapies. Unfortunately, study of cellular signaling pathways remains formidably challenging because sophisticated cascades are involved, and a few molecules are sufficient to trigger signaling responses of a single cell. Here we report the development of far-field photostable-optical-nanoscopy (PHOTON) with photostable single-molecule-nanoparticle-optical-biosensors (SMNOBS) for mapping dynamic cascades of apoptotic signaling pathways of single live cells in real-time at single-molecule (SM) and nanometer (nm) resolutions. We have quantitatively imaged single ligand molecules (tumor necrosis factor α, TNFα) and their binding kinetics with their receptors (TNFR1) on single live cells; tracked formation and internalization of their clusters and their initiation of intracellular signaling pathways in real-time; and studied apoptotic signaling dynamics and mechanisms of single live cells with sufficient temporal and spatial resolutions. This study provides new insights into complex real-time dynamic cascades and molecular mechanisms of apoptotic signaling pathways of single live cells. PHOTON provides superior imaging and sensing capabilities and SMNOBS offer unrivaled biocompatibility and photostability, which enable probing of signaling pathways of single live cells in real-time at SM and nm resolutions.Cellular signaling pathways play crucial roles in cellular functions and design of effective therapies. Unfortunately, study of cellular signaling pathways remains formidably challenging because sophisticated cascades are involved, and a few molecules are sufficient to trigger signaling responses of a single cell. Here we report the development of far-field photostable-optical-nanoscopy (PHOTON) with photostable single-molecule-nanoparticle-optical-biosensors (SMNOBS) for mapping dynamic cascades of apoptotic signaling pathways of single live cells in real-time at single-molecule (SM) and nanometer (nm) resolutions. We have quantitatively imaged single ligand molecules (tumor necrosis factor α, TNFα) and their binding kinetics with their receptors (TNFR1) on single live cells; tracked formation and internalization of their clusters and their initiation of intracellular signaling pathways in real-time; and studied apoptotic signaling dynamics and mechanisms of single live cells with sufficient temporal and spatial resolutions. This study provides new insights into complex real-time dynamic cascades and molecular mechanisms of apoptotic signaling pathways of single live cells. PHOTON provides superior imaging and sensing capabilities and SMNOBS offer unrivaled biocompatibility and photostability, which enable probing of signaling pathways of single live cells in real-time at SM and nm resolutions. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr11739h

The Cellular Automata for modelling of spreading of lava flow on the earth surface

NASA Astrophysics Data System (ADS)

Jarna, A.

2012-12-01

Volcanic risk assessment is a very important scientific, political and economic issue in densely populated areas close to active volcanoes. Development of effective tools for early prediction of a potential volcanic hazard and management of crises are paramount. However, to this date volcanic hazard maps represent the most appropriate way to illustrate the geographical area that can potentially be affected by a volcanic event. Volcanic hazard maps are usually produced by mapping out old volcanic deposits, however dynamic lava flow simulation gaining popularity and can give crucial information to corroborate other methodologies. The methodology which is used here for the generation of volcanic hazard maps is based on numerical simulation of eruptive processes by the principle of Cellular Automata (CA). The python script is integrated into ArcToolbox in ArcMap (ESRI) and the user can select several input and output parameters which influence surface morphology, size and shape of the flow, flow thickness, flow velocity and length of lava flows. Once the input parameters are selected, the software computes and generates hazard maps on the fly. The results can be exported to Google Maps (.klm format) to visualize the results of the computation. For validation of the simulation code are used data from a real lava flow. Comparison of the simulation results with real lava flows mapped out from satellite images will be presented.

Wide-field optical mapping of neural activity and brain haemodynamics: considerations and novel approaches

PubMed Central

Ma, Ying; Shaik, Mohammed A.; Kozberg, Mariel G.; Thibodeaux, David N.; Zhao, Hanzhi T.; Yu, Hang

2016-01-01

Although modern techniques such as two-photon microscopy can now provide cellular-level three-dimensional imaging of the intact living brain, the speed and fields of view of these techniques remain limited. Conversely, two-dimensional wide-field optical mapping (WFOM), a simpler technique that uses a camera to observe large areas of the exposed cortex under visible light, can detect changes in both neural activity and haemodynamics at very high speeds. Although WFOM may not provide single-neuron or capillary-level resolution, it is an attractive and accessible approach to imaging large areas of the brain in awake, behaving mammals at speeds fast enough to observe widespread neural firing events, as well as their dynamic coupling to haemodynamics. Although such wide-field optical imaging techniques have a long history, the advent of genetically encoded fluorophores that can report neural activity with high sensitivity, as well as modern technologies such as light emitting diodes and sensitive and high-speed digital cameras have driven renewed interest in WFOM. To facilitate the wider adoption and standardization of WFOM approaches for neuroscience and neurovascular coupling research, we provide here an overview of the basic principles of WFOM, considerations for implementation of wide-field fluorescence imaging of neural activity, spectroscopic analysis and interpretation of results. This article is part of the themed issue ‘Interpreting BOLD: a dialogue between cognitive and cellular neuroscience’. PMID:27574312

Recent Advances in Superparamagnetic Iron Oxide Nanoparticles for Cellular Imaging and Targeted Therapy Research

PubMed Central

Wang, Yi-Xiang J.; Xuan, Shouhu; Port, Marc; Idee, Jean-Marc

2013-01-01

Advances of nanotechnology have led to the development of nanomaterials with both potential diagnostic and therapeutic applications. Among them, superparamagnetic iron oxide (SPIO) nanoparticles have received particular attention. Over the past decade, various SPIOs with unique physicochemical and biological properties have been designed by modifying the particle structure, size and coating. This article reviews the recent advances in preparing SPIOs with novel properties, the way these physicochemical properties of SPIOs influence their interaction with cells, and the development of SPIOs in liver and lymph nodes magnetic resonance imaging (MRI) contrast. Cellular uptake of SPIO can be exploited in a variety of potential clinical applications, including stem cell and inflammation cell tracking and intra-cellular drug delivery to cancerous cells which offers higher intra-cellular concentration. When SPIOs are used as carrier vehicle, additional advantages can be achieved including magnetic targeting and hyperthermia options, as well as monitoring with MRI. Other potential applications of SPIO include magnetofection and gene delivery, targeted retention of labeled stem cells, sentinel lymph nodes mapping, and magnetic force targeting and cell orientation for tissue engineering. PMID:23621536

CellMap visualizes protein-protein interactions and subcellular localization

PubMed Central

Dallago, Christian; Goldberg, Tatyana; Andrade-Navarro, Miguel Angel; Alanis-Lobato, Gregorio; Rost, Burkhard

2018-01-01

Many tools visualize protein-protein interaction (PPI) networks. The tool introduced here, CellMap, adds one crucial novelty by visualizing PPI networks in the context of subcellular localization, i.e. the location in the cell or cellular component in which a PPI happens. Users can upload images of cells and define areas of interest against which PPIs for selected proteins are displayed (by default on a cartoon of a cell). Annotations of localization are provided by the user or through our in-house database. The visualizer and server are written in JavaScript, making CellMap easy to customize and to extend by researchers and developers. PMID:29497493

Non-imaged based method for matching brains in a common anatomical space for cellular imagery.

PubMed

Midroit, Maëllie; Thevenet, Marc; Fournel, Arnaud; Sacquet, Joelle; Bensafi, Moustafa; Breton, Marine; Chalençon, Laura; Cavelius, Matthias; Didier, Anne; Mandairon, Nathalie

2018-04-22

Cellular imagery using histology sections is one of the most common techniques used in Neuroscience. However, this inescapable technique has severe limitations due to the need to delineate regions of interest on each brain, which is time consuming and variable across experimenters. We developed algorithms based on a vectors field elastic registration allowing fast, automatic realignment of experimental brain sections and associated labeling in a brain atlas with high accuracy and in a streamlined way. Thereby, brain areas of interest can be finely identified without outlining them and different experimental groups can be easily analyzed using conventional tools. This method directly readjusts labeling in the brain atlas without any intermediate manipulation of images. We mapped the expression of cFos, in the mouse brain (C57Bl/6J) after olfactory stimulation or a non-stimulated control condition and found an increased density of cFos-positive cells in the primary olfactory cortex but not in non-olfactory areas of the odor-stimulated animals compared to the controls. Existing methods of matching are based on image registration which often requires expensive material (two-photon tomography mapping or imaging with iDISCO) or are less accurate since they are based on mutual information contained in the images. Our new method is non-imaged based and relies only on the positions of detected labeling and the external contours of sections. We thus provide a new method that permits automated matching of histology sections of experimental brains with a brain reference atlas. Copyright © 2018 Elsevier B.V. All rights reserved.

Statistical parametric mapping of stimuli-evoked changes in quantitative blood flow using extended-focus optical coherence microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Marchand, Paul J.; Bouwens, Arno; Shamaei, Vincent; Nguyen, David; Extermann, Jerome; Bolmont, Tristan; Lasser, Theo

2016-03-01

Magnetic Resonance Imaging has revolutionised our understanding of brain function through its ability to image human cerebral structures non-invasively over the entire brain. By exploiting the different magnetic properties of oxygenated and deoxygenated blood, functional MRI can indirectly map areas undergoing neural activation. Alongside the development of fMRI, powerful statistical tools have been developed in an effort to shed light on the neural pathways involved in processing of sensory and cognitive information. In spite of the major improvements made in fMRI technology, the obtained spatial resolution of hundreds of microns prevents MRI in resolving and monitoring processes occurring at the cellular level. In this regard, Optical Coherence Microscopy is an ideal instrumentation as it can image at high spatio-temporal resolution. Moreover, by measuring the mean and the width of the Doppler spectra of light scattered by moving particles, OCM allows extracting the axial and lateral velocity components of red blood cells. The ability to assess quantitatively total blood velocity, as opposed to classical axial velocity Doppler OCM, is of paramount importance in brain imaging as a large proportion of cortical vascular is oriented perpendicularly to the optical axis. We combine here quantitative blood flow imaging with extended-focus Optical Coherence Microscopy and Statistical Parametric Mapping tools to generate maps of stimuli-evoked cortical hemodynamics at the capillary level.

Mapping cell-specific functional connections in the mouse brain using ChR2-evoked hemodynamics (Conference Presentation)

NASA Astrophysics Data System (ADS)

Bauer, Adam Q.; Kraft, Andrew; Baxter, Grant A.; Bruchas, Michael; Lee, Jin-Moo; Culver, Joseph P.

2017-02-01

Functional magnetic resonance imaging (fMRI) has transformed our understanding of the brain's functional organization. However, mapping subunits of a functional network using hemoglobin alone presents several disadvantages. Evoked and spontaneous hemodynamic fluctuations reflect ensemble activity from several populations of neurons making it difficult to discern excitatory vs inhibitory network activity. Still, blood-based methods of brain mapping remain powerful because hemoglobin provides endogenous contrast in all mammalian brains. To add greater specificity to hemoglobin assays, we integrated optical intrinsic signal(OIS) imaging with optogenetic stimulation to create an Opto-OIS mapping tool that combines the cell-specificity of optogenetics with label-free, hemoglobin imaging. Before mapping, titrated photostimuli determined which stimulus parameters elicited linear hemodynamic responses in the cortex. Optimized stimuli were then scanned over the left hemisphere to create a set of optogenetically-defined effective connectivity (Opto-EC) maps. For many sites investigated, Opto-EC maps exhibited higher spatial specificity than those determined using spontaneous hemodynamic fluctuations. For example, resting-state functional connectivity (RS-FC) patterns exhibited widespread ipsilateral connectivity while Opto-EC maps contained distinct short- and long-range constellations of ipsilateral connectivity. Further, RS-FC maps were usually symmetric about midline while Opto-EC maps displayed more heterogeneous contralateral homotopic connectivity. Both Opto-EC and RS-FC patterns were compared to mouse connectivity data from the Allen Institute. Unlike RS-FC maps, Thy1-based maps collected in awake, behaving mice closely recapitulated the connectivity structure derived using ex vivo anatomical tracer methods. Opto-OIS mapping could be a powerful tool for understanding cellular and molecular contributions to network dynamics and processing in the mouse brain.

Aggregation of gold nanoparticles followed by methotrexate release enables Raman imaging of drug delivery into cancer cells

NASA Astrophysics Data System (ADS)

Durgadas, C. V.; Sharma, C. P.; Paul, W.; Rekha, M. R.; Sreenivasan, K.

2012-09-01

This study refers an aqueous synthesis of methotrexate (MTX)-conjugated gold nanoparticles (GNPs), their interaction with HepG2 cells, and the use of Raman imaging to observe cellular internalization and drug delivery. GNPs of average size 3.5-5 nm were stabilized using the amine terminated bifunctional biocompatible copolymer and amended by conjugating MTX, an anticancer drug. The nanoparticles were released MTX at a faster rate in acidic pH and subsequently found to form aggregates. The Raman signals of cellular components were found to be enhanced by the aggregated particles enabling the mapping to visualize site-specific drug delivery. The methodology seems to have potential in optimizing the characteristics of nanodrug carriers for emptying the cargo precisely at specified sites.

Fluorescence Molecular Tomography: Principles and Potential for Pharmaceutical Research

PubMed Central

Stuker, Florian; Ripoll, Jorge; Rudin, Markus

2011-01-01

Fluorescence microscopic imaging is widely used in biomedical research to study molecular and cellular processes in cell culture or tissue samples. This is motivated by the high inherent sensitivity of fluorescence techniques, the spatial resolution that compares favorably with cellular dimensions, the stability of the fluorescent labels used and the sophisticated labeling strategies that have been developed for selectively labeling target molecules. More recently, two and three-dimensional optical imaging methods have also been applied to monitor biological processes in intact biological organisms such as animals or even humans. These whole body optical imaging approaches have to cope with the fact that biological tissue is a highly scattering and absorbing medium. As a consequence, light propagation in tissue is well described by a diffusion approximation and accurate reconstruction of spatial information is demanding. While in vivo optical imaging is a highly sensitive method, the signal is strongly surface weighted, i.e., the signal detected from the same light source will become weaker the deeper it is embedded in tissue, and strongly depends on the optical properties of the surrounding tissue. Derivation of quantitative information, therefore, requires tomographic techniques such as fluorescence molecular tomography (FMT), which maps the three-dimensional distribution of a fluorescent probe or protein concentration. The combination of FMT with a structural imaging method such as X-ray computed tomography (CT) or Magnetic Resonance Imaging (MRI) will allow mapping molecular information on a high definition anatomical reference and enable the use of prior information on tissue's optical properties to enhance both resolution and sensitivity. Today many of the fluorescent assays originally developed for studies in cellular systems have been successfully translated for experimental studies in animals. The opportunity of monitoring molecular processes non-invasively in the intact organism is highly attractive from a diagnostic point of view but even more so for the drug developer, who can use the techniques for proof-of-mechanism and proof-of-efficacy studies. This review shall elucidate the current status and potential of fluorescence tomography including recent advances in multimodality imaging approaches for preclinical and clinical drug development. PMID:24310495

Cellular resolution circuit mapping with temporal-focused excitation of soma-targeted channelrhodopsin

PubMed Central

Baker, Christopher A; Elyada, Yishai M; Parra, Andres; Bolton, M McLean

2016-01-01

We describe refinements in optogenetic methods for circuit mapping that enable measurements of functional synaptic connectivity with single-neuron resolution. By expanding a two-photon beam in the imaging plane using the temporal focusing method and restricting channelrhodopsin to the soma and proximal dendrites, we are able to reliably evoke action potentials in individual neurons, verify spike generation with GCaMP6s, and determine the presence or absence of synaptic connections with patch-clamp electrophysiological recording. DOI: http://dx.doi.org/10.7554/eLife.14193.001 PMID:27525487

Super-Resolution Imaging of Molecular Emission Spectra and Single Molecule Spectral Fluctuations

PubMed Central

Mlodzianoski, Michael J.; Curthoys, Nikki M.; Gunewardene, Mudalige S.; Carter, Sean; Hess, Samuel T.

2016-01-01

Localization microscopy can image nanoscale cellular details. To address biological questions, the ability to distinguish multiple molecular species simultaneously is invaluable. Here, we present a new version of fluorescence photoactivation localization microscopy (FPALM) which detects the emission spectrum of each localized molecule, and can quantify changes in emission spectrum of individual molecules over time. This information can allow for a dramatic increase in the number of different species simultaneously imaged in a sample, and can create super-resolution maps showing how single molecule emission spectra vary with position and time in a sample. PMID:27002724

Processing the Bouguer anomaly map of Biga and the surrounding area by the cellular neural network: application to the southwestern Marmara region

NASA Astrophysics Data System (ADS)

Aydogan, D.

2007-04-01

An image processing technique called the cellular neural network (CNN) approach is used in this study to locate geological features giving rise to gravity anomalies such as faults or the boundary of two geologic zones. CNN is a stochastic image processing technique based on template optimization using the neighborhood relationships of cells. These cells can be characterized by a functional block diagram that is typical of neural network theory. The functionality of CNN is described in its entirety by a number of small matrices (A, B and I) called the cloning template. CNN can also be considered to be a nonlinear convolution of these matrices. This template describes the strength of the nearest neighbor interconnections in the network. The recurrent perceptron learning algorithm (RPLA) is used in optimization of cloning template. The CNN and standard Canny algorithms were first tested on two sets of synthetic gravity data with the aim of checking the reliability of the proposed approach. The CNN method was compared with classical derivative techniques by applying the cross-correlation method (CC) to the same anomaly map as this latter approach can detect some features that are difficult to identify on the Bouguer anomaly maps. This approach was then applied to the Bouguer anomaly map of Biga and its surrounding area, in Turkey. Structural features in the area between Bandirma, Biga, Yenice and Gonen in the southwest Marmara region are investigated by applying the CNN and CC to the Bouguer anomaly map. Faults identified by these algorithms are generally in accordance with previously mapped surface faults. These examples show that the geologic boundaries can be detected from Bouguer anomaly maps using the cloning template approach. A visual evaluation of the outputs of the CNN and CC approaches is carried out, and the results are compared with each other. This approach provides quantitative solutions based on just a few assumptions, which makes the method more powerful than the classical methods.

In vivo features of melanocytic lesions: multimode hyperspectral dermoscopy, reflectance confocal microscopy, and histopathologic correlates (Conference Presentation)

NASA Astrophysics Data System (ADS)

Vasefi, Fartash; MacKinnon, Nicholas B.; Jain, Manu; Cordova, Miguel A.; Kose, Kivanc; Rajadhyaksha, Milind; Halpern, Allan C.; Farkas, Daniel L.

2017-02-01

Motivation and background: Melanoma, the fastest growing cancer worldwide, kills more than one person every hour in the United States. Determining the depth and distribution of dermal melanin and hemoglobin adds physio-morphologic information to the current diagnostic standard, cellular morphology, to further develop noninvasive methods to discriminate between melanoma and benign skin conditions. Purpose: To compare the performance of a multimode dermoscopy system (SkinSpect), which is designed to quantify and map in three dimensions, in vivo melanin and hemoglobin in skin, and to validate this with histopathology and three dimensional reflectance confocal microscopy (RCM) imaging. Methods: Sequentially capture SkinSpect and RCM images of suspect lesions and nearby normal skin and compare this with histopathology reports, RCM imaging allows noninvasive observation of nuclear, cellular and structural detail in 1-5 μm-thin optical sections in skin, and detection of pigmented skin lesions with sensitivity of 90-95% and specificity of 70-80%. The multimode imaging dermoscope combines polarization (cross and parallel), autofluorescence and hyperspectral imaging to noninvasively map the distribution of melanin, collagen and hemoglobin oxygenation in pigmented skin lesions. Results: We compared in vivo features of ten melanocytic lesions extracted by SkinSpect and RCM imaging, and correlated them to histopathologic results. We present results of two melanoma cases (in situ and invasive), and compare with in vivo features from eight benign lesions. Melanin distribution at different depths and hemodynamics, including abnormal vascularity, detected by both SkinSpect and RCM will be discussed. Conclusion: Diagnostic features such as dermal melanin and hemoglobin concentration provided in SkinSpect skin analysis for melanoma and normal pigmented lesions can be compared and validated using results from RCM and histopathology.

Growth studies at bulk III-Vs by image processing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Donecker, J.; Hempel, G.; Kluge, J.

1996-12-01

The patterns of inhomogeneities in GaAs and InP are studied by scattering and diffraction of light. An adapted version of laser scattering tomography is used for observations with short exposure times and large fields. The information about the three-dimensional distribution of the scatterers in GaAs are evaluated by video travels through the crystal and images of intensities added in interesting directions. Near-infrared transmission and striation distance mapping act like special data compression techniques due to their optical principles. In general, columnar extension of cellular patterns and striations could not be detected in s.i. GaAs. Long-range correlations exist for lineages andmore » slip lines. The comparison with the behavior of striations in doped InP cannot confirm the idea that cellular patterns in GaAs originate from constitutional supercooling during solidification.« less

Noninvasive 3D Visualization of Defects and Crack Propagation in Layered Foam Structures by Phase Contrast Microimaging

NASA Technical Reports Server (NTRS)

Hu, Z. W.; DeCarlo, F.

2006-01-01

Applications of polymeric foams in our modern society continue to grow because of their light weight, high strength, excellent thermal and mechanical insulation, and the ease of engineering. Among others, closed-cell foam has been structurally used for thermally insulating the shuttle external tank. However, internal defects of the foams were difficult to observe non-invasively due to limited sensitivity to the low-density structures possessed by traditional imaging tools such as computed X-ray tomography By combining phase contrast X-ray imaging with pressure loading, we succeeded in precisely mapping intact cellular structure and defects inside the bulk of layered foam and visualizing its subsequent response to the pressure in three-dimensional space. The work demonstrated a powerfir1 approach for yielding insight into underlying problems in lightweight cellular materials otherwise unobtainable.

A "distorted-BODIPY"-based fluorescent probe for imaging of cellular viscosity in live cells.

PubMed

Zhu, Hao; Fan, Jiangli; Li, Miao; Cao, Jianfang; Wang, Jingyun; Peng, Xiaojun

2014-04-14

Cellular viscosity is a critical factor in governing diffusion-mediated cellular processes and is linked to a number of diseases and pathologies. Fluorescent molecular rotors (FMRs) have recently been developed to determine viscosity in solutions or biological fluid. Herein, we report a "distorted-BODIPY"-based probe BV-1 for cellular viscosity, which is different from the conventional "pure rotors". In BV-1, the internal steric hindrance between the meso-CHO group and the 1,7-dimethyl group forced the boron-dipyrrin framework to be distorted, which mainly caused nonradiative deactivation in low-viscosity environment. BV-1 gave high sensitivity (x=0.62) together with stringent selectivity to viscosity, thus enabling viscosity mapping in live cells. Significantly, the increase of cytoplasmic viscosity during apoptosis was observed by BV-1 in real time. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Optical toolkits for in vivo deep tissue laser scanning microscopy: a primer

NASA Astrophysics Data System (ADS)

Lee, Woei Ming; McMenamin, Thomas; Li, Yongxiao

2018-06-01

Life at the microscale is animated and multifaceted. The impact of dynamic in vivo microscopy in small animals has opened up opportunities to peer into a multitude of biological processes at the cellular scale in their native microenvironments. Laser scanning microscopy (LSM) coupled with targeted fluorescent proteins has become an indispensable tool to enable dynamic imaging in vivo at high temporal and spatial resolutions. In the last few decades, the technique has been translated from imaging cells in thin samples to mapping cells in the thick biological tissue of living organisms. Here, we sought to provide a concise overview of the design considerations of a LSM that enables cellular and subcellular imaging in deep tissue. Individual components under review include: long working distance microscope objectives, laser scanning technologies, adaptive optics devices, beam shaping technologies and photon detectors, with an emphasis on more recent advances. The review will conclude with the latest innovations in automated optical microscopy, which would impact tracking and quantification of heterogeneous populations of cells in vivo.

High-throughput dual-colour precision imaging for brain-wide connectome with cytoarchitectonic landmarks at the cellular level

PubMed Central

Gong, Hui; Xu, Dongli; Yuan, Jing; Li, Xiangning; Guo, Congdi; Peng, Jie; Li, Yuxin; Schwarz, Lindsay A.; Li, Anan; Hu, Bihe; Xiong, Benyi; Sun, Qingtao; Zhang, Yalun; Liu, Jiepeng; Zhong, Qiuyuan; Xu, Tonghui; Zeng, Shaoqun; Luo, Qingming

2016-01-01

The precise annotation and accurate identification of neural structures are prerequisites for studying mammalian brain function. The orientation of neurons and neural circuits is usually determined by mapping brain images to coarse axial-sampling planar reference atlases. However, individual differences at the cellular level likely lead to position errors and an inability to orient neural projections at single-cell resolution. Here, we present a high-throughput precision imaging method that can acquire a co-localized brain-wide data set of both fluorescent-labelled neurons and counterstained cell bodies at a voxel size of 0.32 × 0.32 × 2.0 μm in 3 days for a single mouse brain. We acquire mouse whole-brain imaging data sets of multiple types of neurons and projections with anatomical annotation at single-neuron resolution. The results show that the simultaneous acquisition of labelled neural structures and cytoarchitecture reference in the same brain greatly facilitates precise tracing of long-range projections and accurate locating of nuclei. PMID:27374071

A Multi-Modality CMOS Sensor Array for Cell-Based Assay and Drug Screening.

PubMed

Chi, Taiyun; Park, Jong Seok; Butts, Jessica C; Hookway, Tracy A; Su, Amy; Zhu, Chengjie; Styczynski, Mark P; McDevitt, Todd C; Wang, Hua

2015-12-01

In this paper, we present a fully integrated multi-modality CMOS cellular sensor array with four sensing modalities to characterize different cell physiological responses, including extracellular voltage recording, cellular impedance mapping, optical detection with shadow imaging and bioluminescence sensing, and thermal monitoring. The sensor array consists of nine parallel pixel groups and nine corresponding signal conditioning blocks. Each pixel group comprises one temperature sensor and 16 tri-modality sensor pixels, while each tri-modality sensor pixel can be independently configured for extracellular voltage recording, cellular impedance measurement (voltage excitation/current sensing), and optical detection. This sensor array supports multi-modality cellular sensing at the pixel level, which enables holistic cell characterization and joint-modality physiological monitoring on the same cellular sample with a pixel resolution of 80 μm × 100 μm. Comprehensive biological experiments with different living cell samples demonstrate the functionality and benefit of the proposed multi-modality sensing in cell-based assay and drug screening.

3D Time-lapse Imaging and Quantification of Mitochondrial Dynamics

NASA Astrophysics Data System (ADS)

Sison, Miguel; Chakrabortty, Sabyasachi; Extermann, Jérôme; Nahas, Amir; James Marchand, Paul; Lopez, Antonio; Weil, Tanja; Lasser, Theo

2017-02-01

We present a 3D time-lapse imaging method for monitoring mitochondrial dynamics in living HeLa cells based on photothermal optical coherence microscopy and using novel surface functionalization of gold nanoparticles. The biocompatible protein-based biopolymer coating contains multiple functional groups which impart better cellular uptake and mitochondria targeting efficiency. The high stability of the gold nanoparticles allows continuous imaging over an extended time up to 3000 seconds without significant cell damage. By combining temporal autocorrelation analysis with a classical diffusion model, we quantify mitochondrial dynamics and cast these results into 3D maps showing the heterogeneity of diffusion parameters across the whole cell volume.

Object detection via eye tracking and fringe restraint

NASA Astrophysics Data System (ADS)

Pan, Fei; Zhang, Hanming; Zeng, Ying; Tong, Li; Yan, Bin

2017-07-01

Object detection is a computer vision problem which caught a large amount of attention. But the candidate boundingboxes extracted from only image features may end up with false-detection due to the semantic gap between the top-down and the bottom up information. In this paper, we propose a novel method for generating object bounding-boxes proposals using the combination of eye fixation point, saliency detection and edges. The new method obtains a fixation orientated Gaussian map, optimizes the map through single-layer cellular automata, and derives bounding-boxes from the optimized map on three levels. Then we score the boxes by combining all the information above, and choose the box with the highest score to be the final box. We perform an evaluation of our method by comparing with previous state-ofthe art approaches on the challenging POET datasets, the images of which are chosen from PASCAL VOC 2012. Our method outperforms them on small scale objects while comparable to them in general.

Molecular imaging and sensing using plasmonic nanoparticles

NASA Astrophysics Data System (ADS)

Crow, Matthew James

Noble metal nanoparticles exhibit unique optical properties that are beneficial to a variety of applications, including molecular imaging. The large scattering cross sections of nanoparticles provide high contrast necessary for biomarkers. Unlike alternative contrast agents, nanoparticles provide refractive index sensitivity revealing information regarding the local cellular environment. Altering the shape and composition of the nanoparticle shifts the peak resonant wavelength of scattered light, allowing for implementation of multiple spectrally distinct tags. In this project, nanoparticles that scatter in different spectral windows are functionalized with various antibodies recognizing extra-cellular receptors integral to cancer progression. A hyperspectral imaging system is developed, allowing for visualization and spectral characterization of cells labeled with these conjugates. Various molecular imaging and microspectroscopy applications of plasmonic nanoparticles are then investigated. First, anti-EGFR gold nanospheres are shown to quantitatively measure receptor expression with similar performance to fluorescence assays. Second, anti-EGFR gold nanorods and novel anti-IGF-1R silver nanospheres are implemented to indicate local cellular refractive indices. Third, because biosensing capabilities of nanoparticle tags may be limited by plasmonic coupling, polarization mapping is investigated as a method to discern these effects. Fourth, plasmonic coupling is tested to monitor HER-2 dimerization. Experiments reveal the interparticle conformation of proximal HER-2 bound labels, required for plasmonic coupling-enhanced dielectric sensing. Fifth, all three functionalized plasmonic tags are implemented simultaneously to indicate clinically relevant cell immunophenotype information and changes in the cellular dielectric environment. Finally, flow cytometry experiments are conducted utilizing the anti-EGFR nanorod tag to demonstrate profiling of receptor expression distribution and potential increased multiplexing capability.

T1 Mapping by Cardiac Magnetic Resonance and Multidimensional Speckle-Tracking Strain by Echocardiography for the Detection of Acute Cellular Rejection in Cardiac Allograft Recipients.

PubMed

Sade, Leyla Elif; Hazirolan, Tuncay; Kozan, Hatice; Ozdemir, Handan; Hayran, Mutlu; Eroglu, Serpil; Pirat, Bahar; Sezgin, Atilla; Muderrisoglu, Haldun

2018-04-14

The aim of this study was to test the hypothesis that echocardiographic strain imaging, by tracking subtle alterations in myocardial function, and cardiac magnetic resonance T1 mapping, by quantifying tissue properties, are useful and complement each other to detect acute cellular rejection in heart transplant recipients. Noninvasive alternatives to endomyocardial biopsy are highly desirable to monitor acute cellular rejection. Surveillance endomyocardial biopsies, catheterizations, and echocardiograms performed serially according to institutional protocol since transplantation were retrospectively reviewed. Sixteen-segment global longitudinal strain (GLS) and circumferential strain were measured before, during, and after the first rejection and at 2 time points for patients without rejection using Velocity Vector Imaging for the first part of the study. The second part, with cardiac magnetic resonance added to the protocol, served to validate previously derived strain cutoffs, examine the progression of strain over time, and to determine the accuracy of strain and T1 measurements to define acute cellular rejection. All tests were performed within 48 h. Median time to first rejection (16 grade 1 rejection, 15 grade ≥2 rejection) was 3 months (interquartile range: 3 to 36 months) in 49 patients. GLS and global circumferential strain worsened significantly during grade 1 rejection and ≥2 rejection and were independent predictors of any rejection. In the second part of the study, T1 time ≥1,090 ms, extracellular volume ≥32%, GLS >-14%, and global circumferential strain ≥-24% had 100% sensitivity and 100% negative predictive value to define grade ≥2 rejection with 70%, 63%, 55%, and 35% positive predictive values, respectively. The combination of GLS >-16% and T1 time ≥1,060 ms defined grade 1 rejection with 91% sensitivity and 92% negative predictive value. After successful treatment, T1 times decreased significantly. T1 mapping and echocardiographic GLS can serve to guide endomyocardial biopsy selectively. Copyright © 2018 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

Breast MR segmentation and lesion detection with cellular neural networks and 3D template matching.

PubMed

Ertaş, Gökhan; Gülçür, H Ozcan; Osman, Onur; Uçan, Osman N; Tunaci, Mehtap; Dursun, Memduh

2008-01-01

A novel fully automated system is introduced to facilitate lesion detection in dynamic contrast-enhanced, magnetic resonance mammography (DCE-MRM). The system extracts breast regions from pre-contrast images using a cellular neural network, generates normalized maximum intensity-time ratio (nMITR) maps and performs 3D template matching with three layers of 12x12 cells to detect lesions. A breast is considered to be properly segmented when relative overlap >0.85 and misclassification rate <0.10. Sensitivity, false-positive rate per slice and per lesion are used to assess detection performance. The system was tested with a dataset of 2064 breast MR images (344slicesx6 acquisitions over time) from 19 women containing 39 marked lesions. Ninety-seven percent of the breasts were segmented properly and all the lesions were detected correctly (detection sensitivity=100%), however, there were some false-positive detections (31%/lesion, 10%/slice).

Physics, Techniques and Review of Neuroradiological Applications of Diffusion Kurtosis Imaging (DKI).

PubMed

Marrale, M; Collura, G; Brai, M; Toschi, N; Midiri, F; La Tona, G; Lo Casto, A; Gagliardo, C

2016-12-01

In recent years many papers about diagnostic applications of diffusion tensor imaging (DTI) have been published. This is because DTI allows to evaluate in vivo and in a non-invasive way the process of diffusion of water molecules in biological tissues. However, the simplified description of the diffusion process assumed in DTI does not permit to completely map the complex underlying cellular components and structures, which hinder and restrict the diffusion of water molecules. These limitations can be partially overcome by means of diffusion kurtosis imaging (DKI). The aim of this paper is the description of the theory of DKI, a new topic of growing interest in radiology. DKI is a higher order diffusion model that is a straightforward extension of the DTI model. Here, we analyze the physics underlying this method, we report our MRI acquisition protocol with the preprocessing pipeline used and the DKI parametric maps obtained on a 1.5 T scanner, and we review the most relevant clinical applications of this technique in various neurological diseases.

Live Cell Imaging of Viscosity in 3D Tumour Cell Models.

PubMed

Shirmanova, Marina V; Shimolina, Lubov' E; Lukina, Maria M; Zagaynova, Elena V; Kuimova, Marina K

2017-01-01

Abnormal levels of viscosity in tissues and cells are known to be associated with disease and malfunction. While methods to measure bulk macroscopic viscosity of bio-tissues are well developed, imaging viscosity at the microscopic scale remains a challenge, especially in vivo. Molecular rotors are small synthetic viscosity-sensitive fluorophores in which fluorescence parameters are strongly correlated to the microviscosity of their immediate environment. Hence, molecular rotors represent a promising instrument for mapping of viscosity in living cells and tissues at the microscopic level. Quantitative measurements of viscosity can be achieved by recording time-resolved fluorescence decays of molecular rotor using fluorescence lifetime imaging microscopy (FLIM), which is also suitable for dynamic viscosity mapping, both in cellulo and in vivo. Among tools of experimental oncology, 3D tumour cultures, or spheroids, are considered a more adequate in vitro model compared to a cellular monolayer, and represent a less labour-intensive and more unified approach compared to animal tumour models. This chapter describes a methodology for microviscosity imaging in tumour spheroids using BODIPY-based molecular rotors and two photon-excited FLIM.

EPS in Environmental Microbial Biofilms as Examined by Advanced Imaging Techniques

NASA Astrophysics Data System (ADS)

Neu, T. R.; Lawrence, J. R.

2006-12-01

Biofilm communities are highly structured associations of cellular and polymeric components which are involved in biogenic and geogenic environmental processes. Furthermore, biofilms are also important in medical (infection), industrial (biofouling) and technological (biofilm engineering) processes. The interfacial microbial communities in a specific habitat are highly dynamic and change according to the environmental parameters affecting not only the cellular but also the polymeric constituents of the system. Through their EPS biofilms interact with dissolved, colloidal and particulate compounds from the bulk water phase. For a long time the focus in biofilm research was on the cellular constituents in biofilms and the polymer matrix in biofilms has been rather neglected. The polymer matrix is produced not only by different bacteria and archaea but also by eukaryotic micro-organisms such as algae and fungi. The mostly unidentified mixture of EPS compounds is responsible for many biofilm properties and is involved in biofilm functionality. The chemistry of the EPS matrix represents a mixture of polymers including polysaccharides, proteins, nucleic acids, neutral polymers, charged polymers, amphiphilic polymers and refractory microbial polymers. The analysis of the EPS may be done destructively by means of extraction and subsequent chemical analysis or in situ by means of specific probes in combination with advanced imaging. In the last 15 years laser scanning microscopy (LSM) has been established as an indispensable technique for studying microbial communities. LSM with 1-photon and 2-photon excitation in combination with fluorescence techniques allows 3-dimensional investigation of fully hydrated, living biofilm systems. This approach is able to reveal data on biofilm structural features as well as biofilm processes and interactions. The fluorescent probes available allow the quantitative assessment of cellular as well as polymer distribution. For this purpose lectin-binding- analysis has been suggested as a suitable approach to image glycoconjugates within the polymer matrix of biofilm communities. More recently synchrotron radiation is increasingly recognized as a powerful tool for studying biological samples. Hard X-ray excitation can be used to map elemental composition whereas IR imaging allows examination of biological macromolecules. A further technique called soft X-ray scanning transmission microscopy (STXM) has the advantage of both techniques and may be employed to detect elements as well as biomolecules. Using the appropriate spectra, near edge X-ray absorption fine structure (NEXAFS) microscopy allows quantitative chemical mapping at 50 nm resolution. In this presentation the applicability of LSM and STXM will be demonstrated using several examples of different environmental biofilm systems. The techniques in combination provide a new view of complex microbial communities and their interaction with the environment. These advanced imaging techniques offer the possibility to study the spatial structure of cellular and polymeric compounds in biofilms as well as biofilm microhabitats, biofilm functionality and biofilm processes.

MRI-Derived Cellularity Index as a Potential Noninvasive Imaging Biomarker of Prostate Cancer

DTIC Science & Technology

2014-10-01

previously diagnosed with prostate cancer via standard transrectal ultrasound guided prostate biopsy after prostate specific antigen (PSA) elevation or...around 70% and specificity of 55% (72). Functional MR techniques enhance detection, grading, and staging of prostate cancer through the use of dynamic...and specificity in the diagnosis of prostate cancer by increasing tumor conspicuity on DWI or quantitative ADC maps. How- ever, hemorrhage, inflammatory

Direct optical mapping of transcription factor binding sites on field-stretched λ-DNA in nanofluidic devices

PubMed Central

Sriram, K. K.; Yeh, Jia-Wei; Lin, Yii-Lih; Chang, Yi-Ren; Chou, Chia-Fu

2014-01-01

Mapping transcription factor (TF) binding sites along a DNA backbone is crucial in understanding the regulatory circuits that control cellular processes. Here, we deployed a method adopting bioconjugation, nanofluidic confinement and fluorescence single molecule imaging for direct mapping of TF (RNA polymerase) binding sites on field-stretched single DNA molecules. Using this method, we have mapped out five of the TF binding sites of E. coli RNA polymerase to bacteriophage λ-DNA, where two promoter sites and three pseudo-promoter sites are identified with the corresponding binding frequency of 45% and 30%, respectively. Our method is quick, robust and capable of resolving protein-binding locations with high accuracy (∼ 300 bp), making our system a complementary platform to the methods currently practiced. It is advantageous in parallel analysis and less prone to false positive results over other single molecule mapping techniques such as optical tweezers, atomic force microscopy and molecular combing, and could potentially be extended to general mapping of protein–DNA interaction sites. PMID:24753422

Mapping of dendritic lesions in patients with herpes simplex keratitis using in vivo confocal microscopy

PubMed Central

Yokogawa, Hideaki; Kobayashi, Akira; Mori, Natsuko; Sugiyama, Kazuhisa

2015-01-01

Purpose To produce a two-dimensional reconstruction map of dendritic lesions in patients with herpes simplex keratitis (HSK) using in vivo confocal microscopy. Methods Four eyes of four patients (mean 65.8 years) with HSK presenting with a dendritic lesion were enrolled. Slit-lamp biomicroscopy and in vivo laser confocal microscopy were performed. Acquired confocal images at the level of the epithelium were arranged and mapped into subconfluent montages. Changes in the shape and degree of light reflection of abnormal cells and deposits around dendritic lesions as well as other corneal layers were qualitatively evaluated. Results Mapping of dendritic lesion was successful in all cases, and the subconfluent montages clearly showed the larger image of dendritic lesion. In all cases, the dendritic lesion consisted of hyperreflective irregular epithelial cells, and was surrounded by distorted and elongated epithelial cells. In three cases, hyperreflective deposits were noted at the midline of the lesion. The corneal stroma showed a hyperreflective honeycomb pattern. In two cases, inflammatory cells were observed at the level of endothelial cell layer. Conclusion Mapping of dendritic lesions in patients with HSK was successful in all patients using in vivo confocal microscopy. Cellular level observation of dendritic lesion at a relatively larger magnification may help understand the in vivo morphological change of HSK. Further study in more patients with HSK and nonherpetic dendritic lesion is needed to utilize confocal microscopy images in differential diagnosis and follow-up of the epithelial lesions with dendrite. PMID:26445524

Mapping of dendritic lesions in patients with herpes simplex keratitis using in vivo confocal microscopy.

PubMed

Yokogawa, Hideaki; Kobayashi, Akira; Mori, Natsuko; Sugiyama, Kazuhisa

2015-01-01

To produce a two-dimensional reconstruction map of dendritic lesions in patients with herpes simplex keratitis (HSK) using in vivo confocal microscopy. Four eyes of four patients (mean 65.8 years) with HSK presenting with a dendritic lesion were enrolled. Slit-lamp biomicroscopy and in vivo laser confocal microscopy were performed. Acquired confocal images at the level of the epithelium were arranged and mapped into subconfluent montages. Changes in the shape and degree of light reflection of abnormal cells and deposits around dendritic lesions as well as other corneal layers were qualitatively evaluated. Mapping of dendritic lesion was successful in all cases, and the subconfluent montages clearly showed the larger image of dendritic lesion. In all cases, the dendritic lesion consisted of hyperreflective irregular epithelial cells, and was surrounded by distorted and elongated epithelial cells. In three cases, hyperreflective deposits were noted at the midline of the lesion. The corneal stroma showed a hyperreflective honeycomb pattern. In two cases, inflammatory cells were observed at the level of endothelial cell layer. Mapping of dendritic lesions in patients with HSK was successful in all patients using in vivo confocal microscopy. Cellular level observation of dendritic lesion at a relatively larger magnification may help understand the in vivo morphological change of HSK. Further study in more patients with HSK and nonherpetic dendritic lesion is needed to utilize confocal microscopy images in differential diagnosis and follow-up of the epithelial lesions with dendrite.

Color Swapping to Enhance Breast Cancer Digital Images Qualities Using Stain Normalization

NASA Astrophysics Data System (ADS)

Muhimmah, Izzati; Puspasari Wijaya, Dhina; Indrayanti

2017-03-01

Histopathology is the disease diagnosis by means of the visual examination of tissues under the microscope. The virtually transparent tissue sections were prepared using a number of colored histochemical stains bound selectively to the cellular components. A variation of colors comes to be a problem in histopathology based upon the microscope lighting for the range of factors. This research aimed to investigate an image enhancement by applying a nonlinear mapping approach to stain normalization and histogram equalization for contrast enhancement. Validation was carried out in 59 datasets with 96.6% accordance and expert justification.

An image based information system - Architecture for correlating satellite and topological data bases

NASA Technical Reports Server (NTRS)

Bryant, N. A.; Zobrist, A. L.

1978-01-01

The paper describes the development of an image based information system and its use to process a Landsat thematic map showing land use or land cover in conjunction with a census tract polygon file to produce a tabulation of land use acreages per census tract. The system permits the efficient cross-tabulation of two or more geo-coded data sets, thereby setting the stage for the practical implementation of models of diffusion processes or cellular transformation. Characteristics of geographic information systems are considered, and functional requirements, such as data management, geocoding, image data management, and data analysis are discussed. The system is described, and the potentialities of its use are examined.

A 3D Image Filter for Parameter-Free Segmentation of Macromolecular Structures from Electron Tomograms

PubMed Central

Ali, Rubbiya A.; Landsberg, Michael J.; Knauth, Emily; Morgan, Garry P.; Marsh, Brad J.; Hankamer, Ben

2012-01-01

3D image reconstruction of large cellular volumes by electron tomography (ET) at high (≤5 nm) resolution can now routinely resolve organellar and compartmental membrane structures, protein coats, cytoskeletal filaments, and macromolecules. However, current image analysis methods for identifying in situ macromolecular structures within the crowded 3D ultrastructural landscape of a cell remain labor-intensive, time-consuming, and prone to user-bias and/or error. This paper demonstrates the development and application of a parameter-free, 3D implementation of the bilateral edge-detection (BLE) algorithm for the rapid and accurate segmentation of cellular tomograms. The performance of the 3D BLE filter has been tested on a range of synthetic and real biological data sets and validated against current leading filters—the pseudo 3D recursive and Canny filters. The performance of the 3D BLE filter was found to be comparable to or better than that of both the 3D recursive and Canny filters while offering the significant advantage that it requires no parameter input or optimisation. Edge widths as little as 2 pixels are reproducibly detected with signal intensity and grey scale values as low as 0.72% above the mean of the background noise. The 3D BLE thus provides an efficient method for the automated segmentation of complex cellular structures across multiple scales for further downstream processing, such as cellular annotation and sub-tomogram averaging, and provides a valuable tool for the accurate and high-throughput identification and annotation of 3D structural complexity at the subcellular level, as well as for mapping the spatial and temporal rearrangement of macromolecular assemblies in situ within cellular tomograms. PMID:22479430

Time-resolved multicolor two-photon excitation fluorescence microscopy of cells and tissues

NASA Astrophysics Data System (ADS)

Zheng, Wei

2014-11-01

Multilabeling which maps the distribution of different targets is an indispensable technique in many biochemical and biophysical studies. Two-photon excitation fluorescence (TPEF) microscopy of endogenous fluorophores combining with conventional fluorescence labeling techniques such as genetically encoded fluorescent protein (FP) and fluorescent dyes staining could be a powerful tool for imaging living cells. However, the challenge is that the excitation and emission wavelength of these endogenous fluorophores and fluorescent labels are very different. A multi-color ultrafast source is required for the excitation of multiple fluorescence molecules. In this study, we developed a two-photon imaging system with excitations from the pump femtosecond laser and the selected supercontinuum generated from a photonic crystal fiber (PCF). Multiple endogenous fluorophores, fluorescent proteins and fluorescent dyes were excited in their optimal wavelengths simultaneously. A time- and spectral-resolved detection system was used to record the TPEF signals. This detection technique separated the TPEF signals from multiple sources in time and wavelength domains. Cellular organelles such as nucleus, mitochondria, microtubule and endoplasmic reticulum, were clearly revealed in the TPEF images. The simultaneous imaging of multiple fluorophores of cells will greatly aid the study of sub-cellular compartments and protein localization.

Rapid 3D Refractive‐Index Imaging of Live Cells in Suspension without Labeling Using Dielectrophoretic Cell Rotation

PubMed Central

Habaza, Mor; Kirschbaum, Michael; Guernth‐Marschner, Christian; Dardikman, Gili; Barnea, Itay; Korenstein, Rafi; Duschl, Claus

2016-01-01

A major challenge in the field of optical imaging of live cells is achieving rapid, 3D, and noninvasive imaging of isolated cells without labeling. If successful, many clinical procedures involving analysis and sorting of cells drawn from body fluids, including blood, can be significantly improved. A new label‐free tomographic interferometry approach is presented. This approach provides rapid capturing of the 3D refractive‐index distribution of single cells in suspension. The cells flow in a microfluidic channel, are trapped, and then rapidly rotated by dielectrophoretic forces in a noninvasive and precise manner. Interferometric projections of the rotated cell are acquired and processed into the cellular 3D refractive‐index map. Uniquely, this approach provides full (360°) coverage of the rotation angular range around any axis, and knowledge on the viewing angle. The experimental demonstrations presented include 3D, label‐free imaging of cancer cells and three types of white blood cells. This approach is expected to be useful for label‐free cell sorting, as well as for detection and monitoring of pathological conditions resulting in cellular morphology changes or occurrence of specific cell types in blood or other body fluids. PMID:28251046

Probing the brain with molecular fMRI.

PubMed

Ghosh, Souparno; Harvey, Peter; Simon, Jacob C; Jasanoff, Alan

2018-06-01

One of the greatest challenges of modern neuroscience is to incorporate our growing knowledge of molecular and cellular-scale physiology into integrated, organismic-scale models of brain function in behavior and cognition. Molecular-level functional magnetic resonance imaging (molecular fMRI) is a new technology that can help bridge these scales by mapping defined microscopic phenomena over large, optically inaccessible regions of the living brain. In this review, we explain how MRI-detectable imaging probes can be used to sensitize noninvasive imaging to mechanistically significant components of neural processing. We discuss how a combination of innovative probe design, advanced imaging methods, and strategies for brain delivery can make molecular fMRI an increasingly successful approach for spatiotemporally resolved studies of diverse neural phenomena, perhaps eventually in people. Copyright © 2018 Elsevier Ltd. All rights reserved.

Simulating magnetic resonance images based on a model of tumor growth incorporating microenvironment

NASA Astrophysics Data System (ADS)

Jackson, Pamela R.; Hawkins-Daarud, Andrea; Partridge, Savannah C.; Kinahan, Paul E.; Swanson, Kristin R.

2018-03-01

Glioblastoma (GBM), the most aggressive primary brain tumor, is primarily diagnosed and monitored using gadoliniumenhanced T1-weighted and T2-weighted (T2W) magnetic resonance imaging (MRI). Hyperintensity on T2W images is understood to correspond with vasogenic edema and infiltrating tumor cells. GBM's inherent heterogeneity and resulting non-specific MRI image features complicate assessing treatment response. To better understand treatment response, we propose creating a patient-specific untreated virtual imaging control (UVIC), which represents an individual tumor's growth if it had not been treated, for comparison with actual post-treatment images. We generated a T2W MRI UVIC by combining a patient-specific mathematical model of tumor growth with a multi-compartmental MRI signal equation. GBM growth was mathematically modeled using the previously developed Proliferation-Invasion-Hypoxia-Necrosis- Angiogenesis-Edema (PIHNA-E) model, which simulated tumor as being comprised of three cellular phenotypes: normoxic, hypoxic and necrotic cells interacting with a vasculature species, angiogenic factors and extracellular fluid. Within the PIHNA-E model, both hypoxic and normoxic cells emitted angiogenic factors, which recruited additional vessels and caused the vessels to leak, allowing fluid, or edema, to escape into the extracellular space. The model's output was spatial volume fraction maps for each glioma cell type and edema/extracellular space. Volume fraction maps and corresponding T2 values were then incorporated into a multi-compartmental Bloch signal equation to create simulated T2W images. T2 values for individual compartments were estimated from the literature and a normal volunteer. T2 maps calculated from simulated images had normal white matter, normal gray matter, and tumor tissue T2 values within range of literature values.

Quantitative Susceptibility Mapping: Contrast Mechanisms and Clinical Applications

PubMed Central

Liu, Chunlei; Wei, Hongjiang; Gong, Nan-Jie; Cronin, Matthew; Dibb, Russel; Decker, Kyle

2016-01-01

Quantitative susceptibility mapping (QSM) is a recently developed MRI technique for quantifying the spatial distribution of magnetic susceptibility within biological tissues. It first uses the frequency shift in the MRI signal to map the magnetic field profile within the tissue. The resulting field map is then used to determine the spatial distribution of the underlying magnetic susceptibility by solving an inverse problem. The solution is achieved by deconvolving the field map with a dipole field, under the assumption that the magnetic field is a result of the superposition of the dipole fields generated by all voxels and that each voxel has its unique magnetic susceptibility. QSM provides improved contrast to noise ratio for certain tissues and structures compared to its magnitude counterpart. More importantly, magnetic susceptibility is a direct reflection of the molecular composition and cellular architecture of the tissue. Consequently, by quantifying magnetic susceptibility, QSM is becoming a quantitative imaging approach for characterizing normal and pathological tissue properties. This article reviews the mechanism generating susceptibility contrast within tissues and some associated applications. PMID:26844301

Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D)

PubMed Central

Li, Weizhe; Germain, Ronald N.

2017-01-01

Organ homeostasis, cellular differentiation, signal relay, and in situ function all depend on the spatial organization of cells in complex tissues. For this reason, comprehensive, high-resolution mapping of cell positioning, phenotypic identity, and functional state in the context of macroscale tissue structure is critical to a deeper understanding of diverse biological processes. Here we report an easy to use method, clearing-enhanced 3D (Ce3D), which generates excellent tissue transparency for most organs, preserves cellular morphology and protein fluorescence, and is robustly compatible with antibody-based immunolabeling. This enhanced signal quality and capacity for extensive probe multiplexing permits quantitative analysis of distinct, highly intermixed cell populations in intact Ce3D-treated tissues via 3D histo-cytometry. We use this technology to demonstrate large-volume, high-resolution microscopy of diverse cell types in lymphoid and nonlymphoid organs, as well as to perform quantitative analysis of the composition and tissue distribution of multiple cell populations in lymphoid tissues. Combined with histo-cytometry, Ce3D provides a comprehensive strategy for volumetric quantitative imaging and analysis that bridges the gap between conventional section imaging and disassociation-based techniques. PMID:28808033

Phase transitions in coupled map lattices and in associated probabilistic cellular automata.

PubMed

Just, Wolfram

2006-10-01

Analytical tools are applied to investigate piecewise linear coupled map lattices in terms of probabilistic cellular automata. The so-called disorder condition of probabilistic cellular automata is closely related with attracting sets in coupled map lattices. The importance of this condition for the suppression of phase transitions is illustrated by spatially one-dimensional systems. Invariant densities and temporal correlations are calculated explicitly. Ising type phase transitions are found for one-dimensional coupled map lattices acting on repelling sets and for a spatially two-dimensional Miller-Huse-like system with stable long time dynamics. Critical exponents are calculated within a finite size scaling approach. The relevance of detailed balance of the resulting probabilistic cellular automaton for the critical behavior is pointed out.

Left ventricular hypertrophy: The relationship between the electrocardiogram and cardiovascular magnetic resonance imaging.

PubMed

Bacharova, Ljuba; Ugander, Martin

2014-11-01

Conventional assessment of left ventricular hypertrophy (LVH) using the electrocardiogram (ECG), for example, by the Sokolow-Lyon, Romhilt-Estes or Cornell criteria, have relied on assessing changes in the amplitude and/or duration of the QRS complex of the ECG to quantify LV mass. ECG measures of LV mass have typically been validated by imaging with echocardiography or cardiovascular magnetic resonance imaging (CMR). However, LVH can be the result of diverse etiologies, and LVH is also characterized by pathological changes in myocardial tissue characteristics on the genetic, molecular, cellular, and tissue level beyond a pure increase in the number of otherwise normal cardiomyocytes. For example, slowed conduction velocity through the myocardium, which can be due to diffuse myocardial fibrosis, has been shown to be an important determinant of conventional ECG LVH criteria regardless of LV mass. Myocardial tissue characterization by CMR has emerged to not only quantify LV mass, but also detect and quantify the extent and severity of focal or diffuse myocardial fibrosis, edema, inflammation, myocarditis, fatty replacement, myocardial disarray, and myocardial deposition of amyloid proteins (amyloidosis), glycolipids (Fabry disease), or iron (siderosis). This can be undertaken using CMR techniques including late gadolinium enhancement (LGE), T1 mapping, T2 mapping, T2* mapping, extracellular volume fraction (ECV) mapping, fat/water-weighted imaging, and diffusion tensor CMR. This review presents an overview of current and emerging concepts regarding the diagnostic possibilities of both ECG and CMR for LVH in an attempt to narrow gaps in our knowledge regarding the ECG diagnosis of LVH. © 2014 Wiley Periodicals, Inc.

Hyperspectral imaging of endogenous fluorescent metabolic molecules to identify pain states in central nervous system tissue

NASA Astrophysics Data System (ADS)

Staikopoulos, Vasiliki; Gosnell, Martin E.; Anwer, Ayad G.; Mustafa, Sanam; Hutchinson, Mark R.; Goldys, Ewa M.

2016-12-01

Fluorescence-based bio-imaging methods have been extensively used to identify molecular changes occurring in biological samples in various pathological adaptations. Auto-fluorescence generated by endogenous fluorescent molecules within these samples can interfere with signal to background noise making positive antibody based fluorescent staining difficult to resolve. Hyperspectral imaging uses spectral and spatial imaging information for target detection and classification, and can be used to resolve changes in endogenous fluorescent molecules such as flavins, bound and free NADH and retinoids that are involved in cell metabolism. Hyperspectral auto-fluorescence imaging of spinal cord slices was used in this study to detect metabolic differences within pain processing regions of non-pain versus sciatic chronic constriction injury (CCI) animals, an established animal model of peripheral neuropathy. By using an endogenous source of contrast, subtle metabolic variations were detected between tissue samples, making it possible to distinguish between animals from non-injured and injured groups. Tissue maps of native fluorophores, flavins, bound and free NADH and retinoids unveiled subtle metabolic signatures and helped uncover significant tissue regions with compromised mitochondrial function. Taken together, our results demonstrate that hyperspectral imaging provides a new non-invasive method to investigate central changes of peripheral neuropathic injury and other neurodegenerative disease models, and paves the way for novel cellular characterisation in health, disease and during treatment, with proper account of intrinsic cellular heterogeneity.

Mapping of native inorganic elements and injected nanoparticles in a biological organ with laser-induced plasma

NASA Astrophysics Data System (ADS)

Motto-Ros, V.; Sancey, L.; Ma, Q. L.; Lux, F.; Bai, X. S.; Wang, X. C.; Yu, Jin; Panczer, G.; Tillement, O.

2012-11-01

Emission spectroscopy of laser-induced plasma from a thin section of mouse kidney successfully detected inorganic elements, Na, Ca, Cu, and Gd, naturally contained in the organ or artificially injected in the form of Gd-based nanoparticle. A two-dimensional scan of the sample allowed the laser beam to explore its surface with a resolution of 100 μm, resulting in a quantitative elemental mapping of the organ with sub-mM sensitivity. The compatibility of the setup with standard optical microscopy emphasizes the potential to provide multiple images of a same biological tissue with different types of response which can be elemental, molecular, or cellular.

Tomographic phase microscopy: principles and applications in bioimaging [Invited

PubMed Central

Jin, Di; Zhou, Renjie; Yaqoob, Zahid; So, Peter T. C.

2017-01-01

Tomographic phase microscopy (TPM) is an emerging optical microscopic technique for bioimaging. TPM uses digital holographic measurements of complex scattered fields to reconstruct three-dimensional refractive index (RI) maps of cells with diffraction-limited resolution by solving inverse scattering problems. In this paper, we review the developments of TPM from the fundamental physics to its applications in bioimaging. We first provide a comprehensive description of the tomographic reconstruction physical models used in TPM. The RI map reconstruction algorithms and various regularization methods are discussed. Selected TPM applications for cellular imaging, particularly in hematology, are reviewed. Finally, we examine the limitations of current TPM systems, propose future solutions, and envision promising directions in biomedical research. PMID:29386746

Lifespan Trajectories of White Matter Changes in Rhesus Monkeys.

PubMed

Kubicki, M; Baxi, M; Pasternak, O; Tang, Y; Karmacharya, S; Chunga, N; Lyall, A E; Rathi, Y; Eckbo, R; Bouix, S; Mortazavi, F; Papadimitriou, G; Shenton, M E; Westin, C F; Killiany, R; Makris, N; Rosene, D L

2018-04-26

Progress in neurodevelopmental brain research has been achieved through the use of animal models. Such models not only help understanding biological changes that govern brain development, maturation and aging, but are also essential for identifying possible mechanisms of neurodevelopmental and age-related chronic disorders, and to evaluate possible interventions with potential relevance to human disease. Genetic relationship of rhesus monkeys to humans makes those animals a great candidate for such models. With the typical lifespan of 25 years, they undergo cognitive maturation and aging that is similar to this observed in humans. Quantitative structural neuroimaging has been proposed as one of the candidate in vivo biomarkers for tracking white matter brain maturation and aging. While lifespan trajectories of white matter changes have been mapped in humans, such knowledge is not available for nonhuman primates. Here, we analyze and model lifespan trajectories of white matter microstructure using in vivo diffusion imaging in a sample of 44 rhesus monkeys. We report quantitative parameters (including slopes and peaks) of lifespan trajectories for 8 individual white matter tracts. We show different trajectories for cellular and extracellular microstructural imaging components that are associated with white matter maturation and aging, and discuss similarities and differences between those in humans and rhesus monkeys, the importance of our findings, and future directions for the field.Significance Statement: Quantitative structural neuroimaging has been proposed as one of the candidate in vivo biomarkers for tracking brain maturation and aging. While lifespan trajectories of structural white matter changes have been mapped in humans, such knowledge is not available for rhesus monkeys. We present here results of the analysis and modeling of the lifespan trajectories of white matter microstructure using in vivo diffusion imaging in a sample of 44 rhesus monkeys (age 4-27). We report and anatomically map lifespan changes related to cellular and extracellular microstructural components that are associated with white matter maturation and aging.

New method for predicting estrogen receptor status utilizing breast MRI texture kinetic analysis

NASA Astrophysics Data System (ADS)

Chaudhury, Baishali; Hall, Lawrence O.; Goldgof, Dmitry B.; Gatenby, Robert A.; Gillies, Robert; Drukteinis, Jennifer S.

2014-03-01

Magnetic Resonance Imaging (MRI) of breast cancer typically shows that tumors are heterogeneous with spatial variations in blood flow and cell density. Here, we examine the potential link between clinical tumor imaging and the underlying evolutionary dynamics behind heterogeneity in the cellular expression of estrogen receptors (ER) in breast cancer. We assume, in an evolutionary environment, that ER expression will only occur in the presence of significant concentrations of estrogen, which is delivered via the blood stream. Thus, we hypothesize, the expression of ER in breast cancer cells will correlate with blood flow on gadolinium enhanced breast MRI. To test this hypothesis, we performed quantitative analysis of blood flow on dynamic contrast enhanced MRI (DCE-MRI) and correlated it with the ER status of the tumor. Here we present our analytic methods, which utilize a novel algorithm to analyze 20 volumetric DCE-MRI breast cancer tumors. The algorithm generates post initial enhancement (PIE) maps from DCE-MRI and then performs texture features extraction from the PIE map, feature selection, and finally classification of tumors into ER positive and ER negative status. The combined gray level co-occurrence matrices, gray level run length matrices and local binary pattern histogram features allow quantification of breast tumor heterogeneity. The algorithm predicted ER expression with an accuracy of 85% using a Naive Bayes classifier in leave-one-out cross-validation. Hence, we conclude that our data supports the hypothesis that imaging characteristics can, through application of evolutionary principles, provide insights into the cellular and molecular properties of cancer cells.

Is the Cortical Deficit in Amblyopia Due to Reduced Cortical Magnification, Loss of Neural Resolution, or Neural Disorganization?

PubMed

Clavagnier, Simon; Dumoulin, Serge O; Hess, Robert F

2015-11-04

The neural basis of amblyopia is a matter of debate. The following possibilities have been suggested: loss of foveal cells, reduced cortical magnification, loss of spatial resolution of foveal cells, and topographical disarray in the cellular map. To resolve this we undertook a population receptive field (pRF) functional magnetic resonance imaging analysis in the central field in humans with moderate-to-severe amblyopia. We measured the relationship between averaged pRF size and retinal eccentricity in retinotopic visual areas. Results showed that cortical magnification is normal in the foveal field of strabismic amblyopes. However, the pRF sizes are enlarged for the amblyopic eye. We speculate that the pRF enlargement reflects loss of cellular resolution or an increased cellular positional disarray within the representation of the amblyopic eye. The neural basis of amblyopia, a visual deficit affecting 3% of the human population, remains a matter of debate. We undertook the first population receptive field functional magnetic resonance imaging analysis in participants with amblyopia and compared the projections from the amblyopic and fellow normal eye in the visual cortex. The projection from the amblyopic eye was found to have a normal cortical magnification factor, enlarged population receptive field sizes, and topographic disorganization in all early visual areas. This is consistent with an explanation of amblyopia as an immature system with a normal complement of cells whose spatial resolution is reduced and whose topographical map is disordered. This bears upon a number of competing theories for the psychophysical defect and affects future treatment therapies. Copyright © 2015 the authors 0270-6474/15/3514740-16$15.00/0.

Iterons, fractals and computations of automata

NASA Astrophysics Data System (ADS)

Siwak, Paweł

1999-03-01

Processing of strings by some automata, when viewed on space-time (ST) diagrams, reveals characteristic soliton-like coherent periodic objects. They are inherently associated with iterations of automata mappings thus we call them the iterons. In the paper we present two classes of one-dimensional iterons: particles and filtrons. The particles are typical for parallel (cellular) processing, while filtrons, introduced in (32) are specific for serial processing of strings. In general, the images of iterated automata mappings exhibit not only coherent entities but also the fractals, and quasi-periodic and chaotic dynamics. We show typical images of such computations: fractals, multiplication by a number, and addition of binary numbers defined by a Turing machine. Then, the particles are presented as iterons generated by cellular automata in three computations: B/U code conversion (13, 29), majority classification (9), and in discrete version of the FPU (Fermi-Pasta-Ulam) dynamics (7, 23). We disclose particles by a technique of combinational recoding of ST diagrams (as opposed to sequential recoding). Subsequently, we recall the recursive filters based on FCA (filter cellular automata) window operators, and considered by Park (26), Ablowitz (1), Fokas (11), Fuchssteiner (12), Bruschi (5) and Jiang (20). We present the automata equivalents to these filters (33). Some of them belong to the class of filter automata introduced in (30). We also define and illustrate some properties of filtrons. Contrary to particles, the filtrons interact nonlocally in the sense that distant symbols may influence one another. Thus their interactions are very unusual. Some examples have been given in (32). Here we show new examples of filtron phenomena: multifiltron solitonic collisions, attracting and repelling filtrons, trapped bouncing filtrons (which behave like a resonance cavity) and quasi filtrons.

Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping.

PubMed

Treweek, Jennifer B; Chan, Ken Y; Flytzanis, Nicholas C; Yang, Bin; Deverman, Benjamin E; Greenbaum, Alon; Lignell, Antti; Xiao, Cheng; Cai, Long; Ladinsky, Mark S; Bjorkman, Pamela J; Fowlkes, Charless C; Gradinaru, Viviana

2015-11-01

To facilitate fine-scale phenotyping of whole specimens, we describe here a set of tissue fixation-embedding, detergent-clearing and staining protocols that can be used to transform excised organs and whole organisms into optically transparent samples within 1-2 weeks without compromising their cellular architecture or endogenous fluorescence. PACT (passive CLARITY technique) and PARS (perfusion-assisted agent release in situ) use tissue-hydrogel hybrids to stabilize tissue biomolecules during selective lipid extraction, resulting in enhanced clearing efficiency and sample integrity. Furthermore, the macromolecule permeability of PACT- and PARS-processed tissue hybrids supports the diffusion of immunolabels throughout intact tissue, whereas RIMS (refractive index matching solution) grants high-resolution imaging at depth by further reducing light scattering in cleared and uncleared samples alike. These methods are adaptable to difficult-to-image tissues, such as bone (PACT-deCAL), and to magnified single-cell visualization (ePACT). Together, these protocols and solutions enable phenotyping of subcellular components and tracing cellular connectivity in intact biological networks.

Mapping and Quantification of Vascular Branching in Plants, Animals and Humans by VESGEN Software

NASA Technical Reports Server (NTRS)

Parsons-Wingerter, Patricia A.; Vickerman, Mary B.; Keith, Patricia A.

2010-01-01

Humans face daunting challenges in the successful exploration and colonization of space, including adverse alterations in gravity and radiation. The Earth-determined biology of humans, animals and plants is significantly modified in such extraterrestrial environments. One physiological requirement shared by humans with larger plants and animals is a complex, highly branching vascular system that is dynamically responsive to cellular metabolism, immunological protection and specialized cellular/tissue function. The VESsel GENeration (VESGEN) Analysis has been developed as a mature beta version, pre-release research software for mapping and quantification of the fractal-based complexity of vascular branching. Alterations in vascular branching pattern can provide informative read-outs of altered vascular regulation. Originally developed for biomedical applications in angiogenesis, VESGEN 2D has provided novel insights into the cytokine, transgenic and therapeutic regulation of angiogenesis, lymphangiogenesis and other microvascular remodeling phenomena. Vascular trees, networks and tree-network composites are mapped and quantified. Applications include disease progression from clinical ophthalmic images of the human retina; experimental regulation of vascular remodeling in the mouse retina; avian and mouse coronary vasculature, and other experimental models in vivo. We envision that altered branching in the leaves of plants studied on ISS such as Arabidopsis thaliana cans also be analyzed.

Mapping and Quantification of Vascular Branching in Plants, Animals and Humans by VESGEN Software

NASA Technical Reports Server (NTRS)

Parsons-Wingerter, P. A.; Vickerman, M. B.; Keith, P. A.

2010-01-01

Humans face daunting challenges in the successful exploration and colonization of space, including adverse alterations in gravity and radiation. The Earth-determined biology of plants, animals and humans is significantly modified in such extraterrestrial environments. One physiological requirement shared by larger plants and animals with humans is a complex, highly branching vascular system that is dynamically responsive to cellular metabolism, immunological protection and specialized cellular/tissue function. VESsel GENeration (VESGEN) Analysis has been developed as a mature beta version, pre-release research software for mapping and quantification of the fractal-based complexity of vascular branching. Alterations in vascular branching pattern can provide informative read-outs of altered vascular regulation. Originally developed for biomedical applications in angiogenesis, VESGEN 2D has provided novel insights into the cytokine, transgenic and therapeutic regulation of angiogenesis, lymphangiogenesis and other microvascular remodeling phenomena. Vascular trees, networks and tree-network composites are mapped and quantified. Applications include disease progression from clinical ophthalmic images of the human retina; experimental regulation of vascular remodeling in the mouse retina; avian and mouse coronary vasculature, and other experimental models in vivo. We envision that altered branching in the leaves of plants studied on ISS such as Arabidopsis thaliana cans also be analyzed.

Long-term imaging in awake mice using removable cranial windows

PubMed Central

Glickfeld, Lindsey L.; Kerlin, Aaron M.; Reid, R. Clay; Bonin, Vincent; Schafer, Dorothy P.; Andermann, Mark L.

2015-01-01

Cranial window implants in head-fixed rodents are becoming a preparation of choice for stable optical access to large areas of cortex over extended periods of time. Here, we provide a highly detailed and reliable surgical protocol for a cranial window implantation procedure for chronic widefield and cellular imaging in awake, head-fixed mice, which enables subsequent window removal and replacement in the weeks and months following the initial craniotomy. This protocol has facilitated awake, chronic imaging in adolescent as well as adult mice over several months from a large number of cortical brain regions; targeted virus and tracer injections from data obtained using prior awake functional mapping; and functionally-targeted two-photon imaging across all cortical layers in awake mice using a microprism attachment to the cranial window. Collectively, these procedures extend the reach of chronic imaging of cortical function and dysfunction in behaving animals. PMID:25275789

A Method for Imaging Oxygen Distribution and Respiration at a Microscopic Level of Resolution.

PubMed

Rolletschek, Hardy; Liebsch, Gregor

2017-01-01

Conventional oxygen (micro-) sensors assess oxygen concentration within a particular region or across a transect of tissue, but provide no information regarding its bidimensional distribution. Here, a novel imaging technology is presented, in which an optical sensor foil (i.e., the planar optode) is attached to the surface of the sample. The sensor converts a fluorescent signal into an oxygen value. Since each single image captures an entire area of the sample surface, the system is able to deduce the distribution of oxygen at a resolution level of few micrometers. It can be deployed to dynamically monitor oxygen consumption, thereby providing a detailed respiration map at close to cellular resolution. Here, we demonstrate the application of the imaging tool to developing plant seeds; the protocol is explained step by step and some potential pitfalls are discussed.

Field emission scanning electron microscopy (FE-SEM) as an approach for nanoparticle detection inside cells.

PubMed

Havrdova, M; Polakova, K; Skopalik, J; Vujtek, M; Mokdad, A; Homolkova, M; Tucek, J; Nebesarova, J; Zboril, R

2014-12-01

When developing new nanoparticles for bio-applications, it is important to fully characterize the nanoparticle's behavior in biological systems. The most common techniques employed for mapping nanoparticles inside cells include transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM). These techniques entail passing an electron beam through a thin specimen. STEM or TEM imaging is often used for the detection of nanoparticles inside cellular organelles. However, lengthy sample preparation is required (i.e., fixation, dehydration, drying, resin embedding, and cutting). In the present work, a new matrix (FTO glass) for biological samples was used and characterized by field emission scanning electron microscopy (FE-SEM) to generate images comparable to those obtained by TEM. Using FE-SEM, nanoparticle images were acquired inside endo/lysosomes without disruption of the cellular shape. Furthermore, the initial steps of nanoparticle incorporation into the cells were captured. In addition, the conductive FTO glass endowed the sample with high stability under the required accelerating voltage. Owing to these features of the sample, further analyses could be performed (material contrast and energy-dispersive X-ray spectroscopy (EDS)), which confirmed the presence of nanoparticles inside the cells. The results showed that FE-SEM can enable detailed characterization of nanoparticles in endosomes without the need for contrast staining or metal coating of the sample. Images showing the intracellular distribution of nanoparticles together with cellular morphology can give important information on the biocompatibility and demonstrate the potential of nanoparticle utilization in medicine. Copyright © 2014 Elsevier Ltd. All rights reserved.

Multimodal imaging of lymph nodes and tumors using glycol-chitosan-coated gold nanoparticles (Conference Presentation)

NASA Astrophysics Data System (ADS)

Sun, In-Cheol; Dumani, Diego S.; Emelianov, Stanislav Y.

2017-03-01

A key step in staging cancer is the diagnosis of metastasis that spreads through lymphatic system. For this reason, researchers develop various methods of sentinel lymph node mapping that often use a radioactive tracer. This study introduces a safe, cost-effective, high-resolution, high-sensitivity, and real-time method of visualizing the sentinel lymph node: ultrasound-guided photoacoustic (US/PA) imaging augmented by a contrast agent. In this work, we use clearable gold nanoparticles covered by a biocompatible polymer (glycol chitosan) to enhance cellular uptake by macrophages abundant in lymph nodes. We incubate macrophages with glycol-chitosan-coated gold nanoparticles (0.05 mg Au/ml), and then fix them with paraformaldehyde solution for an analysis of in vitro dark-field microscopy and cell phantom. The analysis shows enhanced cellular uptake of nanoparticles by macrophages and strong photoacoustic signal from labeled cells in tissue-mimicking cell phantoms consisting gelatin solution (6 %) with silica gel (25 μm, 0.3%) and fixed macrophages. The in-vivo US/PA imaging of cervical lymph nodes in healthy mice (nu/nu, female, 5 weeks) indicates a strong photoacoustic signal from a lymph node 10 minutes post-injection (2.5 mg Au/ml, 80 μl). The signal intensity and the nanoparticle-labeled volume of tissue within the lymph node continues to increase until 4 h post-injection. Histological analysis further confirms the accumulation of gold nanoparticles within the lymph nodes. This work suggests the feasibility of molecular/cellular US/PA imaging with biocompatible gold nanoparticles as a photoacoustic contrast agent in the diagnosis of lymph-node-related diseases.

Metabolic Mapping: Quantitative Enzyme Cytochemistry and Histochemistry to Determine the Activity of Dehydrogenases in Cells and Tissues.

PubMed

Molenaar, Remco J; Khurshed, Mohammed; Hira, Vashendriya V V; Van Noorden, Cornelis J F

2018-05-26

Altered cellular metabolism is a hallmark of many diseases, including cancer, cardiovascular diseases and infection. The metabolic motor units of cells are enzymes and their activity is heavily regulated at many levels, including the transcriptional, mRNA stability, translational, post-translational and functional level. This complex regulation means that conventional quantitative or imaging assays, such as quantitative mRNA experiments, Western Blots and immunohistochemistry, yield incomplete information regarding the ultimate activity of enzymes, their function and/or their subcellular localization. Quantitative enzyme cytochemistry and histochemistry (i.e., metabolic mapping) show in-depth information on in situ enzymatic activity and its kinetics, function and subcellular localization in an almost true-to-nature situation. We describe a protocol to detect the activity of dehydrogenases, which are enzymes that perform redox reactions to reduce cofactors such as NAD(P) + and FAD. Cells and tissue sections are incubated in a medium that is specific for the enzymatic activity of one dehydrogenase. Subsequently, the dehydrogenase that is the subject of investigation performs its enzymatic activity in its subcellular site. In a chemical reaction with the reaction medium, this ultimately generates blue-colored formazan at the site of the dehydrogenase's activity. The formazan's absorbance is therefore a direct measure of the dehydrogenase's activity and can be quantified using monochromatic light microscopy and image analysis. The quantitative aspect of this protocol enables researchers to draw statistical conclusions from these assays. Besides observational studies, this technique can be used for inhibition studies of specific enzymes. In this context, studies benefit from the true-to-nature advantages of metabolic mapping, giving in situ results that may be physiologically more relevant than in vitro enzyme inhibition studies. In all, metabolic mapping is an indispensable technique to study metabolism at the cellular or tissue level. The technique is easy to adopt, provides in-depth, comprehensive and integrated metabolic information and enables rapid quantitative analysis.

Vibrational imaging of glucose uptake activity in live cells and tissues by stimulated Raman scattering microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Hu, Fanghao; Chen, Zhixing; Zhang, Luyuan; Shen, Yihui; Wei, Lu; Min, Wei

2016-03-01

Glucose is consumed as an energy source by virtually all living organisms, from bacteria to humans. Its uptake activity closely reflects the cellular metabolic status in various pathophysiological transformations, such as diabetes and cancer. Extensive efforts such as positron emission tomography, magnetic resonance imaging and fluorescence microscopy have been made to specifically image glucose uptake activity but all with technical limitations. Here, we report a new platform to visualize glucose uptake activity in live cells and tissues with subcellular resolution and minimal perturbation. A novel glucose analogue with a small alkyne tag (carbon-carbon triple bond) is developed to mimic natural glucose for cellular uptake, which can be imaged with high sensitivity and specificity by targeting the strong and characteristic alkyne vibration on stimulated Raman scattering (SRS) microscope to generate a quantitative three dimensional concentration map. Cancer cells with differing metabolic characteristics can be distinguished. Heterogeneous uptake patterns are observed in tumor xenograft tissues, neuronal culture and mouse brain tissues with clear cell-cell variations. Therefore, by offering the distinct advantage of optical resolution but without the undesirable influence of bulky fluorophores, our method of coupling SRS with alkyne labeled glucose will be an attractive tool to study energy demands of living systems at the single cell level.

The Contribution of Conceptual Change Texts Accompanied by Concept Mapping to Eleventh-Grade Students Understanding of Cellular Respiration Concepts

ERIC Educational Resources Information Center

Al khawaldeh, Salem A.; Al Olaimat, Ali M.

2010-01-01

The present study conducted to investigate the contribution of conceptual change texts, accompanied by concept mapping instruction to eleventh-grade students' understanding of cellular respiration concepts, and their retention of this understanding. Cellular respiration concepts test was developed as a result of examination of related literature…

In Vivo Confocal Intrinsic Optical Signal Identification of Localized Retinal Dysfunction

PubMed Central

Zhang, Qiu-Xiang; Lu, Rong-Wen; Curcio, Christine A.; Yao, Xin-Cheng

2012-01-01

Purpose. The purposes of this study were to investigate the physiological mechanism of stimulus-evoked fast intrinsic optical signals (IOSs) recorded in dynamic confocal imaging of the retina, and to demonstrate the feasibility of in vivo confocal IOS mapping of localized retinal dysfunctions. Methods. A rapid line-scan confocal ophthalmoscope was constructed to achieve in vivo confocal IOS imaging of frog (Rana pipiens) retinas at cellular resolution. In order to investigate the physiological mechanism of confocal IOS, comparative IOS and electroretinography (ERG) measurements were made using normal frog eyes activated by variable-intensity stimuli. A dynamic spatiotemporal filtering algorithm was developed to reject the contamination of hemodynamic changes on fast IOS recording. Laser-injured frog eyes were employed to test the potential of confocal IOS mapping of localized retinal dysfunctions. Results. Comparative IOS and ERG experiments revealed a close correlation between the confocal IOS and retinal ERG, particularly the ERG a-wave, which has been widely used to evaluate photoreceptor function. IOS imaging of laser-injured frog eyes indicated that the confocal IOS could unambiguously detect localized (30 μm) functional lesions in the retina before a morphological abnormality is detectable. Conclusions. The confocal IOS predominantly results from retinal photoreceptors, and can be used to map localized photoreceptor lesion in laser-injured frog eyes. We anticipate that confocal IOS imaging can provide applications in early detection of age-related macular degeneration, retinitis pigmentosa, and other retinal diseases that can cause pathological changes in the photoreceptors. PMID:23150616

Fluorescent BODIPY Rotor: Viscometer for Cellular Organelles and Membrane-Mimicking Vesicles

NASA Astrophysics Data System (ADS)

Kimball, J.; Raut, S.; Fudala, R.; Doan, H.; Maliwal, B.; Sabnis, N.; Lacko, A.; Gryczynski, I.; Dzyuba, S.; Gryczynski, Z.

2015-03-01

Many cellular processes, such as mass and signal transport, metabolism and protein-protein interactions are governed in part by diffusion, and thus affected by their local microviscosity. Changes in this microviscosity has also been linked to various diseases, including atherosclerosis, Alzheimer's disease and diabetes. Therefore, directly measuring the heterogeneous viscosity of cellular constitutes can lead to greater understanding of these processes. To this effect, a novel homodiemeric BODIPY dye was evaluated as a fluorescent rotor probe for this application. A linear dependence on viscosity in the range of typical cellular microviscosity was established for steady-state and time-resolved properties of the dye. It was then embedded in vitro to membrane-mimicking lipid vesicles (DPPC, POPC, and POPC plus cholesterol) and results indicated it to be a viable sensor for lifetime-based determination of microviscosity. The BODIPY dye was lastly endocytosed by SKOV3 cells and Fluorescence Lifetime Imaging Microscopy (FLIM) was performed, successfully mapping the viscosity of internal cell components. This work was supported by the NIH Grant R01EB12003, the NSF Grant CBET-1264608, and the INFOR Grant from TCU.

Metabolic autofluorescence imaging of head and neck cancer organoids quantifies cellular heterogeneity and treatment response (Conference Presentation)

NASA Astrophysics Data System (ADS)

Shah, Amy T.; Heaster, Tiffany M.; Skala, Melissa C.

2017-02-01

Treatment options for head and neck cancer are limited, and can cause an impaired ability to eat, talk, and breathe. Therefore, optimized and personalized therapies could reduce unnecessary toxicities from ineffective treatments. Organoids are generated from primary tumor tissue and provide a physiologically-relevant in vitro model to measure drug response. Additionally, multiphoton fluorescence lifetime imaging (FLIM) of the metabolic cofactors NAD(P)H and FAD can resolve dynamic cellular response to anti-cancer treatment. This study applies FLIM of NAD(P)H and FAD to head and neck cancer organoids. Head and neck cancer tissue was digested and grown in culture as three-dimensional organoids. Gold standard measures of therapeutic response in vivo indicate stable disease after treatment with cetuximab (antibody therapy) or cisplatin (chemotherapy), and treatment response after combination treatment. In parallel, organoids were treated with cetuximab, cisplatin, or combination therapy for 24 hours. Treated organoids exhibit decreased NAD(P)H lifetime (p<0.05) and increased FAD lifetime (p<0.05) compared with control organoids. Additionally, analysis of cellular heterogeneity identifies distinct subpopulations of cells in response to treatment. A quantitative heterogeneity index predicts in vivo treatment response and demonstrates increased cellular heterogeneity in organoids treated with cetuximab or cisplatin compared with combination treatment. Mapping of cell subpopulations enables characterization of spatial relationships between cell subpopulations. Ultimately, an organoid model combined with metabolic fluorescence imaging could provide a high-throughput platform for drug discovery. Organoids grown from patient tissue could enable individualized treatment planning. These achievements could optimize quality of life and treatment outcomes for head and neck cancer patients.

MapA, an iron-regulated, cytoplasmic membrane protein in the cyanobacterium Synechococcus sp. strain PCC7942.

PubMed Central

Webb, R; Troyan, T; Sherman, D; Sherman, L A

1994-01-01

Growth of Synechococcus sp. strain PCC 7942 in iron-deficient media leads to the accumulation of an approximately 34-kDa protein. The gene encoding this protein, mapA (membrane-associated protein A), has been cloned and sequenced (GenBank accession number, L01621). The mapA transcript is not detectable in normally grown cultures but is stably accumulated by cells grown in iron-deficient media. However, the promoter sequence for this gene does not resemble other bacterial iron-regulated promoters described to date. The carboxyl-terminal region of the derived amino acid sequence of MapA resembles bacterial proteins involved in iron acquisition, whereas the amino-terminal end of MapA has a high degree of amino acid identity with the abundant, chloroplast envelope protein E37. An approach employing improved cellular fractionation techniques as well as electron microscopy and immunocytochemistry was essential in localizing MapA protein to the cytoplasmic membrane of Synechococcus sp. strain PCC 7942. When these cells were grown under iron-deficient conditions, a significant fraction of MapA could also be localized to the thylakoid membranes. Images PMID:8051004

Mapping cortical mesoscopic networks of single spiking cortical or sub-cortical neurons

PubMed Central

Xiao, Dongsheng; Vanni, Matthieu P; Mitelut, Catalin C; Chan, Allen W; LeDue, Jeffrey M; Xie, Yicheng; Chen, Andrew CN; Swindale, Nicholas V; Murphy, Timothy H

2017-01-01

Understanding the basis of brain function requires knowledge of cortical operations over wide-spatial scales, but also within the context of single neurons. In vivo, wide-field GCaMP imaging and sub-cortical/cortical cellular electrophysiology were used in mice to investigate relationships between spontaneous single neuron spiking and mesoscopic cortical activity. We make use of a rich set of cortical activity motifs that are present in spontaneous activity in anesthetized and awake animals. A mesoscale spike-triggered averaging procedure allowed the identification of motifs that are preferentially linked to individual spiking neurons by employing genetically targeted indicators of neuronal activity. Thalamic neurons predicted and reported specific cycles of wide-scale cortical inhibition/excitation. In contrast, spike-triggered maps derived from single cortical neurons yielded spatio-temporal maps expected for regional cortical consensus function. This approach can define network relationships between any point source of neuronal spiking and mesoscale cortical maps. DOI: http://dx.doi.org/10.7554/eLife.19976.001 PMID:28160463

Mapping of Brain Activity by Automated Volume Analysis of Immediate Early Genes.

PubMed

Renier, Nicolas; Adams, Eliza L; Kirst, Christoph; Wu, Zhuhao; Azevedo, Ricardo; Kohl, Johannes; Autry, Anita E; Kadiri, Lolahon; Umadevi Venkataraju, Kannan; Zhou, Yu; Wang, Victoria X; Tang, Cheuk Y; Olsen, Olav; Dulac, Catherine; Osten, Pavel; Tessier-Lavigne, Marc

2016-06-16

Understanding how neural information is processed in physiological and pathological states would benefit from precise detection, localization, and quantification of the activity of all neurons across the entire brain, which has not, to date, been achieved in the mammalian brain. We introduce a pipeline for high-speed acquisition of brain activity at cellular resolution through profiling immediate early gene expression using immunostaining and light-sheet fluorescence imaging, followed by automated mapping and analysis of activity by an open-source software program we term ClearMap. We validate the pipeline first by analysis of brain regions activated in response to haloperidol. Next, we report new cortical regions downstream of whisker-evoked sensory processing during active exploration. Last, we combine activity mapping with axon tracing to uncover new brain regions differentially activated during parenting behavior. This pipeline is widely applicable to different experimental paradigms, including animal species for which transgenic activity reporters are not readily available. Copyright © 2016 Elsevier Inc. All rights reserved.

Mapping of brain activity by automated volume analysis of immediate early genes

PubMed Central

Renier, Nicolas; Adams, Eliza L.; Kirst, Christoph; Wu, Zhuhao; Azevedo, Ricardo; Kohl, Johannes; Autry, Anita E.; Kadiri, Lolahon; Venkataraju, Kannan Umadevi; Zhou, Yu; Wang, Victoria X.; Tang, Cheuk Y.; Olsen, Olav; Dulac, Catherine; Osten, Pavel; Tessier-Lavigne, Marc

2016-01-01

Summary Understanding how neural information is processed in physiological and pathological states would benefit from precise detection, localization and quantification of the activity of all neurons across the entire brain, which has not to date been achieved in the mammalian brain. We introduce a pipeline for high speed acquisition of brain activity at cellular resolution through profiling immediate early gene expression using immunostaining and light-sheet fluorescence imaging, followed by automated mapping and analysis of activity by an open-source software program we term ClearMap. We validate the pipeline first by analysis of brain regions activated in response to Haloperidol. Next, we report new cortical regions downstream of whisker-evoked sensory processing during active exploration. Lastly, we combine activity mapping with axon tracing to uncover new brain regions differentially activated during parenting behavior. This pipeline is widely applicable to different experimental paradigms, including animal species for which transgenic activity reporters are not readily available. PMID:27238021

Cardiac T1 Imaging

PubMed Central

Jerosch-Herold, Michael; Kwong, Raymond Y.

2014-01-01

T1 mapping of the heart has evolved into a valuable tool to evaluate myocardial tissue properties, with or without contrast injection, including assessment of myocardial edema and free water content, extra-cellular volume (expansion), and most recently cardiomyocyte hypertrophy. The MRI pulse sequence techniques developed for these applications have had to address at least two important considerations for cardiac applications: measure magnetization inversion recoveries during cardiac motion with sufficient temporal resolution for the shortest expected T1 values, and, secondly, obtain these measurements within a time during which a patient can comfortably suspend breathing. So-called Look-Locker techniques, and variants thereof, which all sample multiple points of a magnetization recovery after each magnetization preparation have therefore become a mainstay in this field. The rapid pace of advances and new findings based on cardiac T1 mapping for assessment of diffuse fibrosis, or myocardial edema show that these techniques enrich the capabilities of MRI for myocardial tissue profiling, which is arguably unmatched by other cardiac imaging modalities. PMID:24509619

Mouse chromosomal mapping of a murine leukemia virus integration region (Mis-1) first identified in rat thymic leukemia.

PubMed Central

Jolicoeur, P; Villeneuve, L; Rassart, E; Kozak, C

1985-01-01

We have previously identified a region of genomic DNA which constitutes the site of frequent provirus integration in rat thymomas induced by Moloney murine leukemia virus (Lemay and Jolicoeur, Proc. Natl. Acad. Sci. USA 81:38-42, 1984). This genetic locus is now designated Mis-1 (Moloney integration site). Cellular sequences homologous to Mis-1 are present in mouse DNA. Using a series of hamster-mouse somatic cell hybrids, we mapped the Mis-1 locus to mouse chromosome 15. Frequent chromosome 15 aberrations have been described in mouse thymomas. Mis-1 represents a putative new oncogene which might be involved in the initiation or maintenance or both of these neoplasms. Images PMID:4068142

Spatial reconstruction of single-cell gene expression

PubMed Central

Satija, Rahul; Farrell, Jeffrey A.; Gennert, David; Schier, Alexander F.; Regev, Aviv

2015-01-01

Spatial localization is a key determinant of cellular fate and behavior, but spatial RNA assays traditionally rely on staining for a limited number of RNA species. In contrast, single-cell RNA-seq allows for deep profiling of cellular gene expression, but established methods separate cells from their native spatial context. Here we present Seurat, a computational strategy to infer cellular localization by integrating single-cell RNA-seq data with in situ RNA patterns. We applied Seurat to spatially map 851 single cells from dissociated zebrafish (Danio rerio) embryos, inferring a transcriptome-wide map of spatial patterning. We confirmed Seurat’s accuracy using several experimental approaches, and used it to identify a set of archetypal expression patterns and spatial markers. Additionally, Seurat correctly localizes rare subpopulations, accurately mapping both spatially restricted and scattered groups. Seurat will be applicable to mapping cellular localization within complex patterned tissues in diverse systems. PMID:25867923

Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex

NASA Astrophysics Data System (ADS)

Ohki, Kenichi; Chung, Sooyoung; Ch'ng, Yeang H.; Kara, Prakash; Reid, R. Clay

2005-02-01

Neurons in the cerebral cortex are organized into anatomical columns, with ensembles of cells arranged from the surface to the white matter. Within a column, neurons often share functional properties, such as selectivity for stimulus orientation; columns with distinct properties, such as different preferred orientations, tile the cortical surface in orderly patterns. This functional architecture was discovered with the relatively sparse sampling of microelectrode recordings. Optical imaging of membrane voltage or metabolic activity elucidated the overall geometry of functional maps, but is averaged over many cells (resolution >100µm). Consequently, the purity of functional domains and the precision of the borders between them could not be resolved. Here, we labelled thousands of neurons of the visual cortex with a calcium-sensitive indicator in vivo. We then imaged the activity of neuronal populations at single-cell resolution with two-photon microscopy up to a depth of 400µm. In rat primary visual cortex, neurons had robust orientation selectivity but there was no discernible local structure; neighbouring neurons often responded to different orientations. In area 18 of cat visual cortex, functional maps were organized at a fine scale. Neurons with opposite preferences for stimulus direction were segregated with extraordinary spatial precision in three dimensions, with columnar borders one to two cells wide. These results indicate that cortical maps can be built with single-cell precision.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Mi; University of Chinese Academy of Sciences, Beijing 100049; Liu, Lianqing, E-mail: lqliu@sia.cn

Highlights: •Nanoscale cellular ultra-structures of macrophages were observed. •The binding affinities of FcγRs were measured directly on macrophages. •The nanoscale distributions of FcγRs were mapped on macrophages. -- Abstract: Fc gamma receptors (FcγR), widely expressed on effector cells (e.g., NK cells, macrophages), play an important role in clinical cancer immunotherapy. The binding of FcγRs to the Fc portions of antibodies that are attached to the target cells can activate the antibody-dependent cell-mediated cytotoxicity (ADCC) killing mechanism which leads to the lysis of target cells. In this work, we used atomic force microscopy (AFM) to observe the cellular ultra-structures and measuremore » the biophysical properties (affinity and distribution) of FcγRs on single macrophages in aqueous environments. AFM imaging was used to obtain the topographies of macrophages, revealing the nanoscale cellular fine structures. For molecular interaction recognition, antibody molecules were attached onto AFM tips via a heterobifunctional polyethylene glycol (PEG) crosslinker. With AFM single-molecule force spectroscopy, the binding affinities of FcγRs were quantitatively measured on single macrophages. Adhesion force mapping method was used to localize the FcγRs, revealing the nanoscale distribution of FcγRs on local areas of macrophages. The experimental results can improve our understanding of FcγRs on macrophages; the established approach will facilitate further research on physiological activities involved in antibody-based immunotherapy.« less

A versatile clearing agent for multi-modal brain imaging

PubMed Central

Costantini, Irene; Ghobril, Jean-Pierre; Di Giovanna, Antonino Paolo; Mascaro, Anna Letizia Allegra; Silvestri, Ludovico; Müllenbroich, Marie Caroline; Onofri, Leonardo; Conti, Valerio; Vanzi, Francesco; Sacconi, Leonardo; Guerrini, Renzo; Markram, Henry; Iannello, Giulio; Pavone, Francesco Saverio

2015-01-01

Extensive mapping of neuronal connections in the central nervous system requires high-throughput µm-scale imaging of large volumes. In recent years, different approaches have been developed to overcome the limitations due to tissue light scattering. These methods are generally developed to improve the performance of a specific imaging modality, thus limiting comprehensive neuroanatomical exploration by multi-modal optical techniques. Here, we introduce a versatile brain clearing agent (2,2′-thiodiethanol; TDE) suitable for various applications and imaging techniques. TDE is cost-efficient, water-soluble and low-viscous and, more importantly, it preserves fluorescence, is compatible with immunostaining and does not cause deformations at sub-cellular level. We demonstrate the effectiveness of this method in different applications: in fixed samples by imaging a whole mouse hippocampus with serial two-photon tomography; in combination with CLARITY by reconstructing an entire mouse brain with light sheet microscopy and in translational research by imaging immunostained human dysplastic brain tissue. PMID:25950610

Transparent, Flexible, Low Noise Graphene Electrodes for Simultaneous Electrophysiology and Neuroimaging

PubMed Central

Kuzum, Duygu; Takano, Hajime; Shim, Euijae; Reed, Jason C; Juul, Halvor; Richardson, Andrew G.; de Vries, Julius; Bink, Hank; Dichter, Marc A.; Lucas, Timothy H.; Coulter, Douglas A.; Cubukcu, Ertugrul; Litt, Brian

2014-01-01

Calcium imaging is a versatile experimental approach capable of resolving single neurons with single-cell spatial resolution in the brain. Electrophysiological recordings provide high temporal, but limited spatial resolution, due to the geometrical inaccessibility of the brain. An approach that integrates the advantages of both techniques could provide new insights into functions of neural circuits. Here, we report a transparent, flexible neural electrode technology based on graphene, which enables simultaneous optical imaging and electrophysiological recording. We demonstrate that hippocampal slices can be imaged through transparent graphene electrodes by both confocal and two-photon microscopy without causing any light-induced artifacts in the electrical recordings. Graphene electrodes record high frequency bursting activity and slow synaptic potentials that are hard to resolve by multi-cellular calcium imaging. This transparent electrode technology may pave the way for high spatio-temporal resolution electrooptic mapping of the dynamic neuronal activity. PMID:25327632

Universal map for cellular automata

NASA Astrophysics Data System (ADS)

García-Morales, V.

2012-08-01

A universal map is derived for all deterministic 1D cellular automata (CAs) containing no freely adjustable parameters and valid for any alphabet size and any neighborhood range (including non-symmetrical neighborhoods). The map can be extended to an arbitrary number of dimensions and topologies and to arbitrary order in time. Specific CA maps for the famous Conway's Game of Life and Wolfram's 256 elementary CAs are given. An induction method for CAs, based in the universal map, allows mathematical expressions for the orbits of a wide variety of elementary CAs to be systematically derived.

Magnetic Resonance Microscopy of Human and Porcine Neurons and Cellular Processes

PubMed Central

Flint, Jeremy J.; Hansen, Brian; Portnoy, Sharon; Lee, Choong-Heon; King, Michael A.; Fey, Michael; Vincent, Franck; Stanisz, Greg J; Vestergaard-Poulsen, Peter; Blackband, Stephen J

2012-01-01

With its unparalleled ability to safely generate high-contrast images of soft tissues, magnetic resonance imaging (MRI) has remained at the forefront of diagnostic clinical medicine. Unfortunately due to resolution limitations, clinical scans are most useful for detecting macroscopic structural changes associated with a small number of pathologies. Moreover, due to a longstanding inability to directly observe magnetic resonance (MR) signal behavior at the cellular level, such information is poorly characterized and generally must be inferred. With the advent of the MR microscope in 1986 came the ability to measure MR signal properties of theretofore unobservable tissue structures. Recently, further improvements in hardware technology have made possible the ability to visualize mammalian cellular structure. In the current study, we expand upon previous work by imaging the neuronal cell bodies and processes of human and porcine α-motor neurons. Complimentary imaging studies are conducted in pig tissue in order to demonstrate qualitative similarities to human samples. Also, apparent diffusion coefficient (ADC) maps were generated inside porcine α-motor neuron cell bodies and portions of their largest processes (mean = 1.7±0.5 μm2/ms based on 53 pixels) as well as in areas containing a mixture of extracellular space, microvasculature, and neuropil (0.59±0.37 μm2/ms based on 33 pixels). Three-dimensional reconstruction of MR images containing α-motor neurons shows the spatial arrangement of neuronal projections between adjacent cells. Such advancements in imaging portend the ability to construct accurate models of MR signal behavior based on direct observation and measurement of the components which comprise functional tissues. These tools would not only be useful for improving our interpretation of macroscopic MRI performed in the clinic, but they could potentially be used to develop new methods of differential diagnosis to aid in the early detection of a multitude of neuropathologies. PMID:22281672

Dynamic positional fate map of the primary heart-forming region.

PubMed

Cui, Cheng; Cheuvront, Tracey J; Lansford, Rusty D; Moreno-Rodriguez, Ricardo A; Schultheiss, Thomas M; Rongish, Brenda J

2009-08-15

Here we show the temporal-spatial orchestration of early heart morphogenesis at cellular level resolution, in vivo, and reconcile conflicting positional fate mapping data regarding the primary heart-forming field(s). We determined the positional fates of precardiac cells using a precision electroporation approach in combination with wide-field time-lapse microscopy in the quail embryo, a warm-blooded vertebrate (HH Stages 4 through 10). Contrary to previous studies, the results demonstrate the existence of a "continuous" circle-shaped heart field that spans the midline, appearing at HH Stage 4, which then expands to form a wide arc of progenitors at HH Stages 5-7. Our time-resolved image data show that a subset of these cardiac progenitor cells do not overlap with the expression of common cardiogenic factors, Nkx-2.5 and Bmp-2, until HH Stage 10, when a tubular heart has formed, calling into question when cardiac fate is specified and by which key factors. Sub-groups and anatomical bands (cohorts) of heart precursor cells dramatically change their relative positions in a process largely driven by endodermal folding and other large-scale tissue deformations. Thus, our novel dynamic positional fate maps resolve the origin of cardiac progenitor cells in amniotes. The data also establish the concept that tissue motion contributes significantly to cellular position fate - i.e., much of the cellular displacement that occurs during assembly of a midline heart tube (HH Stage 9) is NOT due to "migration" (autonomous motility), a commonly held belief. Computational analysis of our time-resolved data lays the foundation for more precise analyses of how cardiac gene regulatory networks correlate with early heart tissue morphogenesis in birds and mammals.

Ultrasound-guided photoacoustic imaging of lymph nodes with biocompatible gold nanoparticles as a novel contrast agent (Conference Presentation)

NASA Astrophysics Data System (ADS)

Sun, In-Cheol; Dumani, Diego; Emelianov, Stanislav Y.

2017-02-01

A key step in staging cancer is the diagnosis of metastasis that spreads through lymphatic system. For this reason, researchers develop various methods of sentinel lymph node mapping that often use a radioactive tracer. This study introduces a safe, cost-effective, high-resolution, high-sensitivity, and real-time method of visualizing the sentinel lymph node: ultrasound-guided photoacoustic (US/PA) imaging augmented by a contrast agent. In this work, we use clearable gold nanoparticles covered by a biocompatible polymer (glycol chitosan) to enhance cellular uptake by macrophages abundant in lymph nodes. We incubate macrophages with glycol-chitosan-coated gold nanoparticles (0.05 mg Au/ml), and then fix them with paraformaldehyde solution for an analysis of in vitro dark-field microscopy and cell phantom. The analysis shows enhanced cellular uptake of nanoparticles by macrophages and strong photoacoustic signal from labeled cells in tissue-mimicking cell phantoms consisting gelatin solution (6 %) with silica gel (25 μm, 0.3%) and fixed macrophages (13 X 105 cells). The in-vivo US/PA imaging of cervical lymph nodes in healthy mice (nu/nu, female, 5 weeks) indicates a strong photoacoustic signal from a lymph node 10 minutes post-injection (2.5 mg Au/ml, 80 μl). The signal intensity and the nanoparticle-labeled volume of tissue within the lymph node continues to increase until 4 h post-injection. Histological analysis further confirms the accumulation of gold nanoparticles within the lymph nodes. This work suggests the feasibility of molecular/cellular US/PA imaging with biocompatible gold nanoparticles as a photoacoustic contrast agent in the diagnosis of lymph-node-related diseases.

Multi-scale imaging and informatics pipeline for in situ pluripotent stem cell analysis.

PubMed

Gorman, Bryan R; Lu, Junjie; Baccei, Anna; Lowry, Nathan C; Purvis, Jeremy E; Mangoubi, Rami S; Lerou, Paul H

2014-01-01

Human pluripotent stem (hPS) cells are a potential source of cells for medical therapy and an ideal system to study fate decisions in early development. However, hPS cells cultured in vitro exhibit a high degree of heterogeneity, presenting an obstacle to clinical translation. hPS cells grow in spatially patterned colony structures, necessitating quantitative single-cell image analysis. We offer a tool for analyzing the spatial population context of hPS cells that integrates automated fluorescent microscopy with an analysis pipeline. It enables high-throughput detection of colonies at low resolution, with single-cellular and sub-cellular analysis at high resolutions, generating seamless in situ maps of single-cellular data organized by colony. We demonstrate the tool's utility by analyzing inter- and intra-colony heterogeneity of hPS cell cycle regulation and pluripotency marker expression. We measured the heterogeneity within individual colonies by analyzing cell cycle as a function of distance. Cells loosely associated with the outside of the colony are more likely to be in G1, reflecting a less pluripotent state, while cells within the first pluripotent layer are more likely to be in G2, possibly reflecting a G2/M block. Our multi-scale analysis tool groups colony regions into density classes, and cells belonging to those classes have distinct distributions of pluripotency markers and respond differently to DNA damage induction. Lastly, we demonstrate that our pipeline can robustly handle high-content, high-resolution single molecular mRNA FISH data by using novel image processing techniques. Overall, the imaging informatics pipeline presented offers a novel approach to the analysis of hPS cells that includes not only single cell features but also colony wide, and more generally, multi-scale spatial configuration.

In vivo nanoparticle imaging of innate immune cells can serve as a marker of disease severity in a model of multiple sclerosis.

PubMed

Kirschbaum, Klara; Sonner, Jana K; Zeller, Matthias W; Deumelandt, Katrin; Bode, Julia; Sharma, Rakesh; Krüwel, Thomas; Fischer, Manuel; Hoffmann, Angelika; Costa da Silva, Milene; Muckenthaler, Martina U; Wick, Wolfgang; Tews, Björn; Chen, John W; Heiland, Sabine; Bendszus, Martin; Platten, Michael; Breckwoldt, Michael O

2016-11-15

Innate immune cells play a key role in the pathogenesis of multiple sclerosis and experimental autoimmune encephalomyelitis (EAE). Current clinical imaging is restricted to visualizing secondary effects of inflammation, such as gliosis and blood-brain barrier disruption. Advanced molecular imaging, such as iron oxide nanoparticle imaging, can allow direct imaging of cellular and molecular activity, but the exact cell types that phagocytose nanoparticles in vivo and how phagocytic activity relates to disease severity is not well understood. In this study we used MRI to map inflammatory infiltrates using high-field MRI and fluorescently labeled cross-linked iron oxide nanoparticles for cell tracking. We confirmed nanoparticle uptake and MR detectability ex vivo. Using in vivo MRI, we identified extensive nanoparticle signal in the cerebellar white matter and circumscribed cortical gray matter lesions that developed during the disease course (4.6-fold increase of nanoparticle accumulation in EAE compared with healthy controls, P < 0.001). Nanoparticles showed good cellular specificity for innate immune cells in vivo, labeling activated microglia, infiltrating macrophages, and neutrophils, whereas there was only sparse uptake by adaptive immune cells. Importantly, nanoparticle signal correlated better with clinical disease than conventional gadolinium (Gd) imaging (r, 0.83 for nanoparticles vs. 0.71 for Gd-imaging, P < 0.001). We validated our approach using the Food and Drug Administration-approved iron oxide nanoparticle ferumoxytol. Our results show that noninvasive molecular imaging of innate immune responses can serve as an imaging biomarker of disease activity in autoimmune-mediated neuroinflammation with potential clinical applications in a wide range of inflammatory diseases.

Dynamic mapping of spontaneously produced H2S in the entire cell space and in live animals using a rationally designed molecular switch.

PubMed

Yang, Linlin; Zhao, Jun; Yu, Xinling; Zhang, Ruilong; Han, Guangmei; Liu, Renyong; Liu, Zhengjie; Zhao, Tingting; Han, Ming-Yong; Zhang, Zhongping

2018-04-16

Hydrogen sulfide (H2S) is a key signaling molecule in the cytoprotection, vascular mediation and neurotransmission of living organisms. In-depth understanding of its production, trafficking, and transformation in cells is very important in the way H2S mediates cellular signal transductions and organism functions; it also motivates the development of H2S probes and imaging technologies. A fundamental challenge, however, is how to engineer probes with sensitivity and cellular penetrability that allow detection of spontaneous production of H2S in the entire cell space and live animals. Here, we report a rationally designed molecular switch capable of accessing all intracellular compartments, including the nucleus, lysosomes and mitochondria, for H2S detection. Our probe comprised three functional domains (H2S sensing, fluorescence, and biomembrane penetration), could enter almost all cell types readily, and exhibit a rapid and ultrasensitive response to H2S (≤120-fold fluorescence enhancement) for the dynamic mapping of spontaneously produced H2S as well as its distribution in the whole cell. In particular, the probe traversed blood/tissue/cell barriers to achieve mapping of endogenous H2S in metabolic organs of a live Danio rerio (zebrafish). These results open-up exciting opportunities to investigate H2S physiology and H2S-related diseases.

Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping

PubMed Central

Treweek, Jennifer B; Deverman, Benjamin E; Greenbaum, Alon; Lignell, Antti; Xiao, Cheng; Cai, Long; Ladinsky, Mark S; Bjorkman, Pamela J; Fowlkes, Charless C; Gradinaru, Viviana

2016-01-01

To facilitate fine-scale phenotyping of whole specimens, we describe here a set of tissue fixation-embedding, detergent-clearing and staining protocols that can be used to transform excised organs and whole organisms into optically transparent samples within 1–2 weeks without compromising their cellular architecture or endogenous fluorescence. PACT (passive CLARITY technique) and PARS (perfusion-assisted agent release in situ) use tissue-hydrogel hybrids to stabilize tissue biomolecules during selective lipid extraction, resulting in enhanced clearing efficiency and sample integrity. Furthermore, the macromolecule permeability of PACT- and PARS-processed tissue hybrids supports the diffusion of immunolabels throughout intact tissue, whereas RIMS (refractive index matching solution) grants high-resolution imaging at depth by further reducing light scattering in cleared and uncleared samples alike. These methods are adaptable to difficult-to-image tissues, such as bone (PACT-deCAL), and to magnified single-cell visualization (ePACT). Together, these protocols and solutions enable phenotyping of subcellular components and tracing cellular connectivity in intact biological networks. PMID:26492141

Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM)☆

PubMed Central

Duke, Elizabeth M.H.; Razi, Minoo; Weston, Anne; Guttmann, Peter; Werner, Stephan; Henzler, Katja; Schneider, Gerd; Tooze, Sharon A.; Collinson, Lucy M.

2014-01-01

Cryo-soft X-ray tomography (cryo-SXT) is a powerful imaging technique that can extract ultrastructural information from whole, unstained mammalian cells as close to the living state as possible. Subcellular organelles including the nucleus, the Golgi apparatus and mitochondria have been identified by morphology alone, due to the similarity in contrast to transmission electron micrographs. In this study, we used cryo-SXT to image endosomes and autophagosomes, organelles that are particularly susceptible to chemical fixation artefacts during sample preparation for electron microscopy. We used two approaches to identify these compartments. For early and recycling endosomes, which are accessible to externally-loaded markers, we used an anti-transferrin receptor antibody conjugated to 10 nm gold particles. For autophagosomes, which are not accessible to externally-applied markers, we developed a correlative cryo-fluorescence and cryo-SXT workflow (cryo-CLXM) to localise GFP-LC3 and RFP-Atg9. We used a stand-alone cryo-fluorescence stage in the home laboratory to localise the cloned fluorophores, followed by cryo-soft X-ray tomography at the synchrotron to analyse cellular ultrastructure. We mapped the 3D ultrastructure of the endocytic and autophagic structures, and discovered clusters of omegasomes arising from ‘hotspots’ on the ER. Thus, immunogold markers and cryo-CLXM can be used to analyse cellular processes that are inaccessible using other imaging modalities. PMID:24238600

Geocoded data structures and their applications to Earth science investigations

NASA Technical Reports Server (NTRS)

Goldberg, M.

1984-01-01

A geocoded data structure is a means for digitally representing a geographically referenced map or image. The characteristics of representative cellular, linked, and hybrid geocoded data structures are reviewed. The data processing requirements of Earth science projects at the Goddard Space Flight Center and the basic tools of geographic data processing are described. Specific ways that new geocoded data structures can be used to adapt these tools to scientists' needs are presented. These include: expanding analysis and modeling capabilities; simplifying the merging of data sets from diverse sources; and saving computer storage space.

On-target labeling of intracellular metabolites combined with chemical mapping of individual hyphae revealing cytoplasmic relocation of isotopologues.

PubMed

Hu, Jie-Bi; Chen, Yu-Chie; Urban, Pawel L

2012-06-05

A microscale analytical platform integrating microbial cell culture, isotopic labeling, along with visual and mass spectrometric imaging with single-cell resolution has been developed and applied in the monitoring of cellular metabolism in fungal mycelium. The method implements open chips with a two-dimensional surface pattern composed of hydrophobic and hydrophilic zones. Two hydrophilic islands are used as medium reservoirs, while the hydrophobic area constitutes the support for the growing aerial hyphae, which do not have direct contact with the medium. The first island, containing (12)C(6)-glucose medium, was initially inoculated with the mycelium (Neurospora crassa), and following the initial incubation period, the hyphae progressed toward the second medium island, containing an isotopically labeled substrate ((13)C(6)-glucose). The (13)C atoms were gradually incorporated into cellular metabolites, which was revealed by MALDI-MS. The fate of the chitin-biosynthesis precursor, uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), was monitored by recording mass spectra with characteristic isotopic patterns, which indicated the presence of various (12)C/(13)C isotopologues. The method enabled mapping the (13)C-labeled UDP-GlcNAc in fungal mycelium and recording its redistribution in hyphae, directly on the chip.

Angle-resolved light scattering of individual rod-shaped bacteria based on Fourier transform light scattering

NASA Astrophysics Data System (ADS)

Jo, Youngju; Jung, Jaehwang; Lee, Jee Woong; Shin, Della; Park, Hyunjoo; Nam, Ki Tae; Park, Ji-Ho; Park, Yongkeun

2014-05-01

Two-dimensional angle-resolved light scattering maps of individual rod-shaped bacteria are measured at the single-cell level. Using quantitative phase imaging and Fourier transform light scattering techniques, the light scattering patterns of individual bacteria in four rod-shaped species (Bacillus subtilis, Lactobacillus casei, Synechococcus elongatus, and Escherichia coli) are measured with unprecedented sensitivity in a broad angular range from -70° to 70°. The measured light scattering patterns are analyzed along the two principal axes of rod-shaped bacteria in order to systematically investigate the species-specific characteristics of anisotropic light scattering. In addition, the cellular dry mass of individual bacteria is calculated and used to demonstrate that the cell-to-cell variations in light scattering within bacterial species is related to the cellular dry mass and growth.

Assessment of Different Strategies to Determine MAP-specific Cellular Immune Responses in Cattle

USDA-ARS?s Scientific Manuscript database

Assessment of cellular immunity in cattle against Mycobacterium avium ssp. paratuberculosis (MAP) by established methods remains unsatisfactory for diagnostic purposes. Recent studies conclude that analysis of T-cell subset responsiveness may improve diagnostic outcome. Aim of this study was to iden...

NeuroCa: integrated framework for systematic analysis of spatiotemporal neuronal activity patterns from large-scale optical recording data

PubMed Central

Jang, Min Jee; Nam, Yoonkey

2015-01-01

Abstract. Optical recording facilitates monitoring the activity of a large neural network at the cellular scale, but the analysis and interpretation of the collected data remain challenging. Here, we present a MATLAB-based toolbox, named NeuroCa, for the automated processing and quantitative analysis of large-scale calcium imaging data. Our tool includes several computational algorithms to extract the calcium spike trains of individual neurons from the calcium imaging data in an automatic fashion. Two algorithms were developed to decompose the imaging data into the activity of individual cells and subsequently detect calcium spikes from each neuronal signal. Applying our method to dense networks in dissociated cultures, we were able to obtain the calcium spike trains of ∼1000 neurons in a few minutes. Further analyses using these data permitted the quantification of neuronal responses to chemical stimuli as well as functional mapping of spatiotemporal patterns in neuronal firing within the spontaneous, synchronous activity of a large network. These results demonstrate that our method not only automates time-consuming, labor-intensive tasks in the analysis of neural data obtained using optical recording techniques but also provides a systematic way to visualize and quantify the collective dynamics of a network in terms of its cellular elements. PMID:26229973

Nanoscale imaging of whole cells using a liquid enclosure and a scanning transmission electron microscope.

PubMed

Peckys, Diana B; Veith, Gabriel M; Joy, David C; de Jonge, Niels

2009-12-14

Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory.

Hyperspectral imaging for simultaneous measurements of two FRET biosensors in pancreatic β-cells.

PubMed

Elliott, Amicia D; Bedard, Noah; Ustione, Alessandro; Baird, Michelle A; Davidson, Michael W; Tkaczyk, Tomasz; Piston, David W

2017-01-01

Fluorescent protein (FP) biosensors based on Förster resonance energy transfer (FRET) are commonly used to study molecular processes in living cells. There are FP-FRET biosensors for many cellular molecules, but it remains difficult to perform simultaneous measurements of multiple biosensors. The overlapping emission spectra of the commonly used FPs, including CFP/YFP and GFP/RFP make dual FRET measurements challenging. In addition, a snapshot imaging modality is required for simultaneous imaging. The Image Mapping Spectrometer (IMS) is a snapshot hyperspectral imaging system that collects high resolution spectral data and can be used to overcome these challenges. We have previously demonstrated the IMS's capabilities for simultaneously imaging GFP and CFP/YFP-based biosensors in pancreatic β-cells. Here, we demonstrate a further capability of the IMS to image simultaneously two FRET biosensors with a single excitation band, one for cAMP and the other for Caspase-3. We use these measurements to measure simultaneously cAMP signaling and Caspase-3 activation in pancreatic β-cells during oxidative stress and hyperglycemia, which are essential components in the pathology of diabetes.

Transplantation and tracking of human-induced pluripotent stem cells in a pig model of myocardial infarction: assessment of cell survival, engraftment, and distribution by hybrid single photon emission computed tomography/computed tomography of sodium iodide symporter transgene expression.

PubMed

Templin, Christian; Zweigerdt, Robert; Schwanke, Kristin; Olmer, Ruth; Ghadri, Jelena-Rima; Emmert, Maximilian Y; Müller, Ennio; Küest, Silke M; Cohrs, Susan; Schibli, Roger; Kronen, Peter; Hilbe, Monika; Reinisch, Andreas; Strunk, Dirk; Haverich, Axel; Hoerstrup, Simon; Lüscher, Thomas F; Kaufmann, Philipp A; Landmesser, Ulf; Martin, Ulrich

2012-07-24

Evaluation of novel cellular therapies in large-animal models and patients is currently hampered by the lack of imaging approaches that allow for long-term monitoring of viable transplanted cells. In this study, sodium iodide symporter (NIS) transgene imaging was evaluated as an approach to follow in vivo survival, engraftment, and distribution of human-induced pluripotent stem cell (hiPSC) derivatives in a pig model of myocardial infarction. Transgenic hiPSC lines stably expressing a fluorescent reporter and NIS (NIS(pos)-hiPSCs) were established. Iodide uptake, efflux, and viability of NIS(pos)-hiPSCs were assessed in vitro. Ten (±2) days after induction of myocardial infarction by transient occlusion of the left anterior descending artery, catheter-based intramyocardial injection of NIS(pos)-hiPSCs guided by 3-dimensional NOGA mapping was performed. Dual-isotope single photon emission computed tomographic/computed tomographic imaging was applied with the use of (123)I to follow donor cell survival and distribution and with the use of (99m)TC-tetrofosmin for perfusion imaging. In vitro, iodide uptake in NIS(pos)-hiPSCs was increased 100-fold above that of nontransgenic controls. In vivo, viable NIS(pos)-hiPSCs could be visualized for up to 15 weeks. Immunohistochemistry demonstrated that hiPSC-derived endothelial cells contributed to vascularization. Up to 12 to 15 weeks after transplantation, no teratomas were detected. This study describes for the first time the feasibility of repeated long-term in vivo imaging of viability and tissue distribution of cellular grafts in large animals. Moreover, this is the first report demonstrating vascular differentiation and long-term engraftment of hiPSCs in a large-animal model of myocardial infarction. NIS(pos)-hiPSCs represent a valuable tool to monitor and improve current cellular treatment strategies in clinically relevant animal models.

Whole Tumor Histogram-profiling of Diffusion-Weighted Magnetic Resonance Images Reflects Tumorbiological Features of Primary Central Nervous System Lymphoma.

PubMed

Schob, Stefan; Münch, Benno; Dieckow, Julia; Quäschling, Ulf; Hoffmann, Karl-Titus; Richter, Cindy; Garnov, Nikita; Frydrychowicz, Clara; Krause, Matthias; Meyer, Hans-Jonas; Surov, Alexey

2018-04-01

Diffusion weighted imaging (DWI) quantifies motion of hydrogen nuclei in biological tissues and hereby has been used to assess the underlying tissue microarchitecture. Histogram-profiling of DWI provides more detailed information on diffusion characteristics of a lesion than the standardly calculated values of the apparent diffusion coefficient (ADC)-minimum, mean and maximum. Hence, the aim of our study was to investigate, which parameters of histogram-profiling of DWI in primary central nervous system lymphoma can be used to specifically predict features like cellular density, chromatin content and proliferative activity. Pre-treatment ADC maps of 21 PCNSL patients (8 female, 13 male, 28-89 years) from a 1.5T system were used for Matlab-based histogram profiling. Results of histopathology (H&E staining) and immunohistochemistry (Ki-67 expression) were quantified. Correlations between histogram-profiling parameters and neuropathologic examination were calculated using SPSS 23.0. The lower percentiles (p10 and p25) showed significant correlations with structural parameters of the neuropathologic examination (cellular density, chromatin content). The highest percentile, p90, correlated significantly with Ki-67 expression, resembling proliferative activity. Kurtosis of the ADC histogram correlated significantly with cellular density. Histogram-profiling of DWI in PCNSL provides a comprehensible set of parameters, which reflect distinct tumor-architectural and tumor-biological features, and hence, are promising biomarkers for treatment response and prognosis. Copyright © 2018. Published by Elsevier Inc.

Spatiotemporal Patterns of Tumor Occurrence in Children with Intraocular Retinoblastoma.

PubMed

King, Benjamin A; Parra, Carlos; Li, Yimei; Helton, Kathleen J; Qaddoumi, Ibrahim; Wilson, Matthew W; Ogg, Robert J

2015-01-01

To accurately map the retinal area covered by tumor in a prospectively enrolled cohort of children diagnosed with retinoblastoma. Orbital MRI in 106 consecutive retinoblastoma patients (44 bilateral) was analyzed. For MRI-visible tumors, the polar angle and angle of eccentricity of points defining tumor perimeter on the retina were determined by triangulation from images in three orthogonal planes. The centroid of the mapped area was calculated to approximate tumor origin, and the location and cumulative tumor burden were analyzed in relation to mutation type (germline vs. somatic), tumor area, and patient age at diagnosis. Location of small tumors undetected by MRI was approximated with fundoscopic images. Mapping was successful for 129 tumors in 91 eyes from 67 patients (39 bilateral, 43 germline mutation). Cumulative tumor burden was highest within the macula and posterior pole and was asymmetrically higher within the inferonasal periphery. Tumor incidence was lowest in the superotemporal periphery. Tumor location varied with age at diagnosis in a complex pattern. Tumor location was concentrated in the macula and superonasal periphery in patients <5.6 months, in the inferotemporal quadrant of the posterior pole in patients 5.6-8.8 months, in the inferonasal quadrant in patients 8.8-13.2 months, and in the nasal and superotemporal periphery in patients >13.2 months. The distribution of MRI-invisible tumors was consistent with the asymmetry of mapped tumors. MRI-based mapping revealed a previously unrecognized pattern of retinoblastoma localization that evolves with age at diagnosis. The structured spatiotemporal distribution of tumors may provide valuable clues about cellular or molecular events associated with tumorigenesis in the developing retina.

Uses, misuses, new uses and fundamental limitations of magnetic resonance imaging in cognitive science

PubMed Central

2016-01-01

When blood oxygenation level-dependent (BOLD) contrast functional magnetic resonance imaging (fMRI) was discovered in the early 1990s, it provoked an explosion of interest in exploring human cognition, using brain mapping techniques based on MRI. Standards for data acquisition and analysis were rapidly put in place, in order to assist comparison of results across laboratories. Recently, MRI data acquisition capabilities have improved dramatically, inviting a rethink of strategies for relating functional brain activity at the systems level with its neuronal substrates and functional connections. This paper reviews the established capabilities of BOLD contrast fMRI, the perceived weaknesses of major methods of analysis, and current results that may provide insights into improved brain modelling. These results have inspired the use of in vivo myeloarchitecture for localizing brain activity, individual subject analysis without spatial smoothing and mapping of changes in cerebral blood volume instead of BOLD activation changes. The apparent fundamental limitations of all methods based on nuclear magnetic resonance are also discussed. This article is part of the themed issue ‘Interpreting BOLD: a dialogue between cognitive and cellular neuroscience’. PMID:27574303

Uses, misuses, new uses and fundamental limitations of magnetic resonance imaging in cognitive science.

PubMed

Turner, Robert

2016-10-05

When blood oxygenation level-dependent (BOLD) contrast functional magnetic resonance imaging (fMRI) was discovered in the early 1990s, it provoked an explosion of interest in exploring human cognition, using brain mapping techniques based on MRI. Standards for data acquisition and analysis were rapidly put in place, in order to assist comparison of results across laboratories. Recently, MRI data acquisition capabilities have improved dramatically, inviting a rethink of strategies for relating functional brain activity at the systems level with its neuronal substrates and functional connections. This paper reviews the established capabilities of BOLD contrast fMRI, the perceived weaknesses of major methods of analysis, and current results that may provide insights into improved brain modelling. These results have inspired the use of in vivo myeloarchitecture for localizing brain activity, individual subject analysis without spatial smoothing and mapping of changes in cerebral blood volume instead of BOLD activation changes. The apparent fundamental limitations of all methods based on nuclear magnetic resonance are also discussed.This article is part of the themed issue 'Interpreting BOLD: a dialogue between cognitive and cellular neuroscience'. © 2016 The Authors.

Redox Indicator Mice Stably Expressing Genetically Encoded Neuronal roGFP: Versatile Tools to Decipher Subcellular Redox Dynamics in Neuropathophysiology.

PubMed

Wagener, Kerstin C; Kolbrink, Benedikt; Dietrich, Katharina; Kizina, Kathrin M; Terwitte, Lukas S; Kempkes, Belinda; Bao, Guobin; Müller, Michael

2016-07-01

Reactive oxygen species (ROS) and downstream redox alterations not only mediate physiological signaling but also neuropathology. For long, ROS/redox imaging was hampered by a lack of reliable probes. Genetically encoded redox sensors overcame this gap and revolutionized (sub)cellular redox imaging. Yet, the successful delivery of sensor-coding DNA, which demands transfection/transduction of cultured preparations or stereotaxic microinjections of each subject, remains challenging. By generating transgenic mice, we aimed to overcome limiting cultured preparations, circumvent surgical interventions, and to extend effectively redox imaging to complex and adult preparations. Our redox indicator mice widely express Thy1-driven roGFP1 (reduction-oxidation-sensitive green fluorescent protein 1) in neuronal cytosol or mitochondria. Negative phenotypic effects of roGFP1 were excluded and its proper targeting and functionality confirmed. Redox mapping by ratiometric wide-field imaging reveals most oxidizing conditions in CA3 neurons. Furthermore, mitochondria are more oxidized than cytosol. Cytosolic and mitochondrial roGFP1s reliably report cell endogenous redox dynamics upon metabolic challenge or stimulation. Fluorescence lifetime imaging yields stable, but marginal, response ranges. We therefore developed automated excitation ratiometric 2-photon imaging. It offers superior sensitivity, spatial resolution, and response dynamics. Redox indicator mice enable quantitative analyses of subcellular redox dynamics in a multitude of preparations and at all postnatal stages. This will uncover cell- and compartment-specific cerebral redox signals and their defined alterations during development, maturation, and aging. Cross-breeding with other disease models will reveal molecular details on compartmental redox homeostasis in neuropathology. Combined with ratiometric 2-photon imaging, this will foster our mechanistic understanding of cellular redox signals in their full complexity. Antioxid. Redox Signal. 25, 41-58.

Redox Indicator Mice Stably Expressing Genetically Encoded Neuronal roGFP: Versatile Tools to Decipher Subcellular Redox Dynamics in Neuropathophysiology

PubMed Central

Wagener, Kerstin C.; Kolbrink, Benedikt; Dietrich, Katharina; Kizina, Kathrin M.; Terwitte, Lukas S.; Kempkes, Belinda; Bao, Guobin

2016-01-01

Abstract Aims: Reactive oxygen species (ROS) and downstream redox alterations not only mediate physiological signaling but also neuropathology. For long, ROS/redox imaging was hampered by a lack of reliable probes. Genetically encoded redox sensors overcame this gap and revolutionized (sub)cellular redox imaging. Yet, the successful delivery of sensor-coding DNA, which demands transfection/transduction of cultured preparations or stereotaxic microinjections of each subject, remains challenging. By generating transgenic mice, we aimed to overcome limiting cultured preparations, circumvent surgical interventions, and to extend effectively redox imaging to complex and adult preparations. Results: Our redox indicator mice widely express Thy1-driven roGFP1 (reduction–oxidation-sensitive green fluorescent protein 1) in neuronal cytosol or mitochondria. Negative phenotypic effects of roGFP1 were excluded and its proper targeting and functionality confirmed. Redox mapping by ratiometric wide-field imaging reveals most oxidizing conditions in CA3 neurons. Furthermore, mitochondria are more oxidized than cytosol. Cytosolic and mitochondrial roGFP1s reliably report cell endogenous redox dynamics upon metabolic challenge or stimulation. Fluorescence lifetime imaging yields stable, but marginal, response ranges. We therefore developed automated excitation ratiometric 2-photon imaging. It offers superior sensitivity, spatial resolution, and response dynamics. Innovation and Conclusion: Redox indicator mice enable quantitative analyses of subcellular redox dynamics in a multitude of preparations and at all postnatal stages. This will uncover cell- and compartment-specific cerebral redox signals and their defined alterations during development, maturation, and aging. Cross-breeding with other disease models will reveal molecular details on compartmental redox homeostasis in neuropathology. Combined with ratiometric 2-photon imaging, this will foster our mechanistic understanding of cellular redox signals in their full complexity. Antioxid. Redox Signal. 25, 41–58. PMID:27059697

Microtubule Severing Stymied by Free Tubulin

NASA Astrophysics Data System (ADS)

Ross, Jennifer; Bailey, Megan

2015-03-01

Proper organization of the microtubule cytoskeletal network is required to perform many necessary cellular functions including mitosis, cell development, and cell motility. Network organization is achieved through filament remodeling by microtubule-associated proteins (MAPs) that control microtubule dynamics. MAPs that stabilize are relatively well understood, while less is known about destabilizing MAPs, such as severing enzymes. Katanin, the first-discovered microtubule-severing enzyme, is a AAA + enzyme that oligomerizes into hexamers and uses ATP hydrolysis to sever microtubules. Using quantitative fluorescence imaging on reconstituted microtubule severing assays in vitro we investigate how katanin can regulate microtubule dynamics. Interestingly, we find microtubule dynamics inhibits katanin severing activity; dynamic microtubules are not severed. Using systematic experiments introducing free tubulin into the assays we find that free tubulin can compete for microtubule filaments for the katanin proteins. Our work indicates that katanin could function best on stabile microtubules or stabile regions of microtubules in cells in regions where free tubulin is sequesters, low, or depleted.

A method for quantitative analysis of clump thickness in cervical cytology slides.

PubMed

Fan, Yilun; Bradley, Andrew P

2016-01-01

Knowledge of the spatial distribution and thickness of cytology specimens is critical to the development of digital slide acquisition techniques that minimise both scan times and image file size. In this paper, we evaluate a novel method to achieve this goal utilising an exhaustive high-resolution scan, an over-complete wavelet transform across multi-focal planes and a clump segmentation of all cellular materials on the slide. The method is demonstrated with a quantitative analysis of ten normal, but difficult to scan Pap stained, Thin-prep, cervical cytology slides. We show that with this method the top and bottom of the specimen can be estimated to an accuracy of 1 μm in 88% and 97% of the fields of view respectively. Overall, cellular material can be over 30 μm thick and the distribution of cells is skewed towards the cover-slip (top of the slide). However, the median clump thickness is 10 μm and only 31% of clumps contain more than three nuclei. Therefore, by finding a focal map of the specimen the number of 1 μm spaced focal planes that are required to be scanned to acquire 95% of the in-focus material can be reduced from 25.4 to 21.4 on average. In addition, we show that by considering the thickness of the specimen, an improved focal map can be produced which further reduces the required number of 1 μm spaced focal planes to 18.6. This has the potential to reduce scan times and raw image data by over 25%. Copyright © 2015 Elsevier Ltd. All rights reserved.

Quantitative fluorescence imaging of protein diffusion and interaction in living cells.

PubMed

Capoulade, Jérémie; Wachsmuth, Malte; Hufnagel, Lars; Knop, Michael

2011-08-07

Diffusion processes and local dynamic equilibria inside cells lead to nonuniform spatial distributions of molecules, which are essential for processes such as nuclear organization and signaling in cell division, differentiation and migration. To understand these mechanisms, spatially resolved quantitative measurements of protein abundance, mobilities and interactions are needed, but current methods have limited capabilities to study dynamic parameters. Here we describe a microscope based on light-sheet illumination that allows massively parallel fluorescence correlation spectroscopy (FCS) measurements and use it to visualize the diffusion and interactions of proteins in mammalian cells and in isolated fly tissue. Imaging the mobility of heterochromatin protein HP1α (ref. 4) in cell nuclei we could provide high-resolution diffusion maps that reveal euchromatin areas with heterochromatin-like HP1α-chromatin interactions. We expect that FCS imaging will become a useful method for the precise characterization of cellular reaction-diffusion processes.

Confocal fluorescence microscopy for rapid evaluation of invasive tumor cellularity of inflammatory breast carcinoma core needle biopsies.

PubMed

Dobbs, Jessica; Krishnamurthy, Savitri; Kyrish, Matthew; Benveniste, Ana Paula; Yang, Wei; Richards-Kortum, Rebecca

2015-01-01

Tissue sampling is a problematic issue for inflammatory breast carcinoma, and immediate evaluation following core needle biopsy is needed to evaluate specimen adequacy. We sought to determine if confocal fluorescence microscopy provides sufficient resolution to evaluate specimen adequacy by comparing invasive tumor cellularity estimated from standard histologic images to invasive tumor cellularity estimated from confocal images of breast core needle biopsy specimens. Grayscale confocal fluorescence images of breast core needle biopsy specimens were acquired following proflavine application. A breast-dedicated pathologist evaluated invasive tumor cellularity in histologic images with hematoxylin and eosin staining and in grayscale and false-colored confocal images of cores. Agreement between cellularity estimates was quantified using a kappa coefficient. 23 cores from 23 patients with suspected inflammatory breast carcinoma were imaged. Confocal images were acquired in an average of less than 2 min per core. Invasive tumor cellularity estimated from histologic and grayscale confocal images showed moderate agreement by kappa coefficient: κ = 0.48 ± 0.09 (p < 0.001). Grayscale confocal images require less than 2 min for acquisition and allow for evaluation of invasive tumor cellularity in breast core needle biopsy specimens with moderate agreement to histologic images. We show that confocal fluorescence microscopy can be performed immediately following specimen acquisition and could indicate the need for additional biopsies at the initial visit.

Alterations of the cerebellum and basal ganglia in bipolar disorder mood states detected by quantitative T1ρ mapping.

PubMed

Johnson, Casey P; Christensen, Gary E; Fiedorowicz, Jess G; Mani, Merry; Shaffer, Joseph J; Magnotta, Vincent A; Wemmie, John A

2018-06-01

Quantitative mapping of T1 relaxation in the rotating frame (T1ρ) is a magnetic resonance imaging technique sensitive to pH and other cellular and microstructural factors, and is a potentially valuable tool for identifying brain alterations in bipolar disorder. Recently, this technique identified differences in the cerebellum and cerebral white matter of euthymic patients vs healthy controls that were consistent with reduced pH in these regions, suggesting an underlying metabolic abnormality. The current study built upon this prior work to investigate brain T1ρ differences across euthymic, depressed, and manic mood states of bipolar disorder. Forty participants with bipolar I disorder and 29 healthy control participants matched for age and gender were enrolled. Participants with bipolar disorder were imaged in one or more mood states, yielding 27, 12, and 13 imaging sessions in euthymic, depressed, and manic mood states, respectively. Three-dimensional, whole-brain anatomical images and T1ρ maps were acquired for all participants, enabling voxel-wise evaluation of T1ρ differences between bipolar mood state and healthy control groups. All three mood state groups had increased T1ρ relaxation times in the cerebellum compared to the healthy control group. Additionally, the depressed and manic groups had reduced T1ρ relaxation times in and around the basal ganglia compared to the control and euthymic groups. The study implicated the cerebellum and basal ganglia in the pathophysiology of bipolar disorder and its mood states, the roles of which are relatively unexplored. These findings motivate further investigation of the underlying cause of the abnormalities, and the potential role of altered metabolic activity in these regions. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Genetic Complexity and Quantitative Trait Loci Mapping of Yeast Morphological Traits

PubMed Central

Nogami, Satoru; Ohya, Yoshikazu; Yvert, Gaël

2007-01-01

Functional genomics relies on two essential parameters: the sensitivity of phenotypic measures and the power to detect genomic perturbations that cause phenotypic variations. In model organisms, two types of perturbations are widely used. Artificial mutations can be introduced in virtually any gene and allow the systematic analysis of gene function via mutants fitness. Alternatively, natural genetic variations can be associated to particular phenotypes via genetic mapping. However, the access to genome manipulation and breeding provided by model organisms is sometimes counterbalanced by phenotyping limitations. Here we investigated the natural genetic diversity of Saccharomyces cerevisiae cellular morphology using a very sensitive high-throughput imaging platform. We quantified 501 morphological parameters in over 50,000 yeast cells from a cross between two wild-type divergent backgrounds. Extensive morphological differences were found between these backgrounds. The genetic architecture of the traits was complex, with evidence of both epistasis and transgressive segregation. We mapped quantitative trait loci (QTL) for 67 traits and discovered 364 correlations between traits segregation and inheritance of gene expression levels. We validated one QTL by the replacement of a single base in the genome. This study illustrates the natural diversity and complexity of cellular traits among natural yeast strains and provides an ideal framework for a genetical genomics dissection of multiple traits. Our results did not overlap with results previously obtained from systematic deletion strains, showing that both approaches are necessary for the functional exploration of genomes. PMID:17319748

Draft De Novo Transcriptome of the Rat Kangaroo Potorous tridactylus as a Tool for Cell Biology

PubMed Central

Udy, Dylan B.; Voorhies, Mark; Chan, Patricia P.; Lowe, Todd M.; Dumont, Sophie

2015-01-01

The rat kangaroo (long-nosed potoroo, Potorous tridactylus) is a marsupial native to Australia. Cultured rat kangaroo kidney epithelial cells (PtK) are commonly used to study cell biological processes. These mammalian cells are large, adherent, and flat, and contain large and few chromosomes—and are thus ideal for imaging intra-cellular dynamics such as those of mitosis. Despite this, neither the rat kangaroo genome nor transcriptome have been sequenced, creating a challenge for probing the molecular basis of these cellular dynamics. Here, we present the sequencing, assembly and annotation of the draft rat kangaroo de novo transcriptome. We sequenced 679 million reads that mapped to 347,323 Trinity transcripts and 20,079 Unigenes. We present statistics emerging from transcriptome-wide analyses, and analyses suggesting that the transcriptome covers full-length sequences of most genes, many with multiple isoforms. We also validate our findings with a proof-of-concept gene knockdown experiment. We expect that this high quality transcriptome will make rat kangaroo cells a more tractable system for linking molecular-scale function and cellular-scale dynamics. PMID:26252667

Draft De Novo Transcriptome of the Rat Kangaroo Potorous tridactylus as a Tool for Cell Biology.

PubMed

Udy, Dylan B; Voorhies, Mark; Chan, Patricia P; Lowe, Todd M; Dumont, Sophie

2015-01-01

The rat kangaroo (long-nosed potoroo, Potorous tridactylus) is a marsupial native to Australia. Cultured rat kangaroo kidney epithelial cells (PtK) are commonly used to study cell biological processes. These mammalian cells are large, adherent, and flat, and contain large and few chromosomes-and are thus ideal for imaging intra-cellular dynamics such as those of mitosis. Despite this, neither the rat kangaroo genome nor transcriptome have been sequenced, creating a challenge for probing the molecular basis of these cellular dynamics. Here, we present the sequencing, assembly and annotation of the draft rat kangaroo de novo transcriptome. We sequenced 679 million reads that mapped to 347,323 Trinity transcripts and 20,079 Unigenes. We present statistics emerging from transcriptome-wide analyses, and analyses suggesting that the transcriptome covers full-length sequences of most genes, many with multiple isoforms. We also validate our findings with a proof-of-concept gene knockdown experiment. We expect that this high quality transcriptome will make rat kangaroo cells a more tractable system for linking molecular-scale function and cellular-scale dynamics.

Cellular Imaging With MRI.

PubMed

Makela, Ashley V; Murrell, Donna H; Parkins, Katie M; Kara, Jenna; Gaudet, Jeffrey M; Foster, Paula J

2016-10-01

Cellular magnetic resonance imaging (MRI) is an evolving field of imaging with strong translational and research potential. The ability to detect, track, and quantify cells in vivo and over time allows for studying cellular events related to disease processes and may be used as a biomarker for decisions about treatments and for monitoring responses to treatments. In this review, we discuss methods for labeling cells, various applications for cellular MRI, the existing limitations, strategies to address these shortcomings, and clinical cellular MRI.

Multimodal nonlinear optical imaging of obesity-induced liver steatosis and fibrosis

NASA Astrophysics Data System (ADS)

Lin, Jian; Lu, Fake; Zheng, Wei; Tai, Dean C. S.; Yu, Hanry; Sheppard, Colin; Huang, Zhiwei

2011-03-01

Liver steatosis/fibrosis represents the major conditions and symptoms for many liver diseases. Nonlinear optical microscopy has emerged as a powerful tool for label-free tissue imaging with high sensitivity and chemical specificity for several typical biochemical compounds. Three nonlinear microscopy imaging modalities are implemented on the sectioned tissues from diseased livers induced by high fat diet (HFD). Coherent anti-Stokes Raman scattering (CARS) imaging visualizes and quantifies the lipid droplets accumulated in the liver, Second harmonic generation (SHG) is used to map the distribution of aggregated collagen fibers, and two-photon excitation fluorescence (TPEF) reveals the morphology of hepatic cells based on the autofluorescence signals from NADH and flavins within the hepatocytes. Our results demonstrate that obesity induces liver steatosis in the beginning stage, which may progress into liver fibrosis with high risk. There is a certain correlation between liver steatosis and fibrosis. This study may provide new insights into the understanding of the mechanisms of steatosis/fibrosis transformations at the cellular and molecular levels.

Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data.

PubMed

Shelton, Elijah; Serwane, Friedhelm; Campàs, Otger

2018-03-01

Modern fluorescence microscopy enables fast 3D imaging of biological and inert systems alike. In many studies, it is important to detect the surface of objects and quantitatively characterize its local geometry, including its mean curvature. We present a fully automated algorithm to determine the location and curvatures of an object from 3D fluorescence images, such as those obtained using confocal or light-sheet microscopy. The algorithm aims at reconstructing surface labelled objects with spherical topology and mild deformations from the spherical geometry with high accuracy, rather than reconstructing arbitrarily deformed objects with lower fidelity. Using both synthetic data with known geometrical characteristics and experimental data of spherical objects, we characterize the algorithm's accuracy over the range of conditions and parameters typically encountered in 3D fluorescence imaging. We show that the algorithm can detect the location of the surface and obtain a map of local mean curvatures with relative errors typically below 2% and 20%, respectively, even in the presence of substantial levels of noise. Finally, we apply this algorithm to analyse the shape and curvature map of fluorescently labelled oil droplets embedded within multicellular aggregates and deformed by cellular forces. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Spatial reconstruction of single-cell gene expression data.

PubMed

Satija, Rahul; Farrell, Jeffrey A; Gennert, David; Schier, Alexander F; Regev, Aviv

2015-05-01

Spatial localization is a key determinant of cellular fate and behavior, but methods for spatially resolved, transcriptome-wide gene expression profiling across complex tissues are lacking. RNA staining methods assay only a small number of transcripts, whereas single-cell RNA-seq, which measures global gene expression, separates cells from their native spatial context. Here we present Seurat, a computational strategy to infer cellular localization by integrating single-cell RNA-seq data with in situ RNA patterns. We applied Seurat to spatially map 851 single cells from dissociated zebrafish (Danio rerio) embryos and generated a transcriptome-wide map of spatial patterning. We confirmed Seurat's accuracy using several experimental approaches, then used the strategy to identify a set of archetypal expression patterns and spatial markers. Seurat correctly localizes rare subpopulations, accurately mapping both spatially restricted and scattered groups. Seurat will be applicable to mapping cellular localization within complex patterned tissues in diverse systems.

Advances in molecular labeling, high throughput imaging and machine intelligence portend powerful functional cellular biochemistry tools.

PubMed

Price, Jeffrey H; Goodacre, Angela; Hahn, Klaus; Hodgson, Louis; Hunter, Edward A; Krajewski, Stanislaw; Murphy, Robert F; Rabinovich, Andrew; Reed, John C; Heynen, Susanne

2002-01-01

Cellular behavior is complex. Successfully understanding systems at ever-increasing complexity is fundamental to advances in modern science and unraveling the functional details of cellular behavior is no exception. We present a collection of prospectives to provide a glimpse of the techniques that will aid in collecting, managing and utilizing information on complex cellular processes via molecular imaging tools. These include: 1) visualizing intracellular protein activity with fluorescent markers, 2) high throughput (and automated) imaging of multilabeled cells in statistically significant numbers, and 3) machine intelligence to analyze subcellular image localization and pattern. Although not addressed here, the importance of combining cell-image-based information with detailed molecular structure and ligand-receptor binding models cannot be overlooked. Advanced molecular imaging techniques have the potential to impact cellular diagnostics for cancer screening, clinical correlations of tissue molecular patterns for cancer biology, and cellular molecular interactions for accelerating drug discovery. The goal of finally understanding all cellular components and behaviors will be achieved by advances in both instrumentation engineering (software and hardware) and molecular biochemistry. Copyright 2002 Wiley-Liss, Inc.

Using cellular automata to generate image representation for biological sequences.

PubMed

Xiao, X; Shao, S; Ding, Y; Huang, Z; Chen, X; Chou, K-C

2005-02-01

A novel approach to visualize biological sequences is developed based on cellular automata (Wolfram, S. Nature 1984, 311, 419-424), a set of discrete dynamical systems in which space and time are discrete. By transforming the symbolic sequence codes into the digital codes, and using some optimal space-time evolvement rules of cellular automata, a biological sequence can be represented by a unique image, the so-called cellular automata image. Many important features, which are originally hidden in a long and complicated biological sequence, can be clearly revealed thru its cellular automata image. With biological sequences entering into databanks rapidly increasing in the post-genomic era, it is anticipated that the cellular automata image will become a very useful vehicle for investigation into their key features, identification of their function, as well as revelation of their "fingerprint". It is anticipated that by using the concept of the pseudo amino acid composition (Chou, K.C. Proteins: Structure, Function, and Genetics, 2001, 43, 246-255), the cellular automata image approach can also be used to improve the quality of predicting protein attributes, such as structural class and subcellular location.

Systematic, spatial imaging of large multimolecular assemblies and the emerging principles of supramolecular order in biological systems

PubMed Central

Schubert, Walter

2013-01-01

Understanding biological systems at the level of their relational (emergent) molecular properties in functional protein networks relies on imaging methods, able to spatially resolve a tissue or a cell as a giant, non-random, topologically defined collection of interacting supermolecules executing myriads of subcellular mechanisms. Here, the development and findings of parameter-unlimited functional super-resolution microscopy are described—a technology based on the fluorescence imaging cycler (IC) principle capable of co-mapping thousands of distinct biomolecular assemblies at high spatial resolution and differentiation (<40 nm distances). It is shown that the subcellular and transcellular features of such supermolecules can be described at the compositional and constitutional levels; that the spatial connection, relational stoichiometry, and topology of supermolecules generate hitherto unrecognized functional self-segmentation of biological tissues; that hierarchical features, common to thousands of simultaneously imaged supermolecules, can be identified; and how the resulting supramolecular order relates to spatial coding of cellular functionalities in biological systems. A large body of observations with IC molecular systems microscopy collected over 20 years have disclosed principles governed by a law of supramolecular segregation of cellular functionalities. This pervades phenomena, such as exceptional orderliness, functional selectivity, combinatorial and spatial periodicity, and hierarchical organization of large molecular systems, across all species investigated so far. This insight is based on the high degree of specificity, selectivity, and sensitivity of molecular recognition processes for fluorescence imaging beyond the spectral resolution limit, using probe libraries controlled by ICs. © 2013 The Authors. Journal of Molecular Recognition published by John Wiley & Sons, Ltd. PMID:24375580

Oxygen sensing PLIM together with FLIM of intrinsic cellular fluorophores for metabolic mapping

NASA Astrophysics Data System (ADS)

Kalinina, Sviatlana; Schaefer, Patrick; Breymayer, Jasmin; Bisinger, Dominik; Chakrabortty, Sabyasachi; Rueck, Angelika

2018-02-01

Otical imaging techniques based on time correlated single photon counting (TCSPC) has found wide applications in medicine and biology. Non-invasive and information-rich fluorescence lifetime imaging microscopy (FLIM) is successfully used for monitoring fluorescent intrinsic metabolic coenzymes as NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) and FAD+ (flavin adenine dinucleotide) in living cells and tissues. The ratio between proteinbound and free coenzymes gives an information about the balance between oxidative phosphorylation and glycolysis in the cells. The changes of the ratio reflects major cellular disorders. A correlation exists between metabolic activity, redox ratio and fluorescence lifetime during stem cell differentiation, neurodegenerative diseases, and carcinogenesis. A multichannel FLIM detection system was designed for monitoring the redox state of NAD(P)H and FAD+ and other intrinsic fluorophores as protoporphyrin IX. In addition, the unique upgrade is useful to perform FLIM and PLIM (phosphorescence lifetime imaging microscopy) simultaneously. PLIM is a promising method to investigate oxygen sensing in biomedical samples. In detail, the oxygen-dependent quenching of phosphorescence of some compounds as transition metal complexes enables measuring of oxygen partial pressure (pO2). Using a two-channel FLIM/PLIM system we monitored intrinsic pO2 by PLIM simultaneously with NAD(P)H by FLIM providing complex metabolic and redox imaging of living cells. Physico-chemical properties of oxygen sensitive probes define certain parameters including their localisation. We present results of some ruthenium based complexes including those specifically bound to mitochondria.

Imaging of intracellular fatty acids by scanning X-ray fluorescence microscopy

PubMed Central

Shimura, Mari; Shindou, Hideo; Szyrwiel, Lukasz; Tokuoka, Suzumi M.; Hamano, Fumie; Matsuyama, Satoshi; Okamoto, Mayumi; Matsunaga, Akihiro; Kita, Yoshihiro; Ishizaka, Yukihito; Yamauchi, Kazuto; Kohmura, Yoshiki; Lobinski, Ryszard; Shimizu, Isao; Shimizu, Takao

2016-01-01

Fatty acids are taken up by cells and incorporated into complex lipids such as neutral lipids and glycerophospholipids. Glycerophospholipids are major constituents of cellular membranes. More than 1000 molecular species of glycerophospholipids differ in their polar head groups and fatty acid compositions. They are related to cellular functions and diseases and have been well analyzed by mass spectrometry. However, intracellular imaging of fatty acids and glycerophospholipids has not been successful due to insufficient resolution using conventional methods. Here, we developed a method for labeling fatty acids with bromine (Br) and applied scanning X-ray fluorescence microscopy (SXFM) to obtain intracellular Br mapping data with submicrometer resolution. Mass spectrometry showed that cells took up Br-labeled fatty acids and metabolized them mainly into glycerophospholipids in CHO cells. Most Br signals observed by SXFM were in the perinuclear region. Higher resolution revealed a spot-like distribution of Br in the cytoplasm. The current method enabled successful visualization of intracellular Br-labeled fatty acids. Single-element labeling combined with SXFM technology facilitates the intracellular imaging of fatty acids, which provides a new tool to determine dynamic changes in fatty acids and their derivatives at the single-cell level.—Shimura, M., Shindou, H., Szyrwiel, L., Tokuoka, S. M., Hamano, F., Matsuyama, S., Okamoto, M., Matsunaga, A., Kita, Y., Ishizaka, Y., Yamauchi, K., Kohmura, Y., Lobinski, R., Shimizu, I., Shimizu, T. Imaging of intracellular fatty acids by scanning X-ray fluorescence microscopy. PMID:27601443

Planar implantable sensor for in vivo measurement of cellular oxygen metabolism in brain tissue.

PubMed

Tsytsarev, Vassiliy; Akkentli, Fatih; Pumbo, Elena; Tang, Qinggong; Chen, Yu; Erzurumlu, Reha S; Papkovsky, Dmitri B

2017-04-01

Brain imaging methods are continually improving. Imaging of the cerebral cortex is widely used in both animal experiments and charting human brain function in health and disease. Among the animal models, the rodent cerebral cortex has been widely used because of patterned neural representation of the whiskers on the snout and relative ease of activating cortical tissue with whisker stimulation. We tested a new planar solid-state oxygen sensor comprising a polymeric film with a phosphorescent oxygen-sensitive coating on the working side, to monitor dynamics of oxygen metabolism in the cerebral cortex following sensory stimulation. Sensory stimulation led to changes in oxygenation and deoxygenation processes of activated areas in the barrel cortex. We demonstrate the possibility of dynamic mapping of relative changes in oxygenation in live mouse brain tissue with such a sensor. Oxygenation-based functional magnetic resonance imaging (fMRI) is very effective method for functional brain mapping but have high costs and limited spatial resolution. Optical imaging of intrinsic signal (IOS) does not provide the required sensitivity, and voltage-sensitive dye optical imaging (VSDi) has limited applicability due to significant toxicity of the voltage-sensitive dye. Our planar solid-state oxygen sensor imaging approach circumvents these limitations, providing a simple optical contrast agent with low toxicity and rapid application. The planar solid-state oxygen sensor described here can be used as a tool in visualization and real-time analysis of sensory-evoked neural activity in vivo. Further, this approach allows visualization of local neural activity with high temporal and spatial resolution. Copyright © 2017 Elsevier B.V. All rights reserved.

SU-E-J-217: Multiparametric MR Imaging of Cranial Tumors On a Dedicated 1.0T MR Simulator Prior to Stereotactic Radiosurgery

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wen, N; Glide-Hurst, C; Liu, M

Purpose: Quantitative magnetic resonance imaging (MRI) of cranial lesions prior to stereotactic radiosurgery (SRS) may improve treatment planning and provide potential prognostic value. The practicality and logistics of acquiring advanced multiparametric MRI sequences to measure vascular and cellular properties of cerebral tumors are explored on a 1.0 Tesla MR Simulator. Methods: MR simulation was performed immediately following routine CT simulation on a 1T MR Simulator. MR sequences used were in the order they were performed: T2-Weighted Turbo Spin Echo (T2W-TSE), T2 FLAIR, Diffusion-weighted (DWI, b = 0, 800 to generate an apparent diffusion coefficient (ADC) map), 3D T1-Weighted Fast Fieldmore » Echo (T1W-FFE), Dynamic Contrast Enhanced (DCE) and Post Gadolinium Contrast Enhanced 3D T1W-FFE images. T1 pre-contrast values was generated by acquiring six different flip angles. The arterial input function was derived from arterial pixels in the perfusion images selected manually. The extended Tofts model was used to generate the permeability maps. Routine MRI scans took about 30 minutes to complete; the additional scans added 12 minutes. Results: To date, seven patients with cerebral tumors have been imaged and tumor physiology characterized. For example, on a glioblastoma patient, the volume contoured on T1 Gd images, ADC map and the pharmacokinetic map (Ktrans) were 1.9, 1.4, and 1.5 cc respectively with strong spatial correlation. The mean ADC value of the entire volume was 1141 μm2/s while the value in the white matter was 811 μm2/s. The mean value of Ktrans was 0.02 min-1 in the tumor volume and 0.00 in the normal white matter. Conclusion: Our initial results suggest that multiparametric MRI sequences may provide a more quantitative evaluation of vascular and tumor properties. Implementing functional imaging during MR-SIM may be particularly beneficial in assessing tumor extent, differentiating radiation necrosis from tumor recurrence, and establishing reliable bio-markers for treatment response evaluation. The Department of Radiation Oncology at Henry Ford Health System has research agreement with Varian Medical System and Philips Health Care.« less

CNNEDGEPOT: CNN based edge detection of 2D near surface potential field data

NASA Astrophysics Data System (ADS)

Aydogan, D.

2012-09-01

All anomalies are important in the interpretation of gravity and magnetic data because they indicate some important structural features. One of the advantages of using gravity or magnetic data for searching contacts is to be detected buried structures whose signs could not be seen on the surface. In this paper, a general view of the cellular neural network (CNN) method with a large scale nonlinear circuit is presented focusing on its image processing applications. The proposed CNN model is used consecutively in order to extract body and body edges. The algorithm is a stochastic image processing method based on close neighborhood relationship of the cells and optimization of A, B and I matrices entitled as cloning template operators. Setting up a CNN (continues time cellular neural network (CTCNN) or discrete time cellular neural network (DTCNN)) for a particular task needs a proper selection of cloning templates which determine the dynamics of the method. The proposed algorithm is used for image enhancement and edge detection. The proposed method is applied on synthetic and field data generated for edge detection of near-surface geological bodies that mask each other in various depths and dimensions. The program named as CNNEDGEPOT is a set of functions written in MATLAB software. The GUI helps the user to easily change all the required CNN model parameters. A visual evaluation of the outputs due to DTCNN and CTCNN are carried out and the results are compared with each other. These examples demonstrate that in detecting the geological features the CNN model can be used for visual interpretation of near surface gravity or magnetic anomaly maps.

Integrated segmentation of cellular structures

NASA Astrophysics Data System (ADS)

Ajemba, Peter; Al-Kofahi, Yousef; Scott, Richard; Donovan, Michael; Fernandez, Gerardo

2011-03-01

Automatic segmentation of cellular structures is an essential step in image cytology and histology. Despite substantial progress, better automation and improvements in accuracy and adaptability to novel applications are needed. In applications utilizing multi-channel immuno-fluorescence images, challenges include misclassification of epithelial and stromal nuclei, irregular nuclei and cytoplasm boundaries, and over and under-segmentation of clustered nuclei. Variations in image acquisition conditions and artifacts from nuclei and cytoplasm images often confound existing algorithms in practice. In this paper, we present a robust and accurate algorithm for jointly segmenting cell nuclei and cytoplasm using a combination of ideas to reduce the aforementioned problems. First, an adaptive process that includes top-hat filtering, Eigenvalues-of-Hessian blob detection and distance transforms is used to estimate the inverse illumination field and correct for intensity non-uniformity in the nuclei channel. Next, a minimum-error-thresholding based binarization process and seed-detection combining Laplacian-of-Gaussian filtering constrained by a distance-map-based scale selection is used to identify candidate seeds for nuclei segmentation. The initial segmentation using a local maximum clustering algorithm is refined using a minimum-error-thresholding technique. Final refinements include an artifact removal process specifically targeted at lumens and other problematic structures and a systemic decision process to reclassify nuclei objects near the cytoplasm boundary as epithelial or stromal. Segmentation results were evaluated using 48 realistic phantom images with known ground-truth. The overall segmentation accuracy exceeds 94%. The algorithm was further tested on 981 images of actual prostate cancer tissue. The artifact removal process worked in 90% of cases. The algorithm has now been deployed in a high-volume histology analysis application.

Histomorphometric analysis of collagen architecture of auricular keloids in an Asian population.

PubMed

Chong, Yosep; Park, Tae Hwan; Seo, Sang won; Chang, Choong Hyun

2015-03-01

Keloids are a pathologic condition of the reparative process, which present as excessive scar formation that involves various cells and cytokines. Many studies focusing on the histologic feature of keloids, however, have shown discordant results without consideration of architectural aspect of collagen structure. The purpose of this study was to demonstrate a schematic illustration of collagen architecture of keloids, specifically auricular keloids, and to analyze each part on the histomorphologic and morphometric basis. Thirty-nine surgically excised auricular keloids were retrieved from the file of Kangbuk Samsung Hospital. After exhaustive histomorphologic analysis, 3 distinctive structural parts, keloidal collagen, organizing collagen, and proliferating core collagen, were identified and mapped in every case. Cellularity of fibroblasts, blood vessel density, degree of inflammatory cell infiltration, and mast cells counts using Masson trichrome stain, Van Gieson stain, toluidine blue stain, and immunohistochemical stains for CD31 and smooth muscle actin were analyzed in each part of each case. Morphometric analysis on these parameters using ImageJ software was performed using 3 representative images of each part. Three parts were histomorphologically distinct by shape and array of collagen bundles, fibroblasts cellularity, blood vessel density, degree of inflammatory cells, and mast cell infiltration. Morphometric analysis revealed statistically significant difference between each part in fibroblasts cellularity, blood vessel density, degree of inflammatory cell infiltration, and mast cells count. All parameters were exceedingly high in whorling hypercellular fibrous nodules in proliferating core collagen showing simultaneous changes in other parts. Morphologically and morphometrically, 3 distinctive parts were identified in auricular keloids. Mast cell infiltrations, blood vessel density, and fibroblast cellularity are simultaneously increased or decreased according to these parts. Proliferating core collagen might serve as a proliferating center of keloids and might be a key portion for tumor growth and recurrence.

Near-Atomic Three-Dimensional Mapping for Site-Specific Chemistry of 'Superbugs'.

PubMed

Adineh, Vahid R; Marceau, Ross K W; Velkov, Tony; Li, Jian; Fu, Jing

2016-11-09

Emergence of multidrug resistant Gram-negative bacteria has caused a global health crisis and last-line class of antibiotics such as polymyxins are increasingly used. The chemical composition at the cell surface plays a key role in antibiotic resistance. Unlike imaging the cellular ultrastructure with well-developed electron microscopy, the acquisition of a high-resolution chemical map of the bacterial surface still remains a technological challenge. In this study, we developed an atom probe tomography (APT) analysis approach to acquire mass spectra in the pulsed-voltage mode and reconstructed the 3D chemical distribution of atoms and molecules in the subcellular domain at the near-atomic scale. Using focused ion beam (FIB) milling together with micromanipulation, site-specific samples were retrieved from a single cell of Acinetobacter baumannii prepared as needle-shaped tips with end radii less than 60 nm, followed by a nanoscale coating of silver in the order of 10 nm. The significantly elevated conductivity provided by the metallic coating enabled successful and routine field evaporation of the biological material, with all the benefits of pulsed-voltage APT. In parallel with conventional cryo-TEM imaging, our novel approach was applied to investigate polymyxin-susceptible and -resistant strains of A. baumannii after treatment of polymyxin B. Acquired atom probe mass spectra from the cell envelope revealed characteristic fragments of phosphocholine from the polymyxin-susceptible strain, but limited signals from this molecule were detected in the polymyxin-resistant strain. This study promises unprecedented capacity for 3D nanoscale imaging and chemical mapping of bacterial cells at the ultimate 3D spatial resolution using APT.

Nanoscale Imaging of Whole Cells Using a Liquid Enclosure and a Scanning Transmission Electron Microscope

PubMed Central

Peckys, Diana B.; Veith, Gabriel M.; Joy, David C.; de Jonge, Niels

2009-01-01

Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory. PMID:20020038

A global interaction network maps a wiring diagram of cellular function

PubMed Central

Costanzo, Michael; VanderSluis, Benjamin; Koch, Elizabeth N.; Baryshnikova, Anastasia; Pons, Carles; Tan, Guihong; Wang, Wen; Usaj, Matej; Hanchard, Julia; Lee, Susan D.; Pelechano, Vicent; Styles, Erin B.; Billmann, Maximilian; van Leeuwen, Jolanda; van Dyk, Nydia; Lin, Zhen-Yuan; Kuzmin, Elena; Nelson, Justin; Piotrowski, Jeff S.; Srikumar, Tharan; Bahr, Sondra; Chen, Yiqun; Deshpande, Raamesh; Kurat, Christoph F.; Li, Sheena C.; Li, Zhijian; Usaj, Mojca Mattiazzi; Okada, Hiroki; Pascoe, Natasha; Luis, Bryan-Joseph San; Sharifpoor, Sara; Shuteriqi, Emira; Simpkins, Scott W.; Snider, Jamie; Suresh, Harsha Garadi; Tan, Yizhao; Zhu, Hongwei; Malod-Dognin, Noel; Janjic, Vuk; Przulj, Natasa; Troyanskaya, Olga G.; Stagljar, Igor; Xia, Tian; Ohya, Yoshikazu; Gingras, Anne-Claude; Raught, Brian; Boutros, Michael; Steinmetz, Lars M.; Moore, Claire L.; Rosebrock, Adam P.; Caudy, Amy A.; Myers, Chad L.; Andrews, Brenda; Boone, Charles

2017-01-01

We generated a global genetic interaction network for Saccharomyces cerevisiae, constructing over 23 million double mutants, identifying ~550,000 negative and ~350,000 positive genetic interactions. This comprehensive network maps genetic interactions for essential gene pairs, highlighting essential genes as densely connected hubs. Genetic interaction profiles enabled assembly of a hierarchical model of cell function, including modules corresponding to protein complexes and pathways, biological processes, and cellular compartments. Negative interactions connected functionally related genes, mapped core bioprocesses, and identified pleiotropic genes, whereas positive interactions often mapped general regulatory connections among gene pairs, rather than shared functionality. The global network illustrates how coherent sets of genetic interactions connect protein complex and pathway modules to map a functional wiring diagram of the cell. PMID:27708008

Core needle biopsy guidance based on EMOCT imaging (Conference Presentation)

NASA Astrophysics Data System (ADS)

Iftimia, Nicusor V.; Park, Jesung; Maguluri, Gopi

2016-03-01

We present a novel method, based on encoder mapping OCT imaging, for real-time guidance of core biopsy procedures. This method provides real-time feedback to the interventional radiologist, such that he/she can reorient the needle during the biopsy and sample the most representative area of the suspicious mass that is being investigated. This aspect is very important for tailoring therapy to the specific cancer based on biomarker analysis, which will become one of the next big advances in our search for the optimal cancer therapy. To enable individualized treatment, the genetic constitution and the DNA repair status in the affected areas is needed for each patient. Thus, representative sampling of the tumor is needed for analyzing various biomarkers, which are used as a tool to personalize cancer therapy. The encoder-based OCT enables samping of large size masses and provides full control on the imaging probe, which is passed through the bore of the biopsy guidance needle. The OCT image is built gradually, based on the feedback of an optical encoder which senses the incremental movement of the needle with a few microns resolution. Tissue mapping is independent of the needle speed, while it is advanced through the tissue. The OCT frame is analyzed in real-time and tissue cellularity is reported in a very simple manner (pie chart). Our preliminary study on a rabbit model of cancer has demonstrated the capability of this technology for accurately differentiating between viable cancer and heterogeneous or necrotic tissue.

Carbon Dioxide Laser Ablation of Basal Cell Carcinoma with Visual Guidance by Reflectance Confocal Microscopy: A Proof of Principle Pilot Study

PubMed Central

Hibler, B.P.; Sierra, H.; Cordova, M.; Phillips, W.; Rajadhyaksha, M.; Nehal, K.S.; Rossi, A.M.

2016-01-01

Background Laser ablation is an alternative, non-surgical treatment modality for low-risk basal cell carcinoma (BCC); however, lack of confirmative tumour destruction or residual tumour presence has been a limiting factor to adoption. Reflectance confocal microscopy (RCM) provides non-invasive, cellular-level resolution imaging of the skin and is capable of identifying tumour. Objective To evaluate the use of RCM to guide carbon dioxide (CO2) laser ablation of BCC, confirm destruction, and correlate findings with histology. Methods RCM was used pre-ablation to evaluate for features of BCC. Ablation was performed with a CO2 laser, and the response rapidly assessed using handheld RCM to evaluate for residual tumour. Confirmative pathology was used to verify confocal imaging. Results RCM imaging identified tumour pre-ablation with features not identified on normal, surrounding skin. Post-ablation, RCM documented complete removal of tumour in six cases and residual tumour in two. Histologic examination identified the ablated area and confirmed clearance of tumour in the six aforementioned cases and corroborated confocal findings for residual tumour in the other two cases. Conclusions We report successful treatment of superficial and nodular BCC using CO2 laser ablation augmented by RCM imaging for pre-ablation guidance and verification of tumour removal post-ablation. Akin to complete circumferential and deep margin control techniques, using RCM helps to map peripheral and deep BCC margins to hone in on areas exhibiting persistent tumour after ablation. CO2 laser ablation visually guided by RCM can help circumvent previously cited limiting factors of laser ablation for tumour destruction by providing cellular-level resolution imaging of tumour and margin assessment in between each laser pass and post-ablation. PMID:26800657

Carbon dioxide laser ablation of basal cell carcinoma with visual guidance by reflectance confocal microscopy: a proof-of-principle pilot study.

PubMed

Hibler, B P; Sierra, H; Cordova, M; Phillips, W; Rajadhyaksha, M; Nehal, K S; Rossi, A M

2016-06-01

Laser ablation is an alternative, nonsurgical treatment modality for low-risk basal cell carcinoma (BCC). However, lack of confirmative tumour destruction or residual tumour presence has been a limiting factor to its adoption. Reflectance confocal microscopy (RCM) provides noninvasive, cellular-level resolution imaging of the skin and is capable of identifying tumour. To evaluate the use of RCM to guide carbon dioxide (CO2 ) laser ablation of BCC, confirm destruction and correlate findings with histology. RCM was used preablation to evaluate for features of BCC. Ablation was performed with a CO2 laser, and the response rapidly assessed using handheld RCM to evaluate for residual tumour. Confirmative pathology was used to verify confocal imaging. Preablation RCM imaging identified tumour with features not identified on normal, surrounding skin. Postablation, RCM documented complete removal of tumour in six cases and residual tumour in two. Histological examination identified the ablated area and confirmed clearance of tumour in the six aforementioned cases and corroborated confocal findings for residual tumour in the other two cases. We report successful treatment of superficial and nodular BCC using CO2 laser ablation augmented by RCM imaging for preablation guidance and verification of tumour removal postablation. Akin to complete circumferential and deep margin control techniques, using RCM helps to map peripheral and deep BCC margins to hone in on areas exhibiting persistent tumour after ablation. CO2 laser ablation visually guided by RCM can help circumvent previously cited limiting factors of laser ablation for tumour destruction by providing cellular-level resolution imaging of tumour and margin assessment in between each laser pass and postablation. © 2016 British Association of Dermatologists.

Registration procedure for spatial correlation of physical energy deposition of particle irradiation and cellular response utilizing cell-fluorescent ion track hybrid detectors

NASA Astrophysics Data System (ADS)

Niklas, M.; Zimmermann, F.; Schlegel, J.; Schwager, C.; Debus, J.; Jäkel, O.; Abdollahi, A.; Greilich, S.

2016-09-01

The hybrid technology cell-fluorescent ion track hybrid detector (Cell-Fit-HD) enables the investigation of radiation-related cellular events along single ion tracks on the subcellular scale in clinical ion beams. The Cell-Fit-HD comprises a fluorescent nuclear track detector (FNTD, the physical compartment), a device for individual particle detection and a substrate for viable cell-coating, i.e. the biological compartment. To date both compartments have been imaged sequentially in situ by confocal laser scanning microscopy (CLSM). This is yet in conflict with a functional read-out of the Cell-Fit-HD utilizing a fast live-cell imaging of the biological compartment with low phototoxicity on greater time scales. The read-out of the biological from the physical compartment was uncoupled. A read-out procedure was developed to image the cell layer by conventional widefield microscopy whereas the FNTD was imaged by CLSM. Point mapping registration of the confocal and widefield imaging data was performed. Non-fluorescent crystal defects (spinels) visible in both read-outs were used as control point pairs. The accuracy achieved was on the sub-µm scale. The read-out procedure by widefield microscopy does not impair the unique ability of spatial correlation by the Cell-Fit-HD. The uncoupling will enlarge the application potential of the hybrid technology significantly. The registration allows for an ultimate correlation of microscopic physical beam parameters and cell kinetics on greater time scales. The method reported herein will be instrumental for the introduction of a novel generation of compact detectors facilitating biodosimetric research towards high-throughput analysis.

Asymmetric-detection time-stretch optical microscopy (ATOM) for ultrafast high-contrast cellular imaging in flow

PubMed Central

Wong, Terence T. W.; Lau, Andy K. S.; Ho, Kenneth K. Y.; Tang, Matthew Y. H.; Robles, Joseph D. F.; Wei, Xiaoming; Chan, Antony C. S.; Tang, Anson H. L.; Lam, Edmund Y.; Wong, Kenneth K. Y.; Chan, Godfrey C. F.; Shum, Ho Cheung; Tsia, Kevin K.

2014-01-01

Accelerating imaging speed in optical microscopy is often realized at the expense of image contrast, image resolution, and detection sensitivity – a common predicament for advancing high-speed and high-throughput cellular imaging. We here demonstrate a new imaging approach, called asymmetric-detection time-stretch optical microscopy (ATOM), which can deliver ultrafast label-free high-contrast flow imaging with well delineated cellular morphological resolution and in-line optical image amplification to overcome the compromised imaging sensitivity at high speed. We show that ATOM can separately reveal the enhanced phase-gradient and absorption contrast in microfluidic live-cell imaging at a flow speed as high as ~10 m/s, corresponding to an imaging throughput of ~100,000 cells/sec. ATOM could thus be the enabling platform to meet the pressing need for intercalating optical microscopy in cellular assay, e.g. imaging flow cytometry – permitting high-throughput access to the morphological information of the individual cells simultaneously with a multitude of parameters obtained in the standard assay. PMID:24413677

A novel 3D micron-scale DPTV (Defocused Particle Tracking Velocimetry) and its applications in microfluidic devices

NASA Astrophysics Data System (ADS)

Roberts, John

2005-11-01

The rapid advancements in micro/nano biotechnology demand quantitative tools for characterizing microfluidic flows in lab-on-a-chip applications, validation of computational results for fully 3D flows in complex micro-devices, and efficient observation of cellular dynamics in 3D. We present a novel 3D micron-scale DPTV (defocused particle tracking velocimetry) that is capable of mapping out 3D Lagrangian, as well as 3D Eulerian velocity flow fields at sub-micron resolution and with one camera. The main part of the imaging system is an epi-fluorescent microscope (Olympus IX 51), and the seeding particles are fluorescent particles with diameter range 300nm - 10um. A software package has been developed for identifying (x,y,z,t) coordinates of the particles using the defocused images. Using the imaging system, we successfully mapped the pressure driven flow fields in microfluidic channels. In particular, we measured the Laglangian flow fields in a microfluidic channel with a herring bone pattern at the bottom, the later is used to enhance fluid mixing in lateral directions. The 3D particle tracks revealed the flow structure that has only been seen in numerical computation. This work is supported by the National Science Foundation (CTS - 0514443), the Nanobiotechnology Center at Cornell, and The New York State Center for Life Science Enterprise.

ADC histogram analysis of muscle lymphoma - Correlation with histopathology in a rare entity.

PubMed

Meyer, Hans-Jonas; Pazaitis, Nikolaos; Surov, Alexey

2018-06-21

Diffusion weighted imaging (DWI) is able to reflect histopathology architecture. A novel imaging approach, namely histogram analysis, is used to further characterize lesion on MRI. The purpose of this study is to correlate histogram parameters derived from apparent diffusion coefficient- (ADC) maps with histopathology parameters in muscle lymphoma. Eight patients (mean age 64.8 years, range 45-72 years) with histopathologically confirmed muscle lymphoma were retrospectively identified. Cell count, total nucleic and average nucleic areas were estimated using ImageJ. Additionally, Ki67-index was calculated. DWI was obtained on a 1.5T scanner by using the b values of 0 and 1000 s/mm2. Histogram analysis was performed as a whole lesion measurement by using a custom-made Matlabbased application. The correlation analysis revealed statistically significant correlation between cell count and ADCmean (p=-0.76, P=0.03) as well with ADCp75 (p=-0.79, P=0.02). Kurtosis and entropy correlated with average nucleic area (p=-0.81, P=0.02, p=0.88, P=0.007, respectively). None of the analyzed ADC parameters correlated with total nucleic area and with Ki67-index. This study identified significant correlations between cellularity and histogram parameters derived from ADC maps in muscle lymphoma. Thus, histogram analysis parameters reflect histopathology in muscle tumors. Advances in knowledge: Whole lesion ADC histogram analysis is able to reflect histopathology parameters in muscle lymphomas.

Comparison of divided and full pupil configurations for line-scanning confocal microscopy in human skin and oral mucosa

NASA Astrophysics Data System (ADS)

Larson, Bjorg; Abeytunge, Sanjeewa; Glazowski, Chris; Rajadhyaksha, Milind

2012-02-01

Confocal point-scanning microscopy has been showing promise in the detection, diagnosing and mapping of skin lesions in clinical settings. The noninvasive technique allows provides optical sectioning and cellular resolution for in vivo diagnosis of melanoma and basal cell carcinoma and pre-operative and intra-operative mapping of margins. The imaging has also enabled more accurate "guided" biopsies while minimizing the otherwise large number of "blind" biopsies. Despite these translational advances, however, point-scanning technology remains relatively complex and expensive. Line-scanning technology may offer an alternative approach to accelerate translation to the clinic. Line-scanning, using fewer optical components, inexpensive linear-array detectors and custom electronics, may enable smaller, simpler and lower-cost confocal microscopes. A line is formed using a cylindrical lens and scanned through the back focal plane of the objective with a galvanometric scanner. A linear CCD is used for detection. Two pupil configurations were compared for performance in imaging human tissue. In the full-pupil configuration, illumination and detection is made through the full objective pupil. In the divided pupil approach, half the pupil is illuminated and the other half is used for detection. The divided pupil configuration loses spatial and axial resolution due to a diminished NA, but the sectioning capability and rejection of background is improved. Imaging in skin and oral mucosa illustrate the performance of the two configurations.

Neuronal networks and mediators of cortical neurovascular coupling responses in normal and altered brain states.

PubMed

Lecrux, C; Hamel, E

2016-10-05

Brain imaging techniques that use vascular signals to map changes in neuronal activity, such as blood oxygenation level-dependent functional magnetic resonance imaging, rely on the spatial and temporal coupling between changes in neurophysiology and haemodynamics, known as 'neurovascular coupling (NVC)'. Accordingly, NVC responses, mapped by changes in brain haemodynamics, have been validated for different stimuli under physiological conditions. In the cerebral cortex, the networks of excitatory pyramidal cells and inhibitory interneurons generating the changes in neural activity and the key mediators that signal to the vascular unit have been identified for some incoming afferent pathways. The neural circuits recruited by whisker glutamatergic-, basal forebrain cholinergic- or locus coeruleus noradrenergic pathway stimulation were found to be highly specific and discriminative, particularly when comparing the two modulatory systems to the sensory response. However, it is largely unknown whether or not NVC is still reliable when brain states are altered or in disease conditions. This lack of knowledge is surprising since brain imaging is broadly used in humans and, ultimately, in conditions that deviate from baseline brain function. Using the whisker-to-barrel pathway as a model of NVC, we can interrogate the reliability of NVC under enhanced cholinergic or noradrenergic modulation of cortical circuits that alters brain states.This article is part of the themed issue 'Interpreting BOLD: a dialogue between cognitive and cellular neuroscience'. © 2016 The Author(s).

High content image analysis for human H4 neuroglioma cells exposed to CuO nanoparticles.

PubMed

Li, Fuhai; Zhou, Xiaobo; Zhu, Jinmin; Ma, Jinwen; Huang, Xudong; Wong, Stephen T C

2007-10-09

High content screening (HCS)-based image analysis is becoming an important and widely used research tool. Capitalizing this technology, ample cellular information can be extracted from the high content cellular images. In this study, an automated, reliable and quantitative cellular image analysis system developed in house has been employed to quantify the toxic responses of human H4 neuroglioma cells exposed to metal oxide nanoparticles. This system has been proved to be an essential tool in our study. The cellular images of H4 neuroglioma cells exposed to different concentrations of CuO nanoparticles were sampled using IN Cell Analyzer 1000. A fully automated cellular image analysis system has been developed to perform the image analysis for cell viability. A multiple adaptive thresholding method was used to classify the pixels of the nuclei image into three classes: bright nuclei, dark nuclei, and background. During the development of our image analysis methodology, we have achieved the followings: (1) The Gaussian filtering with proper scale has been applied to the cellular images for generation of a local intensity maximum inside each nucleus; (2) a novel local intensity maxima detection method based on the gradient vector field has been established; and (3) a statistical model based splitting method was proposed to overcome the under segmentation problem. Computational results indicate that 95.9% nuclei can be detected and segmented correctly by the proposed image analysis system. The proposed automated image analysis system can effectively segment the images of human H4 neuroglioma cells exposed to CuO nanoparticles. The computational results confirmed our biological finding that human H4 neuroglioma cells had a dose-dependent toxic response to the insult of CuO nanoparticles.

Atomic force microscopy-based characterization and design of biointerfaces

NASA Astrophysics Data System (ADS)

Alsteens, David; Gaub, Hermann E.; Newton, Richard; Pfreundschuh, Moritz; Gerber, Christoph; Müller, Daniel J.

2017-03-01

Atomic force microscopy (AFM)-based methods have matured into a powerful nanoscopic platform, enabling the characterization of a wide range of biological and synthetic biointerfaces ranging from tissues, cells, membranes, proteins, nucleic acids and functional materials. Although the unprecedented signal-to-noise ratio of AFM enables the imaging of biological interfaces from the cellular to the molecular scale, AFM-based force spectroscopy allows their mechanical, chemical, conductive or electrostatic, and biological properties to be probed. The combination of AFM-based imaging and spectroscopy structurally maps these properties and allows their 3D manipulation with molecular precision. In this Review, we survey basic and advanced AFM-related approaches and evaluate their unique advantages and limitations in imaging, sensing, parameterizing and designing biointerfaces. It is anticipated that in the next decade these AFM-related techniques will have a profound influence on the way researchers view, characterize and construct biointerfaces, thereby helping to solve and address fundamental challenges that cannot be addressed with other techniques.

Hyperspectral imaging solutions for brain tissue metabolic and hemodynamic monitoring: past, current and future developments

NASA Astrophysics Data System (ADS)

Giannoni, Luca; Lange, Frédéric; Tachtsidis, Ilias

2018-04-01

Hyperspectral imaging (HSI) technologies have been used extensively in medical research, targeting various biological phenomena and multiple tissue types. Their high spectral resolution over a wide range of wavelengths enables acquisition of spatial information corresponding to different light-interacting biological compounds. This review focuses on the application of HSI to monitor brain tissue metabolism and hemodynamics in life sciences. Different approaches involving HSI have been investigated to assess and quantify cerebral activity, mainly focusing on: (1) mapping tissue oxygen delivery through measurement of changes in oxygenated (HbO2) and deoxygenated (HHb) hemoglobin; and (2) the assessment of the cerebral metabolic rate of oxygen (CMRO2) to estimate oxygen consumption by brain tissue. Finally, we introduce future perspectives of HSI of brain metabolism, including its potential use for imaging optical signals from molecules directly involved in cellular energy production. HSI solutions can provide remarkable insight in understanding cerebral tissue metabolism and oxygenation, aiding investigation on brain tissue physiological processes.

Ancestry of a human endogenous retrovirus family.

PubMed Central

Mariani-Costantini, R; Horn, T M; Callahan, R

1989-01-01

The human endogenous retrovirus type II (HERVII) family of HERV genomes has been found by Southern blot analysis to be characteristic of humans, apes, and Old World monkeys. New World monkeys and prosimians lack HERVII proviral genomes. Cellular DNAs of humans, common chimpanzees, gorillas, and orangutans, but not lesser ape lar gibbons, appear to contain the HERVII-related HLM-2 proviral genome integrated at the same site (HLM-2 maps to human chromosome 1). This suggests that the ancestral HERVII retrovirus(es) entered the genomes of Old World anthropoids by infection after the divergence of New World monkeys (platyrrhines) but before the evolutionary radiation of large hominoids. Images PMID:2507793

Polydopamine-based functional composite particles for tumor cell targeting and dual-mode cellular imaging.

PubMed

Zhou, Yalei; Zhou, Jie; Wang, Feng; Yang, Haifeng

2018-05-01

Particles which bear tumor cell targeting and multimode imaging capabilities are promising in tumor diagnosis and cancer therapy. A simple and versatile method to fabricate gold/polydopamine-Methylene Blue@Bovine Serum Albumin-glutaraldehyde-Transferrin composite particles (Au/PDA-MB@BSA-GA-Tf NPs) for tumor cell targeting and fluorescence (FL) / surface-enhanced Raman scattering (SERS) dual-modal imaging were reported in this work. Polydopamine (PDA) spheres played an important role in gold ion reduction, gold nanoparticle (Au NPs) binding and methylene blue (MB) adsorption, MB were employed as both fluorescence label and Raman reporter. In addition, glutaraldehyde (GA) crosslinked bovine serum albumin (BSA) in the outer layer of Au/PDA-MB nanoparticles can prevent MB from dissociation and leakage. The composite nanoparticles were further conjugated with transferrin (Tf) to target transferrin receptor (TfR)-overexpressed cancer cells. The targeting ability as well as the intracellular location of the probe was investigated through SERS mapping and fluorescence imaging. Their excellent biocompatibility was demonstrated by low cytotoxicity against breast cancer cell (4T1 cell). Copyright © 2018 Elsevier B.V. All rights reserved.

A new concept for medical imaging centered on cellular phone technology.

PubMed

Granot, Yair; Ivorra, Antoni; Rubinsky, Boris

2008-04-30

According to World Health Organization reports, some three quarters of the world population does not have access to medical imaging. In addition, in developing countries over 50% of medical equipment that is available is not being used because it is too sophisticated or in disrepair or because the health personnel are not trained to use it. The goal of this study is to introduce and demonstrate the feasibility of a new concept in medical imaging that is centered on cellular phone technology and which may provide a solution to medical imaging in underserved areas. The new system replaces the conventional stand-alone medical imaging device with a new medical imaging system made of two independent components connected through cellular phone technology. The independent units are: a) a data acquisition device (DAD) at a remote patient site that is simple, with limited controls and no image display capability and b) an advanced image reconstruction and hardware control multiserver unit at a central site. The cellular phone technology transmits unprocessed raw data from the patient site DAD and receives and displays the processed image from the central site. (This is different from conventional telemedicine where the image reconstruction and control is at the patient site and telecommunication is used to transmit processed images from the patient site). The primary goal of this study is to demonstrate that the cellular phone technology can function in the proposed mode. The feasibility of the concept is demonstrated using a new frequency division multiplexing electrical impedance tomography system, which we have developed for dynamic medical imaging, as the medical imaging modality. The system is used to image through a cellular phone a simulation of breast cancer tumors in a medical imaging diagnostic mode and to image minimally invasive tissue ablation with irreversible electroporation in a medical imaging interventional mode.

Textural features of dynamic contrast-enhanced MRI derived model-free and model-based parameter maps in glioma grading.

PubMed

Xie, Tian; Chen, Xiao; Fang, Jingqin; Kang, Houyi; Xue, Wei; Tong, Haipeng; Cao, Peng; Wang, Sumei; Yang, Yizeng; Zhang, Weiguo

2018-04-01

Presurgical glioma grading by dynamic contrast-enhanced MRI (DCE-MRI) has unresolved issues. The aim of this study was to investigate the ability of textural features derived from pharmacokinetic model-based or model-free parameter maps of DCE-MRI in discriminating between different grades of gliomas, and their correlation with pathological index. Retrospective. Forty-two adults with brain gliomas. 3.0T, including conventional anatomic sequences and DCE-MRI sequences (variable flip angle T1-weighted imaging and three-dimensional gradient echo volumetric imaging). Regions of interest on the cross-sectional images with maximal tumor lesion. Five commonly used textural features, including Energy, Entropy, Inertia, Correlation, and Inverse Difference Moment (IDM), were generated. All textural features of model-free parameters (initial area under curve [IAUC], maximal signal intensity [Max SI], maximal up-slope [Max Slope]) could effectively differentiate between grade II (n = 15), grade III (n = 13), and grade IV (n = 14) gliomas (P < 0.05). Two textural features, Entropy and IDM, of four DCE-MRI parameters, including Max SI, Max Slope (model-free parameters), vp (Extended Tofts), and vp (Patlak) could differentiate grade III and IV gliomas (P < 0.01) in four measurements. Both Entropy and IDM of Patlak-based K trans and vp could differentiate grade II (n = 15) from III (n = 13) gliomas (P < 0.01) in four measurements. No textural features of any DCE-MRI parameter maps could discriminate between subtypes of grade II and III gliomas (P < 0.05). Both Entropy and IDM of Extended Tofts- and Patlak-based vp showed highest area under curve in discriminating between grade III and IV gliomas. However, intraclass correlation coefficient (ICC) of these features revealed relatively lower inter-observer agreement. No significant correlation was found between microvascular density and textural features, compared with a moderate correlation found between cellular proliferation index and those features. Textural features of DCE-MRI parameter maps displayed a good ability in glioma grading. 3 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2018;47:1099-1111. © 2017 International Society for Magnetic Resonance in Medicine.

High School Biology: A Group Approach to Concept Mapping.

ERIC Educational Resources Information Center

Brown, David S.

2003-01-01

Explains concept mapping as an instructional method in cooperative learning environments, and describes a study investigating the effectiveness of concept mapping on student learning during a photosynthesis and cellular respiration unit. Reports on the positive effects of concept mapping in the experimental group. (Contains 16 references.) (YDS)

Correlating two-photon excited fluorescence imaging of breast cancer cellular redox state with seahorse flux analysis of normalized cellular oxygen consumption

NASA Astrophysics Data System (ADS)

Hou, Jue; Wright, Heather J.; Chan, Nicole; Tran, Richard; Razorenova, Olga V.; Potma, Eric O.; Tromberg, Bruce J.

2016-06-01

Two-photon excited fluorescence (TPEF) imaging of the cellular cofactors nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide is widely used to measure cellular metabolism, both in normal and pathological cells and tissues. When dual-wavelength excitation is used, ratiometric TPEF imaging of the intrinsic cofactor fluorescence provides a metabolic index of cells-the "optical redox ratio" (ORR). With increased interest in understanding and controlling cellular metabolism in cancer, there is a need to evaluate the performance of ORR in malignant cells. We compare TPEF metabolic imaging with seahorse flux analysis of cellular oxygen consumption in two different breast cancer cell lines (MCF-7 and MDA-MB-231). We monitor metabolic index in living cells under both normal culture conditions and, for MCF-7, in response to cell respiration inhibitors and uncouplers. We observe a significant correlation between the TPEF-derived ORR and the flux analyzer measurements (R=0.7901, p<0.001). Our results confirm that the ORR is a valid dynamic index of cell metabolism under a range of oxygen consumption conditions relevant for cancer imaging.

18F-FLT uptake kinetics in head and neck squamous cell carcinoma: a PET imaging study.

PubMed

Liu, Dan; Chalkidou, Anastasia; Landau, David B; Marsden, Paul K; Fenwick, John D

2014-04-01

To analyze the kinetics of 3(')-deoxy-3(')-[F-18]-fluorothymidine (18F-FLT) uptake by head and neck squamous cell carcinomas and involved nodes imaged using positron emission tomography (PET). Two- and three-tissue compartment models were fitted to 12 tumor time-activity-curves (TACs) obtained for 6 structures (tumors or involved nodes) imaged in ten dynamic PET studies of 1 h duration, carried out for five patients. The ability of the models to describe the data was assessed using a runs test, the Akaike information criterion (AIC) and leave-one-out cross-validation. To generate parametric maps the models were also fitted to TACs of individual voxels. Correlations between maps of different parameters were characterized using Pearson'sr coefficient; in particular the phosphorylation rate-constants k3-2tiss and k5 of the two- and three-tissue models were studied alongside the flux parameters KFLT- 2tiss and KFLT of these models, and standardized uptake values (SUV). A methodology based on expectation-maximization clustering and the Bayesian information criterion ("EM-BIC clustering") was used to distil the information from noisy parametric images. Fits of two-tissue models 2C3K and 2C4K and three-tissue models 3C5K and 3C6K comprising three, four, five, and six rate-constants, respectively, pass the runs test for 4, 8, 10, and 11 of 12 tumor TACs. The three-tissue models have lower AIC and cross-validation scores for nine of the 12 tumors. Overall the 3C6K model has the lowest AIC and cross-validation scores and its fitted parameter values are of the same orders of magnitude as literature estimates. Maps of KFLT and KFLT- 2tiss are strongly correlated (r = 0.85) and also correlate closely with SUV maps (r = 0.72 for KFLT- 2tiss, 0.64 for KFLT). Phosphorylation rate-constant maps are moderately correlated with flux maps (r = 0.48 for k3-2tiss vs KFLT- 2tiss and r = 0.68 for k5 vs KFLT); however, neither phosphorylation rate-constant correlates significantly with SUV. EM-BIC clustering reduces the parametric maps to a small number of levels--on average 5.8, 3.5, 3.4, and 1.4 for KFLT- 2tiss, KFLT, k3-2tiss, and k5. This large simplification is potentially useful for radiotherapy dose-painting, but demonstrates the high noise in some maps. Statistical simulations show that voxel level noise degrades TACs generated from the 3C6K model sufficiently that the average AIC score, parameter bias, and total uncertainty of 2C4K model fits are similar to those of 3C6K fits, whereas at the whole tumor level the scores are lower for 3C6K fits. For the patients studied here, whole tumor FLT uptake time-courses are represented better overall by a three-tissue than by a two-tissue model. EM-BIC clustering simplifies noisy parametric maps, providing the best description of the underlying information they contain and is potentially useful for radiotherapy dose-painting. However, the clustering highlights the large degree of noise present in maps of the phosphorylation rate-constantsk5 and k3-2tiss, which are conceptually tightly linked to cellular proliferation. Methods must be found to make these maps more robust-either by constraining other model parameters or modifying dynamic imaging protocols. © 2014 American Association of Physicists in Medicine.

Electron microscopy using the genetically encoded APEX2 tag in cultured mammalian cells

PubMed Central

Martell, Jeffrey D; Deerinck, Thomas J; Lam, Stephanie S; Ellisman, Mark H; Ting, Alice Y

2018-01-01

Electron microscopy (EM) is the premiere technique for high-resolution imaging of cellular ultrastructure. Unambiguous identification of specific proteins or cellular compartments in electron micrographs, however, remains challenging because of difficulties in delivering electron-dense contrast agents to specific subcellular targets within intact cells. We recently reported enhanced ascorbate peroxidase 2 (APEX2) as a broadly applicable genetic tag that generates EM contrast on a specific protein or subcellular compartment of interest. This protocol provides guidelines for designing and validating APEX2 fusion constructs, along with detailed instructions for cell culture, transfection, fixation, heavy-metal staining, embedding in resin, and EM imaging. Although this protocol focuses on EM in cultured mammalian cells, APEX2 is applicable to many cell types and contexts, including intact tissues and organisms, and is useful for numerous applications beyond EM, including live-cell proteomic mapping. This protocol, which describes procedures for sample preparation from cell monolayers and cell pellets, can be completed in 10 d, including time for APEX2 fusion construct validation, cell growth, and solidification of embedding resins. Notably, the only additional steps required relative to a standard EM sample preparation are cell transfection and a 2- to 45-min staining period with 3,3′-diaminobenzidine (DAB) and hydrogen peroxide (H2O2). PMID:28796234

Epidermal keratinocyte polarity and motility require Ca2+ influx through TRPV1

PubMed Central

Graham, David M.; Huang, Ling; Robinson, Kenneth R.; Messerli, Mark A.

2013-01-01

Summary Ca2+ has long been known to play an important role in cellular polarity and guidance. We studied the role of Ca2+ signaling during random and directed cell migration to better understand whether Ca2+ directs cell motility from the leading edge and which ion channels are involved in this function by using primary zebrafish keratinocytes. Rapid line-scan and time-lapse imaging of intracellular Ca2+ (Ca2+i) during migration and automated image alignment enabled us to characterize and map the spatiotemporal changes in Ca2+i. We show that asymmetric distributions of lamellipodial Ca2+ sparks are encoded in frequency, not amplitude, and that they correlate with cellular rotation during migration. Directed migration during galvanotaxis increases the frequency of Ca2+ sparks over the entire lamellipod; however, these events do not give rise to asymmetric Ca2+i signals that correlate with turning. We demonstrate that Ca2+-permeable channels within these cells are mechanically activated and include several transient receptor potential family members, including TRPV1. Last, we demonstrate that cell motility and Ca2+i activity are affected by pharmacological agents that target TRPV1, indicating a novel role for this channel during cell migration. PMID:23943873

The Colorado Lightning Mapping Array

NASA Astrophysics Data System (ADS)

Rison, W.; Krehbiel, P. R.; Thomas, R. J.; Rodeheffer, D.; Fuchs, B.

2012-12-01

A fifteen station Lightning Mapping Array (LMA) was installed in northern Colorado in the spring of 2012. While the driving force for the array was to produce 3-dimensional lightning data to support the Deep Convective Clouds and Chemistry (DC3) Experiment (Barth, this conference), data from the array are being used for several other projects. These include: electrification studies in conjunction with the CSU CHILL radar (Lang et al, this conference); observations of the parent lightning discharges of sprites (Lyons et al, this conference); trying to detect upward discharges triggered by wind turbines, characterizing conditions in which aircraft flying through clouds produce discharges which can be detected by the LMA, and other opportunities, such as observations of lightning in pyrocumulus clouds produced by the High Park Fire west of Fort Collins, CO. All the COLMA stations are solar-powered, and use broadband cellular modems for data communications. This makes the stations completely self-contained and autonomous, allowing a station to be installed anywhere a cellular signal is available. Because most of the stations were installed well away from anthropogenic noise sources, the COLMA is very sensitive. This is evidenced by the numerous plane tracks detected in its the vicinity. The diameter, D, of the COLMA is about 100 km, significantly larger than other LMAs. Because the error in the radial distance r is proportional to (r/D)2, and the error in the altitude z is proportional to (z/D)2, the larger array diameter greatly expands the usable range of the COLMA. The COLMA is able to detect and characterize lighting flashes to a distance of about 350 km from the array center. In addition to a web-based display (lightning.nmt.edu/colma), geo-referenced images are produced and updated at one-minute intervals. These geo-referenced images can be used to overlay the real-time lightning data on Google Earth and other mapping software. These displays were used by the DC3 aircraft operations to guide the research aircraft to the best regions for sampling thunderstorm outflow chemistry, and to provide pilots information on regions to avoid due to lightning hazards.

USING RUNNING DIFFERENCE IMAGES TO TRACK PROPER MOTIONS OF XUV CORONAL INTENSITY ON THE SUN

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sheeley, N. R. Jr.; Warren, H. P.; Lee, J., E-mail: neil.sheeley@nrl.navy.mil, E-mail: harry.warren@nrl.navy.mil

2014-12-20

We have developed a procedure for observing and tracking proper motions of faint XUV coronal intensity on the Sun and have applied this procedure to study the collective motions of cellular plumes and the shorter-period waves in sunspots. Our space/time maps of cellular plumes show a series of tracks with the same 5-8 minute repetition times and ∼100 km s{sup –1} sky-plane speeds found previously in active-region fans and in coronal hole plumes. By synchronizing movies and space/time maps, we find that the tracks are produced by elongated ejections from the unipolar flux concentrations at the bases of the cellular plumes and thatmore » the phases of these ejections are uncorrelated from cell to cell. Thus, the large-scale motion is not a continuous flow, but is more like a system of independent conveyor belts all moving in the same direction along the magnetic field. In contrast, the proper motions in sunspots are clearly waves resulting from periodic disturbances in the sunspot umbras. The periods are ∼2.6 minutes, but the sky-plane speeds and wavelengths depend on the heights of the waves above the sunspot. In the chromosphere, the waves decelerate from 35-45 km s{sup –1} in the umbra to 7-8 km s{sup –1} toward the outer edge of the penumbra, but in the corona, the waves accelerate to ∼60-100 km s{sup –1}. Because chromospheric and coronal tracks originate from the same space/time locations, the coronal waves must emerge from the same umbral flashes that produce the chromospheric waves.« less

The Effect of Substrate Topography on Direct Reprogramming of Fibroblasts to Induced Neurons

PubMed Central

Kulangara, Karina; Adler, Andrew F.; Wang, Hong; Chellappan, Malathi; Hammett, Ellen; Yasuda, Ryohei; Leong, Kam W.

2014-01-01

Cellular reprogramming holds tremendous potential for cell therapy and regenerative medicine. Recently, fibroblasts have been directly converted into induced neurons (iNs) by overexpression of the neuronal transcription factors Ascl1, Brn2 and Myt1L. Hypothesizing that cell-topography interactions could influence the fibroblast-to-neuron reprogramming process, we investigated the effects of various topographies on iNs produced by direct reprogramming. Final iN purity and conversion efficiency were increased on micrograting substrates. Neurite branching was increased on microposts and decreased on microgratings, with a simplified dendritic arbor characterized by the reduction of MAP2+ neurites. Neurite outgrowth increased significantly on various topographies. DNA microarray analysis detected 20 differentially expressed genes in iNs reprogrammed on smooth versus microgratings, and quantitative PCR (qPCR) confirmed the upregulation of Vip and downregulation of Thy1 and Bmp5 on microgratings. Electrophysiology and calcium imaging verified the functionality of these iNs. This study demonstrates the potential of applying topographical cues to optimize cellular reprogramming. PMID:24709523

Oxygen measurement in interstitially perfused cellularized constructs cultured in a miniaturized bioreactor.

PubMed

Raimondi, Manuela T; Giordano, Carmen; Pietrabissa, Riccardo

2015-12-18

The possibility of developing engineered tissue in vitro and maintaining the cell viability and functionality is primarily related to the possibility of controlling key culture parameters such as oxygen concentration and cell-specific oxygen consumption. We measured these parameters in a three-dimensional (3D) cellularized construct maintained under interstitially perfused culture in a miniaturized bioreactor. MG63 osteosarcoma cells were seeded at high density on a 3D polystyrene scaffold. The 3D scaffolds were sensorized with sensor foils made of a polymer, which fluoresce with intensity proportional to the local oxygen tension. Images of the sensor foil in contact with the cellularized construct were acquired with a video camera every four hours for six culture days and were elaborated with analytical imaging software to obtain oxygen concentration maps. The data collected indicate a globally decreasing oxygen concentration profile, with a total drop of 28% after six days of culture and an average drop of 10.5% between the inlet and outlet of the perfused construct. Moreover, by importing the measured oxygen concentration data and the cell counts in a model of mass transport, we calculated the cell-specific oxygen consumption over the whole culture period. The consumption increased with oxygen availability and ranged from 0.1 to 0.7 µmol/h/106 cells. The sensors used here allowed a non-invasive, contamination-free and non-destructive oxygen measurement over the whole culture period. This study is the basis for optimization of the culture parameters involved in oxygen supply, in order to guarantee maintenance of cell viability in our system.

Advancing multiscale structural mapping of the brain through fluorescence imaging and analysis across length scales

PubMed Central

Hogstrom, L. J.; Guo, S. M.; Murugadoss, K.; Bathe, M.

2016-01-01

Brain function emerges from hierarchical neuronal structure that spans orders of magnitude in length scale, from the nanometre-scale organization of synaptic proteins to the macroscopic wiring of neuronal circuits. Because the synaptic electrochemical signal transmission that drives brain function ultimately relies on the organization of neuronal circuits, understanding brain function requires an understanding of the principles that determine hierarchical neuronal structure in living or intact organisms. Recent advances in fluorescence imaging now enable quantitative characterization of neuronal structure across length scales, ranging from single-molecule localization using super-resolution imaging to whole-brain imaging using light-sheet microscopy on cleared samples. These tools, together with correlative electron microscopy and magnetic resonance imaging at the nanoscopic and macroscopic scales, respectively, now facilitate our ability to probe brain structure across its full range of length scales with cellular and molecular specificity. As these imaging datasets become increasingly accessible to researchers, novel statistical and computational frameworks will play an increasing role in efforts to relate hierarchical brain structure to its function. In this perspective, we discuss several prominent experimental advances that are ushering in a new era of quantitative fluorescence-based imaging in neuroscience along with novel computational and statistical strategies that are helping to distil our understanding of complex brain structure. PMID:26855758

Noninvasive amide proton transfer magnetic resonance imaging in evaluating the grading and cellularity of gliomas.

PubMed

Bai, Yan; Lin, Yusong; Zhang, Wei; Kong, Lingfei; Wang, Lifu; Zuo, Panli; Vallines, Ignacio; Schmitt, Benjamin; Tian, Jie; Song, Xiaolei; Zhou, Jinyuan; Wang, Meiyun

2017-01-24

Using noninvasive magnetic resonance imaging techniques to accurately evaluate the grading and cellularity of gliomas is beneficial for improving the patient outcomes. Amide proton transfer imaging is a noninvasive molecular magnetic resonance imaging technique based on chemical exchange saturation transfer mechanism that detects endogenous mobile proteins and peptides in biological tissues. Between August 2012 and November 2015, a total number of 44 patients with pathologically proven gliomas were included in this study. We compared the capability of amide proton transfer magnetic resonance imaging with that of noninvasive diffusion-weighted imaging and noninvasive 3-dimensional pseudo-continuous arterial spin imaging in evaluating the grading and cellularity of gliomas. Our results reveal that amide proton transfer magnetic resonance imaging is a superior imaging technique to diffusion-weighted imaging and 3-dimensional pseudo-continuous arterial spin imaging in the grading of gliomas. In addition, our results showed that the Ki-67 index correlated better with the amide proton transfer-weighted signal intensity than with the apparent diffusion coefficient value or the cerebral blood flow value in the gliomas. Amide proton transfer magnetic resonance imaging is a promising method for predicting the grading and cellularity of gliomas.

Live CLEM imaging to analyze nuclear structures at high resolution.

PubMed

Haraguchi, Tokuko; Osakada, Hiroko; Koujin, Takako

2015-01-01

Fluorescence microscopy (FM) and electron microscopy (EM) are powerful tools for observing molecular components in cells. FM can provide temporal information about cellular proteins and structures in living cells. EM provides nanometer resolution images of cellular structures in fixed cells. We have combined FM and EM to develop a new method of correlative light and electron microscopy (CLEM), called "Live CLEM." In this method, the dynamic behavior of specific molecules of interest is first observed in living cells using fluorescence microscopy (FM) and then cellular structures in the same cell are observed using electron microscopy (EM). Following image acquisition, FM and EM images are compared to enable the fluorescent images to be correlated with the high-resolution images of cellular structures obtained using EM. As this method enables analysis of dynamic events involving specific molecules of interest in the context of specific cellular structures at high resolution, it is useful for the study of nuclear structures including nuclear bodies. Here we describe Live CLEM that can be applied to the study of nuclear structures in mammalian cells.

Image enhancement by non-linear extrapolation in frequency space

NASA Technical Reports Server (NTRS)

Anderson, Charles H. (Inventor); Greenspan, Hayit K. (Inventor)

1998-01-01

An input image is enhanced to include spatial frequency components having frequencies higher than those in an input image. To this end, an edge map is generated from the input image using a high band pass filtering technique. An enhancing map is subsequently generated from the edge map, with the enhanced map having spatial frequencies exceeding an initial maximum spatial frequency of the input image. The enhanced map is generated by applying a non-linear operator to the edge map in a manner which preserves the phase transitions of the edges of the input image. The enhanced map is added to the input image to achieve a resulting image having spatial frequencies greater than those in the input image. Simplicity of computations and ease of implementation allow for image sharpening after enlargement and for real-time applications such as videophones, advanced definition television, zooming, and restoration of old motion pictures.

Implementation of a Synchronized Oscillator Circuit for Fast Sensing and Labeling of Image Objects

PubMed Central

Kowalski, Jacek; Strzelecki, Michal; Kim, Hyongsuk

2011-01-01

We present an application-specific integrated circuit (ASIC) CMOS chip that implements a synchronized oscillator cellular neural network with a matrix size of 32 × 32 for object sensing and labeling in binary images. Networks of synchronized oscillators are a recently developed tool for image segmentation and analysis. Its parallel network operation is based on a “temporary correlation” theory that attempts to describe scene recognition as if performed by the human brain. The synchronized oscillations of neuron groups attract a person’s attention if he or she is focused on a coherent stimulus (image object). For more than one perceived stimulus, these synchronized patterns switch in time between different neuron groups, thus forming temporal maps that code several features of the analyzed scene. In this paper, a new oscillator circuit based on a mathematical model is proposed, and the network architecture and chip functional blocks are presented and discussed. The proposed chip is implemented in AMIS 0.35 μm C035M-D 5M/1P technology. An application of the proposed network chip for the segmentation of insulin-producing pancreatic islets in magnetic resonance liver images is presented. PMID:22163803

Cellular imaging of deep organ using two-photon Bessel light-sheet nonlinear structured illumination microscopy

PubMed Central

Zhao, Ming; Zhang, Han; Li, Yu; Ashok, Amit; Liang, Rongguang; Zhou, Weibin; Peng, Leilei

2014-01-01

In vivo fluorescent cellular imaging of deep internal organs is highly challenging, because the excitation needs to penetrate through strong scattering tissue and the emission signal is degraded significantly by photon diffusion induced by tissue-scattering. We report that by combining two-photon Bessel light-sheet microscopy with nonlinear structured illumination microscopy (SIM), live samples up to 600 microns wide can be imaged by light-sheet microscopy with 500 microns penetration depth, and diffused background in deep tissue light-sheet imaging can be reduced to obtain clear images at cellular resolution in depth beyond 200 microns. We demonstrate in vivo two-color imaging of pronephric glomeruli and vasculature of zebrafish kidney, whose cellular structures located at the center of the fish body are revealed in high clarity by two-color two-photon Bessel light-sheet SIM. PMID:24876996

Raman-based imaging uncovers the effects of alginate hydrogel implants in spinal cord injury

NASA Astrophysics Data System (ADS)

Galli, Roberta; Tamosaityte, Sandra; Koch, Maria; Sitoci-Ficici, Kerim H.; Later, Robert; Uckermann, Ortrud; Beiermeister, Rudolf; Gelinsky, Michael; Schackert, Gabriele; Kirsch, Matthias; Koch, Edmund; Steiner, Gerald

2015-07-01

The treatment of spinal cord injury by using implants that provide a permissive environment for axonal growth is in the focus of the research for regenerative therapies. Here, Raman-based label-free techniques were applied for the characterization of morphochemical properties of surgically induced spinal cord injury in the rat that received an implant of soft unfunctionalized alginate hydrogel. Raman microspectroscopy followed by chemometrics allowed mapping the different degenerative areas, while multimodal multiphoton microscopy (e.g. the combination of coherent anti-Stokes Raman scattering (CARS), endogenous two-photon fluorescence and second harmonic generation on the same platform) enabled to address the morphochemistry of the tissue at cellular level. The regions of injury, characterized by demyelination and scarring, were retrieved and the distribution of key tissue components was evaluated by Raman mapping. The alginate hydrogel was detected in the lesion up to six months after implantation and had positive effects on the nervous tissue. For instance, multimodal multiphoton microscopy complemented the results of Raman mapping, providing the micromorphology of lipid-rich tissue structures by CARS and enabling to discern lipid-rich regions that contained myelinated axons from degenerative regions characterized by myelin fragmentation and presence of foam cells. These findings demonstrate that Raman-based imaging methods provide useful information for the evaluation of alginate implant effects and have therefore the potential to contribute to new strategies for monitoring degenerative and regenerative processes induced in SCI, thereby improving the effectiveness of therapies.

Complex Geometric Models of Diffusion and Relaxation in Healthy and Damaged White Matter

PubMed Central

Farrell, Jonathan A.D.; Smith, Seth A.; Reich, Daniel S.; Calabresi, Peter A.; van Zijl, Peter C.M.

2010-01-01

Which aspects of tissue microstructure affect diffusion weighted MRI signals? Prior models, many of which use Monte-Carlo simulations, have focused on relatively simple models of the cellular microenvironment and have not considered important anatomic details. With the advent of higher-order analysis models for diffusion imaging, such as high-angular-resolution diffusion imaging (HARDI), more realistic models are necessary. This paper presents and evaluates the reproducibility of simulations of diffusion in complex geometries. Our framework is quantitative, does not require specialized hardware, is easily implemented with little programming experience, and is freely available as open-source software. Models may include compartments with different diffusivities, permeabilities, and T2 time constants using both parametric (e.g., spheres and cylinders) and arbitrary (e.g., mesh-based) geometries. Three-dimensional diffusion displacement-probability functions are mapped with high reproducibility, and thus can be readily used to assess reproducibility of diffusion-derived contrasts. PMID:19739233

Macro-/micro-environment-sensitive chemosensing and biological imaging.

PubMed

Yang, Zhigang; Cao, Jianfang; He, Yanxia; Yang, Jung Ho; Kim, Taeyoung; Peng, Xiaojun; Kim, Jong Seung

2014-07-07

Environment-related parameters, including viscosity, polarity, temperature, hypoxia, and pH, play pivotal roles in controlling the physical or chemical behaviors of local molecules. In particular, in a biological environment, such factors predominantly determine the biological properties of the local environment or reflect corresponding status alterations. Abnormal changes in these factors would cause cellular malfunction or become a hallmark of the occurrence of severe diseases. Therefore, in recent years, they have increasingly attracted research interest from the fields of chemistry and biological chemistry. With the emergence of fluorescence sensing and imaging technology, several fluorescent chemosensors have been designed to respond to such parameters and to further map their distributions and variations in vitro/in vivo. In this work, we have reviewed a number of various environment-responsive chemosensors related to fluorescent recognition of viscosity, polarity, temperature, hypoxia, and pH that have been reported thus far.

Advantage of spatial map ion imaging in the study of large molecule photodissociation

NASA Astrophysics Data System (ADS)

Lee, Chin; Lin, Yen-Cheng; Lee, Shih-Huang; Lee, Yin-Yu; Tseng, Chien-Ming; Lee, Yuan-Tseh; Ni, Chi-Kung

2017-07-01

The original ion imaging technique has low velocity resolution, and currently, photodissociation is mostly investigated using velocity map ion imaging. However, separating signals from the background (resulting from undissociated excited parent molecules) is difficult when velocity map ion imaging is used for the photodissociation of large molecules (number of atoms ≥ 10). In this study, we used the photodissociation of phenol at the S1 band origin as an example to demonstrate how our multimass ion imaging technique, based on modified spatial map ion imaging, can overcome this difficulty. The photofragment translational energy distribution obtained when multimass ion imaging was used differed considerably from that obtained when velocity map ion imaging and Rydberg atom tagging were used. We used conventional translational spectroscopy as a second method to further confirm the experimental results, and we conclude that data should be interpreted carefully when velocity map ion imaging or Rydberg atom tagging is used in the photodissociation of large molecules. Finally, we propose a modified velocity map ion imaging technique without the disadvantages of the current velocity map ion imaging technique.

Criteria for the optimal selection of remote sensing optical images to map event landslides

NASA Astrophysics Data System (ADS)

Fiorucci, Federica; Giordan, Daniele; Santangelo, Michele; Dutto, Furio; Rossi, Mauro; Guzzetti, Fausto

2018-01-01

Landslides leave discernible signs on the land surface, most of which can be captured in remote sensing images. Trained geomorphologists analyse remote sensing images and map landslides through heuristic interpretation of photographic and morphological characteristics. Despite a wide use of remote sensing images for landslide mapping, no attempt to evaluate how the image characteristics influence landslide identification and mapping exists. This paper presents an experiment to determine the effects of optical image characteristics, such as spatial resolution, spectral content and image type (monoscopic or stereoscopic), on landslide mapping. We considered eight maps of the same landslide in central Italy: (i) six maps obtained through expert heuristic visual interpretation of remote sensing images, (ii) one map through a reconnaissance field survey, and (iii) one map obtained through a real-time kinematic (RTK) differential global positioning system (dGPS) survey, which served as a benchmark. The eight maps were compared pairwise and to a benchmark. The mismatch between each map pair was quantified by the error index, E. Results show that the map closest to the benchmark delineation of the landslide was obtained using the higher resolution image, where the landslide signature was primarily photographical (in the landslide source and transport area). Conversely, where the landslide signature was mainly morphological (in the landslide deposit) the best mapping result was obtained using the stereoscopic images. Albeit conducted on a single landslide, the experiment results are general, and provide useful information to decide on the optimal imagery for the production of event, seasonal and multi-temporal landslide inventory maps.

A mitochondria-selective near-infrared-emitting fluorescent dye for cellular imaging studies.

PubMed

Choi, Peter; Noguchi, Katsuya; Ishiyama, Munetaka; Denny, William A; Jose, Jiney

2018-05-03

This communication details the synthesis, evaluation of photophysical properties, and cellular imaging studies of cyanine chromophore based fluorescent dye 1 as a selective imaging agent for mitochondria. Copyright © 2018 Elsevier Ltd. All rights reserved.

Pan-neuronal calcium imaging with cellular resolution in freely swimming zebrafish.

PubMed

Kim, Dal Hyung; Kim, Jungsoo; Marques, João C; Grama, Abhinav; Hildebrand, David G C; Gu, Wenchao; Li, Jennifer M; Robson, Drew N

2017-11-01

Calcium imaging with cellular resolution typically requires an animal to be tethered under a microscope, which substantially restricts the range of behaviors that can be studied. To expand the behavioral repertoire amenable to imaging, we have developed a tracking microscope that enables whole-brain calcium imaging with cellular resolution in freely swimming larval zebrafish. This microscope uses infrared imaging to track a target animal in a behavior arena. On the basis of the predicted trajectory of the animal, we applied optimal control theory to a motorized stage system to cancel brain motion in three dimensions. We combined this motion-cancellation system with differential illumination focal filtering, a variant of HiLo microscopy, which enabled us to image the brain of a freely swimming larval zebrafish for more than an hour. This work expands the repertoire of natural behaviors that can be studied with cellular-resolution calcium imaging to potentially include spatial navigation, social behavior, feeding and reward.

Decomposing Oncogenic Transcriptional Signatures to Generate Maps of Divergent Cellular States* | Office of Cancer Genomics

Cancer.gov

The systematic sequencing of the cancer genome has led to the identification of numerous genetic alterations in cancer. However, a deeper understanding of the functional consequences of these alterations is necessary to guide appropriate therapeutic strategies. Here, we describe Onco-GPS (OncoGenic Positioning System), a data-driven analysis framework to organize individual tumor samples with shared oncogenic alterations onto a reference map defined by their underlying cellular states.

Investigation of contrast-enhanced subtracted breast CT images with MAP-EM based on projection-based weighting imaging.

PubMed

Zhou, Zhengdong; Guan, Shaolin; Xin, Runchao; Li, Jianbo

2018-06-01

Contrast-enhanced subtracted breast computer tomography (CESBCT) images acquired using energy-resolved photon counting detector can be helpful to enhance the visibility of breast tumors. In such technology, one challenge is the limited number of photons in each energy bin, thereby possibly leading to high noise in separate images from each energy bin, the projection-based weighted image, and the subtracted image. In conventional low-dose CT imaging, iterative image reconstruction provides a superior signal-to-noise compared with the filtered back projection (FBP) algorithm. In this paper, maximum a posteriori expectation maximization (MAP-EM) based on projection-based weighting imaging for reconstruction of CESBCT images acquired using an energy-resolving photon counting detector is proposed, and its performance was investigated in terms of contrast-to-noise ratio (CNR). The simulation study shows that MAP-EM based on projection-based weighting imaging can improve the CNR in CESBCT images by 117.7%-121.2% compared with FBP based on projection-based weighting imaging method. When compared with the energy-integrating imaging that uses the MAP-EM algorithm, projection-based weighting imaging that uses the MAP-EM algorithm can improve the CNR of CESBCT images by 10.5%-13.3%. In conclusion, MAP-EM based on projection-based weighting imaging shows significant improvement the CNR of the CESBCT image compared with FBP based on projection-based weighting imaging, and MAP-EM based on projection-based weighting imaging outperforms MAP-EM based on energy-integrating imaging for CESBCT imaging.

Brain Injury Lesion Imaging Using Preconditioned Quantitative Susceptibility Mapping without Skull Stripping.

PubMed

Soman, S; Liu, Z; Kim, G; Nemec, U; Holdsworth, S J; Main, K; Lee, B; Kolakowsky-Hayner, S; Selim, M; Furst, A J; Massaband, P; Yesavage, J; Adamson, M M; Spincemallie, P; Moseley, M; Wang, Y

2018-04-01

Identifying cerebral microhemorrhage burden can aid in the diagnosis and management of traumatic brain injury, stroke, hypertension, and cerebral amyloid angiopathy. MR imaging susceptibility-based methods are more sensitive than CT for detecting cerebral microhemorrhage, but methods other than quantitative susceptibility mapping provide results that vary with field strength and TE, require additional phase maps to distinguish blood from calcification, and depict cerebral microhemorrhages as bloom artifacts. Quantitative susceptibility mapping provides universal quantification of tissue magnetic property without these constraints but traditionally requires a mask generated by skull-stripping, which can pose challenges at tissue interphases. We evaluated the preconditioned quantitative susceptibility mapping MR imaging method, which does not require skull-stripping, for improved depiction of brain parenchyma and pathology. Fifty-six subjects underwent brain MR imaging with a 3D multiecho gradient recalled echo acquisition. Mask-based quantitative susceptibility mapping images were created using a commonly used mask-based quantitative susceptibility mapping method, and preconditioned quantitative susceptibility images were made using precondition-based total field inversion. All images were reviewed by a neuroradiologist and a radiology resident. Ten subjects (18%), all with traumatic brain injury, demonstrated blood products on 3D gradient recalled echo imaging. All lesions were visible on preconditioned quantitative susceptibility mapping, while 6 were not visible on mask-based quantitative susceptibility mapping. Thirty-one subjects (55%) demonstrated brain parenchyma and/or lesions that were visible on preconditioned quantitative susceptibility mapping but not on mask-based quantitative susceptibility mapping. Six subjects (11%) demonstrated pons artifacts on preconditioned quantitative susceptibility mapping and mask-based quantitative susceptibility mapping; they were worse on preconditioned quantitative susceptibility mapping. Preconditioned quantitative susceptibility mapping MR imaging can bring the benefits of quantitative susceptibility mapping imaging to clinical practice without the limitations of mask-based quantitative susceptibility mapping, especially for evaluating cerebral microhemorrhage-associated pathologies, such as traumatic brain injury. © 2018 by American Journal of Neuroradiology.

Measurable realistic image-based 3D mapping

NASA Astrophysics Data System (ADS)

Liu, W.; Wang, J.; Wang, J. J.; Ding, W.; Almagbile, A.

2011-12-01

Maps with 3D visual models are becoming a remarkable feature of 3D map services. High-resolution image data is obtained for the construction of 3D visualized models.The3D map not only provides the capabilities of 3D measurements and knowledge mining, but also provides the virtual experienceof places of interest, such as demonstrated in the Google Earth. Applications of 3D maps are expanding into the areas of architecture, property management, and urban environment monitoring. However, the reconstruction of high quality 3D models is time consuming, and requires robust hardware and powerful software to handle the enormous amount of data. This is especially for automatic implementation of 3D models and the representation of complicated surfacesthat still need improvements with in the visualisation techniques. The shortcoming of 3D model-based maps is the limitation of detailed coverage since a user can only view and measure objects that are already modelled in the virtual environment. This paper proposes and demonstrates a 3D map concept that is realistic and image-based, that enables geometric measurements and geo-location services. Additionally, image-based 3D maps provide more detailed information of the real world than 3D model-based maps. The image-based 3D maps use geo-referenced stereo images or panoramic images. The geometric relationships between objects in the images can be resolved from the geometric model of stereo images. The panoramic function makes 3D maps more interactive with users but also creates an interesting immersive circumstance. Actually, unmeasurable image-based 3D maps already exist, such as Google street view, but only provide virtual experiences in terms of photos. The topographic and terrain attributes, such as shapes and heights though are omitted. This paper also discusses the potential for using a low cost land Mobile Mapping System (MMS) to implement realistic image 3D mapping, and evaluates the positioning accuracy that a measureable realistic image-based (MRI) system can produce. The major contribution here is the implementation of measurable images on 3D maps to obtain various measurements from real scenes.

Lossless Compression of Classification-Map Data

NASA Technical Reports Server (NTRS)

Hua, Xie; Klimesh, Matthew

2009-01-01

A lossless image-data-compression algorithm intended specifically for application to classification-map data is based on prediction, context modeling, and entropy coding. The algorithm was formulated, in consideration of the differences between classification maps and ordinary images of natural scenes, so as to be capable of compressing classification- map data more effectively than do general-purpose image-data-compression algorithms. Classification maps are typically generated from remote-sensing images acquired by instruments aboard aircraft (see figure) and spacecraft. A classification map is a synthetic image that summarizes information derived from one or more original remote-sensing image(s) of a scene. The value assigned to each pixel in such a map is the index of a class that represents some type of content deduced from the original image data for example, a type of vegetation, a mineral, or a body of water at the corresponding location in the scene. When classification maps are generated onboard the aircraft or spacecraft, it is desirable to compress the classification-map data in order to reduce the volume of data that must be transmitted to a ground station.

Client-Side Image Maps: Achieving Accessibility and Section 508 Compliance

ERIC Educational Resources Information Center

Beasley, William; Jarvis, Moana

2004-01-01

Image maps are a means of making a picture "clickable", so that different portions of the image can be hyperlinked to different URLS. There are two basic types of image maps: server-side and client-side. Besides requiring access to a CGI on the server, server-side image maps are undesirable from the standpoint of accessibility--creating…

Molecular Imaging Probes for Positron Emission Tomography and Optical Imaging of Sentinel Lymph Node and Tumor

NASA Astrophysics Data System (ADS)

Qin, Zhengtao

Molecular imaging is visualizations and measurements of in vivo biological processes at the molecular or cellular level using specific imaging probes. As an emerging technology, biocompatible macromolecular or nanoparticle based targeted imaging probes have gained increasing popularities. Those complexes consist of a carrier, an imaging reporter, and a targeting ligand. The active targeting ability dramatically increases the specificity. And the multivalency effect may further reduce the dose while providing a decent signal. In this thesis, sentinel lymph node (SLN) mapping and cancer imaging are two research topics. The focus is to develop molecular imaging probes with high specificity and sensitivity, for Positron Emission Tomography (PET) and optical imaging. The objective of this thesis is to explore dextran radiopharmaceuticals and porous silicon nanoparticles based molecular imaging agents. Dextran polymers are excellent carriers to deliver imaging reporters or therapeutic agents due to its well established safety profile and oligosaccharide conjugation chemistry. There is also a wide selection of dextran polymers with different lengths. On the other hand, Silicon nanoparticles represent another class of biodegradable materials for imaging and drug delivery. The success in fluorescence lifetime imaging and enhancements of the immune activation potency was briefly discussed. Chapter 1 begins with an overview on current molecular imaging techniques and imaging probes. Chapter 2 presents a near-IR dye conjugated probe, IRDye 800CW-tilmanocept. Fluorophore density was optimized to generate the maximum brightness. It was labeled with 68Ga and 99mTc and in vivo SLN mapping was successfully performed in different animals, such as mice, rabbits, dogs and pigs. With 99mTc labeled IRDye 800CW-tilmanocept, chapter 3 introduces a two-day imaging protocol with a hand-held imager. Chapter 4 proposed a method to dual radiolabel the IRDye 800CW-tilmanocept with both 68Ga and 99mTc. Chapter 5 introduces a 68Ga metal chelating bioorthogonal tetrazine dextran probe for multistep imaging of a colon cancer. Chapter 6 presents the synthesis and in vivo evaluation of a Hepatocellular Carcinoma targeting PET probe 68Ga-Insulin-Dextran. Chapter 7 discusses a novel method to prepare silicon nanoparticles with great yield and size control. The last chapter 8 concludes all probes developed in this thesis and their clinical relevance.

Luminescent single-walled carbon nanotube-sensitized europium nanoprobes for cellular imaging

PubMed Central

Avti, Pramod K; Sitharaman, Balaji

2012-01-01

Lanthanoid-based optical probes with excitation wavelengths in the ultra-violet (UV) range (300–325 nm) have been widely developed as imaging probes. Efficient cellular imaging requires that lanthanoid optical probes be excited at visible wavelengths, to avoid UV damage to cells. The efficacy of europium-catalyzed single-walled carbon nanotubes (Eu-SWCNTs), as visible nanoprobes for cellular imaging, is reported in this study. Confocal fluorescence microscopy images of breast cancer cells (SK-BR-3 and MCF-7) and normal cells (NIH 3T3), treated with Eu-SWCNT at 0.2 μg/mL concentration, showed bright red luminescence after excitation at 365 nm and 458 nm wavelengths. Cell viability analysis showed no cytotoxic effects after the incubation of cells with Eu-SWCNTs at this concentration. Eu-SWCNT uptake is via the endocytosis mechanism. Labeling efficiency, defined as the percentage of incubated cells that uptake Eu-SWCNT, was 95%–100% for all cell types. The average cellular uptake concentration was 6.68 ng Eu per cell. Intracellular localization was further corroborated by transmission electron microscopy and Raman microscopy. The results indicate that Eu-SWCNT shows potential as a novel cellular imaging probe, wherein SWCNT sensitizes Eu3+ ions to allow excitation at visible wavelengths, and stable time-resolved red emission. The ability to functionalize biomolecules on the exterior surface of Eu-SWCNT makes it an excellent candidate for targeted cellular imaging. PMID:22619533

Scanning ion conductance microscopy: a convergent high-resolution technology for multi-parametric analysis of living cardiovascular cells

PubMed Central

Miragoli, Michele; Moshkov, Alexey; Novak, Pavel; Shevchuk, Andrew; Nikolaev, Viacheslav O.; El-Hamamsy, Ismail; Potter, Claire M. F.; Wright, Peter; Kadir, S.H. Sheikh Abdul; Lyon, Alexander R.; Mitchell, Jane A.; Chester, Adrian H.; Klenerman, David; Lab, Max J.; Korchev, Yuri E.; Harding, Sian E.; Gorelik, Julia

2011-01-01

Cardiovascular diseases are complex pathologies that include alterations of various cell functions at the levels of intact tissue, single cells and subcellular signalling compartments. Conventional techniques to study these processes are extremely divergent and rely on a combination of individual methods, which usually provide spatially and temporally limited information on single parameters of interest. This review describes scanning ion conductance microscopy (SICM) as a novel versatile technique capable of simultaneously reporting various structural and functional parameters at nanometre resolution in living cardiovascular cells at the level of the whole tissue, single cells and at the subcellular level, to investigate the mechanisms of cardiovascular disease. SICM is a multimodal imaging technology that allows concurrent and dynamic analysis of membrane morphology and various functional parameters (cell volume, membrane potentials, cellular contraction, single ion-channel currents and some parameters of intracellular signalling) in intact living cardiovascular cells and tissues with nanometre resolution at different levels of organization (tissue, cellular and subcellular levels). Using this technique, we showed that at the tissue level, cell orientation in the inner and outer aortic arch distinguishes atheroprone and atheroprotected regions. At the cellular level, heart failure leads to a pronounced loss of T-tubules in cardiac myocytes accompanied by a reduction in Z-groove ratio. We also demonstrated the capability of SICM to measure the entire cell volume as an index of cellular hypertrophy. This method can be further combined with fluorescence to simultaneously measure cardiomyocyte contraction and intracellular calcium transients or to map subcellular localization of membrane receptors coupled to cyclic adenosine monophosphate production. The SICM pipette can be used for patch-clamp recordings of membrane potential and single channel currents. In conclusion, SICM provides a highly informative multimodal imaging platform for functional analysis of the mechanisms of cardiovascular diseases, which should facilitate identification of novel therapeutic strategies. PMID:21325316

RECENT DEVELOPMENTS IN THE U. S. GEOLOGICAL SURVEY'S LANDSAT IMAGE MAPPING PROGRAM.

USGS Publications Warehouse

Brownworth, Frederick S.; Rohde, Wayne G.

1986-01-01

At the 1984 ASPRS-ACSM Convention in Washington, D. C. a paper on 'The Emerging U. S. Geological Survey Image Mapping Program' was presented that discussed recent satellite image mapping advancements and published products. Since then Landsat image mapping has become an integral part of the National Mapping Program. The Survey currently produces about 20 Landsat multispectral scanner (MSS) and Thematic Mapper (TM) image map products annually at 1:250,000 and 1:100,000 scales, respectively. These Landsat image maps provide users with a regional or synoptic view of an area. The resultant geographical presentation of the terrain and cultural features will help planners and managers make better decisions regarding the use of our national resources.

Combined lineage mapping and gene expression profiling of embryonic brain patterning using ultrashort pulse microscopy and image registration

NASA Astrophysics Data System (ADS)

Gibbs, Holly C.; Dodson, Colin R.; Bai, Yuqiang; Lekven, Arne C.; Yeh, Alvin T.

2014-12-01

During embryogenesis, presumptive brain compartments are patterned by dynamic networks of gene expression. The spatiotemporal dynamics of these networks, however, have not been characterized with sufficient resolution for us to understand the regulatory logic resulting in morphogenetic cellular behaviors that give the brain its shape. We have developed a new, integrated approach using ultrashort pulse microscopy [a high-resolution, two-photon fluorescence (2PF)-optical coherence microscopy (OCM) platform using 10-fs pulses] and image registration to study brain patterning and morphogenesis in zebrafish embryos. As a demonstration, we used time-lapse 2PF to capture midbrain-hindbrain boundary morphogenesis and a wnt1 lineage map from embryos during brain segmentation. We then performed in situ hybridization to deposit NBT/BCIP, where wnt1 remained actively expressed, and reimaged the embryos with combined 2PF-OCM. When we merged these datasets using morphological landmark registration, we found that the mechanism of boundary formation differs along the dorsoventral axis. Dorsally, boundary sharpening is dominated by changes in gene expression, while ventrally, sharpening may be accomplished by lineage sorting. We conclude that the integrated visualization of lineage reporter and gene expression domains simultaneously with brain morphology will be useful for understanding how changes in gene expression give rise to proper brain compartmentalization and structure.

Combined lineage mapping and gene expression profiling of embryonic brain patterning using ultrashort pulse microscopy and image registration.

PubMed

Gibbs, Holly C; Dodson, Colin R; Bai, Yuqiang; Lekven, Arne C; Yeh, Alvin T

2014-12-01

During embryogenesis, presumptive brain compartments are patterned by dynamic networks of gene expression. The spatiotemporal dynamics of these networks, however, have not been characterized with sufficient resolution for us to understand the regulatory logic resulting in morphogenetic cellular behaviors that give the brain its shape. We have developed a new, integrated approach using ultrashort pulse microscopy [a high-resolution, two-photon fluorescence (2PF)-optical coherence microscopy (OCM) platform using 10-fs pulses] and image registration to study brain patterning and morphogenesis in zebrafish embryos. As a demonstration, we used time-lapse 2PF to capture midbrain-hindbrain boundary morphogenesis and a wnt1 lineage map from embryos during brain segmentation. We then performed in situ hybridization to deposit NBT/BCIP, where wnt1 remained actively expressed, and reimaged the embryos with combined 2PF-OCM. When we merged these datasets using morphological landmark registration, we found that the mechanism of boundary formation differs along the dorsoventral axis. Dorsally, boundary sharpening is dominated by changes in gene expression, while ventrally, sharpening may be accomplished by lineage sorting. We conclude that the integrated visualization of lineage reporter and gene expression domains simultaneously with brain morphology will be useful for understanding how changes in gene expression give rise to proper brain compartmentalization and structure.

A grid matrix-based Raman spectroscopic method to characterize different cell milieu in biopsied axillary sentinel lymph nodes of breast cancer patients.

PubMed

Som, Dipasree; Tak, Megha; Setia, Mohit; Patil, Asawari; Sengupta, Amit; Chilakapati, C Murali Krishna; Srivastava, Anurag; Parmar, Vani; Nair, Nita; Sarin, Rajiv; Badwe, R

2016-01-01

Raman spectroscopy which is based upon inelastic scattering of photons has a potential to emerge as a noninvasive bedside in vivo or ex vivo molecular diagnostic tool. There is a need to improve the sensitivity and predictability of Raman spectroscopy. We developed a grid matrix-based tissue mapping protocol to acquire cellular-specific spectra that also involved digital microscopy for localizing malignant and lymphocytic cells in sentinel lymph node biopsy sample. Biosignals acquired from specific cellular milieu were subjected to an advanced supervised analytical method, i.e., cross-correlation and peak-to-peak ratio in addition to PCA and PC-LDA. We observed decreased spectral intensity as well as shift in the spectral peaks of amides and lipid bands in the completely metastatic (cancer cells) lymph nodes with high cellular density. Spectral library of normal lymphocytes and metastatic cancer cells created using the cellular specific mapping technique can be utilized to create an automated smart diagnostic tool for bench side screening of sampled lymph nodes. Spectral library of normal lymphocytes and metastatic cancer cells created using the cellular specific mapping technique can be utilized to develop an automated smart diagnostic tool for bench side screening of sampled lymph nodes supported by ongoing global research in developing better technology and signal and big data processing algorithms.

Iterative h-minima-based marker-controlled watershed for cell nucleus segmentation.

PubMed

Koyuncu, Can Fahrettin; Akhan, Ece; Ersahin, Tulin; Cetin-Atalay, Rengul; Gunduz-Demir, Cigdem

2016-04-01

Automated microscopy imaging systems facilitate high-throughput screening in molecular cellular biology research. The first step of these systems is cell nucleus segmentation, which has a great impact on the success of the overall system. The marker-controlled watershed is a technique commonly used by the previous studies for nucleus segmentation. These studies define their markers finding regional minima on the intensity/gradient and/or distance transform maps. They typically use the h-minima transform beforehand to suppress noise on these maps. The selection of the h value is critical; unnecessarily small values do not sufficiently suppress the noise, resulting in false and oversegmented markers, and unnecessarily large ones suppress too many pixels, causing missing and undersegmented markers. Because cell nuclei show different characteristics within an image, the same h value may not work to define correct markers for all the nuclei. To address this issue, in this work, we propose a new watershed algorithm that iteratively identifies its markers, considering a set of different h values. In each iteration, the proposed algorithm defines a set of candidates using a particular h value and selects the markers from those candidates provided that they fulfill the size requirement. Working with widefield fluorescence microscopy images, our experiments reveal that the use of multiple h values in our iterative algorithm leads to better segmentation results, compared to its counterparts. © 2016 International Society for Advancement of Cytometry. © 2016 International Society for Advancement of Cytometry.

Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Que, Emily L.; Bleher, Reiner; Duncan, Francesca E.

2014-12-15

Fertilization of a mammalian egg initiates a series of 'zinc sparks' that are necessary to induce the egg-to-embryo transition. Despite the importance of these zinc-efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches that resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy-dispersive spectroscopy, X-ray fluorescence microscopy and three-dimensional elemental tomography for high-resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each ofmore » which contains, on average, 10(6) zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes« less

Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks.

PubMed

Que, Emily L; Bleher, Reiner; Duncan, Francesca E; Kong, Betty Y; Gleber, Sophie C; Vogt, Stefan; Chen, Si; Garwin, Seth A; Bayer, Amanda R; Dravid, Vinayak P; Woodruff, Teresa K; O'Halloran, Thomas V

2015-02-01

Fertilization of a mammalian egg initiates a series of 'zinc sparks' that are necessary to induce the egg-to-embryo transition. Despite the importance of these zinc-efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches that resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy-dispersive spectroscopy, X-ray fluorescence microscopy and three-dimensional elemental tomography for high-resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each of which contains, on average, 10(6) zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes.

Ultrafast imaging of cell elasticity with optical microelastography

PubMed Central

Grasland-Mongrain, Pol; Zorgani, Ali; Nakagawa, Shoma; Bernard, Simon; Paim, Lia Gomes; Fitzharris, Greg; Catheline, Stefan

2018-01-01

Elasticity is a fundamental cellular property that is related to the anatomy, functionality, and pathological state of cells and tissues. However, current techniques based on cell deformation, atomic force microscopy, or Brillouin scattering are rather slow and do not always accurately represent cell elasticity. Here, we have developed an alternative technique by applying shear wave elastography to the micrometer scale. Elastic waves were mechanically induced in live mammalian oocytes using a vibrating micropipette. These audible frequency waves were observed optically at 200,000 frames per second and tracked with an optical flow algorithm. Whole-cell elasticity was then mapped using an elastography method inspired by the seismology field. Using this approach we show that the elasticity of mouse oocytes is decreased when the oocyte cytoskeleton is disrupted with cytochalasin B. The technique is fast (less than 1 ms for data acquisition), precise (spatial resolution of a few micrometers), able to map internal cell structures, and robust and thus represents a tractable option for interrogating biomechanical properties of diverse cell types. PMID:29339488

Nanoscale mapping of the three-dimensional deformation field within commercial nanodiamonds

DOE PAGES

Maqbool, Muhammad Salman; Hoxley, David; Phillips, Nicholas W.; ...

2017-02-21

Here, the unique properties of nanodiamonds make them suitable for use in a wide range of applications, including as biomarkers for cellular tracking in vivo at the molecular level. The sustained fluorescence of nanodiamonds containing nitrogen-vacancy (N-V) centres is related to their internal structure and strain state. Theoretical studies predict that the location of the N-V centre and the nanodiamonds' residual elastic strain state have a major influence on their photoluminescence properties. However, to date there have been no direct measurements made of their spatially resolved deformation fields owing to the challenges that such measurements present. Here we apply themore » recently developed technique of Bragg coherent diffractive imaging (BCDI) to map the three-dimensional deformation field within a single nanodiamond of approximately 0.5 µm diameter. The results indicate that there are high levels of residual elastic strain present in the nanodiamond which could have a critical influence on its optical and electronic properties.« less

Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks

PubMed Central

Que, Emily L.; Bleher, Reiner; Duncan, Francesca E.; Kong, Betty Y.; Gleber, Sophie C.; Vogt, Stefan; Chen, Si; Garwin, Seth A.; Bayer, Amanda R.; Dravid, Vinayak; Woodruff, Teresa K.; O’Halloran, Thomas V.

2015-01-01

Fertilization of a mammalian egg induces a series of ‘zinc sparks’ that are necessary for inducing the egg-to-embryo transition. Despite the importance of these zinc efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches to resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy dispersive spectroscopy, X-ray fluorescence microscopy, and 3D elemental tomography for high resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each of which contains, on average, 106 zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes. PMID:25615666

Nanoscale mapping of the three-dimensional deformation field within commercial nanodiamonds

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maqbool, Muhammad Salman; Hoxley, David; Phillips, Nicholas W.

2017-01-01

The unique properties of nanodiamonds make them suitable for use in a wide range of applications, including as biomarkers for cellular tracking in vivo at the molecular level. The sustained fluorescence of nanodiamonds containing nitrogen-vacancy (N-V) centres is related to their internal structure and strain state. Theoretical studies predict that the location of the N-V centre and the nanodiamonds' residual elastic strain state have a major influence on their photoluminescence properties. However, to date there have been no direct measurements made of their spatially resolved deformation fields owing to the challenges that such measurements present. Here we apply the recentlymore » developed technique of Bragg coherent diffractive imaging (BCDI) to map the three-dimensional deformation field within a single nanodiamond of approximately 0.5 µm diameter. The results indicate that there are high levels of residual elastic strain present in the nanodiamond which could have a critical influence on its optical and electronic properties.« less

In vivo cellular imaging with microscopes enabled by MEMS scanners

NASA Astrophysics Data System (ADS)

Ra, Hyejun

High-resolution optical imaging plays an important role in medical diagnosis and biomedical research. Confocal microscopy is a widely used imaging method for obtaining cellular and sub-cellular images of biological tissue in reflectance and fluorescence modes. Its characteristic optical sectioning capability also enables three-dimensional (3-D) image reconstruction. However, its use has mostly been limited to excised tissues due to the requirement of high numerical aperture (NA) lenses for cellular resolution. Microscope miniaturization can enable in vivo imaging to make possible early cancer diagnosis and biological studies in the innate environment. In this dissertation, microscope miniaturization for in vivo cellular imaging is presented. The dual-axes confocal (DAC) architecture overcomes limitations of the conventional single-axis confocal (SAC) architecture to allow for miniaturization with high resolution. A microelectromechanical systems (MEMS) scanner is the central imaging component that is key in miniaturization of the DAC architecture. The design, fabrication, and characterization of the two-dimensional (2-D) MEMS scanner are presented. The gimbaled MEMS scanner is fabricated on a double silicon-on-insulator (SOI) wafer and is actuated by self-aligned vertical electrostatic combdrives. The imaging performance of the MEMS scanner in a DAC configuration is shown in a breadboard microscope setup, where reflectance and fluorescence imaging is demonstrated. Then, the MEMS scanner is integrated into a miniature DAC microscope. The whole imaging system is integrated into a portable unit for research in small animal models of human biology and disease. In vivo 3-D imaging is demonstrated on mouse skin models showing gene transfer and siRNA silencing. The siRNA silencing process is sequentially imaged in one mouse over time.

Detecting the Extent of Cellular Decomposition after Sub-Eutectoid Annealing in Rolled UMo Foils

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kautz, Elizabeth J.; Jana, Saumyadeep; Devaraj, Arun

2017-07-31

This report presents an automated image processing approach to quantifying microstructure image data, specifically the extent of eutectoid (cellular) decomposition in rolled U-10Mo foils. An image processing approach is used here to be able to quantitatively describe microstructure image data in order to relate microstructure to processing parameters (time, temperature, deformation).

Sparsity-constrained PET image reconstruction with learned dictionaries

NASA Astrophysics Data System (ADS)

Tang, Jing; Yang, Bao; Wang, Yanhua; Ying, Leslie

2016-09-01

PET imaging plays an important role in scientific and clinical measurement of biochemical and physiological processes. Model-based PET image reconstruction such as the iterative expectation maximization algorithm seeking the maximum likelihood solution leads to increased noise. The maximum a posteriori (MAP) estimate removes divergence at higher iterations. However, a conventional smoothing prior or a total-variation (TV) prior in a MAP reconstruction algorithm causes over smoothing or blocky artifacts in the reconstructed images. We propose to use dictionary learning (DL) based sparse signal representation in the formation of the prior for MAP PET image reconstruction. The dictionary to sparsify the PET images in the reconstruction process is learned from various training images including the corresponding MR structural image and a self-created hollow sphere. Using simulated and patient brain PET data with corresponding MR images, we study the performance of the DL-MAP algorithm and compare it quantitatively with a conventional MAP algorithm, a TV-MAP algorithm, and a patch-based algorithm. The DL-MAP algorithm achieves improved bias and contrast (or regional mean values) at comparable noise to what the other MAP algorithms acquire. The dictionary learned from the hollow sphere leads to similar results as the dictionary learned from the corresponding MR image. Achieving robust performance in various noise-level simulation and patient studies, the DL-MAP algorithm with a general dictionary demonstrates its potential in quantitative PET imaging.

Preferential tumor cellular uptake and retention of indocyanine green for in vivo tumor imaging.

PubMed

Onda, Nobuhiko; Kimura, Masayuki; Yoshida, Toshinori; Shibutani, Makoto

2016-08-01

Indocyanine green (ICG) is a fluorescent agent approved for clinical applications by the Food and Drug Administration and European Medicines Agency. This study examined the mechanism of tumor imaging using intravenously administered ICG. The in vivo kinetics of intravenously administered ICG were determined in tumor xenografts using microscopic approaches that enabled both spatio-temporal and high-magnification analyses. The mechanism of ICG-based tumor imaging was examined at the cellular level in six phenotypically different human colon cancer cell lines exhibiting different grades of epithelioid organization. ICG fluorescence imaging detected xenograft tumors, even those < 1 mm in size, based on their preferential cellular uptake and retention of the dye following its rapid tissue-non-specific delivery, in contrast to its rapid clearance by normal tissue. Live-cell imaging revealed that cellular ICG uptake is temperature-dependent and occurs after ICG binding to the cellular membrane, a pattern suggesting endocytic uptake as the mechanism. Cellular ICG uptake correlated inversely with the formation of tight junctions. Intracellular ICG was entrapped in the membrane traffic system, resulting in its slow turnover and prolonged retention by tumor cells. Our results suggest that tumor-specific imaging by ICG involves non-specific delivery of the dye to tissues followed by preferential tumor cellular uptake and retention. The tumor cell-preference of ICG is driven by passive tumor cell-targeting, the inherent ability of ICG to bind to cell membranes, and the high endocytic activity of tumor cells in association with the disruption of their tight junctions. © 2016 UICC.

Exploring the Spatial and Temporal Organization of a Cell’s Proteome

PubMed Central

Beck, Martin; Topf, Maya; Frazier, Zachary; Tjong, Harianto; Xu, Min; Zhang, Shihua; Alber, Frank

2013-01-01

To increase our current understanding of cellular processes, such as cell signaling and division, knowledge is needed about the spatial and temporal organization of the proteome at different organizational levels. These levels cover a wide range of length and time scales: from the atomic structures of macromolecules for inferring their molecular function, to the quantitative description of their abundance, and distribution in the cell. Emerging new experimental technologies are greatly increasing the availability of such spatial information on the molecular organization in living cells. This review addresses three fields that have significantly contributed to our understanding of the proteome’s spatial and temporal organization: first, methods for the structure determination of individual macromolecular assemblies, specifically the fitting of atomic structures into density maps generated from electron microscopy techniques; second, research that visualizes the spatial distributions of these complexes within the cellular context using cryo electron tomography techniques combined with computational image processing; and third, methods for the spatial modeling of the dynamic organization of the proteome, specifically those methods for simulating reaction and diffusion of proteins and complexes in crowded intracellular fluids. The long-term goal is to integrate the varied data about a proteome’s organization into a spatially explicit, predictive model of cellular processes. PMID:21094684

Optical Coherence Microscopy

NASA Astrophysics Data System (ADS)

Aguirre, Aaron D.; Zhou, Chao; Lee, Hsiang-Chieh; Ahsen, Osman O.; Fujimoto, James G.

Cellular imaging of human tissues remains an important advance for many clinical applications of optical coherence tomography (OCT). Imaging cells with traditional OCT systems has not been possible due to the limited transverse resolution of such techniques. Optical coherence microscopy (OCM) refers to OCT methods that achieve high transverse resolution to visualize cells and subcellular features. This chapter provides a comprehensive discussion of the rationale for cellular imaging in human tissues as well as a review of the key technological advances required to achieve it. Time domain and Fourier domain OCM approaches are described with an emphasis on state of the art system designs, including miniaturized endoscopic imaging probes. Clinical applications are discussed and multiple examples of cellular imaging in human tissues are provided.

Comparing orbiter and rover image-based mapping of an ancient sedimentary environment, Aeolis Palus, Gale crater, Mars

NASA Astrophysics Data System (ADS)

Stack, K. M.; Edwards, C. S.; Grotzinger, J. P.; Gupta, S.; Sumner, D. Y.; Calef, F. J.; Edgar, L. A.; Edgett, K. S.; Fraeman, A. A.; Jacob, S. R.; Le Deit, L.; Lewis, K. W.; Rice, M. S.; Rubin, D.; Williams, R. M. E.; Williford, K. H.

2016-12-01

This study provides the first systematic comparison of orbital facies maps with detailed ground-based geology observations from the Mars Science Laboratory (MSL) Curiosity rover to examine the validity of geologic interpretations derived from orbital image data. Orbital facies maps were constructed for the Darwin, Cooperstown, and Kimberley waypoints visited by the Curiosity rover using High Resolution Imaging Science Experiment (HiRISE) images. These maps, which represent the most detailed orbital analysis of these areas to date, were compared with rover image-based geologic maps and stratigraphic columns derived from Curiosity's Mast Camera (Mastcam) and Mars Hand Lens Imager (MAHLI). Results show that bedrock outcrops can generally be distinguished from unconsolidated surficial deposits in high-resolution orbital images and that orbital facies mapping can be used to recognize geologic contacts between well-exposed bedrock units. However, process-based interpretations derived from orbital image mapping are difficult to infer without known regional context or observable paleogeomorphic indicators, and layer-cake models of stratigraphy derived from orbital maps oversimplify depositional relationships as revealed from a rover perspective. This study also shows that fine-scale orbital image-based mapping of current and future Mars landing sites is essential for optimizing the efficiency and science return of rover surface operations.

Comparing orbiter and rover image-based mapping of an ancient sedimentary environment, Aeolis Palus, Gale crater, Mars

USGS Publications Warehouse

Stack, Kathryn M.; Edwards, Christopher; Grotzinger, J. P.; Gupta, S.; Sumner, D.; Edgar, Lauren; Fraeman, A.; Jacob, S.; LeDeit, L.; Lewis, K.W.; Rice, M.S.; Rubin, D.; Calef, F.; Edgett, K.; Williams, R.M.E.; Williford, K.H.

2016-01-01

This study provides the first systematic comparison of orbital facies maps with detailed ground-based geology observations from the Mars Science Laboratory (MSL) Curiosity rover to examine the validity of geologic interpretations derived from orbital image data. Orbital facies maps were constructed for the Darwin, Cooperstown, and Kimberley waypoints visited by the Curiosity rover using High Resolution Imaging Science Experiment (HiRISE) images. These maps, which represent the most detailed orbital analysis of these areas to date, were compared with rover image-based geologic maps and stratigraphic columns derived from Curiosity’s Mast Camera (Mastcam) and Mars Hand Lens Imager (MAHLI). Results show that bedrock outcrops can generally be distinguished from unconsolidated surficial deposits in high-resolution orbital images and that orbital facies mapping can be used to recognize geologic contacts between well-exposed bedrock units. However, process-based interpretations derived from orbital image mapping are difficult to infer without known regional context or observable paleogeomorphic indicators, and layer-cake models of stratigraphy derived from orbital maps oversimplify depositional relationships as revealed from a rover perspective. This study also shows that fine-scale orbital image-based mapping of current and future Mars landing sites is essential for optimizing the efficiency and science return of rover surface operations.

Minimal entropy approximation for cellular automata

NASA Astrophysics Data System (ADS)

Fukś, Henryk

2014-02-01

We present a method for the construction of approximate orbits of measures under the action of cellular automata which is complementary to the local structure theory. The local structure theory is based on the idea of Bayesian extension, that is, construction of a probability measure consistent with given block probabilities and maximizing entropy. If instead of maximizing entropy one minimizes it, one can develop another method for the construction of approximate orbits, at the heart of which is the iteration of finite-dimensional maps, called minimal entropy maps. We present numerical evidence that the minimal entropy approximation sometimes outperforms the local structure theory in characterizing the properties of cellular automata. The density response curve for elementary CA rule 26 is used to illustrate this claim.

Large-scale topology and the default mode network in the mouse connectome

PubMed Central

Stafford, James M.; Jarrett, Benjamin R.; Miranda-Dominguez, Oscar; Mills, Brian D.; Cain, Nicholas; Mihalas, Stefan; Lahvis, Garet P.; Lattal, K. Matthew; Mitchell, Suzanne H.; David, Stephen V.; Fryer, John D.; Nigg, Joel T.; Fair, Damien A.

2014-01-01

Noninvasive functional imaging holds great promise for serving as a translational bridge between human and animal models of various neurological and psychiatric disorders. However, despite a depth of knowledge of the cellular and molecular underpinnings of atypical processes in mouse models, little is known about the large-scale functional architecture measured by functional brain imaging, limiting translation to human conditions. Here, we provide a robust processing pipeline to generate high-resolution, whole-brain resting-state functional connectivity MRI (rs-fcMRI) images in the mouse. Using a mesoscale structural connectome (i.e., an anterograde tracer mapping of axonal projections across the mouse CNS), we show that rs-fcMRI in the mouse has strong structural underpinnings, validating our procedures. We next directly show that large-scale network properties previously identified in primates are present in rodents, although they differ in several ways. Last, we examine the existence of the so-called default mode network (DMN)—a distributed functional brain system identified in primates as being highly important for social cognition and overall brain function and atypically functionally connected across a multitude of disorders. We show the presence of a potential DMN in the mouse brain both structurally and functionally. Together, these studies confirm the presence of basic network properties and functional networks of high translational importance in structural and functional systems in the mouse brain. This work clears the way for an important bridge measurement between human and rodent models, enabling us to make stronger conclusions about how regionally specific cellular and molecular manipulations in mice relate back to humans. PMID:25512496

Correlative two-photon and serial block face scanning electron microscopy in neuronal tissue using 3D near-infrared branding maps.

PubMed

Lees, Robert M; Peddie, Christopher J; Collinson, Lucy M; Ashby, Michael C; Verkade, Paul

2017-01-01

Linking cellular structure and function has always been a key goal of microscopy, but obtaining high resolution spatial and temporal information from the same specimen is a fundamental challenge. Two-photon (2P) microscopy allows imaging deep inside intact tissue, bringing great insight into the structural and functional dynamics of cells in their physiological environment. At the nanoscale, the complex ultrastructure of a cell's environment in tissue can be reconstructed in three dimensions (3D) using serial block face scanning electron microscopy (SBF-SEM). This provides a snapshot of high resolution structural information pertaining to the shape, organization, and localization of multiple subcellular structures at the same time. The pairing of these two imaging modalities in the same specimen provides key information to relate cellular dynamics to the ultrastructural environment. Until recently, approaches to relocate a region of interest (ROI) in tissue from 2P microscopy for SBF-SEM have been inefficient or unreliable. However, near-infrared branding (NIRB) overcomes this by using the laser from a multiphoton microscope to create fiducial markers for accurate correlation of 2P and electron microscopy (EM) imaging volumes. The process is quick and can be user defined for each sample. Here, to increase the efficiency of ROI relocation, multiple NIRB marks are used in 3D to target ultramicrotomy. A workflow is described and discussed to obtain a data set for 3D correlated light and electron microscopy, using three different preparations of brain tissue as examples. Copyright © 2017 Elsevier Inc. All rights reserved.

Simultaneous noncontact topography and electrochemical imaging by SECM/SICM featuring ion current feedback regulation.

PubMed

Takahashi, Yasufumi; Shevchuk, Andrew I; Novak, Pavel; Murakami, Yumi; Shiku, Hitoshi; Korchev, Yuri E; Matsue, Tomokazu

2010-07-28

We described a hybrid system of scanning electrochemical microscopy (SECM) and scanning ion conductance microscopy (SICM) with ion current feedback nanopositioning control for simultaneous imaging of noncontact topography and spatial distribution of electrochemical species. A nanopipette/nanoring electrode probe provided submicrometer resolution of the electrochemical measurement on surfaces with complex topology. The SECM/SICM probe had an aperture radius of 220 nm. The inner and outer radii of the SECM Au nanoring electrode were 330 and 550 nm, respectively. Characterization of the probe was performed with scanning electron microscopy (SEM), cyclic voltammetry (CV), and approach curve measurements. SECM/SICM was applied to simultaneous imaging of topography and electrochemical responses of enzymes (horse radish peroxidase (HRP) and glucose oxidase (GOD)) and single live cells (A6 cells, superior cervical ganglion (SCG) cells, and cardiac myocytes). The measurements revealed the distribution of activity of the enzyme spots on uneven surfaces with submicrometer resolution. SECM/SICM acquired high resolution topographic images of cells together with the map of electrochemical signals. This combined technique was also applied to the evaluation of the permeation property of electroactive species through cellular membranes.

Development and application of Fourier-transform infrared chemical imaging of tumour in human tissue.

PubMed

Petter, C H; Heigl, N; Rainer, M; Bakry, R; Pallua, J; Bonn, G K; Huck, C W

2009-01-01

Fourier-transform infrared (FT-IR) based mapping and imaging is a fast emerging technology which is being increasingly applied to investigate tissues in the high-throughput mode. The high resolution close to the cellular level, the possibility to determine the bio-distribution of molecules of interest (proteins, peptides, lipids, carbohydrates) without any pre-treatment and the offer to yield molecular structure information have brought evidence that this technique allows to gain new insights in cancer pathology. Thus, several individual mainly protein and peptide cancer markers ("biomarkers") can be identified from FT-IR tissue images, enabling accurate discrimination between healthy and tumour areas. Optimal data acquisition (spatial resolution, spectral resolution, signal to noise ratio), classification, and validation are necessary to establish practical protocols that can be translated to the qualitative and quantitative clinical routine analysis. Thereby, the development of modern fast infrared imaging systems has strongly supported its acceptance in clinical histopathology. In this review, the necessity of analysis based on global cancer statistics, instrumental setups and developments, experimental state of the art are summarised and applications to investigate different kinds of cancer (e.g., prostate, breast, cervical, colon, oral cavity) are shown and discussed in detail.

High-yield in vitro recordings from neurons functionally characterized in vivo.

PubMed

Weiler, Simon; Bauer, Joel; Hübener, Mark; Bonhoeffer, Tobias; Rose, Tobias; Scheuss, Volker

2018-06-01

In vivo two-photon calcium imaging provides detailed information about the activity and response properties of individual neurons. However, in vitro methods are often required to study the underlying neuronal connectivity and physiology at the cellular and synaptic levels at high resolution. This protocol provides a fast and reliable workflow for combining the two approaches by characterizing the response properties of individual neurons in mice in vivo using genetically encoded calcium indicators (GECIs), followed by retrieval of the same neurons in brain slices for further analysis in vitro (e.g., circuit mapping). In this approach, a reference frame is provided by fluorescent-bead tracks and sparsely transduced neurons expressing a structural marker in order to re-identify the same neurons. The use of GECIs provides a substantial advancement over previous approaches by allowing for repeated in vivo imaging. This opens the possibility of directly correlating experience-dependent changes in neuronal activity and feature selectivity with changes in neuronal connectivity and physiology. This protocol requires expertise both in in vivo two-photon calcium imaging and in vitro electrophysiology. It takes 3 weeks or more to complete, depending on the time allotted for repeated in vivo imaging of neuronal activity.

Evaluation of a color-coded Landsat 5/6 ratio image for mapping lithologic differences in western South Dakota

USGS Publications Warehouse

Raines, Gary L.; Bretz, R.F.; Shurr, George W.

1979-01-01

From analysis of a color-coded Landsat 5/6 ratio, image, a map of the vegetation density distribution has been produced by Raines of 25,000 sq km of western South Dakota. This 5/6 ratio image is produced digitally calculating the ratios of the bands 5 and 6 of the Landsat data and then color coding these ratios in an image. Bretz and Shurr compared this vegetation density map with published and unpublished data primarily of the U.S. Geological Survey and the South Dakota Geological Survey; good correspondence is seen between this map and existing geologic maps, especially with the soils map. We believe that this Landsat ratio image can be used as a tool to refine existing maps of surficial geology and bedrock, where bedrock is exposed, and to improve mapping accuracy in areas of poor exposure common in South Dakota. In addition, this type of image could be a useful, additional tool in mapping areas that are unmapped.

Close the Textbook & Open "The Cell: An Image Library"

ERIC Educational Resources Information Center

Saunders, Cheston; Taylor, Amy

2014-01-01

Many students leave the biology classroom with misconceptions centered on cellular structure. This article presents an activity in which students utilize images from an online database called "The Cell: An Image Library" (http://www.cellimagelibrary. org/) to gain a greater understanding of the diversity of cellular structure and the…

Software for Verifying Image-Correlation Tie Points

NASA Technical Reports Server (NTRS)

Klimeck, Gerhard; Yagi, Gary

2008-01-01

A computer program enables assessment of the quality of tie points in the image-correlation processes of the software described in the immediately preceding article. Tie points are computed in mappings between corresponding pixels in the left and right images of a stereoscopic pair. The mappings are sometimes not perfect because image data can be noisy and parallax can cause some points to appear in one image but not the other. The present computer program relies on the availability of a left- right correlation map in addition to the usual right left correlation map. The additional map must be generated, which doubles the processing time. Such increased time can now be afforded in the data-processing pipeline, since the time for map generation is now reduced from about 60 to 3 minutes by the parallelization discussed in the previous article. Parallel cluster processing time, therefore, enabled this better science result. The first mapping is typically from a point (denoted by coordinates x,y) in the left image to a point (x',y') in the right image. The second mapping is from (x',y') in the right image to some point (x",y") in the left image. If (x,y) and(x",y") are identical, then the mapping is considered perfect. The perfect-match criterion can be relaxed by introducing an error window that admits of round-off error and a small amount of noise. The mapping procedure can be repeated until all points in each image not connected to points in the other image are eliminated, so that what remains are verified correlation data.

High-throughput isotropic mapping of whole mouse brain using multi-view light-sheet microscopy

NASA Astrophysics Data System (ADS)

Nie, Jun; Li, Yusha; Zhao, Fang; Ping, Junyu; Liu, Sa; Yu, Tingting; Zhu, Dan; Fei, Peng

2018-02-01

Light-sheet fluorescence microscopy (LSFM) uses an additional laser-sheet to illuminate selective planes of the sample, thereby enabling three-dimensional imaging at high spatial-temporal resolution. These advantages make LSFM a promising tool for high-quality brain visualization. However, even by the use of LSFM, the spatial resolution remains insufficient to resolve the neural structures across a mesoscale whole mouse brain in three dimensions. At the same time, the thick-tissue scattering prevents a clear observation from the deep of brain. Here we use multi-view LSFM strategy to solve this challenge, surpassing the resolution limit of standard light-sheet microscope under a large field-of-view (FOV). As demonstrated by the imaging of optically-cleared mouse brain labelled with thy1-GFP, we achieve a brain-wide, isotropic cellular resolution of 3μm. Besides the resolution enhancement, multi-view braining imaging can also recover complete signals from deep tissue scattering and attenuation. The identification of long distance neural projections across encephalic regions can be identified and annotated as a result.

In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy

PubMed Central

Malide, Daniela; Métais, Jean-Yves; Dunbar, Cynthia E.

2014-01-01

We developed and validated a fluorescent marking methodology for clonal tracking of hematopoietic stem and progenitor cells (HSPCs) with high spatial and temporal resolution to study in vivo hematopoiesis using the murine bone marrow transplant experimental model. Genetic combinatorial marking using lentiviral vectors encoding fluorescent proteins (FPs) enabled cell fate mapping through advanced microscopy imaging. Vectors encoding five different FPs: Cerulean, EGFP, Venus, tdTomato, and mCherry were used to concurrently transduce HSPCs, creating a diverse palette of color marked cells. Imaging using confocal/two-photon hybrid microscopy enables simultaneous high resolution assessment of uniquely marked cells and their progeny in conjunction with structural components of the tissues. Volumetric analyses over large areas reveal that spectrally coded HSPC-derived cells can be detected non-invasively in various intact tissues, including the bone marrow (BM), for extensive periods of time following transplantation. Live studies combining video-rate multiphoton and confocal time-lapse imaging in 4D demonstrate the possibility of dynamic cellular and clonal tracking in a quantitative manner. PMID:25145579

Real-Time Three-Dimensional Cell Segmentation in Large-Scale Microscopy Data of Developing Embryos.

PubMed

Stegmaier, Johannes; Amat, Fernando; Lemon, William C; McDole, Katie; Wan, Yinan; Teodoro, George; Mikut, Ralf; Keller, Philipp J

2016-01-25

We present the Real-time Accurate Cell-shape Extractor (RACE), a high-throughput image analysis framework for automated three-dimensional cell segmentation in large-scale images. RACE is 55-330 times faster and 2-5 times more accurate than state-of-the-art methods. We demonstrate the generality of RACE by extracting cell-shape information from entire Drosophila, zebrafish, and mouse embryos imaged with confocal and light-sheet microscopes. Using RACE, we automatically reconstructed cellular-resolution tissue anisotropy maps across developing Drosophila embryos and quantified differences in cell-shape dynamics in wild-type and mutant embryos. We furthermore integrated RACE with our framework for automated cell lineaging and performed joint segmentation and cell tracking in entire Drosophila embryos. RACE processed these terabyte-sized datasets on a single computer within 1.4 days. RACE is easy to use, as it requires adjustment of only three parameters, takes full advantage of state-of-the-art multi-core processors and graphics cards, and is available as open-source software for Windows, Linux, and Mac OS. Copyright © 2016 Elsevier Inc. All rights reserved.

Dual-Modality, Dual-Functional Nanoprobes for Cellular and Molecular Imaging

PubMed Central

Menon, Jyothi U.; Gulaka, Praveen K.; McKay, Madalyn A.; Geethanath, Sairam; Liu, Li; Kodibagkar, Vikram D.

2012-01-01

An emerging need for evaluation of promising cellular therapies is a non-invasive method to image the movement and health of cells following transplantation. However, the use of a single modality to serve this purpose may not be advantageous as it may convey inaccurate or insufficient information. Multi-modal imaging strategies are becoming more popular for in vivo cellular and molecular imaging because of their improved sensitivity, higher resolution and structural/functional visualization. This study aims at formulating Nile Red doped hexamethyldisiloxane (HMDSO) nanoemulsions as dual modality (Magnetic Resonance Imaging/Fluorescence), dual-functional (oximetry/detection) nanoprobes for cellular and molecular imaging. HMDSO nanoprobes were prepared using a HS15-lecithin combination as surfactant and showed an average radius of 71±39 nm by dynamic light scattering and in vitro particle stability in human plasma over 24 hrs. They were found to readily localize in the cytosol of MCF7-GFP cells within 18 minutes of incubation. As proof of principle, these nanoprobes were successfully used for fluorescence imaging and for measuring pO2 changes in cells by magnetic resonance imaging, in vitro, thus showing potential for in vivo applications. PMID:23382776

Analysis of Students' Aptitude to Provide Meaning to Images that Represent Cellular Components at the Molecular Level

PubMed Central

Dahmani, Hassen-Reda; Schneeberger, Patricia

2009-01-01

The number of experimentally derived structures of cellular components is rapidly expanding, and this phenomenon is accompanied by the development of a new semiotic system for teaching. The infographic approach is shifting from a schematic toward a more realistic representation of cellular components. By realistic we mean artist-prepared or computer graphic images that closely resemble experimentally derived structures and are characterized by a low level of styling and simplification. This change brings about a new challenge for teachers: designing course instructions that allow students to interpret these images in a meaningful way. To determine how students deal with this change, we designed several image-based, in-course assessments. The images were highly relevant for the cell biology course but did not resemble any of the images in the teaching documents. We asked students to label the cellular components, describe their function, or both. What we learned from these tests is that realistic images, with a higher apparent level of complexity, do not deter students from investigating their meaning. When given a choice, the students do not necessarily choose the most simplified representation, and they were sensitive to functional indications embedded in realistic images. PMID:19723817

Evaluation of hyperpolarized [1-¹³C]-pyruvate by magnetic resonance to detect ionizing radiation effects in real time.

PubMed

Sandulache, Vlad C; Chen, Yunyun; Lee, Jaehyuk; Rubinstein, Ashley; Ramirez, Marc S; Skinner, Heath D; Walker, Christopher M; Williams, Michelle D; Tailor, Ramesh; Court, Laurence E; Bankson, James A; Lai, Stephen Y

2014-01-01

Ionizing radiation (IR) cytotoxicity is primarily mediated through reactive oxygen species (ROS). Since tumor cells neutralize ROS by utilizing reducing equivalents, we hypothesized that measurements of reducing potential using real-time hyperpolarized (HP) magnetic resonance spectroscopy (MRS) and spectroscopic imaging (MRSI) can serve as a surrogate marker of IR induced ROS. This hypothesis was tested in a pre-clinical model of anaplastic thyroid carcinoma (ATC), an aggressive head and neck malignancy. Human ATC cell lines were utilized to test IR effects on ROS and reducing potential in vitro and [1-¹³C] pyruvate HP-MRS/MRSI imaging of ATC orthotopic xenografts was used to study in vivo effects of IR. IR increased ATC intra-cellular ROS levels resulting in a corresponding decrease in reducing equivalent levels. Exogenous manipulation of cellular ROS and reducing equivalent levels altered ATC radiosensitivity in a predictable manner. Irradiation of ATC xenografts resulted in an acute drop in reducing potential measured using HP-MRS, reflecting the shunting of reducing equivalents towards ROS neutralization. Residual tumor tissue post irradiation demonstrated heterogeneous viability. We have adapted HP-MRS/MRSI to non-invasively measure IR mediated changes in tumor reducing potential in real time. Continued development of this technology could facilitate the development of an adaptive clinical algorithm based on real-time adjustments in IR dose and dose mapping.

Enface Thickness Mapping and Reflectance Imaging of Retinal Layers in Diabetic Retinopathy.

PubMed

Francis, Andrew W; Wanek, Justin; Lim, Jennifer I; Shahidi, Mahnaz

2015-01-01

To present a method for image segmentation and generation of enface thickness maps and reflectance images of retinal layers in healthy and diabetic retinopathy (DR) subjects. High density spectral domain optical coherence tomography (SDOCT) images were acquired in 10 healthy and 4 DR subjects. Customized image analysis software identified 5 retinal cell layer interfaces and generated thickness maps and reflectance images of the total retina (TR), inner retina (IR), outer retina (OR), and the inner segment ellipsoid (ISe) band. Thickness maps in DR subjects were compared to those of healthy subjects by generating deviation maps which displayed retinal locations with thickness below, within, and above the normal 95% confidence interval. In healthy subjects, TR and IR thickness maps displayed the foveal depression and increased thickness in the parafoveal region. OR and ISe thickness maps showed increased thickness at the fovea, consistent with normal retinal anatomy. In DR subjects, thickening and thinning in localized regions were demonstrated on TR, IR, OR, and ISe thickness maps, corresponding to retinal edema and atrophy, respectively. TR and OR reflectance images showed reduced reflectivity in regions of increased thickness. Hard exudates appeared as hyper-reflective spots in IR reflectance images and casted shadows on the deeper OR and ISe reflectance images. The ISe reflectance image clearly showed the presence of focal laser scars. Enface thickness mapping and reflectance imaging of retinal layers is a potentially useful method for quantifying the spatial and axial extent of pathologies due to DR.

Enface Thickness Mapping and Reflectance Imaging of Retinal Layers in Diabetic Retinopathy

PubMed Central

Francis, Andrew W.; Wanek, Justin; Lim, Jennifer I.; Shahidi, Mahnaz

2015-01-01

Purpose To present a method for image segmentation and generation of enface thickness maps and reflectance images of retinal layers in healthy and diabetic retinopathy (DR) subjects. Methods High density spectral domain optical coherence tomography (SDOCT) images were acquired in 10 healthy and 4 DR subjects. Customized image analysis software identified 5 retinal cell layer interfaces and generated thickness maps and reflectance images of the total retina (TR), inner retina (IR), outer retina (OR), and the inner segment ellipsoid (ISe) band. Thickness maps in DR subjects were compared to those of healthy subjects by generating deviation maps which displayed retinal locations with thickness below, within, and above the normal 95% confidence interval. Results In healthy subjects, TR and IR thickness maps displayed the foveal depression and increased thickness in the parafoveal region. OR and ISe thickness maps showed increased thickness at the fovea, consistent with normal retinal anatomy. In DR subjects, thickening and thinning in localized regions were demonstrated on TR, IR, OR, and ISe thickness maps, corresponding to retinal edema and atrophy, respectively. TR and OR reflectance images showed reduced reflectivity in regions of increased thickness. Hard exudates appeared as hyper-reflective spots in IR reflectance images and casted shadows on the deeper OR and ISe reflectance images. The ISe reflectance image clearly showed the presence of focal laser scars. Conclusions Enface thickness mapping and reflectance imaging of retinal layers is a potentially useful method for quantifying the spatial and axial extent of pathologies due to DR. PMID:26699878

User Preferences in Image Map Using

NASA Astrophysics Data System (ADS)

Vondráková, A.; Vozenilek, V.

2016-06-01

In the process of map making, the attention is given to the resulting image map (to be accurate, readable, and suit the primary purpose) and its user aspects. Current cartography understands the user issues as all matters relating to user perception, map use and also user preferences. Most commercial cartographic production is strongly connected to economic circumstances. Companies are discovering user's interests and market demands. However, is it sufficient to focus just on the user's preferences? Recent research on user aspects at Palacký University Olomouc addresses a much wider scope of user aspects. The user's preferences are very often distorting - the users think that the particular image map is kind, beautiful, and useful and they wants to buy it (or use it - it depends on the form of the map production). But when the same user gets the task to use practically this particular map (such as finding the shortest way), so the user concludes that initially preferred map is useless, and uses a map, that was worse evaluated according to his preferences. It is, therefore, necessary to evaluate not only the correctness of image maps and their aesthetics but also to assess the user perception and other user issues. For the accomplishment of such testing, eye-tracking technology is a useful tool. The research analysed how users read image maps, or if they prefer image maps over traditional maps. The eye tracking experiment on the comparison of the conventional and image map reading was conducted. The map readers were asked to solve few simple tasks with either conventional or image map. The readers' choice of the map to solve the task was one of investigated aspect of user preferences. Results demonstrate that the user preferences and user needs are often quite different issues. The research outcomes show that it is crucial to implement map user testing into the cartographic production process.

Autofluorescence-based diagnostic UV imaging of tissues and cells

NASA Astrophysics Data System (ADS)

Renkoski, Timothy E.

Cancer is the second leading cause of death in the United States, and its early diagnosis is critical to improving treatment options and patient outcomes. In autofluorescence (AF) imaging, light of controlled wavelengths is projected onto tissue, absorbed by specific molecules, and re-emitted at longer wavelengths. Images of re-emitted light are used together with spectral information to infer tissue functional information and diagnosis. This dissertation describes AF imaging studies of three different organs using data collected from fresh human surgical specimens. In the ovary study, illumination was at 365 nm, and images were captured at 8 emission wavelengths. Measurements from a multispectral imaging system and fiber optic probe were used to map tissue diagnosis at every image pixel. For the colon and pancreas studies, instrumentation was developed extending AF imaging capability to sub-300 nm excitation. Images excited in the deep UV revealed tryptophan and protein content which are believed to change with disease state. Several excitation wavelength bands from 280 nm to 440 nm were investigated. Microscopic AF images collected in the pancreas study included both cultured and primary cells. Several findings are reported. A method of transforming fiber optic probe spectra for direct comparison with imager spectra was devised. Normalization of AF data by green reflectance data was found useful in correcting hemoglobin absorption. Ratio images, both AF and reflectance, were formulated to highlight growths in the colon. Novel tryptophan AF images were found less useful for colon diagnostics than the new ratio techniques. Microscopic tryptophan AF images produce useful visualization of cellular protein content, but their diagnostic value requires further study.

Integrated adaptive optics optical coherence tomography and adaptive optics scanning laser ophthalmoscope system for simultaneous cellular resolution in vivo retinal imaging

PubMed Central

Zawadzki, Robert J.; Jones, Steven M.; Pilli, Suman; Balderas-Mata, Sandra; Kim, Dae Yu; Olivier, Scot S.; Werner, John S.

2011-01-01

We describe an ultrahigh-resolution (UHR) retinal imaging system that combines adaptive optics Fourier-domain optical coherence tomography (AO-OCT) with an adaptive optics scanning laser ophthalmoscope (AO-SLO) to allow simultaneous data acquisition by the two modalities. The AO-SLO subsystem was integrated into the previously described AO-UHR OCT instrument with minimal changes to the latter. This was done in order to ensure optimal performance and image quality of the AO- UHR OCT. In this design both imaging modalities share most of the optical components including a common AO-subsystem and vertical scanner. One of the benefits of combining Fd-OCT with SLO includes automatic co-registration between two acquisition channels for direct comparison between retinal structures imaged by both modalities (e.g., photoreceptor mosaics or microvasculature maps). Because of differences in the detection scheme of the two systems, this dual imaging modality instrument can provide insight into retinal morphology and potentially function, that could not be accessed easily by a single system. In this paper we describe details of the components and parameters of the combined instrument, including incorporation of a novel membrane magnetic deformable mirror with increased stroke and actuator count used as a single wavefront corrector. We also discuss laser safety calculations for this multimodal system. Finally, retinal images acquired in vivo with this system are presented. PMID:21698028

Multispectral Imaging Broadens Cellular Analysis

NASA Technical Reports Server (NTRS)

2007-01-01

Amnis Corporation, a Seattle-based biotechnology company, developed ImageStream to produce sensitive fluorescence images of cells in flow. The company responded to an SBIR solicitation from Ames Research Center, and proposed to evaluate several methods of extending the depth of field for its ImageStream system and implement the best as an upgrade to its commercial products. This would allow users to view whole cells at the same time, rather than just one section of each cell. Through Phase I and II SBIR contracts, Ames provided Amnis the funding the company needed to develop this extended functionality. For NASA, the resulting high-speed image flow cytometry process made its way into Medusa, a life-detection instrument built to collect, store, and analyze sample organisms from erupting hydrothermal vents, and has the potential to benefit space flight health monitoring. On the commercial end, Amnis has implemented the process in ImageStream, combining high-resolution microscopy and flow cytometry in a single instrument, giving researchers the power to conduct quantitative analyses of individual cells and cell populations at the same time, in the same experiment. ImageStream is also built for many other applications, including cell signaling and pathway analysis; classification and characterization of peripheral blood mononuclear cell populations; quantitative morphology; apoptosis (cell death) assays; gene expression analysis; analysis of cell conjugates; molecular distribution; and receptor mapping and distribution.

ADC texture—An imaging biomarker for high-grade glioma?

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brynolfsson, Patrik; Hauksson, Jón; Karlsson, Mikael

2014-10-15

Purpose: Survival for high-grade gliomas is poor, at least partly explained by intratumoral heterogeneity contributing to treatment resistance. Radiological evaluation of treatment response is in most cases limited to assessment of tumor size months after the initiation of therapy. Diffusion-weighted magnetic resonance imaging (MRI) and its estimate of the apparent diffusion coefficient (ADC) has been widely investigated, as it reflects tumor cellularity and proliferation. The aim of this study was to investigate texture analysis of ADC images in conjunction with multivariate image analysis as a means for identification of pretreatment imaging biomarkers. Methods: Twenty-three consecutive high-grade glioma patients were treatedmore » with radiotherapy (2 Gy/60 Gy) with concomitant and adjuvant temozolomide. ADC maps and T1-weighted anatomical images with and without contrast enhancement were collected prior to treatment, and (residual) tumor contrast enhancement was delineated. A gray-level co-occurrence matrix analysis was performed on the ADC maps in a cuboid encapsulating the tumor in coronal, sagittal, and transversal planes, giving a total of 60 textural descriptors for each tumor. In addition, similar examinations and analyses were performed at day 1, week 2, and week 6 into treatment. Principal component analysis (PCA) was applied to reduce dimensionality of the data, and the five largest components (scores) were used in subsequent analyses. MRI assessment three months after completion of radiochemotherapy was used for classifying tumor progression or regression. Results: The score scatter plots revealed that the first, third, and fifth components of the pretreatment examinations exhibited a pattern that strongly correlated to survival. Two groups could be identified: one with a median survival after diagnosis of 1099 days and one with 345 days, p = 0.0001. Conclusions: By combining PCA and texture analysis, ADC texture characteristics were identified, which seems to hold pretreatment prognostic information, independent of known prognostic factors such as age, stage, and surgical procedure. These findings encourage further studies with a larger patient cohort.« less

T2-weighted images are superior to other MR image types for the determination of diffuse intrinsic pontine glioma intratumoral heterogeneity.

PubMed

Harward, Stephen; Harrison Farber, S; Malinzak, Michael; Becher, Oren; Thompson, Eric M

2018-03-01

Diffuse intrinsic pontine glioma (DIPG) remains the main cause of death in children with brain tumors. Given the inefficacy of numerous peripherally delivered agents to treat DIPG, convection enhanced delivery (CED) of therapeutic agents is a promising treatment modality. The purpose of this study was to determine which MR imaging type provides the best discrimination of intratumoral heterogeneity to guide future stereotactic implantation of CED catheters into the most cellular tumor regions. Patients ages 18 years or younger with a diagnosis of DIPG from 2000 to 2015 were included. Radiographic heterogeneity index (HI) of the tumor was calculated by measuring the standard deviation of signal intensity of the tumor (SD Tumor ) normalized to the genu of the corpus callosum (SD Corpus Callosum ). Four MR image types (T2-weighted, contrast-enhanced T1-weighted, FLAIR, and ADC) were analyzed at several time points both before and after radiotherapy and chemotherapy. HI values across these MR image types were compared and correlated with patient survival. MR images from 18 patients with DIPG were evaluated. The mean survival ± standard deviation was 13.8 ± 13.7 months. T2-weighted images had the highest HI (mean ± SD, 5.1 ± 2.5) followed by contrast-enhanced T1-weighted images (3.7 ± 1.5), FLAIR images (3.0 ± 1.1), and ADC maps (1.6 ± 0.4). ANOVA demonstrated that HI values were significantly higher for T2-weighted images than FLAIR (p < 0.01) and ADC (p < 0.0001). Following radiotherapy, T2-weighted and contrast-enhanced T1-weighted image HI values increased, while FLAIR and ADC HI values decreased. Univariate and multivariate analyses did not reveal a relationship between HI values and patient survival (p > 0.05). For children with DIPG, T2-weighted MRI demonstrates the greatest signal intensity variance suggesting tumor heterogeneity. Within this heterogeneity, T2-weighted signal hypointensity is known to correlate with increased cellularity and thus may represent a putative target for CED catheter placement in future clinical trials.

MR Imaging Biomarkers to Monitor Early Response to Hypoxia-Activated Prodrug TH-302 in Pancreatic Cancer Xenografts.

PubMed

Zhang, Xiaomeng; Wojtkowiak, Jonathan W; Martinez, Gary V; Cornnell, Heather H; Hart, Charles P; Baker, Amanda F; Gillies, Robert

2016-01-01

TH-302 is a hypoxia-activated prodrug known to activate selectively under the hypoxic conditions commonly found in solid tumors. It is currently being evaluated in clinical trials, including two trials in Pancreatic Ductal Adenocarcinomas (PDAC). The current study was undertaken to evaluate imaging biomarkers for prediction and response monitoring of TH-302 efficacy in xenograft models of PDAC. Dynamic contrast-enhanced (DCE) and diffusion weighted (DW) magnetic resonance imaging (MRI) were used to monitor acute effects on tumor vasculature and cellularity, respectively. Three human PDAC xenografts with known differential responses to TH-302 were imaged prior to, and at 24 h and 48 hours following a single dose of TH-302 or vehicle to determine if imaging changes presaged changes in tumor volumes. DW-MRI was performed at five b-values to generate apparent diffusion coefficient of water (ADC) maps. For DCE-MRI, a standard clinically available contrast reagent, Gd-DTPA, was used to determine blood flow into the tumor region of interest. TH-302 induced a dramatic decrease in the DCE transfer constant (Ktrans) within 48 hours after treatment in the sensitive tumors, Hs766t and Mia PaCa-2, whereas TH-302 had no effect on the perfusion behavior of resistant SU.86.86 tumors. Tumor cellularity, estimated from ADC, was significantly increased 24 and 48 hours after treatment in Hs766t, but was not observed in the Mia PaCa-2 and SU.86.86 groups. Notably, growth inhibition of Hs766t was observed immediately (day 3) following initiation of treatment, but was not observed in MiaPaCa-2 tumors until 8 days after initiation of treatment. Based on these preclinical findings, DCE-MRI measures of vascular perfusion dynamics and ADC measures of cell density are suggested as potential TH-302 response biomarkers in clinical trials.

Velocity landscape correlation resolves multiple flowing protein populations from fluorescence image time series.

PubMed

Pandžić, Elvis; Abu-Arish, Asmahan; Whan, Renee M; Hanrahan, John W; Wiseman, Paul W

2018-02-16

Molecular, vesicular and organellar flows are of fundamental importance for the delivery of nutrients and essential components used in cellular functions such as motility and division. With recent advances in fluorescence/super-resolution microscopy modalities we can resolve the movements of these objects at higher spatio-temporal resolutions and with better sensitivity. Previously, spatio-temporal image correlation spectroscopy has been applied to map molecular flows by correlation analysis of fluorescence fluctuations in image series. However, an underlying assumption of this approach is that the sampled time windows contain one dominant flowing component. Although this was true for most of the cases analyzed earlier, in some situations two or more different flowing populations can be present in the same spatio-temporal window. We introduce an approach, termed velocity landscape correlation (VLC), which detects and extracts multiple flow components present in a sampled image region via an extension of the correlation analysis of fluorescence intensity fluctuations. First we demonstrate theoretically how this approach works, test the performance of the method with a range of computer simulated image series with varying flow dynamics. Finally we apply VLC to study variable fluxing of STIM1 proteins on microtubules connected to the plasma membrane of Cystic Fibrosis Bronchial Epithelial (CFBE) cells. Copyright © 2018 Elsevier Inc. All rights reserved.

Imaging technique for real-time temperature monitoring during cryotherapy of lesions.

PubMed

Petrova, Elena; Liopo, Anton; Nadvoretskiy, Vyacheslav; Ermilov, Sergey

2016-11-01

Noninvasive real-time temperature imaging during thermal therapies is able to significantly improve clinical outcomes. An optoacoustic (OA) temperature monitoring method is proposed for noninvasive real-time thermometry of vascularized tissue during cryotherapy. The universal temperature-dependent optoacoustic response (ThOR) of red blood cells (RBCs) is employed to convert reconstructed OA images to temperature maps. To obtain the temperature calibration curve for intensity-normalized OA images, we measured ThOR of 10 porcine blood samples in the range of temperatures from 40°C to ?16°C and analyzed the data for single measurement variations. The nonlinearity (?Tmax) and the temperature of zero OA response (T0) of the calibration curve were found equal to 11.4±0.1°C and ?13.8±0.1°C, respectively. The morphology of RBCs was examined before and after the data collection confirming cellular integrity and intracellular compartmentalization of hemoglobin. For temperatures below 0°C, which are of particular interest for cryotherapy, the accuracy of a single temperature measurement was ±1°C, which is consistent with the clinical requirements. Validation of the proposed OA temperature imaging technique was performed for slow and fast cooling of blood samples embedded in tissue-mimicking phantoms.

Improved Serial Sectioning Techniques for Correlative Light-Electron Microscopy Mapping of Human Langerhans Islets

PubMed Central

Saitoh, Sei; Ohno, Nobuhiko; Saitoh, Yurika; Terada, Nobuo; Shimo, Satoshi; Aida, Kaoru; Fujii, Hideki; Kobayashi, Tetsuro; Ohno, Shinichi

2018-01-01

Combined analysis of immunostaining for various biological molecules coupled with investigations of ultrastructural features of individual cells is a powerful approach for studies of cellular functions in normal and pathological conditions. However, weak antigenicity of tissues fixed by conventional methods poses a problem for immunoassays. This study introduces a method of correlative light and electron microscopy imaging of the same endocrine cells of compact and diffuse islets from human pancreatic tissue specimens. The method utilizes serial sections obtained from Epon-embedded specimens fixed with glutaraldehyde and osmium tetroxide. Double-immunofluorescence staining of thick Epon sections for endocrine hormones (insulin and glucagon) and regenerating islet-derived gene 1 α (REG1α) was performed following the removal of Epoxy resin with sodium ethoxide, antigen retrieval by autoclaving, and de-osmification treatment with hydrogen peroxide. The immunofluorescence images of endocrine cells were superimposed with the electron microscopy images of the same cells obtained from serial ultrathin sections. Immunofluorescence images showed well-preserved secretory granules in endocrine cells, whereas electron microscopy observations demonstrated corresponding secretory granules and intracellular organelles in the same cells. In conclusion, the correlative imaging approach developed by us may be useful for examining ultrastructural features in combination with immunolocalisation of endocrine hormones in the same human pancreatic islets. PMID:29622846

Imaging technique for real-time temperature monitoring during cryotherapy of lesions

PubMed Central

Petrova, Elena; Liopo, Anton; Nadvoretskiy, Vyacheslav; Ermilov, Sergey

2016-01-01

Abstract. Noninvasive real-time temperature imaging during thermal therapies is able to significantly improve clinical outcomes. An optoacoustic (OA) temperature monitoring method is proposed for noninvasive real-time thermometry of vascularized tissue during cryotherapy. The universal temperature-dependent optoacoustic response (ThOR) of red blood cells (RBCs) is employed to convert reconstructed OA images to temperature maps. To obtain the temperature calibration curve for intensity-normalized OA images, we measured ThOR of 10 porcine blood samples in the range of temperatures from 40°C to −16°C and analyzed the data for single measurement variations. The nonlinearity (ΔTmax) and the temperature of zero OA response (T0) of the calibration curve were found equal to 11.4±0.1°C and −13.8±0.1°C, respectively. The morphology of RBCs was examined before and after the data collection confirming cellular integrity and intracellular compartmentalization of hemoglobin. For temperatures below 0°C, which are of particular interest for cryotherapy, the accuracy of a single temperature measurement was ±1°C, which is consistent with the clinical requirements. Validation of the proposed OA temperature imaging technique was performed for slow and fast cooling of blood samples embedded in tissue-mimicking phantoms. PMID:27822579

Impact of cell size on inventory and mapping errors in a cellular geographic information system

NASA Technical Reports Server (NTRS)

Wehde, M. E. (Principal Investigator)

1979-01-01

The author has identified the following significant results. The effect of grid position was found insignificant for maps but highly significant for isolated mapping units. A modelable relationship between mapping error and cell size was observed for the map segment analyzed. Map data structure was also analyzed with an interboundary distance distribution approach. Map data structure and the impact of cell size on that structure were observed. The existence of a model allowing prediction of mapping error based on map structure was hypothesized and two generations of models were tested under simplifying assumptions.

Axial resolution improvement in spectral domain optical coherence tomography using a depth-adaptive maximum-a-posterior framework

NASA Astrophysics Data System (ADS)

Boroomand, Ameneh; Tan, Bingyao; Wong, Alexander; Bizheva, Kostadinka

2015-03-01

The axial resolution of Spectral Domain Optical Coherence Tomography (SD-OCT) images degrades with scanning depth due to the limited number of pixels and the pixel size of the camera, any aberrations in the spectrometer optics and wavelength dependent scattering and absorption in the imaged object [1]. Here we propose a novel algorithm which compensates for the blurring effect of these factors of the depth-dependent axial Point Spread Function (PSF) in SDOCT images. The proposed method is based on a Maximum A Posteriori (MAP) reconstruction framework which takes advantage of a Stochastic Fully Connected Conditional Random Field (SFCRF) model. The aim is to compensate for the depth-dependent axial blur in SD-OCT images and simultaneously suppress the speckle noise which is inherent to all OCT images. Applying the proposed depth-dependent axial resolution enhancement technique to an OCT image of cucumber considerably improved the axial resolution of the image especially at higher imaging depths and allowed for better visualization of cellular membrane and nuclei. Comparing the result of our proposed method with the conventional Lucy-Richardson deconvolution algorithm clearly demonstrates the efficiency of our proposed technique in better visualization and preservation of fine details and structures in the imaged sample, as well as better speckle noise suppression. This illustrates the potential usefulness of our proposed technique as a suitable replacement for the hardware approaches which are often very costly and complicated.

In vivo wide-field multispectral scanning laser ophthalmoscopy–optical coherence tomography mouse retinal imager: longitudinal imaging of ganglion cells, microglia, and Müller glia, and mapping of the mouse retinal and choroidal vasculature

PubMed Central

Zhang, Pengfei; Zam, Azhar; Jian, Yifan; Wang, Xinlei; Li, Yuanpei; Lam, Kit S.; Burns, Marie E.; Sarunic, Marinko V.; Pugh, Edward N.; Zawadzki, Robert J.

2015-01-01

Abstract. Scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT) provide complementary views of the retina, with the former collecting fluorescence data with good lateral but relatively low-axial resolution, and the latter collecting label-free backscattering data with comparable lateral but much higher axial resolution. To take maximal advantage of the information of both modalities in mouse retinal imaging, we have constructed a compact, four-channel, wide-field (∼50  deg) system that simultaneously acquires and automatically coregisters three channels of confocal SLO and Fourier domain OCT data. The scanner control system allows “zoomed” imaging of a region of interest identified in a wide-field image, providing efficient digital sampling and localization of cellular resolution features in longitudinal imaging of individual mice. The SLO is equipped with a “flip-in” spectrometer that enables spectral “fingerprinting” of fluorochromes. Segmentation of retina layers and en face display facilitate spatial comparison of OCT data with SLO fluorescence patterns. We demonstrate that the system can be used to image an individual retinal ganglion cell over many months, to simultaneously image microglia and Müller glia expressing different fluorochromes, to characterize the distinctive spatial distributions and clearance times of circulating fluorochromes with different molecular sizes, and to produce unequivocal images of the heretofore uncharacterized mouse choroidal vasculature. PMID:26677070

Photocrosslinking approaches to interactome mapping

PubMed Central

Pham, Nam D.; Parker, Randy B.; Kohler, Jennifer J.

2012-01-01

Photocrosslinking approaches can be used to map interactome networks within the context of living cells. Photocrosslinking methods rely on use of metabolic engineering or genetic code expansion to incorporate photocrosslinking analogs of amino acids or sugars into cellular biomolecules. Immunological and mass spectrometry techniques are used to analyze crosslinked complexes, thereby defining specific interactomes. Because photocrosslinking can be conducted in native, cellular settings, it can be used to define context-dependent interactions. Photocrosslinking methods are also ideally suited for determining interactome dynamics, mapping interaction interfaces, and identifying transient interactions in which intrinsically disordered proteins and glycoproteins engage. Here we discuss the application of cell-based photocrosslinking to the study of specific problems in immune cell signaling, transcription, membrane protein dynamics, nucleocytoplasmic transport, and chaperone-assisted protein folding. PMID:23149092

Synchrotron radiation induced X-ray emission studies of the antioxidant mechanism of the organoselenium drug ebselen.

PubMed

Aitken, Jade B; Lay, Peter A; Duong, T T Hong; Aran, Roshanak; Witting, Paul K; Harris, Hugh H; Lai, Barry; Vogt, Stefan; Giles, Gregory I

2012-04-01

Synchrotron radiation induced X-ray emission (SRIXE) spectroscopy was used to map the cellular uptake of the organoselenium-based antioxidant drug ebselen using differentiated ND15 cells as a neuronal model. The cellular SRIXE spectra, acquired using a hard X-ray microprobe beam (12.8-keV), showed a large enhancement of fluorescence at the K(α) line for Se (11.2-keV) following treatment with ebselen (10 μM) at time periods from 60 to 240 min. Drug uptake was quantified and ebselen was shown to induce time-dependent changes in cellular elemental content that were characteristic of oxidative stress with the efflux of K, Cl, and Ca species. The SRIXE cellular Se distribution map revealed that ebselen was predominantly localized to a discreet region of the cell which, by comparison with the K and P elemental maps, is postulated to correspond to the endoplasmic reticulum. On the basis of these findings, it is hypothesized that a major outcome of ebselen redox catalysis is the induction of cellular stress. A mechanism of action of ebselen is proposed that involves the cell responding to drug-induced stress by increasing the expression of antioxidant genes. This hypothesis is supported by the observation that ebselen also regulated the homeostasis of the transition metals Mn, Cu, Fe, and Zn, with increases in transition metal uptake paralleling known induction times for the expression of antioxidant metalloenzymes. © SBIC 2012

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM).

PubMed

Tang, Anson H L; Lai, Queenie T K; Chung, Bob M F; Lee, Kelvin C M; Mok, Aaron T Y; Yip, G K; Shum, Anderson H C; Wong, Kenneth K Y; Tsia, Kevin K

2017-06-28

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture - information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization backend), as well as the workflow of imaging flow cytometry based on ATOM, using human cells and micro-algae as the examples.

Higher resolution satellite remote sensing and the impact on image mapping

USGS Publications Warehouse

Watkins, Allen H.; Thormodsgard, June M.

1987-01-01

Recent advances in spatial, spectral, and temporal resolution of civil land remote sensing satellite data are presenting new opportunities for image mapping applications. The U.S. Geological Survey's experimental satellite image mapping program is evolving toward larger scale image map products with increased information content as a result of improved image processing techniques and increased resolution. Thematic mapper data are being used to produce experimental image maps at 1:100,000 scale that meet established U.S. and European map accuracy standards. Availability of high quality, cloud-free, 30-meter ground resolution multispectral data from the Landsat thematic mapper sensor, along with 10-meter ground resolution panchromatic and 20-meter ground resolution multispectral data from the recently launched French SPOT satellite, present new cartographic and image processing challenges.The need to fully exploit these higher resolution data increases the complexity of processing the images into large-scale image maps. The removal of radiometric artifacts and noise prior to geometric correction can be accomplished by using a variety of image processing filters and transforms. Sensor modeling and image restoration techniques allow maximum retention of spatial and radiometric information. An optimum combination of spectral information and spatial resolution can be obtained by merging different sensor types. These processing techniques are discussed and examples are presented.

Confocal Microscopy and Molecular-Specific Optical Contrast Agents for the Detection of Oral Neoplasia

PubMed Central

Carlson, Alicia L.; Gillenwater, Ann M.; Williams, Michelle D.; El-Naggar, Adel K.; Richards-Kortum, R. R.

2009-01-01

Using current clinical diagnostic techniques, it is difficult to visualize tumor morphology and architecture at the cellular level, which is necessary for diagnostic localization of pathologic lesions. Optical imaging techniques have the potential to address this clinical need by providing real-time, sub-cellular resolution images. This paper describes the use of dual mode confocal microscopy and optical molecular-specific contrast agents to image tissue architecture, cellular morphology, and sub-cellular molecular features of normal and neoplastic oral tissues. Fresh tissue slices were prepared from 33 biopsies of clinically normal and abnormal oral mucosa obtained from 14 patients. Reflectance confocal images were acquired after the application of 6% acetic acid, and fluorescence confocal images were acquired after the application of a fluorescence contrast agent targeting the epidermal growth factor receptor (EGFR). The dual imaging modes provided images similar to light microscopy of hematoxylin and eosin and immunohistochemistry staining, but from thick fresh tissue slices. Reflectance images provided information on the architecture of the tissue and the cellular morphology. The nuclear-to-cytoplasmic (N/C) ratio from the reflectance images was at least 7.5 times greater for the carcinoma than the corresponding normal samples, except for one case of highly keratinized carcinoma. Separation of carcinoma from normal and mild dysplasia was achieved using this ratio (p<0.01). Fluorescence images of EGFR expression yielded a mean fluorescence labeling intensity (FLI) that was at least 2.7 times higher for severe dysplasia and carcinoma samples than for the corresponding normal sample, and could be used to distinguish carcinoma from normal and mild dysplasia (p<0.01). Analyzed together, the N/C ratio and the mean FLI may improve the ability to distinguish carcinoma from normal squamous epithelium. PMID:17877424

Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates.

PubMed

Cordina, Nicole M; Sayyadi, Nima; Parker, Lindsay M; Everest-Dass, Arun; Brown, Louise J; Packer, Nicolle H

2018-03-14

Bio-imaging is a key technique in tracking and monitoring important biological processes and fundamental biomolecular interactions, however the interference of background autofluorescence with targeted fluorophores is problematic for many bio-imaging applications. This study reports on two novel methods for reducing interference with cellular autofluorescence for bio-imaging. The first method uses fluorescent nanodiamonds (FNDs), containing nitrogen vacancy centers. FNDs emit at near-infrared wavelengths typically higher than most cellular autofluorescence; and when appropriately functionalized, can be used for background-free imaging of targeted biomolecules. The second method uses europium-chelating tags with long fluorescence lifetimes. These europium-chelating tags enhance background-free imaging due to the short fluorescent lifetimes of cellular autofluorescence. In this study, we used both methods to target E-selectin, a transmembrane glycoprotein that is activated by inflammation, to demonstrate background-free fluorescent staining in fixed endothelial cells. Our findings indicate that both FND and Europium based staining can improve fluorescent bio-imaging capabilities by reducing competition with cellular autofluorescence. 30 nm nanodiamonds coated with the E-selectin antibody was found to enable the most sensitive detective of E-selectin in inflamed cells, with a 40-fold increase in intensity detected.

Survey statistics of automated segmentations applied to optical imaging of mammalian cells.

PubMed

Bajcsy, Peter; Cardone, Antonio; Chalfoun, Joe; Halter, Michael; Juba, Derek; Kociolek, Marcin; Majurski, Michael; Peskin, Adele; Simon, Carl; Simon, Mylene; Vandecreme, Antoine; Brady, Mary

2015-10-15

The goal of this survey paper is to overview cellular measurements using optical microscopy imaging followed by automated image segmentation. The cellular measurements of primary interest are taken from mammalian cells and their components. They are denoted as two- or three-dimensional (2D or 3D) image objects of biological interest. In our applications, such cellular measurements are important for understanding cell phenomena, such as cell counts, cell-scaffold interactions, cell colony growth rates, or cell pluripotency stability, as well as for establishing quality metrics for stem cell therapies. In this context, this survey paper is focused on automated segmentation as a software-based measurement leading to quantitative cellular measurements. We define the scope of this survey and a classification schema first. Next, all found and manually filteredpublications are classified according to the main categories: (1) objects of interests (or objects to be segmented), (2) imaging modalities, (3) digital data axes, (4) segmentation algorithms, (5) segmentation evaluations, (6) computational hardware platforms used for segmentation acceleration, and (7) object (cellular) measurements. Finally, all classified papers are converted programmatically into a set of hyperlinked web pages with occurrence and co-occurrence statistics of assigned categories. The survey paper presents to a reader: (a) the state-of-the-art overview of published papers about automated segmentation applied to optical microscopy imaging of mammalian cells, (b) a classification of segmentation aspects in the context of cell optical imaging, (c) histogram and co-occurrence summary statistics about cellular measurements, segmentations, segmented objects, segmentation evaluations, and the use of computational platforms for accelerating segmentation execution, and (d) open research problems to pursue. The novel contributions of this survey paper are: (1) a new type of classification of cellular measurements and automated segmentation, (2) statistics about the published literature, and (3) a web hyperlinked interface to classification statistics of the surveyed papers at https://isg.nist.gov/deepzoomweb/resources/survey/index.html.

Guided filter-based fusion method for multiexposure images

NASA Astrophysics Data System (ADS)

Hou, Xinglin; Luo, Haibo; Qi, Feng; Zhou, Peipei

2016-11-01

It is challenging to capture a high-dynamic range (HDR) scene using a low-dynamic range camera. A weighted sum-based image fusion (IF) algorithm is proposed so as to express an HDR scene with a high-quality image. This method mainly includes three parts. First, two image features, i.e., gradients and well-exposedness are measured to estimate the initial weight maps. Second, the initial weight maps are refined by a guided filter, in which the source image is considered as the guidance image. This process could reduce the noise in initial weight maps and preserve more texture consistent with the original images. Finally, the fused image is constructed by a weighted sum of source images in the spatial domain. The main contributions of this method are the estimation of the initial weight maps and the appropriate use of the guided filter-based weight maps refinement. It provides accurate weight maps for IF. Compared to traditional IF methods, this algorithm avoids image segmentation, combination, and the camera response curve calibration. Furthermore, experimental results demonstrate the superiority of the proposed method in both subjective and objective evaluations.

Image display device in digital TV

DOEpatents

Choi, Seung Jong [Seoul, KR

2006-07-18

Disclosed is an image display device in a digital TV that is capable of carrying out the conversion into various kinds of resolution by using single bit map data in the digital TV. The image display device includes: a data processing part for executing bit map conversion, compression, restoration and format-conversion for text data; a memory for storing the bit map data obtained according to the bit map conversion and compression in the data processing part and image data inputted from an arbitrary receiving part, the receiving part receiving one of digital image data and analog image data; an image outputting part for reading the image data from the memory; and a display processing part for mixing the image data read from the image outputting part and the bit map data converted in format from the a data processing part. Therefore, the image display device according to the present invention can convert text data in such a manner as to correspond with various resolution, carry out the compression for bit map data, thereby reducing the memory space, and support text data of an HTML format, thereby providing the image with the text data of various shapes.

Discriminative segmentation of microscopic cellular images.

PubMed

Cheng, Li; Ye, Ning; Yu, Weimiao; Cheah, Andre

2011-01-01

Microscopic cellular images segmentation has become an important routine procedure in modern biological research, due to the rapid advancement of fluorescence probes and robotic microscopes in recent years. In this paper we advocate a discriminative learning approach for cellular image segmentation. In particular, three new features are proposed to capture the appearance, shape and context information, respectively. Experiments are conducted on three different cellular image datasets. Despite the significant disparity among these datasets, the proposed approach is demonstrated to perform reasonably well. As expected, for a particular dataset, some features turn out to be more suitable than others. Interestingly, we observe that a further gain can often be obtained on top of using the "good" features, by also retaining those features that perform poorly. This might be due to the complementary nature of these features, as well as the capacity of our approach to better integrate and exploit different sources of information.

Herpes simplex virus 2 VP22 phosphorylation induced by cellular and viral kinases does not influence intracellular localization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Geiss, Brian J.; Cano, Gina L.; Tavis, John E.

2004-12-05

Phosphorylation of the herpes simplex virus (HSV) VP22 protein is regulated by cellular kinases and the UL13 viral kinase, but the sites at which these enzymes induce phosphorylation of HSV-2 VP22 are not known. Using serine-to-alanine mutants to map phosphorylation sites on HSV-2 VP22 in cells, we made three major observations. First, phosphorylation by a cellular kinase mapped to serines 70, 71, and/or 72 within CKII consensus sites analogous to previously identified phosphorylation sites in HSV-1 VP22. Second, we mapped UL13-mediated phosphorylation of HSV-2 VP22 to serines 28 and 34, describing for the first time UL13-dependent phosphorylation sites on VP22.more » Third, previously identified VP22-associated cellular kinase sites in HSV-1 VP22 (serines 292 and 294) were not phosphorylated in HSV-2 VP22 (serines 291 and 293). VP22 expressed alone accumulated in the cytoplasm and to a lesser extent in the nucleus. Phosphorylation by endogenous cellular kinase(s) did not alter the localization of VP22. Co-expression of HSV-2 VP22 with active UL13, but not with enzymatically inactive UL13, resulted in nuclear accumulation of VP22 and altered nuclear morphology. Surprisingly, redistribution of VP22 to the nucleus occurred independently of UL13-induced phosphorylation of VP22. The altered nuclear morphology of UL13-expressing cells was not due to apoptosis. These results demonstrate that phosphorylation of HSV-2 VP22 at multiple serine residues is induced by UL13 and cellular kinase(s), and that the nuclear/cytoplasmic distribution of VP22 is independent of its phosphorylation status but is controlled indirectly by UL13 kinase activity.« less

Cellular and humoral immune responses in sheep vaccinated with candidate antigens MAP2698c and MAP3567 from Mycobacterium avium subspecies paratuberculosis

PubMed Central

Gurung, Ratna B.; Purdie, Auriol C.; Whittington, Richard J.; Begg, Douglas J.

2014-01-01

Control of Johne's disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP) in ruminants using commercially available vaccine reduces production losses, mortality, fecal shedding and histopathological lesions but does not provide complete protection from infection and interferes with serological diagnosis of Johne's disease and bovine tuberculosis. At this time no recombinant antigens have been found to provide superior protection compared to whole killed or live-attenuated MAP vaccines. Therefore, there is a need to evaluate more candidate MAP antigens. In this study recombinant MAP antigens MAP2698c and MAP3567 were formulated with four different MONTANIDE™ (ISA 50V2, 61VG, 71VG, and 201VG) adjuvants and evaluated for their ability to produce specific immune responses in vaccinated sheep. The cellular immune response was measured with an interferon-gamma (IFN-γ) release assay and the humoral immune response was measured by antibody detection enzyme linked immunosorbent assay. Recombinant vaccine formulation with the antigen MAP2698c and MONTANIDE™ ISA 201VG adjuvant produced strong whole-MAP as well as MAP2698c-specific IFN-γ responses in a high proportion of the vaccinated sheep. The formulation caused less severe injection site lesions in comparison to other formulations. The findings from this study suggest that the MAP2698c + 201VG should be evaluated in a challenge trial to determine the efficacy of this vaccine candidate. PMID:25077074

Image processing for optical mapping.

PubMed

Ravindran, Prabu; Gupta, Aditya

2015-01-01

Optical Mapping is an established single-molecule, whole-genome analysis system, which has been used to gain a comprehensive understanding of genomic structure and to study structural variation of complex genomes. A critical component of Optical Mapping system is the image processing module, which extracts single molecule restriction maps from image datasets of immobilized, restriction digested and fluorescently stained large DNA molecules. In this review, we describe robust and efficient image processing techniques to process these massive datasets and extract accurate restriction maps in the presence of noise, ambiguity and confounding artifacts. We also highlight a few applications of the Optical Mapping system.

Planetary maps

USGS Publications Warehouse

,

1992-01-01

An important goal of the USGS planetary mapping program is to systematically map the geology of the Moon, Mars, Venus, and Mercury, and the satellites of the outer planets. These geologic maps are published in the USGS Miscellaneous Investigations (I) Series. Planetary maps on sale at the USGS include shaded-relief maps, topographic maps, geologic maps, and controlled photomosaics. Controlled photomosaics are assembled from two or more photographs or images using a network of points of known latitude and longitude. The images used for most of these planetary maps are electronic images, obtained from orbiting television cameras, various optical-mechanical systems. Photographic film was only used to map Earth's Moon.

{sup 18}F-FLT uptake kinetics in head and neck squamous cell carcinoma: A PET imaging study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Dan, E-mail: dan.liu@oncology.ox.ac.uk; Fenwick, John D.; Chalkidou, Anastasia

2014-04-15

Purpose: To analyze the kinetics of 3{sup ′}-deoxy-3{sup ′}-[F-18]-fluorothymidine (18F-FLT) uptake by head and neck squamous cell carcinomas and involved nodes imaged using positron emission tomography (PET). Methods: Two- and three-tissue compartment models were fitted to 12 tumor time-activity-curves (TACs) obtained for 6 structures (tumors or involved nodes) imaged in ten dynamic PET studies of 1 h duration, carried out for five patients. The ability of the models to describe the data was assessed using a runs test, the Akaike information criterion (AIC) and leave-one-out cross-validation. To generate parametric maps the models were also fitted to TACs of individual voxels.more » Correlations between maps of different parameters were characterized using Pearson'sr coefficient; in particular the phosphorylation rate-constants k{sub 3-2tiss} and k{sub 5} of the two- and three-tissue models were studied alongside the flux parameters K{sub FLT-2tiss} and K{sub FLT} of these models, and standardized uptake values (SUV). A methodology based on expectation-maximization clustering and the Bayesian information criterion (“EM-BIC clustering”) was used to distil the information from noisy parametric images. Results: Fits of two-tissue models 2C3K and 2C4K and three-tissue models 3C5K and 3C6K comprising three, four, five, and six rate-constants, respectively, pass the runs test for 4, 8, 10, and 11 of 12 tumor TACs. The three-tissue models have lower AIC and cross-validation scores for nine of the 12 tumors. Overall the 3C6K model has the lowest AIC and cross-validation scores and its fitted parameter values are of the same orders of magnitude as literature estimates. Maps ofK{sub FLT} and K{sub FLT-2tiss} are strongly correlated (r = 0.85) and also correlate closely with SUV maps (r = 0.72 for K{sub FLT-2tiss}, 0.64 for K{sub FLT}). Phosphorylation rate-constant maps are moderately correlated with flux maps (r = 0.48 for k{sub 3-2tiss} vs K{sub FLT-2tiss} and r = 0.68 for k{sub 5} vs K{sub FLT}); however, neither phosphorylation rate-constant correlates significantly with SUV. EM-BIC clustering reduces the parametric maps to a small number of levels—on average 5.8, 3.5, 3.4, and 1.4 for K{sub FLT-2tiss}, K{sub FLT}, k{sub 3-2tiss}, and k{sub 5.} This large simplification is potentially useful for radiotherapy dose-painting, but demonstrates the high noise in some maps. Statistical simulations show that voxel level noise degrades TACs generated from the 3C6K model sufficiently that the average AIC score, parameter bias, and total uncertainty of 2C4K model fits are similar to those of 3C6K fits, whereas at the whole tumor level the scores are lower for 3C6K fits. Conclusions: For the patients studied here, whole tumor FLT uptake time-courses are represented better overall by a three-tissue than by a two-tissue model. EM-BIC clustering simplifies noisy parametric maps, providing the best description of the underlying information they contain and is potentially useful for radiotherapy dose-painting. However, the clustering highlights the large degree of noise present in maps of the phosphorylation rate-constantsk{sub 5} and k{sub 3-2tiss}, which are conceptually tightly linked to cellular proliferation. Methods must be found to make these maps more robust—either by constraining other model parameters or modifying dynamic imaging protocols.« less

Polymeric nanocomposites loaded with fluoridated hydroxyapatite Ln3+ (Ln = Eu or Tb)/iron oxide for magnetic targeted cellular imaging

PubMed Central

Pan, Jie; Liu, Wei-Jiao; Hua, Chao; Wang, Li-Li; Wan, Dong; Gong, Jun-Bo

2015-01-01

Objective To fabricate polymeric nanocomposites with excellent photoluminescence, magnetic properties, and stability in aqueous solutions, in order to improve specificity and sensitivity of cellular imaging under a magnetic field. Methods Fluoridated Ln3+-doped HAP (Ln3+-HAP) NPs and iron oxides (IOs) can be encapsulated with biocompatible polymers via a modified solvent exaction/evaporation technique to prepare polymeric nanocomposites with fluoridated Ln3+-HAP/iron oxide. The nanocomposites were characterized for surface morphology, fluorescence spectra, magnetic properties and in vitro cytotoxicity. Magnetic targeted cellular imaging of such nanocomposites was also evaluated with confocal laser scanning microscope using A549 cells with or without magnetic field. Results The fabricated nanocomposites showed good stability and excellent luminescent properties, as well as low in vitro cytotoxicity, indicating that the nanocomposites are suitable for biological applications. Nanocomposites under magnetic field achieved much higher cellular uptake via an energy-dependent pathway than those without magnetic field. Conclusion The nanocomposites fabricated in this study will be a promising tool for magnetic targeted cellular imaging with improved specificity and enhanced selection. PMID:26487962

Imaging cellularity in benign and malignant peripheral nerve sheath tumors: Utility of the "target sign" by diffusion weighted imaging.

PubMed

Ahlawat, Shivani; Fayad, Laura M

2018-05-01

To determine the utility of "target sign" on diffusion weighted imaging (DWI) and apparent diffusion coefficient (ADC) mapping for peripheral nerve sheath tumor (PNST) characterization. This IRB-approved, HIPAA-compliant study retrospectively reviewed the MR imaging (comprised of T2- FS, DWI (b-values 50, 400, 800 s/mm 2 and ADC mapping), and static contrast-enhanced (CE) T1-W imaging) of 42 patients (mean age: 40 years (range 8-68 years), 48% (20/42) females) with 15 malignant PNSTs (MPNSTs) and 33 benign PNSTs (BPNSTs). MPNSTs were histologically confirmed while BPNSTs were histologically-proven or with stable clinical and imaging appearance for at least 12 months. Two radiologists assessed imaging characteristics (size, signal intensity, heterogeneity, perilesional edema or enhancement) and the presence or absence of "target sign," on each sequence. A "target sign" was defined as a biphasic pattern of peripheral hyperintensity and homogeneous central hypointensity. Descriptive statistics are reported. Cohen's κ statistic or interclass correlation coefficient (ICC) were used to evaluate interobserver agreement between two observers. Univariate and multiple logistic regression analysis were performed to identify MRI features with predictive values. MPNSTs were larger than BPNSTs (6.3 ± 2.5 cm vs 3.5 ± -2.1 cm, p = 0.0002), had perilesional edema (87%(13/15) vs 18%(6/33), p < 0.0001), heterogeneous enhancement (71%(10/14) vs 13%(4/31), p = 0.0001) and perilesional enhancement (79%(11/14) vs 18%(6/31), p = 0.0001), respectively. The "target sign" was present in: 24%(8/33) BPNSTs vs 0/15 MPNST on T2-FS (p = 0.26); 39%(13/33) BPNSTs vs 20%(3/15) MPNST on DWI using b-value = 50 s/mm 2 (p = 0.5); 55%(18/33) BPNSTs vs 6%(1/15) MPNST on DWI using b-value = 400 s/mm 2 (p = 0.002); 48%(16/33) BPNSTs vs 6%(1/15) MPNST on DWI using b-value = 800 s/mm 2 (p = 0.005) and 64%(21/33) benign vs 0/15 MPNST on ADC mapping(p < 0.0001). By CE-T1 imaging, 32%(10/31) BPNSTs and 7%(1/14) MPNST had a target sign(p = 0.07). The odds of an MPNST in cases with minimum ADC ≤ 1.0 × 10(-3) mm(2)/s are 150 times higher than in cases with ADC > 1.0 × 10(-3). In this explorative study, a "target sign" suggests a benign PNST and is more often visible on DWI using high b-values and ADC maps compared with anatomic sequences. Copyright © 2018 Elsevier B.V. All rights reserved.

Smartphone confocal microscopy for imaging cellular structures in human skin in vivo.

PubMed

Freeman, Esther E; Semeere, Aggrey; Osman, Hany; Peterson, Gary; Rajadhyaksha, Milind; González, Salvador; Martin, Jeffery N; Anderson, R Rox; Tearney, Guillermo J; Kang, Dongkyun

2018-04-01

We report development of a low-cost smartphone confocal microscope and its first demonstration of in vivo human skin imaging. The smartphone confocal microscope uses a slit aperture and diffraction grating to conduct two-dimensional confocal imaging without using any beam scanning devices. Lateral and axial resolutions of the smartphone confocal microscope were measured as 2 and 5 µm, respectively. In vivo confocal images of human skin revealed characteristic cellular structures, including spinous and basal keratinocytes and papillary dermis. Results suggest that the smartphone confocal microscope has a potential to examine cellular details in vivo and may help disease diagnosis in resource-poor settings, where conducting standard histopathologic analysis is challenging.

Smartphone confocal microscopy for imaging cellular structures in human skin in vivo

PubMed Central

Freeman, Esther E.; Semeere, Aggrey; Osman, Hany; Peterson, Gary; Rajadhyaksha, Milind; González, Salvador; Martin, Jeffery N.; Anderson, R. Rox; Tearney, Guillermo J.; Kang, Dongkyun

2018-01-01

We report development of a low-cost smartphone confocal microscope and its first demonstration of in vivo human skin imaging. The smartphone confocal microscope uses a slit aperture and diffraction grating to conduct two-dimensional confocal imaging without using any beam scanning devices. Lateral and axial resolutions of the smartphone confocal microscope were measured as 2 and 5 µm, respectively. In vivo confocal images of human skin revealed characteristic cellular structures, including spinous and basal keratinocytes and papillary dermis. Results suggest that the smartphone confocal microscope has a potential to examine cellular details in vivo and may help disease diagnosis in resource-poor settings, where conducting standard histopathologic analysis is challenging. PMID:29675328

Evaluation of two novel methods for assessing intracellular oxygen

NASA Astrophysics Data System (ADS)

Williams, Catrin F.; Kombrabail, M.; Vijayalakshmi, K.; White, Nick; Krishnamoorthy, G.; Lloyd, David

2012-08-01

The ability to resolve the spatio-temporal complexity of intracellular O2 distribution is the ‘Holy Grail’ of cellular physiology. In an effort to obtain a non-invasive approach of mapping intracellular O2 tensions, two methods of phosphorescent lifetime imaging microscopy were examined in the current study. These were picosecond time-resolved epiphosphorescence microscopy (single 0.5 µm focused spot) and two-photon confocal laser scanning microscopy with pinhole shifting. Both methods utilized nanoparticle-embedded Ru complex (45 nm diameter) as the phosphorescent probe, excited using pulsed outputs of Ti-sapphire Tsunami lasers (710-1050 nm). The former method used a 1 ps pulse width excitation beam with vertical polarization via a dichroic mirror (610 nm, XF43) and a 20× objective (NA 0.55, Nikon). Transmitted luminescence (1-2 × 104 counts s-1) was collected and time-correlated single photon counted decay times measured. Alternatively, an unmodified Zeiss LSM510 Confocal NLO microscope with 40× objective (NA 1.3) used successively shifted pinhole positions to collect image data from the lagging trail of the raster scan. Images obtained from two-photon excitation of a yeast (Schizosaccharomyces pombe) and a flagellate fish parasite (Spironucleus vortens), electroporated with Ru complex, indicated the intracellular location and magnitude of O2 gradients, thus confirming the feasibility of optical mapping under different external O2 concentrations. Both methods gave similar lifetimes for Ru complex phosphorescence under aerobic and anaerobic gas phases. Estimation of O2 tensions within individual fibroblasts (human dermal fibroblast (HDF)) and mammary adenocarcinoma (MCF-7) cells was possible using epiphosphorescence microscopy. MCF-7 cells showed lower intracellular O2 concentrations than HDF cells, possibly due to higher metabolic rates in the former. Future work should involve construction of higher resolution 3D maps of Ru coordinate complex lifetime distribution in cultured cell lines.

An improved image non-blind image deblurring method based on FoEs

NASA Astrophysics Data System (ADS)

Zhu, Qidan; Sun, Lei

2013-03-01

Traditional non-blind image deblurring algorithms always use maximum a posterior(MAP). MAP estimates involving natural image priors can reduce the ripples effectively in contrast to maximum likelihood(ML). However, they have been found lacking in terms of restoration performance. Based on this issue, we utilize MAP with KL penalty to replace traditional MAP. We develop an image reconstruction algorithm that minimizes the KL divergence between the reference distribution and the prior distribution. The approximate KL penalty can restrain over-smooth caused by MAP. We use three groups of images and Harris corner detection to prove our method. The experimental results show that our algorithm of non-blind image restoration can effectively reduce the ringing effect and exhibit the state-of-the-art deblurring results.

Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina.

PubMed

Zawadzki, Robert J; Zhang, Pengfei; Zam, Azhar; Miller, Eric B; Goswami, Mayank; Wang, Xinlei; Jonnal, Ravi S; Lee, Sang-Hyuck; Kim, Dae Yu; Flannery, John G; Werner, John S; Burns, Marie E; Pugh, Edward N

2015-06-01

Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a large-scale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.

Objective quality assessment of tone-mapped images.

PubMed

Yeganeh, Hojatollah; Wang, Zhou

2013-02-01

Tone-mapping operators (TMOs) that convert high dynamic range (HDR) to low dynamic range (LDR) images provide practically useful tools for the visualization of HDR images on standard LDR displays. Different TMOs create different tone-mapped images, and a natural question is which one has the best quality. Without an appropriate quality measure, different TMOs cannot be compared, and further improvement is directionless. Subjective rating may be a reliable evaluation method, but it is expensive and time consuming, and more importantly, is difficult to be embedded into optimization frameworks. Here we propose an objective quality assessment algorithm for tone-mapped images by combining: 1) a multiscale signal fidelity measure on the basis of a modified structural similarity index and 2) a naturalness measure on the basis of intensity statistics of natural images. Validations using independent subject-rated image databases show good correlations between subjective ranking score and the proposed tone-mapped image quality index (TMQI). Furthermore, we demonstrate the extended applications of TMQI using two examples-parameter tuning for TMOs and adaptive fusion of multiple tone-mapped images.

Multi-color electron microscopy by element-guided identification of cells, organelles and molecules.

PubMed

Scotuzzi, Marijke; Kuipers, Jeroen; Wensveen, Dasha I; de Boer, Pascal; Hagen, Kees C W; Hoogenboom, Jacob P; Giepmans, Ben N G

2017-04-07

Cellular complexity is unraveled at nanometer resolution using electron microscopy (EM), but interpretation of macromolecular functionality is hampered by the difficulty in interpreting grey-scale images and the unidentified molecular content. We perform large-scale EM on mammalian tissue complemented with energy-dispersive X-ray analysis (EDX) to allow EM-data analysis based on elemental composition. Endogenous elements, labels (gold and cadmium-based nanoparticles) as well as stains are analyzed at ultrastructural resolution. This provides a wide palette of colors to paint the traditional grey-scale EM images for composition-based interpretation. Our proof-of-principle application of EM-EDX reveals that endocrine and exocrine vesicles exist in single cells in Islets of Langerhans. This highlights how elemental mapping reveals unbiased biomedical relevant information. Broad application of EM-EDX will further allow experimental analysis on large-scale tissue using endogenous elements, multiple stains, and multiple markers and thus brings nanometer-scale 'color-EM' as a promising tool to unravel molecular (de)regulation in biomedicine.

Multi-color electron microscopy by element-guided identification of cells, organelles and molecules

PubMed Central

Scotuzzi, Marijke; Kuipers, Jeroen; Wensveen, Dasha I.; de Boer, Pascal; Hagen, Kees (C.) W.; Hoogenboom, Jacob P.; Giepmans, Ben N. G.

2017-01-01

Cellular complexity is unraveled at nanometer resolution using electron microscopy (EM), but interpretation of macromolecular functionality is hampered by the difficulty in interpreting grey-scale images and the unidentified molecular content. We perform large-scale EM on mammalian tissue complemented with energy-dispersive X-ray analysis (EDX) to allow EM-data analysis based on elemental composition. Endogenous elements, labels (gold and cadmium-based nanoparticles) as well as stains are analyzed at ultrastructural resolution. This provides a wide palette of colors to paint the traditional grey-scale EM images for composition-based interpretation. Our proof-of-principle application of EM-EDX reveals that endocrine and exocrine vesicles exist in single cells in Islets of Langerhans. This highlights how elemental mapping reveals unbiased biomedical relevant information. Broad application of EM-EDX will further allow experimental analysis on large-scale tissue using endogenous elements, multiple stains, and multiple markers and thus brings nanometer-scale ‘color-EM’ as a promising tool to unravel molecular (de)regulation in biomedicine. PMID:28387351

Pathway-Specific Dopamine Abnormalities in Schizophrenia.

PubMed

Weinstein, Jodi J; Chohan, Muhammad O; Slifstein, Mark; Kegeles, Lawrence S; Moore, Holly; Abi-Dargham, Anissa

2017-01-01

In light of the clinical evidence implicating dopamine in schizophrenia and the prominent hypotheses put forth regarding alterations in dopaminergic transmission in this disease, molecular imaging has been used to examine multiple aspects of the dopaminergic system. We review the imaging methods used and compare the findings across the different molecular targets. Findings have converged to suggest early dysregulation in the striatum, especially in the rostral caudate, manifesting as excess synthesis and release. Recent data showed deficit extending to most cortical regions and even to other extrastriatal subcortical regions not previously considered to be "hypodopaminergic" in schizophrenia. These findings yield a new topography for the dopaminergic dysregulation in schizophrenia. We discuss the dopaminergic innervation within the individual projection fields to provide a topographical map of this dual dysregulation and explore potential cellular and circuit-based mechanisms for brain region-dependent alterations in dopaminergic parameters. This refined knowledge is essential to better guide translational studies and efforts in early drug development. Copyright © 2016 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.

Live imaging reveals the progenitors and cell dynamics of limb regeneration

PubMed Central

Alwes, Frederike; Enjolras, Camille; Averof, Michalis

2016-01-01

Regeneration is a complex and dynamic process, mobilizing diverse cell types and remodelling tissues over long time periods. Tracking cell fate and behaviour during regeneration in active adult animals is especially challenging. Here, we establish continuous live imaging of leg regeneration at single-cell resolution in the crustacean Parhyale hawaiensis. By live recordings encompassing the first 4-5 days after amputation, we capture the cellular events that contribute to wound closure and morphogenesis of regenerating legs with unprecedented resolution and temporal detail. Using these recordings we are able to track cell lineages, to generate fate maps of the blastema and to identify the progenitors of regenerated epidermis. We find that there are no specialized stem cells for the epidermis. Most epidermal cells in the distal part of the leg stump proliferate, acquire new positional values and contribute to new segments in the regenerating leg. DOI: http://dx.doi.org/10.7554/eLife.19766.001 PMID:27776632

Assessing Diffusion in the Extra-Cellular Space of Brain Tissue by Dynamic MRI Mapping of Contrast Agent Concentrations

NASA Astrophysics Data System (ADS)

Mériaux, Sébastien; Conti, Allegra; Larrat, Benoît

2018-05-01

The characterization of extracellular space (ECS) architecture represents valuable information for the understanding of transport mechanisms occurring in brain parenchyma. ECS tortuosity reflects the hindrance imposed by cell membranes to molecular diffusion. Numerous strategies have been proposed to measure the diffusion through ECS and to estimate its tortuosity. The first method implies the perfusion for several hours of a radiotracer which effective diffusion coefficient D* is determined after post mortem processing. The most well-established techniques are real-time iontophoresis that measures the concentration of a specific ion at known distance from its release point, and integrative optical imaging that relies on acquiring microscopy images of macromolecules labelled with fluorophore. After presenting these methods, we focus on a recent Magnetic Resonance Imaging (MRI)-based technique that consists in acquiring concentration maps of a contrast agent diffusing within ECS. Thanks to MRI properties, molecular diffusion and tortuosity can be estimated in 3D for deep brain regions. To further discuss the reliability of this technique, we point out the influence of the delivery method on the estimation of D*. We compare the value of D* for a contrast agent intracerebrally injected, with its value when the agent is delivered to the brain after an ultrasound-induced blood-brain barrier (BBB) permeabilization. Several studies have already shown that tortuosity may be modified in pathological conditions. Therefore, we believe that MRI-based techniques could be useful in a clinical context for characterizing the diffusion properties of pathological ECS and thus predicting the drug biodistribution into the targeted area.

Label-Free Analysis of Cellular Lipid Droplet Formation by Non-Linear Microscopy

NASA Astrophysics Data System (ADS)

Schie, Iwan W.

Cellular lipid droplets (LD) are cellular organelles that can be found in every cell type. Recent research indicates that cellular LD are involved in a large number of cellular metabolic functions, such as lipid metabolism, protection from lipotoxicity, protein storage and degradation, and many more. LD formation is frequently associated with adverse health effects, i.e. alcoholic and non-alcoholic fatty liver disease, diabetes type-2, as well as many cardiovascular disorders. Despite their wide presence, LDs are the least studied and most poorly understood cellular organelles. Typically, LDs are investigated using fluorescence-based techniques that require staining with exogenous fluorophores. Other techniques, e.g. biochemical assays, require the destruction of cells that prohibit the analysis of living cells. Therefore, in my thesis research I developed a novel compound fast-scanning nonlinear optical microscope equipped with the ability to also acquire Raman spectra at specific image locations. This system allows us to image label-free cellular LD formation in living cells and analyze the composition of single cellular LDs. Images can be acquired at near video-rate (˜16 frames/s). Furthermore, the system has the ability to acquire very large images of tissue of up to 7.5x15 cm2 total area by stitching together scans with dimensions of 1x1 mm2 in less than 1 minute. The system also enables the user to acquire Raman spectra from points of interest in the multiphoton images and provides chemically-specific data from sample volumes as small as 1 femtoliter. In my thesis I used this setup to determine the effects of VLDL lipolysis products on primary rat hepatocytes. By analyzing the Raman spectra and comparing the peak ratios for saturated and unsaturated fatty acid it was determined that the small cellular LD are highly saturated, while large cellular LDs contain mostly unsaturated lipids. Furthermore, I established a method to determine the specific contribution of each individual fatty acids to a single cellular LD based on non-negative least squares analysis. The calculated quantities for oleic and palmitic acid from 10 individual cellular LDs were compared to results of a gas chromatography (GC) analysis of 2x10 6 cells. The analysis found that the data obtained by Raman spectroscopy of individual LDs closely resemble GC data of a significantly larger number of LDs.

Live-cell imaging.

PubMed

Cole, Richard

2014-01-01

It would be hard to argue that live-cell imaging has not changed our view of biology. The past 10 years have seen an explosion of interest in imaging cellular processes, down to the molecular level. There are now many advanced techniques being applied to live cell imaging. However, cellular health is often under appreciated. For many researchers, if the cell at the end of the experiment has not gone into apoptosis or is blebbed beyond recognition, than all is well. This is simply incorrect. There are many factors that need to be considered when performing live-cell imaging in order to maintain cellular health such as: imaging modality, media, temperature, humidity, PH, osmolality, and photon dose. The wavelength of illuminating light, and the total photon dose that the cells are exposed to, comprise two of the most important and controllable parameters of live-cell imaging. The lowest photon dose that achieves a measureable metric for the experimental question should be used, not the dose that produces cover photo quality images. This is paramount to ensure that the cellular processes being investigated are in their in vitro state and not shifted to an alternate pathway due to environmental stress. The timing of the mitosis is an ideal canary in the gold mine, in that any stress induced from the imaging will result in the increased length of mitosis, thus providing a control model for the current imagining conditions.

Live-cell imaging

PubMed Central

Cole, Richard

2014-01-01

It would be hard to argue that live-cell imaging has not changed our view of biology. The past 10 years have seen an explosion of interest in imaging cellular processes, down to the molecular level. There are now many advanced techniques being applied to live cell imaging. However, cellular health is often under appreciated. For many researchers, if the cell at the end of the experiment has not gone into apoptosis or is blebbed beyond recognition, than all is well. This is simply incorrect. There are many factors that need to be considered when performing live-cell imaging in order to maintain cellular health such as: imaging modality, media, temperature, humidity, PH, osmolality, and photon dose. The wavelength of illuminating light, and the total photon dose that the cells are exposed to, comprise two of the most important and controllable parameters of live-cell imaging. The lowest photon dose that achieves a measureable metric for the experimental question should be used, not the dose that produces cover photo quality images. This is paramount to ensure that the cellular processes being investigated are in their in vitro state and not shifted to an alternate pathway due to environmental stress. The timing of the mitosis is an ideal canary in the gold mine, in that any stress induced from the imaging will result in the increased length of mitosis, thus providing a control model for the current imagining conditions. PMID:25482523

Fully Convolutional Network-Based Multifocus Image Fusion.

PubMed

Guo, Xiaopeng; Nie, Rencan; Cao, Jinde; Zhou, Dongming; Qian, Wenhua

2018-07-01

As the optical lenses for cameras always have limited depth of field, the captured images with the same scene are not all in focus. Multifocus image fusion is an efficient technology that can synthesize an all-in-focus image using several partially focused images. Previous methods have accomplished the fusion task in spatial or transform domains. However, fusion rules are always a problem in most methods. In this letter, from the aspect of focus region detection, we propose a novel multifocus image fusion method based on a fully convolutional network (FCN) learned from synthesized multifocus images. The primary novelty of this method is that the pixel-wise focus regions are detected through a learning FCN, and the entire image, not just the image patches, are exploited to train the FCN. First, we synthesize 4500 pairs of multifocus images by repeatedly using a gaussian filter for each image from PASCAL VOC 2012, to train the FCN. After that, a pair of source images is fed into the trained FCN, and two score maps indicating the focus property are generated. Next, an inversed score map is averaged with another score map to produce an aggregative score map, which take full advantage of focus probabilities in two score maps. We implement the fully connected conditional random field (CRF) on the aggregative score map to accomplish and refine a binary decision map for the fusion task. Finally, we exploit the weighted strategy based on the refined decision map to produce the fused image. To demonstrate the performance of the proposed method, we compare its fused results with several start-of-the-art methods not only on a gray data set but also on a color data set. Experimental results show that the proposed method can achieve superior fusion performance in both human visual quality and objective assessment.

Interactive Map | USDA Plant Hardiness Zone Map

Science.gov Websites

Choose Basemap: Terrain Road Map Satellite Image Turn on Basemap Roads and Labels Zone Color Transparency menu to switch between Terrain, Road Map, and Satellite Image. Turn on Basemap Roads and Labels Click option is available only for Terrain and Satellite Image basemap choices. Zone Color Transparency The

A Probabilistic Feature Map-Based Localization System Using a Monocular Camera.

PubMed

Kim, Hyungjin; Lee, Donghwa; Oh, Taekjun; Choi, Hyun-Taek; Myung, Hyun

2015-08-31

Image-based localization is one of the most widely researched localization techniques in the robotics and computer vision communities. As enormous image data sets are provided through the Internet, many studies on estimating a location with a pre-built image-based 3D map have been conducted. Most research groups use numerous image data sets that contain sufficient features. In contrast, this paper focuses on image-based localization in the case of insufficient images and features. A more accurate localization method is proposed based on a probabilistic map using 3D-to-2D matching correspondences between a map and a query image. The probabilistic feature map is generated in advance by probabilistic modeling of the sensor system as well as the uncertainties of camera poses. Using the conventional PnP algorithm, an initial camera pose is estimated on the probabilistic feature map. The proposed algorithm is optimized from the initial pose by minimizing Mahalanobis distance errors between features from the query image and the map to improve accuracy. To verify that the localization accuracy is improved, the proposed algorithm is compared with the conventional algorithm in a simulation and realenvironments.

A Probabilistic Feature Map-Based Localization System Using a Monocular Camera

PubMed Central

Kim, Hyungjin; Lee, Donghwa; Oh, Taekjun; Choi, Hyun-Taek; Myung, Hyun

2015-01-01

Image-based localization is one of the most widely researched localization techniques in the robotics and computer vision communities. As enormous image data sets are provided through the Internet, many studies on estimating a location with a pre-built image-based 3D map have been conducted. Most research groups use numerous image data sets that contain sufficient features. In contrast, this paper focuses on image-based localization in the case of insufficient images and features. A more accurate localization method is proposed based on a probabilistic map using 3D-to-2D matching correspondences between a map and a query image. The probabilistic feature map is generated in advance by probabilistic modeling of the sensor system as well as the uncertainties of camera poses. Using the conventional PnP algorithm, an initial camera pose is estimated on the probabilistic feature map. The proposed algorithm is optimized from the initial pose by minimizing Mahalanobis distance errors between features from the query image and the map to improve accuracy. To verify that the localization accuracy is improved, the proposed algorithm is compared with the conventional algorithm in a simulation and realenvironments. PMID:26404284

Discovery of a New Cellular Motion and Its Relevance to Breast Cancer and Involution

DTIC Science & Technology

2014-02-01

motion (CAMo), live cell imaging , confocal microscopy Overall Project Summary: During this first year of funding we have concentrated our work to...cell types in 3D cultures and in vivo. Subtask 1.1a: Real time live cell imaging using confocal microscopy will be used to image cellular movement...exciting as they are important steps in understanding behavior of normal myoepithelial cells using live cell imaging in physiologically

Simultaneous imaging of cellular morphology and multiple biomarkers using an acousto-optic tunable filter-based bright field microscope.

PubMed

Wachman, Elliot S; Geyer, Stanley J; Recht, Joel M; Ward, Jon; Zhang, Bill; Reed, Murray; Pannell, Chris

2014-05-01

An acousto-optic tunable filter (AOTF)-based multispectral imaging microscope system allows the combination of cellular morphology and multiple biomarker stainings on a single microscope slide. We describe advances in AOTF technology that have greatly improved spectral purity, field uniformity, and image quality. A multispectral imaging bright field microscope using these advances demonstrates pathology results that have great potential for clinical use.

Measuring spatial and temporal Ca2+ signals in Arabidopsis plants.

PubMed

Zhu, Xiaohong; Taylor, Aaron; Zhang, Shenyu; Zhang, Dayong; Feng, Ying; Liang, Gaimei; Zhu, Jian-Kang

2014-09-02

Developmental and environmental cues induce Ca(2+) fluctuations in plant cells. Stimulus-specific spatial-temporal Ca(2+) patterns are sensed by cellular Ca(2+) binding proteins that initiate Ca(2+) signaling cascades. However, we still know little about how stimulus specific Ca(2+) signals are generated. The specificity of a Ca(2+) signal may be attributed to the sophisticated regulation of the activities of Ca(2+) channels and/or transporters in response to a given stimulus. To identify these cellular components and understand their functions, it is crucial to use systems that allow a sensitive and robust recording of Ca(2+) signals at both the tissue and cellular levels. Genetically encoded Ca(2+) indicators that are targeted to different cellular compartments have provided a platform for live cell confocal imaging of cellular Ca(2+) signals. Here we describe instructions for the use of two Ca(2+) detection systems: aequorin based FAS (film adhesive seedlings) luminescence Ca(2+) imaging and case12 based live cell confocal fluorescence Ca(2+) imaging. Luminescence imaging using the FAS system provides a simple, robust and sensitive detection of spatial and temporal Ca(2+) signals at the tissue level, while live cell confocal imaging using Case12 provides simultaneous detection of cytosolic and nuclear Ca(2+) signals at a high resolution.

Cartographic quality of ERTS-1 images

NASA Technical Reports Server (NTRS)

Welch, R. I.

1973-01-01

Analyses of simulated and operational ERTS images have provided initial estimates of resolution, ground resolution, detectability thresholds and other measures of image quality of interest to earth scientists and cartographers. Based on these values, including an approximate ground resolution of 250 meters for both RBV and MSS systems, the ERTS-1 images appear suited to the production and/or revision of planimetric and photo maps of 1:500,000 scale and smaller for which map accuracy standards are compatible with the imaged detail. Thematic mapping, although less constrained by map accuracy standards, will be influenced by measurement thresholds and errors which have yet to be accurately determined for ERTS images. This study also indicates the desirability of establishing a quantitative relationship between image quality values and map products which will permit both engineers and cartographers/earth scientists to contribute to the design requirements of future satellite imaging systems.

Context Specificity of Stress-activated Mitogen-activated Protein (MAP) Kinase Signaling: The Story as Told by Caenorhabditis elegans*

PubMed Central

Andrusiak, Matthew G.; Jin, Yishi

2016-01-01

Stress-associated p38 and JNK mitogen-activated protein (MAP) kinase signaling cascades trigger specific cellular responses and are involved in multiple disease states. At the root of MAP kinase signaling complexity is the differential use of common components on a context-specific basis. The roundworm Caenorhabditis elegans was developed as a system to study genes required for development and nervous system function. The powerful genetics of C. elegans in combination with molecular and cellular dissections has led to a greater understanding of how p38 and JNK signaling affects many biological processes under normal and stress conditions. This review focuses on the studies revealing context specificity of different stress-activated MAPK components in C. elegans. PMID:26907690

A Mathematical Model for Storage and Recall of Images using Targeted Synchronization of Coupled Maps.

PubMed

Palaniyandi, P; Rangarajan, Govindan

2017-08-21

We propose a mathematical model for storage and recall of images using coupled maps. We start by theoretically investigating targeted synchronization in coupled map systems wherein only a desired (partial) subset of the maps is made to synchronize. A simple method is introduced to specify coupling coefficients such that targeted synchronization is ensured. The principle of this method is extended to storage/recall of images using coupled Rulkov maps. The process of adjusting coupling coefficients between Rulkov maps (often used to model neurons) for the purpose of storing a desired image mimics the process of adjusting synaptic strengths between neurons to store memories. Our method uses both synchronisation and synaptic weight modification, as the human brain is thought to do. The stored image can be recalled by providing an initial random pattern to the dynamical system. The storage and recall of the standard image of Lena is explicitly demonstrated.

Depth map generation using a single image sensor with phase masks.

PubMed

Jang, Jinbeum; Park, Sangwoo; Jo, Jieun; Paik, Joonki

2016-06-13

Conventional stereo matching systems generate a depth map using two or more digital imaging sensors. It is difficult to use the small camera system because of their high costs and bulky sizes. In order to solve this problem, this paper presents a stereo matching system using a single image sensor with phase masks for the phase difference auto-focusing. A novel pattern of phase mask array is proposed to simultaneously acquire two pairs of stereo images. Furthermore, a noise-invariant depth map is generated from the raw format sensor output. The proposed method consists of four steps to compute the depth map: (i) acquisition of stereo images using the proposed mask array, (ii) variational segmentation using merging criteria to simplify the input image, (iii) disparity map generation using the hierarchical block matching for disparity measurement, and (iv) image matting to fill holes to generate the dense depth map. The proposed system can be used in small digital cameras without additional lenses or sensors.

Magnetic resonance imaging findings of cellular angiofibroma of the tunica vaginalis of the testis: a case report.

PubMed

Ntorkou, Alexandra A; Tsili, Athina C; Giannakis, Dimitrios; Batistatou, Anna; Stavrou, Sotirios; Sofikitis, Nikolaos; Argyropoulou, Maria I

2016-03-31

Cellular angiofibroma represents a rare mesenchymal tumor typically involving the inguinoscrotal area in middle-aged men. Although the origin of this benign tumor is unknown, it is histologically classified as an angiomyxoid tumor. Cellular angiofibroma is characterized by a diversity of pathological and imaging features. An accurate preoperative diagnosis is challenging. Magnetic resonance imaging examination of the scrotum has been reported as a valuable adjunct modality in the investigation of scrotal pathology. The technique by providing both structural and functional information is useful in the differentiation between extratesticular and intratesticular diseases and in the preoperative characterization of the histologic nature of various scrotal lesions. There are few reports in the English literature addressing the magnetic resonance imaging findings of cellular angiofibroma of the scrotum and no reports on functional magnetic resonance imaging data. Here we present the first case of a cellular angiofibroma arising from the tunica vaginalis of the testis and we discuss the value of a multiparametric magnetic resonance protocol, including diffusion-weighted imaging, magnetization transfer imaging and dynamic contrast-enhanced magnetic resonance imaging in the preoperative diagnosis of this rare neoplasm. A 47-year Greek man presented with a painless left scrotal swelling, which had gradually enlarged during the last 6 months. Magnetic resonance imaging of his scrotum displayed a left paratesticular mass, in close proximity to the tunica vaginalis, with heterogeneous high signal intensity on T2-weighted images and no areas of restricted diffusion. The tumor was hypointense on magnetization transfer images, suggestive for the presence of macromolecules. On dynamic contrast-enhanced magnetic resonance imaging the mass showed intense heterogeneous enhancement with a type II curve. Magnetic resonance imaging findings were strongly suggestive of a benign paratesticular tumor, which was confirmed on pathology following lesion excision. Magnetic resonance imaging of the scrotum by combining conventional and functional magnetic resonance data provides useful diagnostic information in the preoperative characterization of scrotal masses. A possible diagnosis of a benign paratesticular tumor based on magnetic resonance imaging features may improve patient care and decrease the number of unnecessary radical surgical explorations.

Cellular neural network-based hybrid approach toward automatic image registration

NASA Astrophysics Data System (ADS)

Arun, Pattathal VijayaKumar; Katiyar, Sunil Kumar

2013-01-01

Image registration is a key component of various image processing operations that involve the analysis of different image data sets. Automatic image registration domains have witnessed the application of many intelligent methodologies over the past decade; however, inability to properly model object shape as well as contextual information has limited the attainable accuracy. A framework for accurate feature shape modeling and adaptive resampling using advanced techniques such as vector machines, cellular neural network (CNN), scale invariant feature transform (SIFT), coreset, and cellular automata is proposed. CNN has been found to be effective in improving feature matching as well as resampling stages of registration and complexity of the approach has been considerably reduced using coreset optimization. The salient features of this work are cellular neural network approach-based SIFT feature point optimization, adaptive resampling, and intelligent object modelling. Developed methodology has been compared with contemporary methods using different statistical measures. Investigations over various satellite images revealed that considerable success was achieved with the approach. This system has dynamically used spectral and spatial information for representing contextual knowledge using CNN-prolog approach. This methodology is also illustrated to be effective in providing intelligent interpretation and adaptive resampling.

Development of a multi-scale and multi-modality imaging system to characterize tumours and their microenvironment in vivo

NASA Astrophysics Data System (ADS)

Rouffiac, Valérie; Ser-Leroux, Karine; Dugon, Emilie; Leguerney, Ingrid; Polrot, Mélanie; Robin, Sandra; Salomé-Desnoulez, Sophie; Ginefri, Jean-Christophe; Sebrié, Catherine; Laplace-Builhé, Corinne

2015-03-01

In vivo high-resolution imaging of tumor development is possible through dorsal skinfold chamber implantable on mice model. However, current intravital imaging systems are weakly tolerated along time by mice and do not allow multimodality imaging. Our project aims to develop a new chamber for: 1- long-term micro/macroscopic visualization of tumor (vascular and cellular compartments) and tissue microenvironment; and 2- multimodality imaging (photonic, MRI and sonography). Our new experimental device was patented in March 2014 and was primarily assessed on 75 mouse engrafted with 4T1-Luc tumor cell line, and validated in confocal and multiphoton imaging after staining the mice vasculature using Dextran 155KDa-TRITC or Dextran 2000kDa-FITC. Simultaneously, a universal stage was designed for optimal removal of respiratory and cardiac artifacts during microscopy assays. Experimental results from optical, ultrasound (Bmode and pulse subtraction mode) and MRI imaging (anatomic sequences) showed that our patented design, unlike commercial devices, improves longitudinal monitoring over several weeks (35 days on average against 12 for the commercial chamber) and allows for a better characterization of the early and late tissue alterations due to tumour development. We also demonstrated the compatibility for multimodality imaging and the increase of mice survival was by a factor of 2.9, with our new skinfold chamber. Current developments include: 1- defining new procedures for multi-labelling of cells and tissue (screening of fluorescent molecules and imaging protocols); 2- developing ultrasound and MRI imaging procedures with specific probes; 3- correlating optical/ultrasound/MRI data for a complete mapping of tumour development and microenvironment.

The goya mouse mutant reveals distinct newly identified roles for MAP3K1 in the development and survival of cochlear sensory hair cells.

PubMed

Parker, Andrew; Cross, Sally H; Jackson, Ian J; Hardisty-Hughes, Rachel; Morse, Susan; Nicholson, George; Coghill, Emma; Bowl, Michael R; Brown, Steve D M

2015-12-01

Mitogen-activated protein kinase, MAP3K1, plays an important role in a number of cellular processes, including epithelial migration during eye organogenesis. In addition, studies in keratinocytes indicate that MAP3K1 signalling through JNK is important for actin stress fibre formation and cell migration. However, MAP3K1 can also act independently of JNK in the regulation of cell proliferation and apoptosis. We have identified a mouse mutant, goya, which exhibits the eyes-open-at-birth and microphthalmia phenotypes. In addition, these mice also have hearing loss. The goya mice carry a splice site mutation in the Map3k1 gene. We show that goya and kinase-deficient Map3k1 homozygotes initially develop supernumerary cochlear outer hair cells (OHCs) that subsequently degenerate, and a progressive profound hearing loss is observed by 9 weeks of age. Heterozygote mice also develop supernumerary OHCs, but no cellular degeneration or hearing loss is observed. MAP3K1 is expressed in a number of inner-ear cell types, including outer and inner hair cells, stria vascularis and spiral ganglion. Investigation of targets downstream of MAP3K1 identified an increase in p38 phosphorylation (Thr180/Tyr182) in multiple cochlear tissues. We also show that the extra OHCs do not arise from aberrant control of proliferation via p27KIP1. The identification of the goya mutant reveals a signalling molecule involved with hair-cell development and survival. Mammalian hair cells do not have the ability to regenerate after damage, which can lead to irreversible sensorineural hearing loss. Given the observed goya phenotype, and the many diverse cellular processes that MAP3K1 is known to act upon, further investigation of this model might help to elaborate upon the mechanisms underlying sensory hair cell specification, and pathways important for their survival. In addition, MAP3K1 is revealed as a new candidate gene for human sensorineural hearing loss. © 2015. Published by The Company of Biologists Ltd.

Gamut mapping in a high-dynamic-range color space

NASA Astrophysics Data System (ADS)

Preiss, Jens; Fairchild, Mark D.; Ferwerda, James A.; Urban, Philipp

2014-01-01

In this paper, we present a novel approach of tone mapping as gamut mapping in a high-dynamic-range (HDR) color space. High- and low-dynamic-range (LDR) images as well as device gamut boundaries can simultaneously be represented within such a color space. This enables a unified transformation of the HDR image into the gamut of an output device (in this paper called HDR gamut mapping). An additional aim of this paper is to investigate the suitability of a specific HDR color space to serve as a working color space for the proposed HDR gamut mapping. For the HDR gamut mapping, we use a recent approach that iteratively minimizes an image-difference metric subject to in-gamut images. A psychophysical experiment on an HDR display shows that the standard reproduction workflow of two subsequent transformations - tone mapping and then gamut mapping - may be improved by HDR gamut mapping.

Automated processing of label-free Raman microscope images of macrophage cells with standardized regression for high-throughput analysis.

PubMed

Milewski, Robert J; Kumagai, Yutaro; Fujita, Katsumasa; Standley, Daron M; Smith, Nicholas I

2010-11-19

Macrophages represent the front lines of our immune system; they recognize and engulf pathogens or foreign particles thus initiating the immune response. Imaging macrophages presents unique challenges, as most optical techniques require labeling or staining of the cellular compartments in order to resolve organelles, and such stains or labels have the potential to perturb the cell, particularly in cases where incomplete information exists regarding the precise cellular reaction under observation. Label-free imaging techniques such as Raman microscopy are thus valuable tools for studying the transformations that occur in immune cells upon activation, both on the molecular and organelle levels. Due to extremely low signal levels, however, Raman microscopy requires sophisticated image processing techniques for noise reduction and signal extraction. To date, efficient, automated algorithms for resolving sub-cellular features in noisy, multi-dimensional image sets have not been explored extensively. We show that hybrid z-score normalization and standard regression (Z-LSR) can highlight the spectral differences within the cell and provide image contrast dependent on spectral content. In contrast to typical Raman imaging processing methods using multivariate analysis, such as single value decomposition (SVD), our implementation of the Z-LSR method can operate nearly in real-time. In spite of its computational simplicity, Z-LSR can automatically remove background and bias in the signal, improve the resolution of spatially distributed spectral differences and enable sub-cellular features to be resolved in Raman microscopy images of mouse macrophage cells. Significantly, the Z-LSR processed images automatically exhibited subcellular architectures whereas SVD, in general, requires human assistance in selecting the components of interest. The computational efficiency of Z-LSR enables automated resolution of sub-cellular features in large Raman microscopy data sets without compromise in image quality or information loss in associated spectra. These results motivate further use of label free microscopy techniques in real-time imaging of live immune cells.

Large-scale Scanning Transmission Electron Microscopy (Nanotomy) of Healthy and Injured Zebrafish Brain.

PubMed

Kuipers, Jeroen; Kalicharan, Ruby D; Wolters, Anouk H G; van Ham, Tjakko J; Giepmans, Ben N G

2016-05-25

Large-scale 2D electron microscopy (EM), or nanotomy, is the tissue-wide application of nanoscale resolution electron microscopy. Others and we previously applied large scale EM to human skin pancreatic islets, tissue culture and whole zebrafish larvae(1-7). Here we describe a universally applicable method for tissue-scale scanning EM for unbiased detection of sub-cellular and molecular features. Nanotomy was applied to investigate the healthy and a neurodegenerative zebrafish brain. Our method is based on standardized EM sample preparation protocols: Fixation with glutaraldehyde and osmium, followed by epoxy-resin embedding, ultrathin sectioning and mounting of ultrathin-sections on one-hole grids, followed by post staining with uranyl and lead. Large-scale 2D EM mosaic images are acquired using a scanning EM connected to an external large area scan generator using scanning transmission EM (STEM). Large scale EM images are typically ~ 5 - 50 G pixels in size, and best viewed using zoomable HTML files, which can be opened in any web browser, similar to online geographical HTML maps. This method can be applied to (human) tissue, cross sections of whole animals as well as tissue culture(1-5). Here, zebrafish brains were analyzed in a non-invasive neuronal ablation model. We visualize within a single dataset tissue, cellular and subcellular changes which can be quantified in various cell types including neurons and microglia, the brain's macrophages. In addition, nanotomy facilitates the correlation of EM with light microscopy (CLEM)(8) on the same tissue, as large surface areas previously imaged using fluorescent microscopy, can subsequently be subjected to large area EM, resulting in the nano-anatomy (nanotomy) of tissues. In all, nanotomy allows unbiased detection of features at EM level in a tissue-wide quantifiable manner.

Large-scale Scanning Transmission Electron Microscopy (Nanotomy) of Healthy and Injured Zebrafish Brain

PubMed Central

Kuipers, Jeroen; Kalicharan, Ruby D.; Wolters, Anouk H. G.

2016-01-01

Large-scale 2D electron microscopy (EM), or nanotomy, is the tissue-wide application of nanoscale resolution electron microscopy. Others and we previously applied large scale EM to human skin pancreatic islets, tissue culture and whole zebrafish larvae1-7. Here we describe a universally applicable method for tissue-scale scanning EM for unbiased detection of sub-cellular and molecular features. Nanotomy was applied to investigate the healthy and a neurodegenerative zebrafish brain. Our method is based on standardized EM sample preparation protocols: Fixation with glutaraldehyde and osmium, followed by epoxy-resin embedding, ultrathin sectioning and mounting of ultrathin-sections on one-hole grids, followed by post staining with uranyl and lead. Large-scale 2D EM mosaic images are acquired using a scanning EM connected to an external large area scan generator using scanning transmission EM (STEM). Large scale EM images are typically ~ 5 - 50 G pixels in size, and best viewed using zoomable HTML files, which can be opened in any web browser, similar to online geographical HTML maps. This method can be applied to (human) tissue, cross sections of whole animals as well as tissue culture1-5. Here, zebrafish brains were analyzed in a non-invasive neuronal ablation model. We visualize within a single dataset tissue, cellular and subcellular changes which can be quantified in various cell types including neurons and microglia, the brain's macrophages. In addition, nanotomy facilitates the correlation of EM with light microscopy (CLEM)8 on the same tissue, as large surface areas previously imaged using fluorescent microscopy, can subsequently be subjected to large area EM, resulting in the nano-anatomy (nanotomy) of tissues. In all, nanotomy allows unbiased detection of features at EM level in a tissue-wide quantifiable manner. PMID:27285162

In Vivo Confocal Microscopy of the Ocular Surface: From Bench to Bedside

PubMed Central

Villani, Edoardo; Baudouin, Christophe; Efron, Nathan; Hamrah, Pedram; Kojima, Takashi; Patel, Sanjay V.; Pflugfelder, Stephen C.; Zhivov, Andrey; Dogru, Murat

2014-01-01

In vivo confocal microscopy (IVCM) is an emerging technology that provides minimally invasive, high resolution, steady-state assessment of the ocular surface at the cellular level. Several challenges still remain but, at present, IVCM may be considered a promising technique for clinical diagnosis and management. This mini-review summarizes some key findings in IVCM of the ocular surface, focusing on recent and promising attempts to move “from bench to bedside”. IVCM allows prompt diagnosis, disease course follow-up, and management of potentially blinding atypical forms of infectious processes, such as acanthamoeba and fungal keratitis. This technology has improved our knowledge of corneal alterations and some of the processes that affect the visual outcome after lamellar keratoplasty and excimer keratorefractive surgery. In dry eye disease, IVCM has provided new information on the whole-ocular surface morphofunctional unit. It has also improved understanding of pathophysiologic mechanisms and helped in the assessment of prognosis and treatment. IVCM is particularly useful in the study of corneal nerves, enabling description of the morphology, density, and disease- or surgically induced alterations of nerves, particularly the subbasal nerve plexus. In glaucoma, IVCM constitutes an important aid to evaluate filtering blebs, to better understand the conjunctival wound healing process, and to assess corneal changes induced by topical antiglaucoma medications and their preservatives. IVCM has significantly enhanced our understanding of the ocular response to contact lens wear. It has provided new perspectives at a cellular level on a wide range of contact lens complications, revealing findings that were not previously possible to image in the living human eye. The final section of this mini-review provides a focus on advances in confocal microscopy imaging. These include 2D wide-field mapping, 3D reconstruction of the cornea and automated image analysis. PMID:24215436

Measuring the Autocorrelation Function of Nanoscale Three-Dimensional Density Distribution in Individual Cells Using Scanning Transmission Electron Microscopy, Atomic Force Microscopy, and a New Deconvolution Algorithm.

PubMed

Li, Yue; Zhang, Di; Capoglu, Ilker; Hujsak, Karl A; Damania, Dhwanil; Cherkezyan, Lusik; Roth, Eric; Bleher, Reiner; Wu, Jinsong S; Subramanian, Hariharan; Dravid, Vinayak P; Backman, Vadim

2017-06-01

Essentially all biological processes are highly dependent on the nanoscale architecture of the cellular components where these processes take place. Statistical measures, such as the autocorrelation function (ACF) of the three-dimensional (3D) mass-density distribution, are widely used to characterize cellular nanostructure. However, conventional methods of reconstruction of the deterministic 3D mass-density distribution, from which these statistical measures can be calculated, have been inadequate for thick biological structures, such as whole cells, due to the conflict between the need for nanoscale resolution and its inverse relationship with thickness after conventional tomographic reconstruction. To tackle the problem, we have developed a robust method to calculate the ACF of the 3D mass-density distribution without tomography. Assuming the biological mass distribution is isotropic, our method allows for accurate statistical characterization of the 3D mass-density distribution by ACF with two data sets: a single projection image by scanning transmission electron microscopy and a thickness map by atomic force microscopy. Here we present validation of the ACF reconstruction algorithm, as well as its application to calculate the statistics of the 3D distribution of mass-density in a region containing the nucleus of an entire mammalian cell. This method may provide important insights into architectural changes that accompany cellular processes.

Measuring the Autocorrelation Function of Nanoscale Three-Dimensional Density Distribution in Individual Cells Using Scanning Transmission Electron Microscopy, Atomic Force Microscopy, and a New Deconvolution Algorithm

PubMed Central

Li, Yue; Zhang, Di; Capoglu, Ilker; Hujsak, Karl A.; Damania, Dhwanil; Cherkezyan, Lusik; Roth, Eric; Bleher, Reiner; Wu, Jinsong S.; Subramanian, Hariharan; Dravid, Vinayak P.; Backman, Vadim

2018-01-01

Essentially all biological processes are highly dependent on the nanoscale architecture of the cellular components where these processes take place. Statistical measures, such as the autocorrelation function (ACF) of the three-dimensional (3D) mass–density distribution, are widely used to characterize cellular nanostructure. However, conventional methods of reconstruction of the deterministic 3D mass–density distribution, from which these statistical measures can be calculated, have been inadequate for thick biological structures, such as whole cells, due to the conflict between the need for nanoscale resolution and its inverse relationship with thickness after conventional tomographic reconstruction. To tackle the problem, we have developed a robust method to calculate the ACF of the 3D mass–density distribution without tomography. Assuming the biological mass distribution is isotropic, our method allows for accurate statistical characterization of the 3D mass–density distribution by ACF with two data sets: a single projection image by scanning transmission electron microscopy and a thickness map by atomic force microscopy. Here we present validation of the ACF reconstruction algorithm, as well as its application to calculate the statistics of the 3D distribution of mass–density in a region containing the nucleus of an entire mammalian cell. This method may provide important insights into architectural changes that accompany cellular processes. PMID:28416035

Time-resolved spectroscopic imaging reveals the fundamentals of cellular NADH fluorescence.

PubMed

Li, Dong; Zheng, Wei; Qu, Jianan Y

2008-10-15

A time-resolved spectroscopic imaging system is built to study the fluorescence characteristics of nicotinamide adenine dinucleotide (NADH), an important metabolic coenzyme and endogenous fluorophore in cells. The system provides a unique approach to measure fluorescence signals in different cellular organelles and cytoplasm. The ratios of free over protein-bound NADH signals in cytosol and nucleus are slightly higher than those in mitochondria. The mitochondrial fluorescence contributes about 70% of overall cellular fluorescence and is not a completely dominant signal. Furthermore, NADH signals in mitochondria, cytosol, and the nucleus respond to the changes of cellular activity differently, suggesting that cytosolic and nuclear fluorescence may complicate the well-known relationship between mitochondrial fluorescence and cellular metabolism.

Image Maps in the World-Wide Web: The Uses and Limitations.

ERIC Educational Resources Information Center

Cochenour, John J.; And Others

A study of nine different image maps from World Wide Web home pages was conducted to evaluate their effectiveness in information display and access, relative to visual, navigational, and practical characteristics. Nine independent viewers completed 20-question surveys on the image maps, in which they evaluated the characteristics of the maps on a…

Dynamic maps of UV damage formation and repair for the human genome

PubMed Central

Hu, Jinchuan; Adebali, Ogun; Adar, Sheera; Sancar, Aziz

2017-01-01

Formation and repair of UV-induced DNA damage in human cells are affected by cellular context. To study factors influencing damage formation and repair genome-wide, we developed a highly sensitive single-nucleotide resolution damage mapping method [high-sensitivity damage sequencing (HS–Damage-seq)]. Damage maps of both cyclobutane pyrimidine dimers (CPDs) and pyrimidine-pyrimidone (6-4) photoproducts [(6-4)PPs] from UV-irradiated cellular and naked DNA revealed that the effect of transcription factor binding on bulky adducts formation varies, depending on the specific transcription factor, damage type, and strand. We also generated time-resolved UV damage maps of both CPDs and (6-4)PPs by HS–Damage-seq and compared them to the complementary repair maps of the human genome obtained by excision repair sequencing to gain insight into factors that affect UV-induced DNA damage and repair and ultimately UV carcinogenesis. The combination of the two methods revealed that, whereas UV-induced damage is virtually uniform throughout the genome, repair is affected by chromatin states, transcription, and transcription factor binding, in a manner that depends on the type of DNA damage. PMID:28607063

Dynamic maps of UV damage formation and repair for the human genome.

PubMed

Hu, Jinchuan; Adebali, Ogun; Adar, Sheera; Sancar, Aziz

2017-06-27

Formation and repair of UV-induced DNA damage in human cells are affected by cellular context. To study factors influencing damage formation and repair genome-wide, we developed a highly sensitive single-nucleotide resolution damage mapping method [high-sensitivity damage sequencing (HS-Damage-seq)]. Damage maps of both cyclobutane pyrimidine dimers (CPDs) and pyrimidine-pyrimidone (6-4) photoproducts [(6-4)PPs] from UV-irradiated cellular and naked DNA revealed that the effect of transcription factor binding on bulky adducts formation varies, depending on the specific transcription factor, damage type, and strand. We also generated time-resolved UV damage maps of both CPDs and (6-4)PPs by HS-Damage-seq and compared them to the complementary repair maps of the human genome obtained by excision repair sequencing to gain insight into factors that affect UV-induced DNA damage and repair and ultimately UV carcinogenesis. The combination of the two methods revealed that, whereas UV-induced damage is virtually uniform throughout the genome, repair is affected by chromatin states, transcription, and transcription factor binding, in a manner that depends on the type of DNA damage.

A natural-color mapping for single-band night-time image based on FPGA

NASA Astrophysics Data System (ADS)

Wang, Yilun; Qian, Yunsheng

2018-01-01

A natural-color mapping for single-band night-time image method based on FPGA can transmit the color of the reference image to single-band night-time image, which is consistent with human visual habits and can help observers identify the target. This paper introduces the processing of the natural-color mapping algorithm based on FPGA. Firstly, the image can be transformed based on histogram equalization, and the intensity features and standard deviation features of reference image are stored in SRAM. Then, the real-time digital images' intensity features and standard deviation features are calculated by FPGA. At last, FPGA completes the color mapping through matching pixels between images using the features in luminance channel.

Using image mapping towards biomedical and biological data sharing

PubMed Central

2013-01-01

Image-based data integration in eHealth and life sciences is typically concerned with the method used for anatomical space mapping, needed to retrieve, compare and analyse large volumes of biomedical data. In mapping one image onto another image, a mechanism is used to match and find the corresponding spatial regions which have the same meaning between the source and the matching image. Image-based data integration is useful for integrating data of various information structures. Here we discuss a broad range of issues related to data integration of various information structures, review exemplary work on image representation and mapping, and discuss the challenges that these techniques may bring. PMID:24059352

Modification of measurement methods for evaluation of tissue-engineered cartilage function and biochemical properties using nanosecond pulsed laser

NASA Astrophysics Data System (ADS)

Ishihara, Miya; Sato, Masato; Kutsuna, Toshiharu; Ishihara, Masayuki; Mochida, Joji; Kikuchi, Makoto

2008-02-01

There is a demand in the field of regenerative medicine for measurement technology that enables determination of functions and components of engineered tissue. To meet this demand, we developed a method for extracellular matrix characterization using time-resolved autofluorescence spectroscopy, which enabled simultaneous measurements with mechanical properties using relaxation of laser-induced stress wave. In this study, in addition to time-resolved fluorescent spectroscopy, hyperspectral sensor, which enables to capture both spectral and spatial information, was used for evaluation of biochemical characterization of tissue-engineered cartilage. Hyperspectral imaging system provides spectral resolution of 1.2 nm and image rate of 100 images/sec. The imaging system consisted of the hyperspectral sensor, a scanner for x-y plane imaging, magnifying optics and Xenon lamp for transmmissive lighting. Cellular imaging using the hyperspectral image system has been achieved by improvement in spatial resolution up to 9 micrometer. The spectroscopic cellular imaging could be observed using cultured chondrocytes as sample. At early stage of culture, the hyperspectral imaging offered information about cellular function associated with endogeneous fluorescent biomolecules.

Imaging deep skeletal muscle structure using a high-sensitivity ultrathin side-viewing optical coherence tomography needle probe

PubMed Central

Yang, Xiaojie; Lorenser, Dirk; McLaughlin, Robert A.; Kirk, Rodney W.; Edmond, Matthew; Simpson, M. Cather; Grounds, Miranda D.; Sampson, David D.

2013-01-01

We have developed an extremely miniaturized optical coherence tomography (OCT) needle probe (outer diameter 310 µm) with high sensitivity (108 dB) to enable minimally invasive imaging of cellular structure deep within skeletal muscle. Three-dimensional volumetric images were acquired from ex vivo mouse tissue, examining both healthy and pathological dystrophic muscle. Individual myofibers were visualized as striations in the images. Degradation of cellular structure in necrotic regions was seen as a loss of these striations. Tendon and connective tissue were also visualized. The observed structures were validated against co-registered hematoxylin and eosin (H&E) histology sections. These images of internal cellular structure of skeletal muscle acquired with an OCT needle probe demonstrate the potential of this technique to visualize structure at the microscopic level deep in biological tissue in situ. PMID:24466482

Preliminary Image Map of the 2007 Harris Fire Perimeter, Barrett Lake Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Santiago Fire Perimeter, Santiago Peak Quadrangle, Orange and Riverside Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Buckweed Fire Perimeter, Green Valley Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Warners Ranch Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Otay Mesa Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Rice Fire Perimeter, Bonsall Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Poomacha Fire Perimeter, Pechanga Quadrangle, Riverside and San Diego Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Tecate Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Poomacha Fire Perimeter, Temecula Quadrangle, Riverside and San Diego Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Buckweed Fire Perimeter, Agua Dulce Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, San Pasqual Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Buckweed Fire Perimeter, Mint Canyon Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Escondido Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Poomacha Fire Perimeter, Boucher Hill Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ammo Fire Perimeter, Margarita Peak Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Ramona Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ammo Fire Perimeter, San Onofre Bluff Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Santiago Fire Perimeter, Orange Quadrangle, Orange County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Otay Mountain Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ranch Fire Perimeter, Cobblestone Mountain Quadrangle, Los Angeles and Ventura Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Poomacha Fire Perimeter, Palomar Observatory Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, El Cajon Mountain Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch and Poomacha Fire Perimeters, Rodriguez Mountain Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Santa Ysabel Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ammo Fire Perimeter, Las Pulgas Canyon Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Jamul Mountains Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Santiago Fire Perimeter, Lake Forest Quadrangle, Orange County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Cajon Fire Perimeter, San Bernardino North Quadrangle, San Bernardino County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Slide Fire Perimeter, Butler Peak Quadrangle, San Bernardino County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, San Vicente Reservoir Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ammo Fire Perimeter, San Clemente Quadrangle, Orange and San Diego Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Cajon Fire Perimeter, Devore Quadrangle, San Bernardino County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ranch Fire Perimeter, Fillmore Quadrangle, Ventura County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ranch Fire Perimeter, Piru Quadrangle, Ventura County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Magic and Buckweed Fire Perimeters, Newhall Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Dulzura Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Grass Valley Fire Perimeter, Lake Arrowhead Quadrangle, San Bernardino County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Potrero Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch and Poomacha Fire Perimeters, Mesa Grande Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Canyon Fire Perimeter, Malibu Beach Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Santiago Fire Perimeter, Black Star Canyon Quadrangle, Orange, Riverside, and San Bernardino Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Buckweed Fire Perimeter, Warm Springs Mountain Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ranch Fire Perimeter, Whitaker Peak Quadrangle, Los Angeles and Ventura Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Poomacha Fire Perimeter, Vail Lake Quadrangle, Riverside and San Diego Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Valley Center Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Santiago Fire Perimeter, Tustin Quadrangle, Orange County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Rancho Santa Fe Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Slide Fire Perimeter, Harrison Mountain Quadrangle, San Bernardino County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Buckweed Fire Perimeter, Sleepy Valley Quadrangle, Los Angeles County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Ranch and Magic Fire Perimeters, Val Verde Quadrangle, Los Angeles and Ventura Counties, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Poway Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Poomacha Fire Perimeter, Pala Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Witch Fire Perimeter, Tule Springs Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Harris Fire Perimeter, Morena Reservoir Quadrangle, San Diego County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Preliminary Image Map of the 2007 Slide Fire Perimeter, Keller Peak Quadrangle, San Bernardino County, California

USGS Publications Warehouse

Clark, Perry S.; Scratch, Wendy S.; Bias, Gaylord W.; Stander, Gregory B.; Sexton, Jenne L.; Krawczak, Bridgette J.

2008-01-01

In the fall of 2007, wildfires burned out of control in southern California. The extent of these fires encompassed large geographic areas that included a variety of landscapes from urban to wilderness. The U.S. Geological Survey National Geospatial Technical Operations Center (NGTOC) is currently (2008) developing a quadrangle-based 1:24,000-scale image map product. One of the concepts behind the image map product is to provide an updated map in electronic format to assist with emergency response. This image map is one of 55 preliminary image map quadrangles covering the areas burned by the southern California wildfires. Each map is a layered, geo-registered Portable Document Format (.pdf) file. For more information about the layered geo-registered .pdf, see the readme file (http://pubs.usgs.gov/of/2008/1029/downloads/CA_Agua_Dulce_of2008-1029_README.txt). To view the areas affected and the quadrangles mapped in this preliminary project, see the map index (http://pubs.usgs.gov/of/2008/1029/downloads/CA_of2008_1029-1083_index.pdf) provided with this report.

Profiling pleural effusion cells by a diffraction imaging method

NASA Astrophysics Data System (ADS)

Al-Qaysi, Safaa; Hong, Heng; Wen, Yuhua; Lu, Jun Q.; Feng, Yuanming; Hu, Xin-Hua

2018-02-01

Assay of cells in pleural effusion (PE) is an important means of disease diagnosis. Conventional cytology of effusion samples, however, has low sensitivity and depends heavily on the expertise of cytopathologists. We applied a polarization diffraction imaging flow cytometry method on effusion cells to investigate their features. Diffraction imaging of the PE cell samples has been performed on 6000 to 12000 cells for each effusion cell sample of three patients. After prescreening to remove images by cellular debris and aggregated non-cellular particles, the image textures were extracted with a gray level co-occurrence matrix (GLCM) algorithm. The distribution of the imaged cells in the GLCM parameters space was analyzed by a Gaussian Mixture Model (GMM) to determine the number of clusters among the effusion cells. These results yield insight on textural features of diffraction images and related cellular morphology in effusion samples and can be used toward the development of a label-free method for effusion cells assay.

Intelligent Interfaces for Mining Large-Scale RNAi-HCS Image Databases

PubMed Central

Lin, Chen; Mak, Wayne; Hong, Pengyu; Sepp, Katharine; Perrimon, Norbert

2010-01-01

Recently, High-content screening (HCS) has been combined with RNA interference (RNAi) to become an essential image-based high-throughput method for studying genes and biological networks through RNAi-induced cellular phenotype analyses. However, a genome-wide RNAi-HCS screen typically generates tens of thousands of images, most of which remain uncategorized due to the inadequacies of existing HCS image analysis tools. Until now, it still requires highly trained scientists to browse a prohibitively large RNAi-HCS image database and produce only a handful of qualitative results regarding cellular morphological phenotypes. For this reason we have developed intelligent interfaces to facilitate the application of the HCS technology in biomedical research. Our new interfaces empower biologists with computational power not only to effectively and efficiently explore large-scale RNAi-HCS image databases, but also to apply their knowledge and experience to interactive mining of cellular phenotypes using Content-Based Image Retrieval (CBIR) with Relevance Feedback (RF) techniques. PMID:21278820

In vivo confocal microscopy of the cornea: New developments in image acquisition, reconstruction and analysis using the HRT-Rostock Corneal Module

PubMed Central

Petroll, W. Matthew; Robertson, Danielle M.

2015-01-01

The optical sectioning ability of confocal microscopy allows high magnification images to be obtained from different depths within a thick tissue specimen, and is thus ideally suited to the study of intact tissue in living subjects. In vivo confocal microscopy has been used in a variety of corneal research and clinical applications since its development over 25 years ago. In this article we review the latest developments in quantitative corneal imaging with the Heidelberg Retinal Tomograph with Rostock Corneal Module (HRT-RCM). We provide an overview of the unique strengths and weaknesses of the HRT-RCM. We discuss techniques for performing 3-D imaging with the HRT-RCM, including hardware and software modifications that allow full thickness confocal microscopy through focusing (CMTF) of the cornea, which can provide quantitative measurements of corneal sublayer thicknesses, stromal cell and extracellular matrix backscatter, and depth dependent changes in corneal keratocyte density. We also review current approaches for quantitative imaging of the subbasal nerve plexus, which require a combination of advanced image acquisition and analysis procedures, including wide field mapping and 3-D reconstruction of nerve structures. The development of new hardware, software, and acquisition techniques continues to expand the number of applications of the HRT-RCM for quantitative in vivo corneal imaging at the cellular level. Knowledge of these rapidly evolving strategies should benefit corneal clinicians and basic scientists alike. PMID:25998608

Mapping Radiation Injury and Recovery in Bone Marrow Using 18F-FLT PET/CT and USPIO MRI in a Rat Model.

PubMed

Rendon, David A; Kotedia, Khushali; Afshar, Solmaz F; Punia, Jyotinder N; Sabek, Omaima M; Shirkey, Beverly A; Zawaski, Janice A; Gaber, M Waleed

2016-02-01

We present and test the use of multimodality imaging as a topological tool to map the amount of the body exposed to ionizing radiation and the location of exposure, which are important indicators of survival and recovery. To achieve our goal, PET/CT imaging with 3'-deoxy-3'-(18)F-fluorothymidine ((18)F-FLT) was used to measure cellular proliferation in bone marrow (BM), whereas MRI using ultra-small superparamagnetic iron oxide (USPIO) particles provided noninvasive information on radiation-induced vascular damage. Animals were x-ray-irradiated at a dose of 7.5 Gy with 1 of 3 radiation schemes-whole-body irradiation, half-body shielding (HBS), or 1-leg shielding (1LS)-and imaged repeatedly. The spatial information from the CT scan was used to segment the region corresponding to BM from the PET scan using algorithms developed in-house, allowing for quantification of proliferating cells, and BM blood volume was estimated by measuring the changes in the T2 relaxation rates (ΔR2) collected from MR scans. (18)F-FLT PET/CT imaging differentiated irradiated from unirradiated BM regions. Two days after irradiation, proliferation of 1LS animals was significantly lower than sham (P = 0.0001, femurs; P < 0.0001, tibias) and returned to sham levels by day 10 (P = 0.6344, femurs; P = 0.3962, tibias). The degree of shielding affected proliferation recovery, showing an increase in the irradiated BM of the femurs, but not the tibias, of HBS animals when compared with 1LS (P = 0.0310, femurs; P = 0.5832, tibias). MRI of irradiated spines detected radiation-induced BM vascular damage, measured by the significant increase in ΔR2 2 d after whole-body irradiation (P = 0.0022) and HBS (P = 0.0003) with a decreasing trend of values, returning to levels close to baseline over 10 d. Our data were corroborated using γ-counting and histopathology. We demonstrated that (18)F-FLT PET/CT and USPIO MRI are valuable tools in mapping regional radiation exposure and the effects of radiation on BM. Analysis of the (18)F-FLT signal allowed for a clear demarcation of exposed BM regions and elucidated the kinetics of BM recovery, whereas USPIO MRI was used to assess vascular damage and recovery. © 2016 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

An Integrated Tone Mapping for High Dynamic Range Image Visualization

NASA Astrophysics Data System (ADS)

Liang, Lei; Pan, Jeng-Shyang; Zhuang, Yongjun

2018-01-01

There are two type tone mapping operators for high dynamic range (HDR) image visualization. HDR image mapped by perceptual operators have strong sense of reality, but will lose local details. Empirical operators can maximize local detail information of HDR image, but realism is not strong. A common tone mapping operator suitable for all applications is not available. This paper proposes a novel integrated tone mapping framework which can achieve conversion between empirical operators and perceptual operators. In this framework, the empirical operator is rendered based on improved saliency map, which simulates the visual attention mechanism of the human eye to the natural scene. The results of objective evaluation prove the effectiveness of the proposed solution.

Functional reorganization of visual cortex maps after ischemic lesions is accompanied by changes in expression of cytoskeletal proteins and NMDA and GABA(A) receptor subunits.

PubMed

Zepeda, Angelica; Sengpiel, Frank; Guagnelli, Miguel Angel; Vaca, Luis; Arias, Clorinda

2004-02-25

Reorganization of cortical representations after focal visual cortex lesions has been documented. It has been suggested that functional reorganization may rely on cellular mechanisms involving modifications in the excitatory/inhibitory neurotransmission balance and on morphological changes of neurons peripheral to the lesion. We explored functional reorganization of cortical retinotopic maps after a focal ischemic lesion in primary visual cortex of kittens using optical imaging of intrinsic signals. After 1, 2, and 5 weeks postlesion (wPL), we addressed whether functional reorganization correlated in time with changes in the expression of MAP-2, GAP-43, GFAP, GABA(A) receptor subunit alpha1 (GABA(A)alpha1), subunit 1 of the NMDA receptor (NMDAR1), and in neurotransmitter levels at the border of the lesion. Our results show that: (1) retinotopic maps reorganize with time after an ischemic lesion; (2) MAP-2 levels increase gradually from 1wPL to 5wPL; (3) MAP-2 upregulation is associated with an increase in dendritic-like structures surrounding the lesion and a decrease in GFAP-positive cells; (4) GAP-43 levels reach the highest point at 2wPL; (5) NMDAR1 and glutamate contents increase in parallel from 1wPL to 5wPL; (6) GABA(A)alpha1 levels increase from 1wPL to 2wPL but do not change after this time point; and (7) GABA contents remain low from 1wPL to 5wPL. This is a comprehensive study showing for the first time that functional reorganization correlates in time with dendritic sprouting and with changes in the excitatory/inhibitory neurotransmission systems previously proposed to participate in cortical remodeling and suggests mechanisms by which plasticity of cortical representations may occur.

Rice crop mapping and change prediction using multi-temporal satellite images in the Mekong Delta, Vietnam

NASA Astrophysics Data System (ADS)

Chen, C. R.; Chen, C. F.; Nguyen, S. T.

2014-12-01

The rice cropping systems in the Vietnamese Mekong Delta (VMD) has been undergoing major changes to cope with developing agro-economics, increasing population and changing climate. Information on rice cropping practices and changes in cropping systems is critical for policymakers to devise successful strategies to ensure food security and rice grain exports for the country. The primary objective of this research is to map rice cropping systems and predict future dynamics of rice cropping systems using the MODIS time-series data of 2002, 2006, and 2010. First, a phenology-based classification approach was applied for the classification and assessment of rice cropping systems in study region. Second, the Cellular Automata-Markov (CA-Markov) models was used to simulate the rice-cropping system map of VMD for 2010. The comparisons between the classification maps and the ground reference data indicated satisfactory results with overall accuracies and Kappa coefficients, respectively, of 81.4% and 0.75 for 2002, 80.6% and 0.74 for 2006 and 85.5% and 0.81 for 2010. The simulated map of rice cropping system for 2010 was extrapolated by CA-Markov model based on the trend of rice cropping systems during 2002~2006. The comparison between predicted scenario and classification map for 2010 presents a reasonably closer agreement. In conclusion, the CA-Markov model performs a powerful tool for the dynamic modeling of changes in rice cropping systems, and the results obtained demonstrate that the approach produces satisfactory results in terms of accuracy, quantitative forecast and spatial pattern changes. Meanwhile, the projections of the future changes would provide useful inputs to the agricultural policy for effective management of the rice cropping practices in VMD.

Computed inverse resonance imaging for magnetic susceptibility map reconstruction.

PubMed

Chen, Zikuan; Calhoun, Vince

2012-01-01

This article reports a computed inverse magnetic resonance imaging (CIMRI) model for reconstructing the magnetic susceptibility source from MRI data using a 2-step computational approach. The forward T2*-weighted MRI (T2*MRI) process is broken down into 2 steps: (1) from magnetic susceptibility source to field map establishment via magnetization in the main field and (2) from field map to MR image formation by intravoxel dephasing average. The proposed CIMRI model includes 2 inverse steps to reverse the T2*MRI procedure: field map calculation from MR-phase image and susceptibility source calculation from the field map. The inverse step from field map to susceptibility map is a 3-dimensional ill-posed deconvolution problem, which can be solved with 3 kinds of approaches: the Tikhonov-regularized matrix inverse, inverse filtering with a truncated filter, and total variation (TV) iteration. By numerical simulation, we validate the CIMRI model by comparing the reconstructed susceptibility maps for a predefined susceptibility source. Numerical simulations of CIMRI show that the split Bregman TV iteration solver can reconstruct the susceptibility map from an MR-phase image with high fidelity (spatial correlation ≈ 0.99). The split Bregman TV iteration solver includes noise reduction, edge preservation, and image energy conservation. For applications to brain susceptibility reconstruction, it is important to calibrate the TV iteration program by selecting suitable values of the regularization parameter. The proposed CIMRI model can reconstruct the magnetic susceptibility source of T2*MRI by 2 computational steps: calculating the field map from the phase image and reconstructing the susceptibility map from the field map. The crux of CIMRI lies in an ill-posed 3-dimensional deconvolution problem, which can be effectively solved by the split Bregman TV iteration algorithm.

Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks

DOE PAGES

Que, Emily L.; Bleher, Reiner; Duncan, Francesca E.; ...

2014-12-15

Fertilization of a mammalian egg induces a series of ‘zinc sparks’ that are necessary for inducing the egg-to-embryo transition. Despite the importance of these zinc efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches to resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy dispersive spectroscopy, X-ray fluorescence microscopy, and 3D elemental tomography for high resolution elemental mapping. Here we show that the zinc spark arises from a system of thousands ofmore » zinc-loaded vesicles, each of which contains, on average, 106 zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. We conclude that the discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes.« less

Distribution of trace levels of therapeutic gallium in bone as mapped by synchrotron x-ray microscopy.

PubMed Central

Bockman, R S; Repo, M A; Warrell, R P; Pounds, J G; Schidlovsky, G; Gordon, B M; Jones, K W

1990-01-01

Gallium nitrate, a drug that inhibits calcium release from bone, has been proven a safe and effective treatment for the accelerated bone resorption associated with cancer. Though bone is a target organ for gallium, the kinetics, sites, and effects of gallium accumulation in bone are not known. We have used synchrotron x-ray microscopy to map the distribution of trace levels of gallium in bone. After short-term in vivo administration of gallium nitrate to rats, trace (nanogram) amounts of gallium preferentially localized to the metabolically active regions in the metaphysis as well as the endosteal and periosteal surfaces of diaphyseal bone, regions where new bone formation and modeling were occurring. The amounts measured were well below the levels known to be cytotoxic. Iron and zinc, trace elements normally found in bone, were decreased in amount after in vivo administration of gallium. These studies represent a first step toward understanding the mechanism(s) of action of gallium in bone by suggesting the possible cellular, structural, and elemental "targets" of gallium. Images PMID:2349224

Impaired hippocampal place cell dynamics in a mouse model of the 22q11.2 deletion

PubMed Central

Zaremba, Jeffrey D; Diamantopoulou, Anastasia; Danielson, Nathan B; Grosmark, Andres D; Kaifosh, Patrick W; Bowler, John C; Liao, Zhenrui; Sparks, Fraser T; Gogos, Joseph A; Losonczy, Attila

2018-01-01

Hippocampal place cells represent the cellular substrate of episodic memory. Place cell ensembles reorganize to support learning but must also maintain stable representations to facilitate memory recall. Despite extensive research, the learning-related role of place cell dynamics in health and disease remains elusive. Using chronic two-photon Ca2+ imaging in hippocampal area CA1 of wild-type and Df(16)A+/− mice, an animal model of 22q11.2 deletion syndrome, one of the most common genetic risk factors for cognitive dysfunction and schizophrenia, we found that goal-oriented learning in wild-type mice was supported by stable spatial maps and robust remapping of place fields toward the goal location. Df(16)A+/− mice showed a significant learning deficit accompanied by reduced spatial map stability and the absence of goal-directed place cell reorganization. These results expand our understanding of the hippocampal ensemble dynamics supporting cognitive flexibility and demonstrate their importance in a model of 22q11.2-associated cognitive dysfunction. PMID:28869582

Quantitative fluorescence loss in photobleaching for analysis of protein transport and aggregation

PubMed Central

2012-01-01

Background Fluorescence loss in photobleaching (FLIP) is a widely used imaging technique, which provides information about protein dynamics in various cellular regions. In FLIP, a small cellular region is repeatedly illuminated by an intense laser pulse, while images are taken with reduced laser power with a time lag between the bleaches. Despite its popularity, tools are lacking for quantitative analysis of FLIP experiments. Typically, the user defines regions of interest (ROIs) for further analysis which is subjective and does not allow for comparing different cells and experimental settings. Results We present two complementary methods to detect and quantify protein transport and aggregation in living cells from FLIP image series. In the first approach, a stretched exponential (StrExp) function is fitted to fluorescence loss (FL) inside and outside the bleached region. We show by reaction–diffusion simulations, that the StrExp function can describe both, binding/barrier–limited and diffusion-limited FL kinetics. By pixel-wise regression of that function to FL kinetics of enhanced green fluorescent protein (eGFP), we determined in a user-unbiased manner from which cellular regions eGFP can be replenished in the bleached area. Spatial variation in the parameters calculated from the StrExp function allow for detecting diffusion barriers for eGFP in the nucleus and cytoplasm of living cells. Polyglutamine (polyQ) disease proteins like mutant huntingtin (mtHtt) can form large aggregates called inclusion bodies (IB’s). The second method combines single particle tracking with multi-compartment modelling of FL kinetics in moving IB’s to determine exchange rates of eGFP-tagged mtHtt protein (eGFP-mtHtt) between aggregates and the cytoplasm. This method is self-calibrating since it relates the FL inside and outside the bleached regions. It makes it therefore possible to compare release kinetics of eGFP-mtHtt between different cells and experiments. Conclusions We present two complementary methods for quantitative analysis of FLIP experiments in living cells. They provide spatial maps of exchange dynamics and absolute binding parameters of fluorescent molecules to moving intracellular entities, respectively. Our methods should be of great value for quantitative studies of intracellular transport. PMID:23148417

Factors associated with success of image-guided tumour biopsies: Results from a prospective molecular triage study (MOSCATO-01).

PubMed

Tacher, Vania; Le Deley, Marie-Cécile; Hollebecque, Antoine; Deschamps, Frederic; Vielh, Philippe; Hakime, Antoine; Ileana, Ecaterina; Abedi-Ardekani, Behnoush; Charpy, Cécile; Massard, Christophe; Rosellini, Silvia; Gajda, Dorota; Celebic, Aljosa; Ferté, Charles; Ngo-Camus, Maud; Gouissem, Siham; Koubi-Pick, Valérie; Andre, Fabrice; Vassal, Gilles; Deandreis, Désirée; Lacroix, Ludovic; Soria, Jean-Charles; De Baère, Thierry

2016-05-01

MOSCATO-01 is a molecular triage trial based on on-purpose tumour biopsies to perform molecular portraits. We aimed at identifying factors associated with high tumour cellularity. Tumour cellularity (percentage of tumour cells in samples defined at pathology) was evaluated according to patient characteristics, target lesion characteristics, operators' experience and biopsy approach. Among 460 patients enrolled between November, 2011 and March, 2014, 334 patients (73%) had an image-guided needle biopsy of the primary tumour (N = 38) or a metastatic lesion (N = 296). Biopsies were performed on liver (N = 127), lung (N = 72), lymph nodes (N = 71), bone (N = 11), or another tumour site (N = 53). Eighteen patients (5%) experienced a complication: pneumothorax in 10 patients treated medically, and haemorrhage in 8, requiring embolisation in 3 cases. Median tumour cellularity was 50% (interquartile range, 30-70%). The molecular analysis was successful in 291/334 cases (87%). On-going chemotherapy, tumour origin (primary versus metastatic), lesion size, tumour growth rate, presence of necrosis on imaging, standardised uptake value, and needle size were not statistically associated with cellularity. Compared to liver or lung biopsies, cellularity was significantly lower in bone and higher in other sites (P < 0.0001). Cellularity significantly increased with the number of collected samples (P < 0.0001) and was higher in contrast-enhanced ultrasound-guided biopsies (P < 0.02). In paired samples, cellularity in central samples was lower than in peripheral samples in 85, equal in 68 and higher in 89 of the cases. Image-guided biopsy is feasible and safe in cancer patients for molecular screening. Imaging modality, multiple sampling of the lesion, and the organ chosen for biopsy were associated with higher tumour cellularity. Copyright © 2016 Elsevier Ltd. All rights reserved.

Voyager Cartography

NASA Technical Reports Server (NTRS)

Batson, R. M.; Bridges, P. M.; Mullins, K. F.

1985-01-01

The Jovian and Saturnian satellites are being mapped at several scales from Voyager 1 and 2 data. The maps include specially formatted color mosaics, controlled photomosaics, and airbrush maps. More than 500 Voyager images of the Jovian and Saturnian satellites were radiometrically processed in preparation for cartographic processing. Of these images, 235 were geometrically transformed to map projections for base mosaic compilations. Special techniques for producing hybrid photomosaic/airbrush maps of Callisto are under investigation. The techniques involve making controlled computer mosaics of all available images with highest resolution images superimposed on lowest resolution images. The mosaics are then improved by airbrushing: seams and artifacts are removed, and image details enhanced that had been lost by saturation in some images. A controlled mosaic of the northern hemisphere of Rhea is complete, as is all processing for a similar mosaic of the equatorial region. Current plans and status of the various series are shown in a table.

Infrared and visible image fusion method based on saliency detection in sparse domain

NASA Astrophysics Data System (ADS)

Liu, C. H.; Qi, Y.; Ding, W. R.

2017-06-01

Infrared and visible image fusion is a key problem in the field of multi-sensor image fusion. To better preserve the significant information of the infrared and visible images in the final fused image, the saliency maps of the source images is introduced into the fusion procedure. Firstly, under the framework of the joint sparse representation (JSR) model, the global and local saliency maps of the source images are obtained based on sparse coefficients. Then, a saliency detection model is proposed, which combines the global and local saliency maps to generate an integrated saliency map. Finally, a weighted fusion algorithm based on the integrated saliency map is developed to achieve the fusion progress. The experimental results show that our method is superior to the state-of-the-art methods in terms of several universal quality evaluation indexes, as well as in the visual quality.

Advances in high-resolution imaging--techniques for three-dimensional imaging of cellular structures.

PubMed

Lidke, Diane S; Lidke, Keith A

2012-06-01

A fundamental goal in biology is to determine how cellular organization is coupled to function. To achieve this goal, a better understanding of organelle composition and structure is needed. Although visualization of cellular organelles using fluorescence or electron microscopy (EM) has become a common tool for the cell biologist, recent advances are providing a clearer picture of the cell than ever before. In particular, advanced light-microscopy techniques are achieving resolutions below the diffraction limit and EM tomography provides high-resolution three-dimensional (3D) images of cellular structures. The ability to perform both fluorescence and electron microscopy on the same sample (correlative light and electron microscopy, CLEM) makes it possible to identify where a fluorescently labeled protein is located with respect to organelle structures visualized by EM. Here, we review the current state of the art in 3D biological imaging techniques with a focus on recent advances in electron microscopy and fluorescence super-resolution techniques.

High-Throughput Method for Automated Colony and Cell Counting by Digital Image Analysis Based on Edge Detection

PubMed Central

Choudhry, Priya

2016-01-01

Counting cells and colonies is an integral part of high-throughput screens and quantitative cellular assays. Due to its subjective and time-intensive nature, manual counting has hindered the adoption of cellular assays such as tumor spheroid formation in high-throughput screens. The objective of this study was to develop an automated method for quick and reliable counting of cells and colonies from digital images. For this purpose, I developed an ImageJ macro Cell Colony Edge and a CellProfiler Pipeline Cell Colony Counting, and compared them to other open-source digital methods and manual counts. The ImageJ macro Cell Colony Edge is valuable in counting cells and colonies, and measuring their area, volume, morphology, and intensity. In this study, I demonstrate that Cell Colony Edge is superior to other open-source methods, in speed, accuracy and applicability to diverse cellular assays. It can fulfill the need to automate colony/cell counting in high-throughput screens, colony forming assays, and cellular assays. PMID:26848849

3D visualization of subcellular structures of Schizosaccharomyces pombe by hard X-ray tomography.

PubMed

Yang, Y; Li, W; Liu, G; Zhang, X; Chen, J; Wu, W; Guan, Y; Xiong, Y; Tian, Y; Wu, Z

2010-10-01

Cellular structures of the fission yeast, Schizosaccharomyces pombe, were examined by using hard X-ray tomography. Since cells are nearly transparent to hard X-rays, Zernike phase contrast and heavy metal staining were introduced to improve image contrast. Through using such methods, images taken at 8 keV displayed sufficient contrast for observing cellular structures. The cell wall, the intracellular organelles and the entire structural organization of the whole cells were visualized in three-dimensional at a resolution better than 100 nm. Comparison between phase contrast and absorption contrast was also made, indicating the obvious advantage of phase contrast for cellular imaging at this energy. Our results demonstrate that hard X-ray tomography with Zernike phase contrast is suitable for cellular imaging. Its unique abilities make it have potential to become a useful tool for revealing structural information from cells, especially thick eukaryotic cells. © 2010 The Authors Journal compilation © 2010 The Royal Microscopical Society.

Theranostic Gold Nanoantennas for Simultaneous Multiplexed Raman Imaging of Immunomarkers and Photothermal Therapy.

PubMed

Webb, Joseph A; Ou, Yu-Chuan; Faley, Shannon; Paul, Eden P; Hittinger, Joseph P; Cutright, Camden C; Lin, Eugene C; Bellan, Leon M; Bardhan, Rizia

2017-07-31

In this study, we demonstrate the theranostic capability of actively targeted, site-specific multibranched gold nanoantennas (MGNs) in triple-negative breast cancer (TNBC) cells in vitro. By utilizing multiplexed surface-enhanced Raman scattering (SERS) imaging, enabled by the narrow peak widths of Raman signatures, we simultaneously targeted immune checkpoint receptor programmed death ligand 1 (PDL1) and the epidermal growth factor receptor (EGFR) overexpressed in TNBC cells. A 1:1 mixture of MGNs functionalized with anti-PDL1 antibodies and Raman tag 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB) and MGNs functionalized with anti-EGFR antibodies and Raman tag para -mercaptobenzoic acid ( p MBA) were incubated with the cells. SERS imaging revealed a cellular traffic map of MGN localization by surface binding and receptor-mediated endocytosis, enabling targeted diagnosis of both biomarkers. Furthermore, cells incubated with anti-EGFR- p MBA-MGNs and illuminated with an 808 nm laser for 15 min at 4.7 W/cm 2 exhibited photothermal cell death only within the laser spot (indicated by live/dead cell fluorescence assay). Therefore, this study not only provides an optical imaging platform that can track immunomarkers with spatiotemporal control but also demonstrates an externally controlled light-triggered therapeutic approach enabling receptor-specific treatment with biocompatible theranostic nanoprobes.

Theranostic Gold Nanoantennas for Simultaneous Multiplexed Raman Imaging of Immunomarkers and Photothermal Therapy

PubMed Central

2017-01-01

In this study, we demonstrate the theranostic capability of actively targeted, site-specific multibranched gold nanoantennas (MGNs) in triple-negative breast cancer (TNBC) cells in vitro. By utilizing multiplexed surface-enhanced Raman scattering (SERS) imaging, enabled by the narrow peak widths of Raman signatures, we simultaneously targeted immune checkpoint receptor programmed death ligand 1 (PDL1) and the epidermal growth factor receptor (EGFR) overexpressed in TNBC cells. A 1:1 mixture of MGNs functionalized with anti-PDL1 antibodies and Raman tag 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB) and MGNs functionalized with anti-EGFR antibodies and Raman tag para-mercaptobenzoic acid (pMBA) were incubated with the cells. SERS imaging revealed a cellular traffic map of MGN localization by surface binding and receptor-mediated endocytosis, enabling targeted diagnosis of both biomarkers. Furthermore, cells incubated with anti-EGFR–pMBA–MGNs and illuminated with an 808 nm laser for 15 min at 4.7 W/cm2 exhibited photothermal cell death only within the laser spot (indicated by live/dead cell fluorescence assay). Therefore, this study not only provides an optical imaging platform that can track immunomarkers with spatiotemporal control but also demonstrates an externally controlled light-triggered therapeutic approach enabling receptor-specific treatment with biocompatible theranostic nanoprobes. PMID:28782050

Five Micron High Resolution MALDI Mass Spectrometry Imaging with Simple, Interchangeable, Multi-Resolution Optical System

DOE PAGES

Feenstra, Adam D.; Dueñas, Maria Emilia; Lee, Young Jin

2017-01-03

High-spatial resolution mass spectrometry imaging (MSI) is crucial for the mapping of chemical distributions at the cellular and subcellular level. Here in this work, we improved our previous laser optical system for matrix-assisted laser desorption ionization (MALDI)-MSI, from ~9 μm practical laser spot size to a practical laser spot size of ~4 μm, thereby allowing for 5 μm resolution imaging without oversampling. This is accomplished through a combination of spatial filtering, beam expansion, and reduction of the final focal length. Most importantly, the new laser optics system allows for simple modification of the spot size solely through the interchanging ofmore » the beam expander component. Using 10×, 5×, and no beam expander, we could routinely change between ~4, ~7, and ~45 μm laser spot size, in less than 5 min. We applied this multi-resolution MALDI-MSI system to a single maize root tissue section with three different spatial resolutions of 5, 10, and 50 μm and compared the differences in imaging quality and signal sensitivity. Lastly, we also demonstrated the difference in depth of focus between the optical systems with 10× and 5× beam expanders.« less

Five Micron High Resolution MALDI Mass Spectrometry Imaging with Simple, Interchangeable, Multi-Resolution Optical System

DOE Office of Scientific and Technical Information (OSTI.GOV)

Feenstra, Adam D.; Dueñas, Maria Emilia; Lee, Young Jin

High-spatial resolution mass spectrometry imaging (MSI) is crucial for the mapping of chemical distributions at the cellular and subcellular level. Here in this work, we improved our previous laser optical system for matrix-assisted laser desorption ionization (MALDI)-MSI, from ~9 μm practical laser spot size to a practical laser spot size of ~4 μm, thereby allowing for 5 μm resolution imaging without oversampling. This is accomplished through a combination of spatial filtering, beam expansion, and reduction of the final focal length. Most importantly, the new laser optics system allows for simple modification of the spot size solely through the interchanging ofmore » the beam expander component. Using 10×, 5×, and no beam expander, we could routinely change between ~4, ~7, and ~45 μm laser spot size, in less than 5 min. We applied this multi-resolution MALDI-MSI system to a single maize root tissue section with three different spatial resolutions of 5, 10, and 50 μm and compared the differences in imaging quality and signal sensitivity. Lastly, we also demonstrated the difference in depth of focus between the optical systems with 10× and 5× beam expanders.« less

Analysis of the changes in the tarcrete layer on the desert surface of Kuwait using satellite imagery and cell-based modeling

NASA Astrophysics Data System (ADS)

Al-Doasari, Ahmad E.

The 1991 Gulf War caused massive environmental damage in Kuwait. Deposition of oil and soot droplets from hundreds of burning oil-wells created a layer of tarcrete on the desert surface covering over 900 km2. This research investigates the spatial change in the tarcrete extent from 1991 to 1998 using Landsat Thematic Mapper (TM) imagery and statistical modeling techniques. The pixel structure of TM data allows the spatial analysis of the change in tarcrete extent to be conducted at the pixel (cell) level within a geographical information system (GIS). There are two components to this research. The first is a comparison of three remote sensing classification techniques used to map the tarcrete layer. The second is a spatial-temporal analysis and simulation of tarcrete changes through time. The analysis focuses on an area of 389 km2 located south of the Al-Burgan oil field. Five TM images acquired in 1991, 1993, 1994, 1995, and 1998 were geometrically and atmospherically corrected. These images were classified into six classes: oil lakes; heavy, intermediate, light, and traces of tarcrete; and sand. The classification methods tested were unsupervised, supervised, and neural network supervised (fuzzy ARTMAP). Field data of tarcrete characteristics were collected to support the classification process and to evaluate the classification accuracies. Overall, the neural network method is more accurate (60 percent) than the other two methods; both the unsupervised and the supervised classification accuracy assessments resulted in 46 percent accuracy. The five classifications were used in a lagged autologistic model to analyze the spatial changes of the tarcrete through time. The autologistic model correctly identified overall tarcrete contraction between 1991--1993 and 1995--1998. However, tarcrete contraction between 1993--1994 and 1994--1995 was less well marked, in part because of classification errors in the maps from these time periods. Initial simulations of tarcrete contraction with a cellular automaton model were not very successful. However, more accurate classifications could improve the simulations. This study illustrates how an empirical investigation using satellite images, field data, GIS, and spatial statistics can simulate dynamic land-cover change through the use of a discrete statistical and cellular automaton model.

Histogram Analysis of Diffusion Weighted Imaging at 3T is Useful for Prediction of Lymphatic Metastatic Spread, Proliferative Activity, and Cellularity in Thyroid Cancer.

PubMed

Schob, Stefan; Meyer, Hans Jonas; Dieckow, Julia; Pervinder, Bhogal; Pazaitis, Nikolaos; Höhn, Anne Kathrin; Garnov, Nikita; Horvath-Rizea, Diana; Hoffmann, Karl-Titus; Surov, Alexey

2017-04-12

Pre-surgical diffusion weighted imaging (DWI) is increasingly important in the context of thyroid cancer for identification of the optimal treatment strategy. It has exemplarily been shown that DWI at 3T can distinguish undifferentiated from well-differentiated thyroid carcinoma, which has decisive implications for the magnitude of surgery. This study used DWI histogram analysis of whole tumor apparent diffusion coefficient (ADC) maps. The primary aim was to discriminate thyroid carcinomas which had already gained the capacity to metastasize lymphatically from those not yet being able to spread via the lymphatic system. The secondary aim was to reflect prognostically important tumor-biological features like cellularity and proliferative activity with ADC histogram analysis. Fifteen patients with follicular-cell derived thyroid cancer were enrolled. Lymph node status, extent of infiltration of surrounding tissue, and Ki-67 and p53 expression were assessed in these patients. DWI was obtained in a 3T system using b values of 0, 400, and 800 s/mm². Whole tumor ADC volumes were analyzed using a histogram-based approach. Several ADC parameters showed significant correlations with immunohistopathological parameters. Most importantly, ADC histogram skewness and ADC histogram kurtosis were able to differentiate between nodal negative and nodal positive thyroid carcinoma. histogram analysis of whole ADC tumor volumes has the potential to provide valuable information on tumor biology in thyroid carcinoma. However, further studies are warranted.

Histogram Analysis of Diffusion Weighted Imaging at 3T is Useful for Prediction of Lymphatic Metastatic Spread, Proliferative Activity, and Cellularity in Thyroid Cancer

PubMed Central

Schob, Stefan; Meyer, Hans Jonas; Dieckow, Julia; Pervinder, Bhogal; Pazaitis, Nikolaos; Höhn, Anne Kathrin; Garnov, Nikita; Horvath-Rizea, Diana; Hoffmann, Karl-Titus; Surov, Alexey

2017-01-01

Pre-surgical diffusion weighted imaging (DWI) is increasingly important in the context of thyroid cancer for identification of the optimal treatment strategy. It has exemplarily been shown that DWI at 3T can distinguish undifferentiated from well-differentiated thyroid carcinoma, which has decisive implications for the magnitude of surgery. This study used DWI histogram analysis of whole tumor apparent diffusion coefficient (ADC) maps. The primary aim was to discriminate thyroid carcinomas which had already gained the capacity to metastasize lymphatically from those not yet being able to spread via the lymphatic system. The secondary aim was to reflect prognostically important tumor-biological features like cellularity and proliferative activity with ADC histogram analysis. Fifteen patients with follicular-cell derived thyroid cancer were enrolled. Lymph node status, extent of infiltration of surrounding tissue, and Ki-67 and p53 expression were assessed in these patients. DWI was obtained in a 3T system using b values of 0, 400, and 800 s/mm2. Whole tumor ADC volumes were analyzed using a histogram-based approach. Several ADC parameters showed significant correlations with immunohistopathological parameters. Most importantly, ADC histogram skewness and ADC histogram kurtosis were able to differentiate between nodal negative and nodal positive thyroid carcinoma. Conclusions: histogram analysis of whole ADC tumor volumes has the potential to provide valuable information on tumor biology in thyroid carcinoma. However, further studies are warranted. PMID:28417929

Geologic map of Ophir and central Candor Chasmata (MTM -05072) of Mars

USGS Publications Warehouse

Lucchitta, Baerbel K.

1999-01-01

The geologic map of Ophir and central Candor Chasmata is one of a series of 1:500,000 scale maps prepared for areas on Mars that are of particular scientific interest and may serve as potential future landing sites. This map is also part of a set that includes east Candor Chasma, west Candor Chasma, and Melas Chasma. The geologic interpretations are based dominantly on medium- and high-resolution Viking images, many of them stereoscopic, and supplemented by lower resolution apoapsis and other color images. A strip of very high resolution stereoscopic images (~20 m/pixel) crosses the central part of the quadrangle from northwest to southeast and served to clarify detailed relations not obvious on other images. A topographic map with contour intervals of 200 m was also used, as were multidirectional oblique images derived from merged image mosaics and topography (see fig. 1) (Bertolini and McEwen, 1990). Geologic relations and interpretations are based on the entire central Valles Marineris map set. The map area is included in the Valles Marineris map of Witbeck and others (1991), but units were defined independently. Age assignments, however, were integrated with those by Witbeck and others and Scott and Tanaka (1986).

Multispectral Landsat images of Antartica

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lucchitta, B.K.; Bowell, J.A.; Edwards, K.L.

1988-01-01

The U.S. Geological Survey has a program to map Antarctica by using colored, digitally enhanced Landsat multispectral scanner images to increase existing map coverage and to improve upon previously published Landsat maps. This report is a compilation of images and image mosaic that covers four complete and two partial 1:250,000-scale quadrangles of the McMurdo Sound region.

Saliency detection using mutual consistency-guided spatial cues combination

NASA Astrophysics Data System (ADS)

Wang, Xin; Ning, Chen; Xu, Lizhong

2015-09-01

Saliency detection has received extensive interests due to its remarkable contribution to wide computer vision and pattern recognition applications. However, most existing computational models are designed for detecting saliency in visible images or videos. When applied to infrared images, they may suffer from limitations in saliency detection accuracy and robustness. In this paper, we propose a novel algorithm to detect visual saliency in infrared images by mutual consistency-guided spatial cues combination. First, based on the luminance contrast and contour characteristics of infrared images, two effective saliency maps, i.e., the luminance contrast saliency map and contour saliency map are constructed, respectively. Afterwards, an adaptive combination scheme guided by mutual consistency is exploited to integrate these two maps to generate the spatial saliency map. This idea is motivated by the observation that different maps are actually related to each other and the fusion scheme should present a logically consistent view of these maps. Finally, an enhancement technique is adopted to incorporate spatial saliency maps at various scales into a unified multi-scale framework to improve the reliability of the final saliency map. Comprehensive evaluations on real-life infrared images and comparisons with many state-of-the-art saliency models demonstrate the effectiveness and superiority of the proposed method for saliency detection in infrared images.

Lineage mapping and characterization of the native progenitor population in cellular allograft.

PubMed

Neman, Josh; Duenas, Vincent; Kowolik, Claudia; Hambrecht, Amanda; Chen, Mike; Jandial, Rahul

2013-02-01

The gold standard for bone grafting remains the autograft. However, the attractiveness of autograft is counterbalanced by donor site morbidity. To mimic autograft-and its fundamental properties of osteoconductivity, osteoinductivity, and osteogenicity-novel bone grafting materials such as cellular allograft (Osteocel Plus) are composed of allograft in which the progenitor cells are preserved. However, the true identity of these cells remains obscure largely due to the lack of specific bona fide antigenic markers for stem versus progenitor cells. To characterize the stem and progenitor population in cellular allograft, Osteocel Plus. To determine whether cells endogenous to a cellular allograft undergo extensive self-renewal (a functional hallmark of stem cells), we employed a novel use of lineage mapping using a modern and refined replication incompetent lentiviral library with high complexity to uniquely label single cells with indelible genetic tags faithfully passed on to all progeny, allowing identification of highly proliferative clones. We used genetic and proteomic profiling as well as functional assays to show that these cells are capable of multipotential differentiation (the second functional hallmark of stem cells). Use of these two functional hallmarks enabled us to establish the existence of a stem and progenitor cell population in cellular allografts. Specifically, we employed (1) cellular dissociation and (2) in vitro expansion and differentiation capacity of cells released from cellular allograft. We determined differential gene expression profiling of a bona fide human mesenchymal stem cell line and cells from cellular allograft using focused PCR arrays mesenchymal stem cell (MSC) and osteogenesis associated. Proteomic profiling of cells from cellular allograft was performed using (1) immunofluorescence for BMP-2, Runx2 SMADs, CD44, Stro-1, Collagen, RANKL, Osterix Osteocalcin, and Ki67; (2) flow cytometry for Ki67, CD44, Stro-1, Thy1, CD146, and Osteocalcin; and (3) enzyme-linked immunosorbent assays (ELISA) for BMP-2, Osteocalcin, RANKL, Osteoprotegrin, and Osteocalcin. Clonal analysis of cells from cellular allograft was performed utilizing advance lentivirus lineage mapping techniques and massive parallel sequencing. Alizarin Red, Alcian Blue, and Oil red O staining assessed tripotential differentiation capacity. Serial trypsinization of allograft cellular bone matrix yielded approximately 1×105 cells per mL with viability greater than 90%. Cells expressed a panel of 84 MSC-associated genes in a pattern similar to but not identical to pure MSCs; specifically, 59 of 84 genes showed less than a 2.5-fold change in both cell types. Protein analysis showed that cellular allograft -derived cells maintained in nondifferentiation media expressed the early osteo-progenitor markers BMP-2, SMADs, and Runx2. Corresponding flow cytometry data for MSC markers revealed the presence of Stro-1 (49%), CD44 (99%), CD90 (42%), and CD146 (97%). Lineage mapping indicated that 62% of clones persisted and generated progeny through 10 passages, strongly suggesting the presence of bona fide stem cells. Passage 10 clones also exhibited tri-lineage differentiation capacity into osteogenic (Alizarin Red with H&E counterstain), chondrogenic (Alcian Blue), and adipogenic (Oil red O). Cells that did not proliferate through 10 passages presumably differentiated along an osteo-progenitor lineage. These data indicate that cellular allograft (Osteocel Plus) contains a heterogeneous population of cells with most cells demonstrating the capacity for extensive self-renewal and multipotential differentiation, which are hallmarks of stem cells. Whether stem cell-enriched allografts function comparably to autograft will require further studies, and their efficacy in facilitating arthrodesis will depend on randomized clinical studies. Copyright © 2013 Elsevier Inc. All rights reserved.

High-throughput microscopy must re-invent the microscope rather than speed up its functions

PubMed Central

Oheim, M

2007-01-01

Knowledge gained from the revolutions in genomics and proteomics has helped to identify many of the key molecules involved in cellular signalling. Researchers, both in academia and in the pharmaceutical industry, now screen, at a sub-cellular level, where and when these proteins interact. Fluorescence imaging and molecular labelling combine to provide a powerful tool for real-time functional biochemistry with molecular resolution. However, they traditionally have been work-intensive, required trained personnel, and suffered from low through-put due to sample preparation, loading and handling. The need for speeding up microscopy is apparent from the tremendous complexity of cellular signalling pathways, the inherent biological variability, as well as the possibility that the same molecule plays different roles in different sub-cellular compartments. Research institutes and companies have teamed up to develop imaging cytometers of ever-increasing complexity. However, to truly go high-speed, sub-cellular imaging must free itself from the rigid framework of current microscopes. PMID:17603553

EnGeoMAP - geological applications within the EnMAP hyperspectral satellite science program

NASA Astrophysics Data System (ADS)

Boesche, N. K.; Mielke, C.; Rogass, C.; Guanter, L.

2016-12-01

Hyperspectral investigations from near field to space substantially contribute to geological exploration and mining monitoring of raw material and mineral deposits. Due to their spectral characteristics, large mineral occurrences and minefields can be identified from space and the spatial distribution of distinct proxy minerals be mapped. In the frame of the EnMAP hyperspectral satellite science program a mineral and elemental mapping tool was developed - the EnGeoMAP. It contains a basic mineral mapping and a rare earth element mapping approach. This study shows the performance of EnGeoMAP based on simulated EnMAP data of the rare earth element bearing Mountain Pass Carbonatite Complex, USA, and the Rodalquilar and Lomilla Calderas, Spain, which host the economically relevant gold-silver, lead-zinc-silver-gold and alunite deposits. The mountain pass image data was simulated on the basis of AVIRIS Next Generation images, while the Rodalquilar data is based on HyMap images. The EnGeoMAP - Base approach was applied to both images, while the mountain pass image data were additionally analysed using the EnGeoMAP - REE software tool. The results are mineral and elemental maps that serve as proxies for the regional lithology and deposit types. The validation of the maps is based on chemical analyses of field samples. Current airborne sensors meet the spatial and spectral requirements for detailed mineral mapping and future hyperspectral space borne missions will additionally provide a large coverage. For those hyperspectral missions, EnGeoMAP is a rapid data analysis tool that is provided to spectral geologists working in mineral exploration.

Psoriasis skin biopsy image segmentation using Deep Convolutional Neural Network.

PubMed

Pal, Anabik; Garain, Utpal; Chandra, Aditi; Chatterjee, Raghunath; Senapati, Swapan

2018-06-01

Development of machine assisted tools for automatic analysis of psoriasis skin biopsy image plays an important role in clinical assistance. Development of automatic approach for accurate segmentation of psoriasis skin biopsy image is the initial prerequisite for developing such system. However, the complex cellular structure, presence of imaging artifacts, uneven staining variation make the task challenging. This paper presents a pioneering attempt for automatic segmentation of psoriasis skin biopsy images. Several deep neural architectures are tried for segmenting psoriasis skin biopsy images. Deep models are used for classifying the super-pixels generated by Simple Linear Iterative Clustering (SLIC) and the segmentation performance of these architectures is compared with the traditional hand-crafted feature based classifiers built on popularly used classifiers like K-Nearest Neighbor (KNN), Support Vector Machine (SVM) and Random Forest (RF). A U-shaped Fully Convolutional Neural Network (FCN) is also used in an end to end learning fashion where input is the original color image and the output is the segmentation class map for the skin layers. An annotated real psoriasis skin biopsy image data set of ninety (90) images is developed and used for this research. The segmentation performance is evaluated with two metrics namely, Jaccard's Coefficient (JC) and the Ratio of Correct Pixel Classification (RCPC) accuracy. The experimental results show that the CNN based approaches outperform the traditional hand-crafted feature based classification approaches. The present research shows that practical system can be developed for machine assisted analysis of psoriasis disease. Copyright © 2018 Elsevier B.V. All rights reserved.

Making a georeferenced mosaic of historical map series using constrained polynomial fit

NASA Astrophysics Data System (ADS)

Molnár, G.

2009-04-01

Present day GIS software packages make it possible to handle several hundreds of rasterised map sheets. For proper usage of such datasets we usually have two requirements: First these map sheets should be georeferenced, secondly these georeferenced maps should fit properly together, without overlap and short. Both requirements can be fulfilled easily, if the geodetic background for the map series is accurate, and the projection of the map series is known. In this case the individual map sheets should be georeferenced in the projected coordinate system of the map series. This means every individual map sheets are georeferenced using overprinted coordinate grid or image corner projected coordinates as ground control points (GCPs). If after this georeferencing procedure the map sheets do not fit together (for example because of using different projection for every map sheet, as it is in the case of Third Military Survey) a common projection can be chosen, and all the georeferenced maps should be transformed to this common projection using a map-to-map transformation. If the geodetic background is not so strong, ie. there are distortions inside the map sheets, a polynomial (linear quadratic or cubic) polynomial fit can be used for georeferencing the map sheets. Finding identical surface objects (as GCPs) on the historical map and on a present day cartographic map, let us to determine a transformation between raw image coordinates (x,y) and the projected coordinates (Easting, Northing, E,N). This means, for all the map sheets, several GCPs should be found, (for linear, quadratic of cubic transformations at least 3, 5 or 10 respectively) and every map sheets should be transformed to a present day coordinate system individually using these GCPs. The disadvantage of this method is that, after the transformation, the individual transformed map sheets not necessarily fit together properly any more. To overcome this problem neither the reverse order of procedure helps: if we make the mosaic first (eg. graphically) and we try the polynomial fit of this mosaic afterwards, neither using this can we reduce the error of internal inaccuracy of the map-sheets. We can overcome this problem by calculating the transformation parameters of polynomial fit with constrains (Mikhail, 1976). The constrain is that the common edge of two neighboring map-sheets should be transformed identically, ie. the right edge of the left image and the left edge of the right image should fit together after the transformation. This condition should fulfill for all the internal (not only the vertical, but also for the horizontal) edges of the mosaic. Constrains are expressed as a relationship between parameters: The parameters of the polynomial transformation should fulfill not only the least squares adjustment criteria but also the constrain: the transformed coordinates should be identical on the image edges. (With the example mentioned above, for image points of the rightmost column of the left image the transformed coordinates should be the same a for the image points of the leftmost column of the right image, and these transformed coordinates can depend on the line number image coordinate of the raster point.) The normal equation system can be calculated with Lagrange-multipliers. The resulting set of parameters for all map-sheets should be applied on the transformation of the images. This parameter set can not been directly applied in GIS software for the transformation. The simplest solution applying this parameters is ‘simulating' GCPs for every image, and applying these simulated GCPs for the georeferencing of the individual map sheets. This method is applied on a set of map-sheets of the First military Survey of the Habsburg Empire with acceptable results. Reference: Mikhail, E. M.: Observations and Least Squares. IEP—A Dun-Donnelley Publisher, New York, 1976. 497 pp.

Spatial and temporal in vivo analysis of circulating and sessile immune cells in mosquitoes: hemocyte mitosis following infection

PubMed Central

2013-01-01

Background Mosquitoes respond to infection by mounting immune responses. The primary regulators of these immune responses are cells called hemocytes, which kill pathogens via phagocytosis and via the production of soluble antimicrobial factors. Mosquito hemocytes are circulated throughout the hemocoel (body cavity) by the swift flow of hemolymph (blood), and data show that some hemocytes also exist as sessile cells that are attached to tissues. The purpose of this study was to create a quantitative physical map of hemocyte distribution in the mosquito, Anopheles gambiae, and to describe the cellular immune response in an organismal context. Results Using correlative imaging methods we found that the number of hemocytes in a mosquito decreases with age, but that regardless of age, approximately 75% of the hemocytes occur in circulation and 25% occur as sessile cells. Infection induces an increase in the number of hemocytes, and tubulin and nuclear staining showed that this increase is primarily due to mitosis and, more specifically, autonomous cell division, by circulating granulocytes. The majority of sessile hemocytes are present on the abdominal wall, although significant numbers of hemocytes are also present in the thorax, head, and several of the appendages. Within the abdominal wall, the areas of highest hemocyte density are the periostial regions (regions surrounding the valves of the heart, or ostia), which are ideal locations for pathogen capture as these are areas of high hemolymph flow. Conclusions These data describe the spatial and temporal distribution of mosquito hemocytes, and map the cellular response to infection throughout the hemocoel. PMID:23631603

Cell Motility Dynamics: A Novel Segmentation Algorithm to Quantify Multi-Cellular Bright Field Microscopy Images

PubMed Central

Zaritsky, Assaf; Natan, Sari; Horev, Judith; Hecht, Inbal; Wolf, Lior; Ben-Jacob, Eshel; Tsarfaty, Ilan

2011-01-01

Confocal microscopy analysis of fluorescence and morphology is becoming the standard tool in cell biology and molecular imaging. Accurate quantification algorithms are required to enhance the understanding of different biological phenomena. We present a novel approach based on image-segmentation of multi-cellular regions in bright field images demonstrating enhanced quantitative analyses and better understanding of cell motility. We present MultiCellSeg, a segmentation algorithm to separate between multi-cellular and background regions for bright field images, which is based on classification of local patches within an image: a cascade of Support Vector Machines (SVMs) is applied using basic image features. Post processing includes additional classification and graph-cut segmentation to reclassify erroneous regions and refine the segmentation. This approach leads to a parameter-free and robust algorithm. Comparison to an alternative algorithm on wound healing assay images demonstrates its superiority. The proposed approach was used to evaluate common cell migration models such as wound healing and scatter assay. It was applied to quantify the acceleration effect of Hepatocyte growth factor/scatter factor (HGF/SF) on healing rate in a time lapse confocal microscopy wound healing assay and demonstrated that the healing rate is linear in both treated and untreated cells, and that HGF/SF accelerates the healing rate by approximately two-fold. A novel fully automated, accurate, zero-parameters method to classify and score scatter-assay images was developed and demonstrated that multi-cellular texture is an excellent descriptor to measure HGF/SF-induced cell scattering. We show that exploitation of textural information from differential interference contrast (DIC) images on the multi-cellular level can prove beneficial for the analyses of wound healing and scatter assays. The proposed approach is generic and can be used alone or alongside traditional fluorescence single-cell processing to perform objective, accurate quantitative analyses for various biological applications. PMID:22096600

Cell motility dynamics: a novel segmentation algorithm to quantify multi-cellular bright field microscopy images.

PubMed

Zaritsky, Assaf; Natan, Sari; Horev, Judith; Hecht, Inbal; Wolf, Lior; Ben-Jacob, Eshel; Tsarfaty, Ilan

2011-01-01

Confocal microscopy analysis of fluorescence and morphology is becoming the standard tool in cell biology and molecular imaging. Accurate quantification algorithms are required to enhance the understanding of different biological phenomena. We present a novel approach based on image-segmentation of multi-cellular regions in bright field images demonstrating enhanced quantitative analyses and better understanding of cell motility. We present MultiCellSeg, a segmentation algorithm to separate between multi-cellular and background regions for bright field images, which is based on classification of local patches within an image: a cascade of Support Vector Machines (SVMs) is applied using basic image features. Post processing includes additional classification and graph-cut segmentation to reclassify erroneous regions and refine the segmentation. This approach leads to a parameter-free and robust algorithm. Comparison to an alternative algorithm on wound healing assay images demonstrates its superiority. The proposed approach was used to evaluate common cell migration models such as wound healing and scatter assay. It was applied to quantify the acceleration effect of Hepatocyte growth factor/scatter factor (HGF/SF) on healing rate in a time lapse confocal microscopy wound healing assay and demonstrated that the healing rate is linear in both treated and untreated cells, and that HGF/SF accelerates the healing rate by approximately two-fold. A novel fully automated, accurate, zero-parameters method to classify and score scatter-assay images was developed and demonstrated that multi-cellular texture is an excellent descriptor to measure HGF/SF-induced cell scattering. We show that exploitation of textural information from differential interference contrast (DIC) images on the multi-cellular level can prove beneficial for the analyses of wound healing and scatter assays. The proposed approach is generic and can be used alone or alongside traditional fluorescence single-cell processing to perform objective, accurate quantitative analyses for various biological applications.

Organ specific mapping of in vivo redox state in control and cigarette smoke-exposed mice using EPR/NMR co-imaging

PubMed Central

Caia, George L.; Efimova, Olga V.; Velayutham, Murugesan; El-Mahdy, Mohamed A.; Abdelghany, Tamer M.; Kesselring, Eric; Petryakov, Sergey; Sun, Ziqi; Samouilov, Alexandre; Zweier, Jay L.

2014-01-01

In vivo mapping of alterations in redox status is important for understanding organ specific pathology and disease. While electron paramagnetic resonance imaging (EPRI) enables spatial mapping of free radicals, it does not provide anatomic visualization of the body. Proton MRI is well suited to provide anatomical visualization. We applied EPR/NMR co-imaging instrumentation to map and monitor the redox state of living mice under normal or oxidative stress conditions induced by secondhand cigarette smoke (SHS) exposure. A hybrid co-imaging instrument, EPRI (1.2 GHz) / proton MRI (16.18 MHz), suitable for whole-body co-imaging of mice was utilized with common magnet and gradients along with dual EPR/NMR resonators that enable co-imaging without sample movement. The metabolism of the nitroxide probe, 3–carbamoyl–proxyl (3-CP), was used to map the redox state of control and SHS-exposed mice. Co-imaging allowed precise 3D mapping of radical distribution and reduction in major organs such as the heart, lungs, liver, bladder and kidneys. Reductive metabolism was markedly decreased in SHS-exposed mice and EPR/NMR co-imaging allowed quantitative assessment of this throughout the body. Thus, in vivo EPR/NMR co-imaging enables in vivo organ specific mapping of free radical metabolism and redox stress and the alterations that occur in the pathogenesis of disease. PMID:22296801

Organ specific mapping of in vivo redox state in control and cigarette smoke-exposed mice using EPR/NMR co-imaging

NASA Astrophysics Data System (ADS)

Caia, George L.; Efimova, Olga V.; Velayutham, Murugesan; El-Mahdy, Mohamed A.; Abdelghany, Tamer M.; Kesselring, Eric; Petryakov, Sergey; Sun, Ziqi; Samouilov, Alexandre; Zweier, Jay L.

2012-03-01

In vivo mapping of alterations in redox status is important for understanding organ specific pathology and disease. While electron paramagnetic resonance imaging (EPRI) enables spatial mapping of free radicals, it does not provide anatomic visualization of the body. Proton MRI is well suited to provide anatomical visualization. We applied EPR/NMR co-imaging instrumentation to map and monitor the redox state of living mice under normal or oxidative stress conditions induced by secondhand cigarette smoke (SHS) exposure. A hybrid co-imaging instrument, EPRI (1.2 GHz)/proton MRI (16.18 MHz), suitable for whole-body co-imaging of mice was utilized with common magnet and gradients along with dual EPR/NMR resonators that enable co-imaging without sample movement. The metabolism of the nitroxide probe, 3-carbamoyl-proxyl (3-CP), was used to map the redox state of control and SHS-exposed mice. Co-imaging allowed precise 3D mapping of radical distribution and reduction in major organs such as the heart, lungs, liver, bladder and kidneys. Reductive metabolism was markedly decreased in SHS-exposed mice and EPR/NMR co-imaging allowed quantitative assessment of this throughout the body. Thus, in vivo EPR/NMR co-imaging enables in vivo organ specific mapping of free radical metabolism and redox stress and the alterations that occur in the pathogenesis of disease.

A novel algorithm for thermal image encryption.

PubMed

Hussain, Iqtadar; Anees, Amir; Algarni, Abdulmohsen

2018-04-16

Thermal images play a vital character at nuclear plants, Power stations, Forensic labs biological research, and petroleum products extraction. Safety of thermal images is very important. Image data has some unique features such as intensity, contrast, homogeneity, entropy and correlation among pixels that is why somehow image encryption is trickier as compare to other encryptions. With conventional image encryption schemes it is normally hard to handle these features. Therefore, cryptographers have paid attention to some attractive properties of the chaotic maps such as randomness and sensitivity to build up novel cryptosystems. That is why, recently proposed image encryption techniques progressively more depends on the application of chaotic maps. This paper proposed an image encryption algorithm based on Chebyshev chaotic map and S8 Symmetric group of permutation based substitution boxes. Primarily, parameters of chaotic Chebyshev map are chosen as a secret key to mystify the primary image. Then, the plaintext image is encrypted by the method generated from the substitution boxes and Chebyshev map. By this process, we can get a cipher text image that is perfectly twisted and dispersed. The outcomes of renowned experiments, key sensitivity tests and statistical analysis confirm that the proposed algorithm offers a safe and efficient approach for real-time image encryption.

Live-Cell Imaging of Mitochondria and the Actin Cytoskeleton in Budding Yeast.

PubMed

Higuchi-Sanabria, Ryo; Swayne, Theresa C; Boldogh, Istvan R; Pon, Liza A

2016-01-01

Maintenance and regulation of proper mitochondrial dynamics and functions are necessary for cellular homeostasis. Numerous diseases, including neurodegeneration and muscle myopathies, and overall cellular aging are marked by declining mitochondrial function and subsequent loss of multiple other cellular functions. For these reasons, optimized protocols are needed for visualization and quantification of mitochondria and their function and fitness. In budding yeast, mitochondria are intimately associated with the actin cytoskeleton and utilize actin for their movement and inheritance. This chapter describes optimal approaches for labeling mitochondria and the actin cytoskeleton in living budding yeast cells, for imaging the labeled cells, and for analyzing the resulting images.

Multimodal Image Alignment via Linear Mapping between Feature Modalities.

PubMed

Jiang, Yanyun; Zheng, Yuanjie; Hou, Sujuan; Chang, Yuchou; Gee, James

2017-01-01

We propose a novel landmark matching based method for aligning multimodal images, which is accomplished uniquely by resolving a linear mapping between different feature modalities. This linear mapping results in a new measurement on similarity of images captured from different modalities. In addition, our method simultaneously solves this linear mapping and the landmark correspondences by minimizing a convex quadratic function. Our method can estimate complex image relationship between different modalities and nonlinear nonrigid spatial transformations even in the presence of heavy noise, as shown in our experiments carried out by using a variety of image modalities.

Supporting performance and configuration management of GTE cellular networks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tan, Ming; Lafond, C.; Jakobson, G.

GTE Laboratories, in cooperation with GTE Mobilnet, has developed and deployed PERFFEX (PERFormance Expert), an intelligent system for performance and configuration management of cellular networks. PERFEX assists cellular network performance and radio engineers in the analysis of large volumes of cellular network performance and configuration data. It helps them locate and determine the probable causes of performance problems, and provides intelligent suggestions about how to correct them. The system combines an expert cellular network performance tuning capability with a map-based graphical user interface, data visualization programs, and a set of special cellular engineering tools. PERFEX is in daily use atmore » more than 25 GTE Mobile Switching Centers. Since the first deployment of the system in late 1993, PERFEX has become a major GTE cellular network performance optimization tool.« less

Automated classification of immunostaining patterns in breast tissue from the human protein atlas.

PubMed

Swamidoss, Issac Niwas; Kårsnäs, Andreas; Uhlmann, Virginie; Ponnusamy, Palanisamy; Kampf, Caroline; Simonsson, Martin; Wählby, Carolina; Strand, Robin

2013-01-01

The Human Protein Atlas (HPA) is an effort to map the location of all human proteins (http://www.proteinatlas.org/). It contains a large number of histological images of sections from human tissue. Tissue micro arrays (TMA) are imaged by a slide scanning microscope, and each image represents a thin slice of a tissue core with a dark brown antibody specific stain and a blue counter stain. When generating antibodies for protein profiling of the human proteome, an important step in the quality control is to compare staining patterns of different antibodies directed towards the same protein. This comparison is an ultimate control that the antibody recognizes the right protein. In this paper, we propose and evaluate different approaches for classifying sub-cellular antibody staining patterns in breast tissue samples. The proposed methods include the computation of various features including gray level co-occurrence matrix (GLCM) features, complex wavelet co-occurrence matrix (CWCM) features, and weighted neighbor distance using compound hierarchy of algorithms representing morphology (WND-CHARM)-inspired features. The extracted features are used into two different multivariate classifiers (support vector machine (SVM) and linear discriminant analysis (LDA) classifier). Before extracting features, we use color deconvolution to separate different tissue components, such as the brownly stained positive regions and the blue cellular regions, in the immuno-stained TMA images of breast tissue. We present classification results based on combinations of feature measurements. The proposed complex wavelet features and the WND-CHARM features have accuracy similar to that of a human expert. Both human experts and the proposed automated methods have difficulties discriminating between nuclear and cytoplasmic staining patterns. This is to a large extent due to mixed staining of nucleus and cytoplasm. Methods for quantification of staining patterns in histopathology have many applications, ranging from antibody quality control to tumor grading.

Targeted functional imaging of estrogen receptors with 99mTc-GAP-EDL.

PubMed

Takahashi, Nobukazu; Yang, David J; Kohanim, Saady; Oh, Chang-Sok; Yu, Dong-Fang; Azhdarinia, Ali; Kurihara, Hiroaki; Zhang, Xiaochun; Chang, Joe Y; Kim, E Edmund

2007-03-01

To evaluate the feasibility of using (99m)Tc-glutamate peptide-estradiol in functional imaging of estrogen receptor-positive [ER(+)] diseases. 3-Aminoethyl estradiol (EDL) was conjugated to glutamate peptide (GAP) to yield GAP-EDL. Cellular uptake studies of (99m)Tc-GAP-EDL were conducted in ER(+) cell lines (MCF-7, 13762 and T47D). To demonstrate whether GAP-EDL increases MAP kinase activation, Western blot analysis of GAP-EDL was performed in 13762 cells. Biodistribution was conducted in nine rats with 13762 breast tumors at 0.5-4 h. Each rat was administered (99m)Tc-GAP-EDL. Two animal models (rats and rabbits) were created to ascertain whether tumor uptake of (99m)Tc-GAP-EDL was via an ER-mediated process. In the tumor model, breast tumor-bearing rats were pretreated with diethylstilbestrol (DES) 1 h prior to receiving (99m)Tc-GAP-EDL. In the endometriosis model, part of the rabbit uterine tissue was dissected and grafted to the peritoneal wall. The rabbit was administered with (99m)Tc-GAP-EDL. There was a 10-40% reduction in uptake of (99m)Tc-GAP-EDL in cells treated with DES or tamoxifen compared with untreated cells. Western blot analysis showed an ERK1/2 phosphorylation process with GAP-EDL. Biodistribution studies showed that tumor uptake and tumor-to-muscle count density ratio in (99m)Tc-GAP-EDL groups were significantly higher than those in (99m)Tc-GAP groups at 4 h. Among (99m)Tc-GAP-EDL groups, region of interest analysis of images showed that tumor-to muscle ratios were decreased in blocking groups. In the endometriosis model, the grafted uterine tissue could be visualized by (99m)Tc-GAP-EDL. Cellular or tumor uptake of (99m)Tc-GAP-EDL occurs via an ER-mediated process. (99m)Tc-GAP-EDL is a useful agent for imaging functional ER(+) disease.

Quantitative phase-digital holographic microscopy: a new imaging modality to identify original cellular biomarkers of diseases

NASA Astrophysics Data System (ADS)

Marquet, P.; Rothenfusser, K.; Rappaz, B.; Depeursinge, C.; Jourdain, P.; Magistretti, P. J.

2016-03-01

Quantitative phase microscopy (QPM) has recently emerged as a powerful label-free technique in the field of living cell imaging allowing to non-invasively measure with a nanometric axial sensitivity cell structure and dynamics. Since the phase retardation of a light wave when transmitted through the observed cells, namely the quantitative phase signal (QPS), is sensitive to both cellular thickness and intracellular refractive index related to the cellular content, its accurate analysis allows to derive various cell parameters and monitor specific cell processes, which are very likely to identify new cell biomarkers. Specifically, quantitative phase-digital holographic microscopy (QP-DHM), thanks to its numerical flexibility facilitating parallelization and automation processes, represents an appealing imaging modality to both identify original cellular biomarkers of diseases as well to explore the underlying pathophysiological processes.

Configuration of a high-content imaging platform for hit identification and pharmacological assessment of JMJD3 demethylase enzyme inhibitors.

PubMed

Mulji, Alpa; Haslam, Carl; Brown, Fiona; Randle, Rebecca; Karamshi, Bhumika; Smith, Julia; Eagle, Robert; Munoz-Muriedas, Jordi; Taylor, Joanna; Sheikh, Arshad; Bridges, Angela; Gill, Kirsty; Jepras, Rob; Smee, Penny; Barker, Mike; Woodrow, Mike; Liddle, John; Thomas, Pamela; Jones, Emma; Gordon, Laurie; Tanner, Rob; Leveridge, Melanie; Hutchinson, Sue; Martin, Margaret; Brown, Murray; Kruidenier, Laurens; Katso, Roy

2012-01-01

The biological complexity associated with the regulation of histone demethylases makes it desirable to configure a cellular mechanistic assay format that simultaneously encompasses as many of the relevant cellular processes as possible. In this report, the authors describe the configuration of a JMJD3 high-content cellular mechanistic imaging assay that uses single-cell multiparameter measurements to accurately assess cellular viability and the enzyme-dependent demethylation of the H3K27(Me)3 mark by exogenously expressed JMJD3. This approach couples robust statistical analyses with the spatial resolving power of cellular imaging. This enables segregation of expressing and nonexpressing cells into discrete subpopulations and consequently pharmacological quantification of compounds of interest in the expressing population at varying JMJD3 expression levels. Moreover, the authors demonstrate the utility of this hit identification strategy through the successful prosecution of a medium-throughput focused campaign of an 87 500-compound file, which has enabled the identification of JMJD3 cellular-active chemotypes. This study represents the first report of a demethylase high-content imaging assay with the ability to capture a repertoire of pharmacological tools, which are likely both to inform our mechanistic understanding of how JMJD3 is modulated and, more important, to contribute to the identification of novel therapeutic modalities for this demethylase enzyme.

Direct chemical evidence for sphingolipid domains in the plasma membranes of fibroblasts [High-Resolution Chemical Imaging of Sphingolipid Distribution in the Plasma Membrane

DOE PAGES

Frisz, Jessica F.; Lou, Kaiyan; Klitzing, Haley A.; ...

2013-01-28

Sphingolipids play important roles in plasma membrane structure and cell signaling. Yet, their lateral distribution in the plasma membrane is poorly understood. Here we quantitatively analyzed the sphingolipid organization on the entire dorsal surface of intact cells by mapping the distribution of 15N-enriched ions from metabolically labeled 15N-sphingolipids in the plasma membrane using high-resolution imaging mass spectrometry. Many types of control experiments (internal, positive, negative, and fixation temperature), along with parallel experiments involving the imaging of fluorescent sphingolipids$-$both in living cells and during fixation of living cells$-$exclude potential artifacts. Micrometer-scale sphingolipid patches consisting of numerous 15Nsphingolipid microdomains with mean diametersmore » of ~200 nm are always present in the plasma membrane. Depletion of 30% of the cellular cholesterol did not eliminate the sphingolipid domains, but did reduce their abundance and long range organization in the plasma membrane. In contrast, disruption of the cytoskeleton eliminated the sphingolipid domains. These results indicate that these sphingolipid assemblages are not lipid rafts, and are instead a distinctly different type of sphingolipid-enriched plasma membrane domain that depends upon cortical actin.« less

Laser-emission imaging of nuclear biomarkers for high-contrast cancer screening and immunodiagnosis

PubMed Central

Chen, Yu-Cheng; Tan, Xiaotian; Sun, Qihan; Chen, Qiushu; Wang, Wenjie; Fan, Xudong

2017-01-01

Detection of nuclear biomarkers such as nucleic acids and nuclear proteins is critical for early-stage cancer diagnosis and prognosis. Conventional methods relying on morphological assessment of cell nuclei in histopathology slides may be subjective, whereas colorimetric immunohistochemical and fluorescence-based imaging are limited by strong light absorption, broad-emission bands and low contrast. Here, we describe the development and use of a scanning laser-emission-based microscope that maps lasing emissions from nuclear biomarkers in human tissues. 41 tissue samples from 35 patients labelled with site-specific and biomarker-specific antibody-conjugated dyes were sandwiched in a Fabry-Pérot microcavity while an excitation laser beam built a laser-emission image. We observed multiple sub-cellular lasing emissions from cancer cell nuclei, with a threshold of tens of μJ/mm2, sub-micron resolution (<700 nm), and a lasing band in the few-nanometre range. Different lasing thresholds of nuclei in cancer and normal tissues enabled the identification and multiplexed detection of nuclear proteomic biomarkers, with a high sensitivity for early-stage cancer diagnosis. Laser-emission-based cancer screening and immunodiagnosis might find use in precision medicine and facilitate research in cell biology. PMID:29204310

Toxicological Tipping Points: Learning Boolean Networks from High-Content Imaging Data. (BOSC meeting)

EPA Science Inventory

The objective of this work is to elucidate biological networks underlying cellular tipping points using time-course data. We discretized the high-content imaging (HCI) data and inferred Boolean networks (BNs) that could accurately predict dynamic cellular trajectories. We found t...

Glucose-functionalized Au nanoprisms for optoacoustic imaging and near-infrared photothermal therapy

NASA Astrophysics Data System (ADS)

Han, Jishu; Zhang, Jingjing; Yang, Meng; Cui, Daxiang; de La Fuente, Jesus M.

2015-12-01

Targeted imaging and tumor therapy using nanomaterials has stimulated research interest recently, but the high cytotoxicity and low cellular uptake of nanomaterials limit their bioapplication. In this paper, glucose (Glc) was chosen to functionalize Au nanoprisms (NPrs) for improving the cytotoxicity and cellular uptake of Au@PEG-Glc NPrs into cancer cells. Glucose is a primary source of energy at the cellular level and at cellular membranes for cell recognition. A coating of glucose facilitates the accumulation of Au@PEG-Glc NPrs in a tumor region much more than Au@PEG NPrs. Due to the high accumulation and excellent photoabsorbing property of Au@PEG-Glc NPrs, enhanced optoacoustic imaging of a tumor in vivo was achieved, and visualization of the tumor further guided cancer treatment. Based on the optical-thermal conversion performance of Au@PEG-Glc NPrs, the tumor in vivo was effectively cured through photothermal therapy. The current work demonstrates the great potential of Au@PEG-Glc NPrs in optoacoustic imaging and photothermal cancer therapy in future.Targeted imaging and tumor therapy using nanomaterials has stimulated research interest recently, but the high cytotoxicity and low cellular uptake of nanomaterials limit their bioapplication. In this paper, glucose (Glc) was chosen to functionalize Au nanoprisms (NPrs) for improving the cytotoxicity and cellular uptake of Au@PEG-Glc NPrs into cancer cells. Glucose is a primary source of energy at the cellular level and at cellular membranes for cell recognition. A coating of glucose facilitates the accumulation of Au@PEG-Glc NPrs in a tumor region much more than Au@PEG NPrs. Due to the high accumulation and excellent photoabsorbing property of Au@PEG-Glc NPrs, enhanced optoacoustic imaging of a tumor in vivo was achieved, and visualization of the tumor further guided cancer treatment. Based on the optical-thermal conversion performance of Au@PEG-Glc NPrs, the tumor in vivo was effectively cured through photothermal therapy. The current work demonstrates the great potential of Au@PEG-Glc NPrs in optoacoustic imaging and photothermal cancer therapy in future. Electronic supplementary information (ESI) available: The evolution of the UV-vis absorption of Au NPrs by centrifugation, TEM image of PEG-capped Au NPrs, the UV-vis absorption of glucose, cytotoxicity of Au@PEG-Glc NPrs, gastric cell viabilities versus the concentration of Au@PEG-Glc NPrs and gastric cell viabilities filled with 80 μg Au@PEG-Glc NPrs versus the irradiation time, optoacoustic signals of Au NPr solution and Au@PEG NPrs. See DOI: 10.1039/c5nr06261f

Context Specificity of Stress-activated Mitogen-activated Protein (MAP) Kinase Signaling: The Story as Told by Caenorhabditis elegans.

PubMed

Andrusiak, Matthew G; Jin, Yishi

2016-04-08

Stress-associated p38 and JNK mitogen-activated protein (MAP) kinase signaling cascades trigger specific cellular responses and are involved in multiple disease states. At the root of MAP kinase signaling complexity is the differential use of common components on a context-specific basis. The roundwormCaenorhabditis eleganswas developed as a system to study genes required for development and nervous system function. The powerful genetics ofC. elegansin combination with molecular and cellular dissections has led to a greater understanding of how p38 and JNK signaling affects many biological processes under normal and stress conditions. This review focuses on the studies revealing context specificity of different stress-activated MAPK components inC. elegans. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Variable Delay Multi-Pulse Train for Fast Chemical Exchange Saturation Transfer and Relayed-Nuclear Overhauser Enhancement MRI

PubMed Central

Xu, Jiadi; Yadav, Nirbhay N.; Bar-Shir, Amnon; Jones, Craig K.; Chan, Kannie W. Y.; Zhang, Jiangyang; Walczak, P.; McMahon, Michael T.; van Zijl, Peter C. M.

2013-01-01

Purpose Chemical exchange saturation transfer (CEST) imaging is a new MRI technology allowing the detection of low concentration endogenous cellular proteins and metabolites indirectly through their exchangeable protons. A new technique, variable delay multi-pulse CEST (VDMP-CEST), is proposed to eliminate the need for recording full Z-spectra and performing asymmetry analysis to obtain CEST contrast. Methods The VDMP-CEST scheme involves acquiring images with two (or more) delays between radiofrequency saturation pulses in pulsed CEST, producing a series of CEST images sensitive to the speed of saturation transfer. Subtracting two images or fitting a time series produces CEST and relayed-nuclear Overhauser enhancement CEST maps without effects of direct water saturation and, when using low radiofrequency power, minimal magnetization transfer contrast interference. Results When applied to several model systems (bovine serum albumin, crosslinked bovine serum albumin, l-glutamic acid) and in vivo on healthy rat brain, VDMP-CEST showed sensitivity to slow to intermediate range magnetization transfer processes (rate < 100–150 Hz), such as amide proton transfer and relayed nuclear Overhauser enhancement-CEST. Images for these contrasts could be acquired in short scan times by using a single radiofrequency frequency. Conclusions VDMP-CEST provides an approach to detect CEST effect by sensitizing saturation experiments to slower exchange processes without interference of direct water saturation and without need to acquire Z-spectra and perform asymmetry analysis. PMID:23813483

Surface Chemistry Manipulation of Gold Nanorods Displays High Cellular Uptake In Vitro While Preserving Optical Properties for Bio-Imaging and Photo-Thermal Applications

DTIC Science & Technology

2016-03-28

PROPERTIES FOR BIO -IMAGING AND PHOTO-THERMAL APPLICATIONS ANTHONY B. POLITO III, Maj, USAF, BSC, PhD, MT(ASCP)SBB March 2016 Final Report for March...HIGH CELLULAR UPTAKE IN VITRO WHILE PRESERVING OPTICAL PROPERTIES FOR BIO -IMAGING AND PHOTO-THERMAL APPLICATIONS. 5a. CONTRACT NUMBER 5b...These findings identify MTAB-TA GNRs as prime candidates for use in nano-based bio -imaging and photo-thermal applications. 15. SUBJECT TERMS

Automated and Adaptable Quantification of Cellular Alignment from Microscopic Images for Tissue Engineering Applications

PubMed Central

Xu, Feng; Beyazoglu, Turker; Hefner, Evan; Gurkan, Umut Atakan

2011-01-01

Cellular alignment plays a critical role in functional, physical, and biological characteristics of many tissue types, such as muscle, tendon, nerve, and cornea. Current efforts toward regeneration of these tissues include replicating the cellular microenvironment by developing biomaterials that facilitate cellular alignment. To assess the functional effectiveness of the engineered microenvironments, one essential criterion is quantification of cellular alignment. Therefore, there is a need for rapid, accurate, and adaptable methodologies to quantify cellular alignment for tissue engineering applications. To address this need, we developed an automated method, binarization-based extraction of alignment score (BEAS), to determine cell orientation distribution in a wide variety of microscopic images. This method combines a sequenced application of median and band-pass filters, locally adaptive thresholding approaches and image processing techniques. Cellular alignment score is obtained by applying a robust scoring algorithm to the orientation distribution. We validated the BEAS method by comparing the results with the existing approaches reported in literature (i.e., manual, radial fast Fourier transform-radial sum, and gradient based approaches). Validation results indicated that the BEAS method resulted in statistically comparable alignment scores with the manual method (coefficient of determination R2=0.92). Therefore, the BEAS method introduced in this study could enable accurate, convenient, and adaptable evaluation of engineered tissue constructs and biomaterials in terms of cellular alignment and organization. PMID:21370940

Cellular resolution functional imaging in behaving rats using voluntary head restraint

PubMed Central

Scott, Benjamin B.; Brody, Carlos D.; Tank, David W.

2013-01-01

SUMMARY High-throughput operant conditioning systems for rodents provide efficient training on sophisticated behavioral tasks. Combining these systems with technologies for cellular resolution functional imaging would provide a powerful approach to study neural dynamics during behavior. Here we describe an integrated two-photon microscope and behavioral apparatus that allows cellular resolution functional imaging of cortical regions during epochs of voluntary head restraint. Rats were trained to initiate periods of restraint up to 8 seconds in duration, which provided the mechanical stability necessary for in vivo imaging while allowing free movement between behavioral trials. A mechanical registration system repositioned the head to within a few microns, allowing the same neuronal populations to be imaged on each trial. In proof-of-principle experiments, calcium dependent fluorescence transients were recorded from GCaMP-labeled cortical neurons. In contrast to previous methods for head restraint, this system can also be incorporated into high-throughput operant conditioning systems. PMID:24055015

Quantitative X-ray mapping, scatter diagrams and the generation of correction maps to obtain more information about your material

NASA Astrophysics Data System (ADS)

Wuhrer, R.; Moran, K.

2014-03-01

Quantitative X-ray mapping with silicon drift detectors and multi-EDS detector systems have become an invaluable analysis technique and one of the most useful methods of X-ray microanalysis today. The time to perform an X-ray map has reduced considerably with the ability to map minor and trace elements very accurately due to the larger detector area and higher count rate detectors. Live X-ray imaging can now be performed with a significant amount of data collected in a matter of minutes. A great deal of information can be obtained from X-ray maps. This includes; elemental relationship or scatter diagram creation, elemental ratio mapping, chemical phase mapping (CPM) and quantitative X-ray maps. In obtaining quantitative x-ray maps, we are able to easily generate atomic number (Z), absorption (A), fluorescence (F), theoretical back scatter coefficient (η), and quantitative total maps from each pixel in the image. This allows us to generate an image corresponding to each factor (for each element present). These images allow the user to predict and verify where they are likely to have problems in our images, and are especially helpful to look at possible interface artefacts. The post-processing techniques to improve the quantitation of X-ray map data and the development of post processing techniques for improved characterisation are covered in this paper.

Earth Resources Technology Satellite data collection project, ERTS - Bolivia. [thematic mapping

NASA Technical Reports Server (NTRS)

Brockmann, C. E.

1974-01-01

The Earth Resources Technology Satellite program of Bolivia has developed a multidisciplinary project to carry out investigations in cartography and to prepare various thematic maps. In cartography, investigations are being carried out with the ERTS-1 images and with existing maps, to determine their application to the preparation of new cartographic products on one hand and on the other to map those regions where the cartography is still deficient. The application of the MSS images to the geological mapping has given more than satisfactory results. Working with conventional photointerpretation, it has been possible to prepare regional geological maps, tectonic maps, studies relative to mining, geomorphological maps, studies relative to petroleum exploration, volcanological maps and maps of hydrologic basins. In agriculture, the ERTS images are used to study land classification and forest and soils mapping.

A stochastic approach to estimate the uncertainty of dose mapping caused by uncertainties in b-spline registration

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hub, Martina; Thieke, Christian; Kessler, Marc L.

2012-04-15

Purpose: In fractionated radiation therapy, image guidance with daily tomographic imaging becomes more and more clinical routine. In principle, this allows for daily computation of the delivered dose and for accumulation of these daily dose distributions to determine the actually delivered total dose to the patient. However, uncertainties in the mapping of the images can translate into errors of the accumulated total dose, depending on the dose gradient. In this work, an approach to estimate the uncertainty of mapping between medical images is proposed that identifies areas bearing a significant risk of inaccurate dose accumulation. Methods: This method accounts formore » the geometric uncertainty of image registration and the heterogeneity of the dose distribution, which is to be mapped. Its performance is demonstrated in context of dose mapping based on b-spline registration. It is based on evaluation of the sensitivity of dose mapping to variations of the b-spline coefficients combined with evaluation of the sensitivity of the registration metric with respect to the variations of the coefficients. It was evaluated based on patient data that was deformed based on a breathing model, where the ground truth of the deformation, and hence the actual true dose mapping error, is known. Results: The proposed approach has the potential to distinguish areas of the image where dose mapping is likely to be accurate from other areas of the same image, where a larger uncertainty must be expected. Conclusions: An approach to identify areas where dose mapping is likely to be inaccurate was developed and implemented. This method was tested for dose mapping, but it may be applied in context of other mapping tasks as well.« less

Testing random forest classification for identifying lava flows and mapping age groups on a single Landsat 8 image

NASA Astrophysics Data System (ADS)

Li, Long; Solana, Carmen; Canters, Frank; Kervyn, Matthieu

2017-10-01

Mapping lava flows using satellite images is an important application of remote sensing in volcanology. Several volcanoes have been mapped through remote sensing using a wide range of data, from optical to thermal infrared and radar images, using techniques such as manual mapping, supervised/unsupervised classification, and elevation subtraction. So far, spectral-based mapping applications mainly focus on the use of traditional pixel-based classifiers, without much investigation into the added value of object-based approaches and into advantages of using machine learning algorithms. In this study, Nyamuragira, characterized by a series of > 20 overlapping lava flows erupted over the last century, was used as a case study. The random forest classifier was tested to map lava flows based on pixels and objects. Image classification was conducted for the 20 individual flows and for 8 groups of flows of similar age using a Landsat 8 image and a DEM of the volcano, both at 30-meter spatial resolution. Results show that object-based classification produces maps with continuous and homogeneous lava surfaces, in agreement with the physical characteristics of lava flows, while lava flows mapped through the pixel-based classification are heterogeneous and fragmented including much "salt and pepper noise". In terms of accuracy, both pixel-based and object-based classification performs well but the former results in higher accuracies than the latter except for mapping lava flow age groups without using topographic features. It is concluded that despite spectral similarity, lava flows of contrasting age can be well discriminated and mapped by means of image classification. The classification approach demonstrated in this study only requires easily accessible image data and can be applied to other volcanoes as well if there is sufficient information to calibrate the mapping.

A stochastic approach to estimate the uncertainty of dose mapping caused by uncertainties in b-spline registration

PubMed Central

Hub, Martina; Thieke, Christian; Kessler, Marc L.; Karger, Christian P.

2012-01-01

Purpose: In fractionated radiation therapy, image guidance with daily tomographic imaging becomes more and more clinical routine. In principle, this allows for daily computation of the delivered dose and for accumulation of these daily dose distributions to determine the actually delivered total dose to the patient. However, uncertainties in the mapping of the images can translate into errors of the accumulated total dose, depending on the dose gradient. In this work, an approach to estimate the uncertainty of mapping between medical images is proposed that identifies areas bearing a significant risk of inaccurate dose accumulation. Methods: This method accounts for the geometric uncertainty of image registration and the heterogeneity of the dose distribution, which is to be mapped. Its performance is demonstrated in context of dose mapping based on b-spline registration. It is based on evaluation of the sensitivity of dose mapping to variations of the b-spline coefficients combined with evaluation of the sensitivity of the registration metric with respect to the variations of the coefficients. It was evaluated based on patient data that was deformed based on a breathing model, where the ground truth of the deformation, and hence the actual true dose mapping error, is known. Results: The proposed approach has the potential to distinguish areas of the image where dose mapping is likely to be accurate from other areas of the same image, where a larger uncertainty must be expected. Conclusions: An approach to identify areas where dose mapping is likely to be inaccurate was developed and implemented. This method was tested for dose mapping, but it may be applied in context of other mapping tasks as well. PMID:22482640

Landsat Image Map Production Methods at the U. S. Geological Survey

USGS Publications Warehouse

Kidwell, R.D.; Binnie, D.R.; Martin, S.

1987-01-01

To maintain consistently high quality in satellite image map production, the U. S. Geological Survey (USGS) has developed standard procedures for the photographic and digital production of Landsat image mosaics, and for lithographic printing of multispectral imagery. This paper gives a brief review of the photographic, digital, and lithographic procedures currently in use for producing image maps from Landsat data. It is shown that consistency in the printing of image maps is achieved by standardizing the materials and procedures that affect the image detail and color balance of the final product. Densitometric standards are established by printing control targets using the pressplates, inks, pre-press proofs, and paper to be used for printing.

Wide Swath Stereo Mapping from Gaofen-1 Wide-Field-View (WFV) Images Using Calibration

PubMed Central

Chen, Shoubin; Liu, Jingbin; Huang, Wenchao

2018-01-01

The development of Earth observation systems has changed the nature of survey and mapping products, as well as the methods for updating maps. Among optical satellite mapping methods, the multiline array stereo and agile stereo modes are the most common methods for acquiring stereo images. However, differences in temporal resolution and spatial coverage limit their application. In terms of this issue, our study takes advantage of the wide spatial coverage and high revisit frequencies of wide swath images and aims at verifying the feasibility of stereo mapping with the wide swath stereo mode and reaching a reliable stereo accuracy level using calibration. In contrast with classic stereo modes, the wide swath stereo mode is characterized by both a wide spatial coverage and high-temporal resolution and is capable of obtaining a wide range of stereo images over a short period. In this study, Gaofen-1 (GF-1) wide-field-view (WFV) images, with total imaging widths of 800 km, multispectral resolutions of 16 m and revisit periods of four days, are used for wide swath stereo mapping. To acquire a high-accuracy digital surface model (DSM), the nonlinear system distortion in the GF-1 WFV images is detected and compensated for in advance. The elevation accuracy of the wide swath stereo mode of the GF-1 WFV images can be improved from 103 m to 30 m for a DSM with proper calibration, meeting the demands for 1:250,000 scale mapping and rapid topographic map updates and showing improved efficacy for satellite imaging. PMID:29494540

Acrylamide inhibits cellular differentiation of human neuroblastoma and glioblastoma cells.

PubMed

Chen, Jong-Hang; Chou, Chin-Cheng

2015-08-01

This study explores human neuroblastoma (SH-SY5Y) and human glioblastoma (U-1240 MG) cellular differentiation changes under exposure to acrylamide (ACR). Differentiation of SH-SY5Y and U-1240 MG cells were induced by retinoic acid (RA) and butyric acid (BA), respectively. Morphological observations and MTT assay showed that the induced cellular differentiation and cell proliferation were inhibited by ACR in a time- and dose-dependent manner. ACR co-treatment with RA attenuated SH-SY5Y expressions of neurofilament protein-L (NF-L), microtubule-associated protein 1b (MAP1b; 1.2 to 0.7, p < 0.001), MAP2c (2.2 to 0.8, p < 0.05), and Janus kinase1 (JAK1; 1.9 to 0.6, p < 0.001), while ACR co-treatment with BA attenuated U-1240 MG expressions of glial fibrillary acidic protein (GFAP), MAP1b (1.2 to 0.6, p < 0.001), MAP2c (1.5 to 0.7, p < 0.01), and JAK1 (2.1 to 0.5, p < 0.001), respectively. ACR also decreased the phosphorylation of extracellular-signal-regulated kinases (ERK) and c-Jun N-terminal kinases (JNK) in U-1240 MG cells, while caffeine reversed this suppression of ERK and JNK phosphorylation caused by ACR treatment. These results showed that RA-induced neurogenesis of SH-SY5Y and BA-induced astrogliogenesis of U-1240 MG cells were attenuated by ACR and were associated with down-regulation of MAPs expression and JAK-STAT signaling. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enhanced visual perception through tone mapping

NASA Astrophysics Data System (ADS)

Harrison, Andre; Mullins, Linda L.; Raglin, Adrienne; Etienne-Cummings, Ralph

2016-05-01

Tone mapping operators compress high dynamic range images to improve the picture quality on a digital display when the dynamic range of the display is lower than that of the image. However, tone mapping operators have been largely designed and evaluated based on the aesthetic quality of the resulting displayed image or how perceptually similar the compressed image appears relative to the original scene. They also often require per image tuning of parameters depending on the content of the image. In military operations, however, the amount of information that can be perceived is more important than the aesthetic quality of the image and any parameter adjustment needs to be as automated as possible regardless of the content of the image. We have conducted two studies to evaluate the perceivable detail of a set of tone mapping algorithms, and we apply our findings to develop and test an automated tone mapping algorithm that demonstrates a consistent improvement in the amount of perceived detail. An automated, and thereby predictable, tone mapping method enables a consistent presentation of perceivable features, can reduce the bandwidth required to transmit the imagery, and can improve the accessibility of the data by reducing the needed expertise of the analyst(s) viewing the imagery.

A novel false color mapping model-based fusion method of visual and infrared images

NASA Astrophysics Data System (ADS)

Qi, Bin; Kun, Gao; Tian, Yue-xin; Zhu, Zhen-yu

2013-12-01

A fast and efficient image fusion method is presented to generate near-natural colors from panchromatic visual and thermal imaging sensors. Firstly, a set of daytime color reference images are analyzed and the false color mapping principle is proposed according to human's visual and emotional habits. That is, object colors should remain invariant after color mapping operations, differences between infrared and visual images should be enhanced and the background color should be consistent with the main scene content. Then a novel nonlinear color mapping model is given by introducing the geometric average value of the input visual and infrared image gray and the weighted average algorithm. To determine the control parameters in the mapping model, the boundary conditions are listed according to the mapping principle above. Fusion experiments show that the new fusion method can achieve the near-natural appearance of the fused image, and has the features of enhancing color contrasts and highlighting the infrared brilliant objects when comparing with the traditional TNO algorithm. Moreover, it owns the low complexity and is easy to realize real-time processing. So it is quite suitable for the nighttime imaging apparatus.

Postprocessing classification images

NASA Technical Reports Server (NTRS)

Kan, E. P.

1979-01-01

Program cleans up remote-sensing maps. It can be used with existing image-processing software. Remapped images closely resemble familiar resource information maps and can replace or supplement classification images not postprocessed by this program.

Evidence for network evolution in an arabidopsis interactome map

USDA-ARS?s Scientific Manuscript database

Plants have unique features that evolved in response to their environments and ecosystems. A full account of the complex cellular networks that underlie plant-specific functions is still missing. We describe a proteome-wide binary protein-protein interaction map for the interactome network of the pl...

Urban Land Cover/use Change Detection Using High Resolution SPOT 5 and SPOT 6 Images and Urban Atlas Nomenclature

NASA Astrophysics Data System (ADS)

Akay, S. S.; Sertel, E.

2016-06-01

Urban land cover/use changes like urbanization and urban sprawl have been impacting the urban ecosystems significantly therefore determination of urban land cover/use changes is an important task to understand trends and status of urban ecosystems, to support urban planning and to aid decision-making for urban-based projects. High resolution satellite images could be used to accurately, periodically and quickly map urban land cover/use and their changes by time. This paper aims to determine urban land cover/use changes in Gaziantep city centre between 2010 and 2105 using object based images analysis and high resolution SPOT 5 and SPOT 6 images. 2.5 m SPOT 5 image obtained in 5th of June 2010 and 1.5 m SPOT 6 image obtained in 7th of July 2015 were used in this research to precisely determine land changes in five-year period. In addition to satellite images, various ancillary data namely Normalized Difference Vegetation Index (NDVI), Difference Water Index (NDWI) maps, cadastral maps, OpenStreetMaps, road maps and Land Cover maps, were integrated into the classification process to produce high accuracy urban land cover/use maps for these two years. Both images were geometrically corrected to fulfil the 1/10,000 scale geometric accuracy. Decision tree based object oriented classification was applied to identify twenty different urban land cover/use classes defined in European Urban Atlas project. Not only satellite images and satellite image-derived indices but also different thematic maps were integrated into decision tree analysis to create rule sets for accurate mapping of each class. Rule sets of each satellite image for the object based classification involves spectral, spatial and geometric parameter to automatically produce urban map of the city centre region. Total area of each class per related year and their changes in five-year period were determined and change trend in terms of class transformation were presented. Classification accuracy assessment was conducted by creating a confusion matrix to illustrate the thematic accuracy of each class.

Iterative Refinement of Transmission Map for Stereo Image Defogging Using a Dual Camera Sensor.

PubMed

Kim, Heegwang; Park, Jinho; Park, Hasil; Paik, Joonki

2017-12-09

Recently, the stereo imaging-based image enhancement approach has attracted increasing attention in the field of video analysis. This paper presents a dual camera-based stereo image defogging algorithm. Optical flow is first estimated from the stereo foggy image pair, and the initial disparity map is generated from the estimated optical flow. Next, an initial transmission map is generated using the initial disparity map. Atmospheric light is then estimated using the color line theory. The defogged result is finally reconstructed using the estimated transmission map and atmospheric light. The proposed method can refine the transmission map iteratively. Experimental results show that the proposed method can successfully remove fog without color distortion. The proposed method can be used as a pre-processing step for an outdoor video analysis system and a high-end smartphone with a dual camera system.

Iterative Refinement of Transmission Map for Stereo Image Defogging Using a Dual Camera Sensor

PubMed Central

Park, Jinho; Park, Hasil

2017-01-01

Recently, the stereo imaging-based image enhancement approach has attracted increasing attention in the field of video analysis. This paper presents a dual camera-based stereo image defogging algorithm. Optical flow is first estimated from the stereo foggy image pair, and the initial disparity map is generated from the estimated optical flow. Next, an initial transmission map is generated using the initial disparity map. Atmospheric light is then estimated using the color line theory. The defogged result is finally reconstructed using the estimated transmission map and atmospheric light. The proposed method can refine the transmission map iteratively. Experimental results show that the proposed method can successfully remove fog without color distortion. The proposed method can be used as a pre-processing step for an outdoor video analysis system and a high-end smartphone with a dual camera system. PMID:29232826

Mapping cell populations in flow cytometry data for cross-sample comparison using the Friedman-Rafsky test statistic as a distance measure.

PubMed

Hsiao, Chiaowen; Liu, Mengya; Stanton, Rick; McGee, Monnie; Qian, Yu; Scheuermann, Richard H

2016-01-01

Flow cytometry (FCM) is a fluorescence-based single-cell experimental technology that is routinely applied in biomedical research for identifying cellular biomarkers of normal physiological responses and abnormal disease states. While many computational methods have been developed that focus on identifying cell populations in individual FCM samples, very few have addressed how the identified cell populations can be matched across samples for comparative analysis. This article presents FlowMap-FR, a novel method for cell population mapping across FCM samples. FlowMap-FR is based on the Friedman-Rafsky nonparametric test statistic (FR statistic), which quantifies the equivalence of multivariate distributions. As applied to FCM data by FlowMap-FR, the FR statistic objectively quantifies the similarity between cell populations based on the shapes, sizes, and positions of fluorescence data distributions in the multidimensional feature space. To test and evaluate the performance of FlowMap-FR, we simulated the kinds of biological and technical sample variations that are commonly observed in FCM data. The results show that FlowMap-FR is able to effectively identify equivalent cell populations between samples under scenarios of proportion differences and modest position shifts. As a statistical test, FlowMap-FR can be used to determine whether the expression of a cellular marker is statistically different between two cell populations, suggesting candidates for new cellular phenotypes by providing an objective statistical measure. In addition, FlowMap-FR can indicate situations in which inappropriate splitting or merging of cell populations has occurred during gating procedures. We compared the FR statistic with the symmetric version of Kullback-Leibler divergence measure used in a previous population matching method with both simulated and real data. The FR statistic outperforms the symmetric version of KL-distance in distinguishing equivalent from nonequivalent cell populations. FlowMap-FR was also employed as a distance metric to match cell populations delineated by manual gating across 30 FCM samples from a benchmark FlowCAP data set. An F-measure of 0.88 was obtained, indicating high precision and recall of the FR-based population matching results. FlowMap-FR has been implemented as a standalone R/Bioconductor package so that it can be easily incorporated into current FCM data analytical workflows. © The Authors. Published by Wiley Periodicals, Inc. on behalf of ISAC.

Solution of the problem of superposing image and digital map for detection of new objects

NASA Astrophysics Data System (ADS)

Rizaev, I. S.; Miftakhutdinov, D. I.; Takhavova, E. G.

2018-01-01

The problem of superposing the map of the terrain with the image of the terrain is considered. The image of the terrain may be represented in different frequency bands. Further analysis of the results of collation the digital map with the image of the appropriate terrain is described. Also the approach to detection of differences between information represented on the digital map and information of the image of the appropriate area is offered. The algorithm for calculating the values of brightness of the converted image area on the original picture is offered. The calculation is based on using information about the navigation parameters and information according to arranged bench marks. For solving the posed problem the experiments were performed. The results of the experiments are shown in this paper. The presented algorithms are applicable to the ground complex of remote sensing data to assess differences between resulting images and accurate geopositional data. They are also suitable for detecting new objects in the image, based on the analysis of the matching the digital map and the image of corresponding locality.

Label-free imaging of gold nanoparticles in single live cells by photoacoustic microscopy

NASA Astrophysics Data System (ADS)

Tian, Chao; Qian, Wei; Shao, Xia; Xie, Zhixing; Cheng, Xu; Liu, Shengchun; Cheng, Qian; Liu, Bing; Wang, Xueding

2016-03-01

Gold nanoparticles (AuNPs) have been extensively explored as a model nanostructure in nanomedicine and have been widely used to provide advanced biomedical research tools in diagnostic imaging and therapy. Due to the necessity of targeting AuNPs to individual cells, evaluation and visualization of AuNPs in the cellular level is critical to fully understand their interaction with cellular environment. Currently imaging technologies, such as fluorescence microscopy and transmission electron microscopy all have advantages and disadvantages. In this paper, we synthesized AuNPs by femtosecond pulsed laser ablation, modified their surface chemistry through sequential bioconjugation, and targeted the functionalized AuNPs with individual cancer cells. Based on their high optical absorption contrast, we developed a novel, label-free imaging method to evaluate and visualize intracellular AuNPs using photoacoustic microscopy (PAM). Preliminary study shows that the PAM imaging technique is capable of imaging cellular uptake of AuNPs in vivo at single-cell resolution, which provide an important tool for the study of AuNPs in nanomedicine.

Preoperative Cerebral Oxygen Extraction Fraction Imaging Generated from 7T MR Quantitative Susceptibility Mapping Predicts Development of Cerebral Hyperperfusion following Carotid Endarterectomy.

PubMed

Nomura, J-I; Uwano, I; Sasaki, M; Kudo, K; Yamashita, F; Ito, K; Fujiwara, S; Kobayashi, M; Ogasawara, K

2017-12-01

Preoperative hemodynamic impairment in the affected cerebral hemisphere is associated with the development of cerebral hyperperfusion following carotid endarterectomy. Cerebral oxygen extraction fraction images generated from 7T MR quantitative susceptibility mapping correlate with oxygen extraction fraction images on positron-emission tomography. The present study aimed to determine whether preoperative oxygen extraction fraction imaging generated from 7T MR quantitative susceptibility mapping could identify patients at risk for cerebral hyperperfusion following carotid endarterectomy. Seventy-seven patients with unilateral internal carotid artery stenosis (≥70%) underwent preoperative 3D T2*-weighted imaging using a multiple dipole-inversion algorithm with a 7T MR imager. Quantitative susceptibility mapping images were then obtained, and oxygen extraction fraction maps were generated. Quantitative brain perfusion single-photon emission CT was also performed before and immediately after carotid endarterectomy. ROIs were automatically placed in the bilateral middle cerebral artery territories in all images using a 3D stereotactic ROI template, and affected-to-contralateral ratios in the ROIs were calculated on quantitative susceptibility mapping-oxygen extraction fraction images. Ten patients (13%) showed post-carotid endarterectomy hyperperfusion (cerebral blood flow increases of ≥100% compared with preoperative values in the ROIs on brain perfusion SPECT). Multivariate analysis showed that a high quantitative susceptibility mapping-oxygen extraction fraction ratio was significantly associated with the development of post-carotid endarterectomy hyperperfusion (95% confidence interval, 33.5-249.7; P = .002). Sensitivity, specificity, and positive- and negative-predictive values of the quantitative susceptibility mapping-oxygen extraction fraction ratio for the prediction of the development of post-carotid endarterectomy hyperperfusion were 90%, 84%, 45%, and 98%, respectively. Preoperative oxygen extraction fraction imaging generated from 7T MR quantitative susceptibility mapping identifies patients at risk for cerebral hyperperfusion following carotid endarterectomy. © 2017 by American Journal of Neuroradiology.

Geologic Mapping of Athabasca Valles

NASA Technical Reports Server (NTRS)

Keszthelyi, L. P.; Jaeger, W. L.; Tanaka, K.; Hare, T.

2008-01-01

Two factors drive us to map the Athabasca Valles area in unusual detail: (1) the extremely well-preserved and exposed surface morphologies and (2) the extensive high resolution imaging. In particular, the near-complete CTX coverage of Athabasca Valles proper and the extensive coverage of its surroundings have been invaluable. The mapping has been done exclusively in ArcGIS, using individual CTX, THEMIS VIS, and MOC frames overlying the THEMIS IR daytime basemap. MOLA shot points and gridded DTMs are also included. It was found that CTX images processed through ISIS are almost always within 300 m of the MOLA derived locations, and usually within tens of meters, with no adjustments to camera pointing. THEMIS VIS images appear to be systematically shifted to the southwest of their correct positions and MOC images are often kilometers off. The good SNR and minimal artifacts make the CTX images vastly more useful than the THEMIS VIS or MOC images. The bulk of the mapping was done at 1:50,000 scale on CTX images. In more complex areas, mapping at 1:24,000 proved necessary. The CTX images were usually simultaneously viewed on a second monitor using the ISIS3 qview program to display the full dynamic range of the CTX data. Where CTX data was not available, mapping was often done at 1:100,000 and most contacts are mapped as approximate.

Mapped Landmark Algorithm for Precision Landing

NASA Technical Reports Server (NTRS)

Johnson, Andrew; Ansar, Adnan; Matthies, Larry

2007-01-01

A report discusses a computer vision algorithm for position estimation to enable precision landing during planetary descent. The Descent Image Motion Estimation System for the Mars Exploration Rovers has been used as a starting point for creating code for precision, terrain-relative navigation during planetary landing. The algorithm is designed to be general because it handles images taken at different scales and resolutions relative to the map, and can produce mapped landmark matches for any planetary terrain of sufficient texture. These matches provide a measurement of horizontal position relative to a known landing site specified on the surface map. Multiple mapped landmarks generated per image allow for automatic detection and elimination of bad matches. Attitude and position can be generated from each image; this image-based attitude measurement can be used by the onboard navigation filter to improve the attitude estimate, which will improve the position estimates. The algorithm uses normalized correlation of grayscale images, producing precise, sub-pixel images. The algorithm has been broken into two sub-algorithms: (1) FFT Map Matching (see figure), which matches a single large template by correlation in the frequency domain, and (2) Mapped Landmark Refinement, which matches many small templates by correlation in the spatial domain. Each relies on feature selection, the homography transform, and 3D image correlation. The algorithm is implemented in C++ and is rated at Technology Readiness Level (TRL) 4.

Detection of endometrial lesions by degree of linear polarization maps

NASA Astrophysics Data System (ADS)

Kim, Jihoon; Fazleabas, Asgerally; Walsh, Joseph T.

2010-02-01

Endometriosis is one of the most common causes of chronic pelvic pain and infertility and is characterized by the presence of endometrial glands and stroma outside of the uterine cavity. A novel laparoscopic polarization imaging system was designed to detect endometriosis by imaging endometrial lesions. Linearly polarized light with varying incident polarization angles illuminated endometrial lesions. Degree of linear polarization image maps of endometrial lesions were constructed by using remitted polarized light. The image maps were compared with regular laparoscopy image. The degree of linear polarization map contributed to the detection of endometriosis by revealing structures inside the lesion. The utilization of rotating incident polarization angle (IPA) for the linearly polarized light provides extended understanding of endometrial lesions. The developed polarization system with varying IPA and the collected image maps could provide improved characterization of endometrial lesions via higher visibility of the structure of the lesions and thereby improve diagnosis of endometriosis.

Spatial data software integration - Merging CAD/CAM/mapping with GIS and image processing

NASA Technical Reports Server (NTRS)

Logan, Thomas L.; Bryant, Nevin A.

1987-01-01

The integration of CAD/CAM/mapping with image processing using geographic information systems (GISs) as the interface is examined. Particular emphasis is given to the development of software interfaces between JPL's Video Image Communication and Retrieval (VICAR)/Imaged Based Information System (IBIS) raster-based GIS and the CAD/CAM/mapping system. The design and functions of the VICAR and IBIS are described. Vector data capture and editing are studied. Various software programs for interfacing between the VICAR/IBIS and CAD/CAM/mapping are presented and analyzed.

Mapping broom snakeweed through image analysis of color-infrared photography and digital imagery.

PubMed

Everitt, J H; Yang, C

2007-11-01

A study was conducted on a south Texas rangeland area to evaluate aerial color-infrared (CIR) photography and CIR digital imagery combined with unsupervised image analysis techniques to map broom snakeweed [Gutierrezia sarothrae (Pursh.) Britt. and Rusby]. Accuracy assessments performed on computer-classified maps of photographic images from two sites had mean producer's and user's accuracies for broom snakeweed of 98.3 and 88.3%, respectively; whereas, accuracy assessments performed on classified maps from digital images of the same two sites had mean producer's and user's accuracies for broom snakeweed of 98.3 and 92.8%, respectively. These results indicate that CIR photography and CIR digital imagery combined with image analysis techniques can be used successfully to map broom snakeweed infestations on south Texas rangelands.

Northern Everglades, Florida, satellite image map

USGS Publications Warehouse

Thomas, Jean-Claude; Jones, John W.

2002-01-01

These satellite image maps are one product of the USGS Land Characteristics from Remote Sensing project, funded through the USGS Place-Based Studies Program with support from the Everglades National Park. The objective of this project is to develop and apply innovative remote sensing and geographic information system techniques to map the distribution of vegetation, vegetation characteristics, and related hydrologic variables through space and over time. The mapping and description of vegetation characteristics and their variations are necessary to accurately simulate surface hydrology and other surface processes in South Florida and to monitor land surface changes. As part of this research, data from many airborne and satellite imaging systems have been georeferenced and processed to facilitate data fusion and analysis. These image maps were created using image fusion techniques developed as part of this project.

New segmentation-based tone mapping algorithm for high dynamic range image

NASA Astrophysics Data System (ADS)

Duan, Weiwei; Guo, Huinan; Zhou, Zuofeng; Huang, Huimin; Cao, Jianzhong

2017-07-01

The traditional tone mapping algorithm for the display of high dynamic range (HDR) image has the drawback of losing the impression of brightness, contrast and color information. To overcome this phenomenon, we propose a new tone mapping algorithm based on dividing the image into different exposure regions in this paper. Firstly, the over-exposure region is determined using the Local Binary Pattern information of HDR image. Then, based on the peak and average gray of the histogram, the under-exposure and normal-exposure region of HDR image are selected separately. Finally, the different exposure regions are mapped by differentiated tone mapping methods to get the final result. The experiment results show that the proposed algorithm achieve the better performance both in visual quality and objective contrast criterion than other algorithms.

A novel color image encryption scheme using alternate chaotic mapping structure

NASA Astrophysics Data System (ADS)

Wang, Xingyuan; Zhao, Yuanyuan; Zhang, Huili; Guo, Kang

2016-07-01

This paper proposes an color image encryption algorithm using alternate chaotic mapping structure. Initially, we use the R, G and B components to form a matrix. Then one-dimension logistic and two-dimension logistic mapping is used to generate a chaotic matrix, then iterate two chaotic mappings alternately to permute the matrix. For every iteration, XOR operation is adopted to encrypt plain-image matrix, then make further transformation to diffuse the matrix. At last, the encrypted color image is obtained from the confused matrix. Theoretical analysis and experimental results has proved the cryptosystem is secure and practical, and it is suitable for encrypting color images.

Determination of skeleton and sign map for phase obtaining from a single ESPI image

NASA Astrophysics Data System (ADS)

Yang, Xia; Yu, Qifeng; Fu, Sihua

2009-06-01

A robust method of determining the sign map and skeletons for ESPI images is introduced in this paper. ESPI images have high speckle noise which makes it difficult to obtain the fringe information, especially from a single image. To overcome the effects of high speckle noise, local directional computing windows are designed according to the fringe directions. Then by calculating the gradients from the filtered image in directional windows, sign map and good skeletons can be determined robustly. Based on the sign map, single image phase-extracting methods such as quadrature transform can be improved. And based on skeletons, fringe phases can be obtained directly by normalization methods. Experiments show that this new method is robust and effective for extracting phase from a single ESPI fringe image.

Surface mineral maps of Afghanistan derived from HyMap imaging spectrometer data, version 2

USGS Publications Warehouse

Kokaly, Raymond F.; King, Trude V.V.; Hoefen, Todd M.

2013-01-01

This report presents a new version of surface mineral maps derived from HyMap imaging spectrometer data collected over Afghanistan in the fall of 2007. This report also describes the processing steps applied to the imaging spectrometer data. The 218 individual flight lines composing the Afghanistan dataset, covering more than 438,000 square kilometers, were georeferenced to a mosaic of orthorectified Landsat images. The HyMap data were converted from radiance to reflectance using a radiative transfer program in combination with ground-calibration sites and a network of cross-cutting calibration flight lines. The U.S. Geological Survey Material Identification and Characterization Algorithm (MICA) was used to generate two thematic maps of surface minerals: a map of iron-bearing minerals and other materials, which have their primary absorption features at the shorter wavelengths of the reflected solar wavelength range, and a map of carbonates, phyllosilicates, sulfates, altered minerals, and other materials, which have their primary absorption features at the longer wavelengths of the reflected solar wavelength range. In contrast to the original version, version 2 of these maps is provided at full resolution of 23-meter pixel size. The thematic maps, MICA summary images, and the material fit and depth images are distributed in digital files linked to this report, in a format readable by remote sensing software and Geographic Information Systems (GIS). The digital files can be downloaded from http://pubs.usgs.gov/ds/787/downloads/.

The Circumpolar Arctic Vegetation Map: AVHRR-derived base maps, environmental controls, and integrated mapping procedures

Treesearch

D. A. WALKER; W. A. GOULD; MAIERH. A.; M. K. RAYNOLDS

2002-01-01

A new false-colour-infrared image derived from biweekly 1993 and 1995 Advanced Very High Resolution Radiometer (AVHRR) data provides a snow-free and cloud-free base image for the interpretation of vegetation as part of a 1:7.5M-scale Circumpolar Arctic Vegetation Map (CAVM). A maximum-NDVI (Normalized DiVerence Vegetation Index) image prepared from the same data...

Evaluation of imaging biomarkers for identification of single cancer cells in blood

NASA Astrophysics Data System (ADS)

Odaka, Masao; Kim, Hyonchol; Girault, Mathias; Hattori, Akihiro; Terazono, Hideyuki; Matsuura, Kenji; Yasuda, Kenji

2015-06-01

A method of discriminating single cancer cells from whole blood cells based on their morphological visual characteristics (i.e., “imaging biomarker”) was examined. Cells in healthy rat blood, a cancer cell line (MAT-LyLu), and cells in cancer-cell-implanted rat blood were chosen as models, and their bright-field (BF, whole-cell morphology) and fluorescence (FL, nucleus morphology) images were taken by an on-chip multi-imaging flow cytometry system and compared. Eight imaging biomarker indices, i.e., cellular area in a BF image, nucleus area in an FL image, area ratio of a whole cell and its nucleus, distance of the mass center between a whole cell and nucleus, cellular and nucleus perimeter, and perimeter ratios were calculated and analyzed using the BF and FL images taken. Results show that cancer cells can be clearly distinguished from healthy blood cells using correlation diagrams for cellular and nucleus areas as two different categories. Moreover, a portion of cancer cells showed a low nucleus perimeter ratio less than 0.9 because of the irregular nucleus morphologies of cancer cells. These results indicate that the measurements of imaging biomarkers are practically applicable to identifying cancer cells in blood.

Accurate Construction of Photoactivated Localization Microscopy (PALM) Images for Quantitative Measurements

PubMed Central

Coltharp, Carla; Kessler, Rene P.; Xiao, Jie

2012-01-01

Localization-based superresolution microscopy techniques such as Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) have allowed investigations of cellular structures with unprecedented optical resolutions. One major obstacle to interpreting superresolution images, however, is the overcounting of molecule numbers caused by fluorophore photoblinking. Using both experimental and simulated images, we determined the effects of photoblinking on the accurate reconstruction of superresolution images and on quantitative measurements of structural dimension and molecule density made from those images. We found that structural dimension and relative density measurements can be made reliably from images that contain photoblinking-related overcounting, but accurate absolute density measurements, and consequently faithful representations of molecule counts and positions in cellular structures, require the application of a clustering algorithm to group localizations that originate from the same molecule. We analyzed how applying a simple algorithm with different clustering thresholds (tThresh and dThresh) affects the accuracy of reconstructed images, and developed an easy method to select optimal thresholds. We also identified an empirical criterion to evaluate whether an imaging condition is appropriate for accurate superresolution image reconstruction with the clustering algorithm. Both the threshold selection method and imaging condition criterion are easy to implement within existing PALM clustering algorithms and experimental conditions. The main advantage of our method is that it generates a superresolution image and molecule position list that faithfully represents molecule counts and positions within a cellular structure, rather than only summarizing structural properties into ensemble parameters. This feature makes it particularly useful for cellular structures of heterogeneous densities and irregular geometries, and allows a variety of quantitative measurements tailored to specific needs of different biological systems. PMID:23251611

System and method for image mapping and visual attention

NASA Technical Reports Server (NTRS)

Peters, II, Richard A. (Inventor)

2010-01-01

A method is described for mapping dense sensory data to a Sensory Ego Sphere (SES). Methods are also described for finding and ranking areas of interest in the images that form a complete visual scene on an SES. Further, attentional processing of image data is best done by performing attentional processing on individual full-size images from the image sequence, mapping each attentional location to the nearest node, and then summing attentional locations at each node.

System and method for image mapping and visual attention

NASA Technical Reports Server (NTRS)

Peters, II, Richard A. (Inventor)

2011-01-01

A method is described for mapping dense sensory data to a Sensory Ego Sphere (SES). Methods are also described for finding and ranking areas of interest in the images that form a complete visual scene on an SES. Further, attentional processing of image data is best done by performing attentional processing on individual full-size images from the image sequence, mapping each attentional location to the nearest node, and then summing all attentional locations at each node.

NeuroSeg: automated cell detection and segmentation for in vivo two-photon Ca2+ imaging data.

PubMed

Guan, Jiangheng; Li, Jingcheng; Liang, Shanshan; Li, Ruijie; Li, Xingyi; Shi, Xiaozhe; Huang, Ciyu; Zhang, Jianxiong; Pan, Junxia; Jia, Hongbo; Zhang, Le; Chen, Xiaowei; Liao, Xiang

2018-01-01

Two-photon Ca 2+ imaging has become a popular approach for monitoring neuronal population activity with cellular or subcellular resolution in vivo. This approach allows for the recording of hundreds to thousands of neurons per animal and thus leads to a large amount of data to be processed. In particular, manually drawing regions of interest is the most time-consuming aspect of data analysis. However, the development of automated image analysis pipelines, which will be essential for dealing with the likely future deluge of imaging data, remains a major challenge. To address this issue, we developed NeuroSeg, an open-source MATLAB program that can facilitate the accurate and efficient segmentation of neurons in two-photon Ca 2+ imaging data. We proposed an approach using a generalized Laplacian of Gaussian filter to detect cells and weighting-based segmentation to separate individual cells from the background. We tested this approach on an in vivo two-photon Ca 2+ imaging dataset obtained from mouse cortical neurons with differently sized view fields. We show that this approach exhibits superior performance for cell detection and segmentation compared with the existing published tools. In addition, we integrated the previously reported, activity-based segmentation into our approach and found that this combined method was even more promising. The NeuroSeg software, including source code and graphical user interface, is freely available and will be a useful tool for in vivo brain activity mapping.

Coating barium titanate nanoparticles with polyethylenimine improves cellular uptake and allows for coupled imaging and gene delivery

PubMed Central

Dempsey, Christopher; Lee, Isac; Cowan, Katie; Suh, Junghae

2015-01-01

Barium titanate nanoparticles (BT NP) belong to a class of second harmonic generating (SHG) nanoprobes that have recently demonstrated promise in biological imaging. Unfortunately, BT NPs display low cellular uptake efficiencies, which may be a problem if cellular internalization is desired or required for a particular application. To overcome this issue, while concomitantly developing a particle platform that can also deliver nucleic acids into cells, we coated the BT NPs with the cationic polymer polyethylenimine (PEI) – one of the most effective nonviral gene delivery agents. Coating of BT with PEI yielded complexes with positive zeta potentials and resulted in an 8-fold increase in cellular uptake of the BT NPs. Importantly, we were able to achieve high levels of gene delivery with the BT-PEI/DNA complexes, supporting further efforts to generate BT platforms for coupled imaging and gene therapy. PMID:23973999

A Double Perturbation Method for Reducing Dynamical Degradation of the Digital Baker Map

NASA Astrophysics Data System (ADS)

Liu, Lingfeng; Lin, Jun; Miao, Suoxia; Liu, Bocheng

2017-06-01

The digital Baker map is widely used in different kinds of cryptosystems, especially for image encryption. However, any chaotic map which is realized on the finite precision device (e.g. computer) will suffer from dynamical degradation, which refers to short cycle lengths, low complexity and strong correlations. In this paper, a novel double perturbation method is proposed for reducing the dynamical degradation of the digital Baker map. Both state variables and system parameters are perturbed by the digital logistic map. Numerical experiments show that the perturbed Baker map can achieve good statistical and cryptographic properties. Furthermore, a new image encryption algorithm is provided as a simple application. With a rather simple algorithm, the encrypted image can achieve high security, which is competitive to the recently proposed image encryption algorithms.

OdorMapComparer: an application for quantitative analyses and comparisons of fMRI brain odor maps.

PubMed

Liu, Nian; Xu, Fuqiang; Miller, Perry L; Shepherd, Gordon M

2007-01-01

Brain odor maps are reconstructed flat images that describe the spatial activity patterns in the glomerular layer of the olfactory bulbs in animals exposed to different odor stimuli. We have developed a software application, OdorMapComparer, to carry out quantitative analyses and comparisons of the fMRI odor maps. This application is an open-source window program that first loads two odor map images being compared. It allows image transformations including scaling, flipping, rotating, and warping so that the two images can be appropriately aligned to each other. It performs simple subtraction, addition, and average of signals in the two images. It also provides comparative statistics including the normalized correlation (NC) and spatial correlation coefficient. Experimental studies showed that the rodent fMRI odor maps for aliphatic aldehydes displayed spatial activity patterns that are similar in gross outlines but somewhat different in specific subregions. Analyses with OdorMapComparer indicate that the similarity between odor maps decreases with increasing difference in the length of carbon chains. For example, the map of butanal is more closely related to that of pentanal (with a NC = 0.617) than to that of octanal (NC = 0.082), which is consistent with animal behavioral studies. The study also indicates that fMRI odor maps are statistically odor-specific and repeatable across both the intra- and intersubject trials. OdorMapComparer thus provides a tool for quantitative, statistical analyses and comparisons of fMRI odor maps in a fashion that is integrated with the overall odor mapping techniques.

Defining an Analytic Framework to Evaluate Quantitative MRI Markers of Traumatic Axonal Injury: Preliminary Results in a Mouse Closed Head Injury Model

PubMed Central

Sadeghi, N.; Namjoshi, D.; Irfanoglu, M. O.; Wellington, C.; Diaz-Arrastia, R.

2017-01-01

Diffuse axonal injury (DAI) is a hallmark of traumatic brain injury (TBI) pathology. Recently, the Closed Head Injury Model of Engineered Rotational Acceleration (CHIMERA) was developed to generate an experimental model of DAI in a mouse. The characterization of DAI using diffusion tensor magnetic resonance imaging (MRI; diffusion tensor imaging, DTI) may provide a useful set of outcome measures for preclinical and clinical studies. The objective of this study was to identify the complex neurobiological underpinnings of DTI features following DAI using a comprehensive and quantitative evaluation of DTI and histopathology in the CHIMERA mouse model. A consistent neuroanatomical pattern of pathology in specific white matter tracts was identified across ex vivo DTI maps and photomicrographs of histology. These observations were confirmed by voxelwise and regional analysis of DTI maps, demonstrating reduced fractional anisotropy (FA) in distinct regions such as the optic tract. Similar regions were identified by quantitative histology and exhibited axonal damage as well as robust gliosis. Additional analysis using a machine-learning algorithm was performed to identify regions and metrics important for injury classification in a manner free from potential user bias. This analysis found that diffusion metrics were able to identify injured brains almost with the same degree of accuracy as the histology metrics. Good agreement between regions detected as abnormal by histology and MRI was also found. The findings of this work elucidate the complexity of cellular changes that give rise to imaging abnormalities and provide a comprehensive and quantitative evaluation of the relative importance of DTI and histological measures to detect brain injury. PMID:28966972

Spatially modulated ephrinA1:EphA2 signaling increases local contractility and global focal adhesion dynamics to promote cell motility.

PubMed

Chen, Zhongwen; Oh, Dongmyung; Biswas, Kabir H; Yu, Cheng-Han; Zaidel-Bar, Ronen; Groves, Jay T

2018-06-19

Recent studies have revealed pronounced effects of the spatial distribution of EphA2 receptors on cellular response to receptor activation. However, little is known about molecular mechanisms underlying this spatial sensitivity, in part due to lack of experimental systems. Here, we introduce a hybrid live-cell patterned supported lipid bilayer experimental platform in which the sites of EphA2 activation and integrin adhesion are spatially controlled. Using a series of live-cell imaging and single-molecule tracking experiments, we map the transmission of signals from ephrinA1:EphA2 complexes. Results show that ligand-dependent EphA2 activation induces localized myosin-dependent contractions while simultaneously increasing focal adhesion dynamics throughout the cell. Mechanistically, Src kinase is activated at sites of ephrinA1:EphA2 clustering and subsequently diffuses on the membrane to focal adhesions, where it up-regulates FAK and paxillin tyrosine phosphorylation. EphrinA1:EphA2 signaling triggers multiple cellular responses with differing spatial dependencies to enable a directed migratory response to spatially resolved contact with ephrinA1 ligands.

2D and 3D visualization methods of endoscopic panoramic bladder images

NASA Astrophysics Data System (ADS)

Behrens, Alexander; Heisterklaus, Iris; Müller, Yannick; Stehle, Thomas; Gross, Sebastian; Aach, Til

2011-03-01

While several mosaicking algorithms have been developed to compose endoscopic images of the internal urinary bladder wall into panoramic images, the quantitative evaluation of these output images in terms of geometrical distortions have often not been discussed. However, the visualization of the distortion level is highly desired for an objective image-based medical diagnosis. Thus, we present in this paper a method to create quality maps from the characteristics of transformation parameters, which were applied to the endoscopic images during the registration process of the mosaicking algorithm. For a global first view impression, the quality maps are laid over the panoramic image and highlight image regions in pseudo-colors according to their local distortions. This illustration supports then surgeons to identify geometrically distorted structures easily in the panoramic image, which allow more objective medical interpretations of tumor tissue in shape and size. Aside from introducing quality maps in 2-D, we also discuss a visualization method to map panoramic images onto a 3-D spherical bladder model. Reference points are manually selected by the surgeon in the panoramic image and the 3-D model. Then the panoramic image is mapped by the Hammer-Aitoff equal-area projection onto the 3-D surface using texture mapping. Finally the textured bladder model can be freely moved in a virtual environment for inspection. Using a two-hemisphere bladder representation, references between panoramic image regions and their corresponding space coordinates within the bladder model are reconstructed. This additional spatial 3-D information thus assists the surgeon in navigation, documentation, as well as surgical planning.

A self-trained classification technique for producing 30 m percent-water maps from Landsat data

USGS Publications Warehouse

Rover, Jennifer R.; Wylie, Bruce K.; Ji, Lei

2010-01-01

Small bodies of water can be mapped with moderate-resolution satellite data using methods where water is mapped as subpixel fractions using field measurements or high-resolution images as training datasets. A new method, developed from a regression-tree technique, uses a 30 m Landsat image for training the regression tree that, in turn, is applied to the same image to map subpixel water. The self-trained method was evaluated by comparing the percent-water map with three other maps generated from established percent-water mapping methods: (1) a regression-tree model trained with a 5 m SPOT 5 image, (2) a regression-tree model based on endmembers and (3) a linear unmixing classification technique. The results suggest that subpixel water fractions can be accurately estimated when high-resolution satellite data or intensively interpreted training datasets are not available, which increases our ability to map small water bodies or small changes in lake size at a regional scale.

Segmentation of radiologic images with self-organizing maps: the segmentation problem transformed into a classification task

NASA Astrophysics Data System (ADS)

Pelikan, Erich; Vogelsang, Frank; Tolxdorff, Thomas

1996-04-01

The texture-based segmentation of x-ray images of focal bone lesions using topological maps is introduced. Texture characteristics are described by image-point correlation of feature images to feature vectors. For the segmentation, the topological map is labeled using an improved labeling strategy. Results of the technique are demonstrated on original and synthetic x-ray images and quantified with the aid of quality measures. In addition, a classifier-specific contribution analysis is applied for assessing the feature space.

A Method of Spatial Mapping and Reclassification for High-Spatial-Resolution Remote Sensing Image Classification

PubMed Central

Wang, Guizhou; Liu, Jianbo; He, Guojin

2013-01-01

This paper presents a new classification method for high-spatial-resolution remote sensing images based on a strategic mechanism of spatial mapping and reclassification. The proposed method includes four steps. First, the multispectral image is classified by a traditional pixel-based classification method (support vector machine). Second, the panchromatic image is subdivided by watershed segmentation. Third, the pixel-based multispectral image classification result is mapped to the panchromatic segmentation result based on a spatial mapping mechanism and the area dominant principle. During the mapping process, an area proportion threshold is set, and the regional property is defined as unclassified if the maximum area proportion does not surpass the threshold. Finally, unclassified regions are reclassified based on spectral information using the minimum distance to mean algorithm. Experimental results show that the classification method for high-spatial-resolution remote sensing images based on the spatial mapping mechanism and reclassification strategy can make use of both panchromatic and multispectral information, integrate the pixel- and object-based classification methods, and improve classification accuracy. PMID:24453808

Facilitating in vivo tumor localization by principal component analysis based on dynamic fluorescence molecular imaging

NASA Astrophysics Data System (ADS)

Gao, Yang; Chen, Maomao; Wu, Junyu; Zhou, Yuan; Cai, Chuangjian; Wang, Daliang; Luo, Jianwen

2017-09-01

Fluorescence molecular imaging has been used to target tumors in mice with xenograft tumors. However, tumor imaging is largely distorted by the aggregation of fluorescent probes in the liver. A principal component analysis (PCA)-based strategy was applied on the in vivo dynamic fluorescence imaging results of three mice with xenograft tumors to facilitate tumor imaging, with the help of a tumor-specific fluorescent probe. Tumor-relevant features were extracted from the original images by PCA and represented by the principal component (PC) maps. The second principal component (PC2) map represented the tumor-related features, and the first principal component (PC1) map retained the original pharmacokinetic profiles, especially of the liver. The distribution patterns of the PC2 map of the tumor-bearing mice were in good agreement with the actual tumor location. The tumor-to-liver ratio and contrast-to-noise ratio were significantly higher on the PC2 map than on the original images, thus distinguishing the tumor from its nearby fluorescence noise of liver. The results suggest that the PC2 map could serve as a bioimaging marker to facilitate in vivo tumor localization, and dynamic fluorescence molecular imaging with PCA could be a valuable tool for future studies of in vivo tumor metabolism and progression.

Harvesting geographic features from heterogeneous raster maps

NASA Astrophysics Data System (ADS)

Chiang, Yao-Yi

2010-11-01

Raster maps offer a great deal of geospatial information and are easily accessible compared to other geospatial data. However, harvesting geographic features locked in heterogeneous raster maps to obtain the geospatial information is challenging. This is because of the varying image quality of raster maps (e.g., scanned maps with poor image quality and computer-generated maps with good image quality), the overlapping geographic features in maps, and the typical lack of metadata (e.g., map geocoordinates, map source, and original vector data). Previous work on map processing is typically limited to a specific type of map and often relies on intensive manual work. In contrast, this thesis investigates a general approach that does not rely on any prior knowledge and requires minimal user effort to process heterogeneous raster maps. This approach includes automatic and supervised techniques to process raster maps for separating individual layers of geographic features from the maps and recognizing geographic features in the separated layers (i.e., detecting road intersections, generating and vectorizing road geometry, and recognizing text labels). The automatic technique eliminates user intervention by exploiting common map properties of how road lines and text labels are drawn in raster maps. For example, the road lines are elongated linear objects and the characters are small connected-objects. The supervised technique utilizes labels of road and text areas to handle complex raster maps, or maps with poor image quality, and can process a variety of raster maps with minimal user input. The results show that the general approach can handle raster maps with varying map complexity, color usage, and image quality. By matching extracted road intersections to another geospatial dataset, we can identify the geocoordinates of a raster map and further align the raster map, separated feature layers from the map, and recognized features from the layers with the geospatial dataset. The road vectorization and text recognition results outperform state-of-art commercial products, and with considerably less user input. The approach in this thesis allows us to make use of the geospatial information of heterogeneous maps locked in raster format.

High-resolution maps of real and illusory tactile activation in primary somatosensory cortex in individual monkeys with functional magnetic resonance imaging and optical imaging.

PubMed

Chen, Li M; Turner, Gregory H; Friedman, Robert M; Zhang, Na; Gore, John C; Roe, Anna W; Avison, Malcolm J

2007-08-22

Although blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI) has been widely used to explore human brain function, questions remain regarding the ultimate spatial resolution of positive BOLD fMRI, and indeed the extent to which functional maps revealed by positive BOLD correlate spatially with maps obtained with other high-spatial-resolution mapping techniques commonly used in animals, such as optical imaging of intrinsic signal (OIS) and single-unit electrophysiology. Here, we demonstrate that the positive BOLD signal at 9.4T can reveal the fine topography of individual fingerpads in single-condition activation maps in nonhuman primates. These digit maps are similar to maps obtained from the same animal using intrinsic optical imaging. Furthermore, BOLD fMRI reliably resolved submillimeter spatial shifts in activation in area 3b previously identified with OIS (Chen et al., 2003) as neural correlates of the "funneling illusion." These data demonstrate that at high field, high-spatial-resolution topographic maps can be achieved using the positive BOLD signal, weakening previous notions regarding the spatial specificity of the positive BOLD signal.

Susceptibility-based functional brain mapping by 3D deconvolution of an MR-phase activation map.

PubMed

Chen, Zikuan; Liu, Jingyu; Calhoun, Vince D

2013-05-30

The underlying source of T2*-weighted magnetic resonance imaging (T2*MRI) for brain imaging is magnetic susceptibility (denoted by χ). T2*MRI outputs a complex-valued MR image consisting of magnitude and phase information. Recent research has shown that both the magnitude and the phase images are morphologically different from the source χ, primarily due to 3D convolution, and that the source χ can be reconstructed from complex MR images by computed inverse MRI (CIMRI). Thus, we can obtain a 4D χ dataset from a complex 4D MR dataset acquired from a brain functional MRI study by repeating CIMRI to reconstruct 3D χ volumes at each timepoint. Because the reconstructed χ is a more direct representation of neuronal activity than the MR image, we propose a method for χ-based functional brain mapping, which is numerically characterised by a temporal correlation map of χ responses to a stimulant task. Under the linear imaging conditions used for T2*MRI, we show that the χ activation map can be calculated from the MR phase map by CIMRI. We validate our approach using numerical simulations and Gd-phantom experiments. We also analyse real data from a finger-tapping visuomotor experiment and show that the χ-based functional mapping provides additional activation details (in the form of positive and negative correlation patterns) beyond those generated by conventional MR-magnitude-based mapping. Copyright © 2013 Elsevier B.V. All rights reserved.

Phase imaging of mechanical properties of live cells (Conference Presentation)

NASA Astrophysics Data System (ADS)

Wax, Adam

2017-02-01

The mechanisms by which cells respond to mechanical stimuli are essential for cell function yet not well understood. Many rheological tools have been developed to characterize cellular viscoelastic properties but these typically require direct mechanical contact, limiting their throughput. We have developed a new approach for characterizing the organization of subcellular structures using a label free, noncontact, single-shot phase imaging method that correlates to measured cellular mechanical stiffness. The new analysis approach measures refractive index variance and relates it to disorder strength. These measurements are compared to cellular stiffness, measured using the same imaging tool to visualize nanoscale responses to flow shear stimulus. The utility of the technique is shown by comparing shear stiffness and phase disorder strength across five cellular populations with varying mechanical properties. An inverse relationship between disorder strength and shear stiffness is shown, suggesting that cell mechanical properties can be assessed in a format amenable to high throughput studies using this novel, non-contact technique. Further studies will be presented which include examination of mechanical stiffness in early carcinogenic events and investigation of the role of specific cellular structural proteins in mechanotransduction.

Avoiding Stair-Step Artifacts in Image Registration for GOES-R Navigation and Registration Assessment

NASA Technical Reports Server (NTRS)

Grycewicz, Thomas J.; Tan, Bin; Isaacson, Peter J.; De Luccia, Frank J.; Dellomo, John

2016-01-01

In developing software for independent verification and validation (IVV) of the Image Navigation and Registration (INR) capability for the Geostationary Operational Environmental Satellite R Series (GOES-R) Advanced Baseline Imager (ABI), we have encountered an image registration artifact which limits the accuracy of image offset estimation at the subpixel scale using image correlation. Where the two images to be registered have the same pixel size, subpixel image registration preferentially selects registration values where the image pixel boundaries are close to lined up. Because of the shape of a curve plotting input displacement to estimated offset, we call this a stair-step artifact. When one image is at a higher resolution than the other, the stair-step artifact is minimized by correlating at the higher resolution. For validating ABI image navigation, GOES-R images are correlated with Landsat-based ground truth maps. To create the ground truth map, the Landsat image is first transformed to the perspective seen from the GOES-R satellite, and then is scaled to an appropriate pixel size. Minimizing processing time motivates choosing the map pixels to be the same size as the GOES-R pixels. At this pixel size image processing of the shift estimate is efficient, but the stair-step artifact is present. If the map pixel is very small, stair-step is not a problem, but image correlation is computation-intensive. This paper describes simulation-based selection of the scale for truth maps for registering GOES-R ABI images.

Prospective Elementary Science Teachers' Understanding of Photosynthesis and Cellular Respiration in the Context of Multiple Biological Levels as Nested Systems

ERIC Educational Resources Information Center

Akçay, Süleyman

2017-01-01

In this study, Turkish prospective elementary science teachers' understanding of photosynthesis and cellular respiration has been analysed within the contexts of ecosystem knowledge, organism knowledge and interconnection knowledge (IK). In the analysis, concept maps developed by 74 prospective teachers were used. The study was carried out with…

South Florida Everglades: satellite image map

USGS Publications Warehouse

Jones, John W.; Thomas, Jean-Claude; Desmond, G.B.

2001-01-01

These satellite image maps are one product of the USGS Land Characteristics from Remote Sensing project, funded through the USGS Place-Based Studies Program (http://access.usgs.gov/) with support from the Everglades National Park (http://www.nps.gov/ever/). The objective of this project is to develop and apply innovative remote sensing and geographic information system techniques to map the distribution of vegetation, vegetation characteristics, and related hydrologic variables through space and over time. The mapping and description of vegetation characteristics and their variations are necessary to accurately simulate surface hydrology and other surface processes in South Florida and to monitor land surface changes. As part of this research, data from many airborne and satellite imaging systems have been georeferenced and processed to facilitate data fusion and analysis. These image maps were created using image fusion techniques developed as part of this project.

Mapping biomass for a northern forest ecosystem using multi-frequency SAR data

NASA Technical Reports Server (NTRS)

Ranson, K. J.; Sun, Guoqing

1992-01-01

Image processing methods for mapping standing biomass for a forest in Maine, using NASA/JPL airborne synthetic aperture radar (AIRSAR) polarimeter data, are presented. By examining the dependence of backscattering on standing biomass, it is determined that the ratio of HV backscattering from a longer wavelength (P- or L-band) to a shorter wavelength (C) is a good combination for mapping total biomass. This ratio enhances the correlation of the image signature to the standing biomass and compensates for a major part of the variations in backscattering attributed to radar incidence angle. The image processing methods used include image calibration, ratioing, filtering, and segmentation. The image segmentation algorithm uses both means and variances of the image, and it is combined with the image filtering process. Preliminary assessment of the resultant biomass maps suggests that this is a promising method.

Molecular genetic characterization of the RD-114 gene family of endogenous feline retroviral sequences.

PubMed Central

Reeves, R H; O'Brien, S J

1984-01-01

RD-114 is a replication-competent, xenotropic retrovirus which is homologous to a family of moderately repetitive DNA sequences present at ca. 20 copies in the normal cellular genome of domestic cats. To examine the extent and character of genomic divergence of the RD-114 gene family as well as to assess their positional association within the cat genome, we have prepared a series of molecular clones of endogenous RD-114 DNA segments from a genomic library of cat cellular DNA. Their restriction endonuclease maps were compared with each other as well as to that of the prototype-inducible RD-114 which was molecularly cloned from a chronically infected human cell line. The endogenous sequences analyzed were similar to each other in that they were colinear with RD-114 proviral DNA, were bounded by long terminal redundancies, and conserved many restriction sites in the gag and pol regions. However, the env regions of many of the sequences examined were substantially deleted. Several of the endogenous RD-114 genomes contained a novel envelope sequence which was unrelated to the env gene of the prototype RD-114 env gene but which, like RD-114 and endogenous feline leukemia virus provirus, was found only in species of the genus Felis, and not in other closely related Felidae genera. The endogenous RD-114 sequences each had a distinct cellular flank which indicates that these sequences are not tandem but dispersed nonspecifically throughout the genome. Southern analysis of cat cellular DNA confirmed the conclusions about conserved restriction sites in endogenous sequences and indicated that a single locus may be responsible for the production of the major inducible form of RD-114. Images PMID:6090693

Multi-focus image fusion using a guided-filter-based difference image.

PubMed

Yan, Xiang; Qin, Hanlin; Li, Jia; Zhou, Huixin; Yang, Tingwu

2016-03-20

The aim of multi-focus image fusion technology is to integrate different partially focused images into one all-focused image. To realize this goal, a new multi-focus image fusion method based on a guided filter is proposed and an efficient salient feature extraction method is presented in this paper. Furthermore, feature extraction is primarily the main objective of the present work. Based on salient feature extraction, the guided filter is first used to acquire the smoothing image containing the most sharpness regions. To obtain the initial fusion map, we compose a mixed focus measure by combining the variance of image intensities and the energy of the image gradient together. Then, the initial fusion map is further processed by a morphological filter to obtain a good reprocessed fusion map. Lastly, the final fusion map is determined via the reprocessed fusion map and is optimized by a guided filter. Experimental results demonstrate that the proposed method does markedly improve the fusion performance compared to previous fusion methods and can be competitive with or even outperform state-of-the-art fusion methods in terms of both subjective visual effects and objective quality metrics.

Bedrock and structural geologic maps of eastern Candor Sulci, western Ceti Mensa, and southeastern Ceti Mensa, Candor Chasma, Valles Marineris region of Mars

USGS Publications Warehouse

Okubo, Chris H.; Gaither, Tenielle A.

2017-05-12

This map product contains a set of three 1:18,000-scale maps showing the geology and structure of study areas in the western Candor Chasma region of Valles Marineris, Mars. These maps are part of an informal series of large-scale maps and map-based topical studies aimed at refining current understanding of the geologic history of western Candor Chasma. The map bases consist of digital elevation models and orthorectified images derived from High Resolution Imaging Science Experiment (HiRISE) data. These maps are accompanied by geologic cross sections, colorized elevation maps, and cutouts of HiRISE images showing key superposition relations. Also included in this product is a Correlation of Map Units that integrates units across all three map areas, as well as an integrated Description of Map Units and an integrated Explanation of Map Symbols. The maps were assembled using ArcGIS software produced by Environmental Systems Research Institute (http://www.esri.com). The ArcGIS projects and databases associated with each map are included online as supplemental data.

Laser speckle micro-rheology for biomechanical evaluation of breast tumors (Conference Presentation)

NASA Astrophysics Data System (ADS)

Hajjarian Kashany, Zeinab; Nadkarni, Seemantini K.

2016-03-01

The stiffness of the extra cellular matrix (ECM) is recognized as a key regulator of cancer cell proliferation, migration and invasion. Therefore technologies that quantify ECM stiffness with micro-scale scale resolution will likely provide important insights into neoplastic progression. Laser Speckle Micro-Rheology (LSM) is a novel optical tool for measuring tissue viscoelastic properties with micro-scale resolution. In LSM, speckle images are collected through an objective lens by a high-speed camera. Spatio-temporal correlation analysis of speckle frames yields the intensity autocorrelation function, g2(t), for each pixel, and subsequently a 2D map of viscoelastic modulus, G*(ω) is reconstructed. Here, we investigate the utility of LSM for micro-mechanical evaluation of the ECM in human breast lesions. Specimens collected 18 women undergoing lumpectomy or mastectomy were evaluated with LSM. Because collagen is the key protein associated with ECM stiffness, G*(ω) maps obtained from LSM were compared with collagen content measured by second harmonic generation (SHG) microscopy. Regions of low G*(ω), identified by LSM, corresponded to low-intensity SHG signal and adipose tissue. Likewise, regions with high G*(ω) in LSM images matched high intensity SHG signal caused by desmoplastic collagen accumulation. Quantitative regression analysis demonstrated a strong, statistically significant correlation between G*(ω) and SHG signal intensity (R=0.66 p< 0.01). These findings highlight the capability of LSM for quantifying the ECM micro-mechanics, potentially providing important insights into the biomechanical regulators of breast cancer progression.

STrategically Acquired Gradient Echo (STAGE) imaging, part I: Creating enhanced T1 contrast and standardized susceptibility weighted imaging and quantitative susceptibility mapping.

PubMed

Chen, Yongsheng; Liu, Saifeng; Wang, Yu; Kang, Yan; Haacke, E Mark

2018-02-01

To provide whole brain grey matter (GM) to white matter (WM) contrast enhanced T1W (T1WE) images, multi-echo quantitative susceptibility mapping (QSM), proton density (PD) weighted images, T1 maps, PD maps, susceptibility weighted imaging (SWI), and R2* maps with minimal misregistration in scanning times <5min. Strategically acquired gradient echo (STAGE) imaging includes two fully flow compensated double echo gradient echo acquisitions with a resolution of 0.67×1.33×2.0mm 3 acquired in 5min for 64 slices. Ten subjects were recruited and scanned at 3 Tesla. The optimum pair of flip angles (6° and 24° with TR=25ms at 3T) were used for both T1 mapping with radio frequency (RF) transmit field correction and creating enhanced GM/WM contrast (the T1WE). The proposed T1WE image was created from a combination of the proton density weighted (6°, PDW) and T1W (24°) images and corrected for RF transmit field variations. Prior to the QSM calculation, a multi-echo phase unwrapping strategy was implemented using the unwrapped short echo to unwrap the longer echo to speed up computation. R2* maps were used to mask deep grey matter and veins during the iterative QSM calculation. A weighted-average sum of susceptibility maps was generated to increase the signal-to-noise ratio (SNR) and the contrast-to-noise ratio (CNR). The proposed T1WE image has a significantly improved CNR both for WM to deep GM and WM to cortical GM compared to the acquired T1W image (the first echo of 24° scan) and the T1MPRAGE image. The weighted-average susceptibility maps have 80±26%, 55±22%, 108±33% SNR increases across the ten subjects compared to the single echo result of 17.5ms for the putamen, caudate nucleus, and globus pallidus, respectively. STAGE imaging offers the potential to create a standardized brain imaging protocol providing four pieces of quantitative tissue property information and multiple types of qualitative information in just 5min. Published by Elsevier Inc.

Using satellite data in map design and production

USGS Publications Warehouse

Hutchinson, John A.

2002-01-01

Satellite image maps have been produced by the U.S. Geological Survey (USGS) since shortly after the launch of the first Landsat satellite in 1972. Over the years, the use of image data to design and produce maps has developed from a manual and photographic process to one that incorporates geographic information systems, desktop publishing, and digital prepress techniques. At the same time, the content of most image-based maps produced by the USGS has shifted from raw image data to land cover or other information layers derived from satellite imagery, often portrayed in combination with shaded relief.

Topographic map of the western region of Dao Vallis in Hellas Planitia, Mars; MTM 500k -40/082E OMKT

USGS Publications Warehouse

Rosiek, Mark R.; Redding, Bonnie L.; Galuszka, Donna M.

2006-01-01

This map, compiled photogrammetrically from Viking Orbiter stereo image pairs, is part of a series of topographic maps of areas of special scientific interest on Mars. Contours were derived from a digital terrain model (DTM) compiled on a digital photogrammetric workstation using Viking Orbiter stereo image pairs with orientation parameters derived from an analytic aerotriangulation. The image base for this map employs Viking Orbiter images from orbits 406 and 363. An orthophotomosaic was created on the digital photogrammetric workstation using the DTM compiled from stereo models.

Sequential Superresolution Imaging of Multiple Targets Using a Single Fluorophore

PubMed Central

Lidke, Diane S.; Lidke, Keith A.

2015-01-01

Fluorescence superresolution (SR) microscopy, or fluorescence nanoscopy, provides nanometer scale detail of cellular structures and allows for imaging of biological processes at the molecular level. Specific SR imaging methods, such as localization-based imaging, rely on stochastic transitions between on (fluorescent) and off (dark) states of fluorophores. Imaging multiple cellular structures using multi-color imaging is complicated and limited by the differing properties of various organic dyes including their fluorescent state duty cycle, photons per switching event, number of fluorescent cycles before irreversible photobleaching, and overall sensitivity to buffer conditions. In addition, multiple color imaging requires consideration of multiple optical paths or chromatic aberration that can lead to differential aberrations that are important at the nanometer scale. Here, we report a method for sequential labeling and imaging that allows for SR imaging of multiple targets using a single fluorophore with negligible cross-talk between images. Using brightfield image correlation to register and overlay multiple image acquisitions with ~10 nm overlay precision in the x-y imaging plane, we have exploited the optimal properties of AlexaFluor647 for dSTORM to image four distinct cellular proteins. We also visualize the changes in co-localization of the epidermal growth factor (EGF) receptor and clathrin upon EGF addition that are consistent with clathrin-mediated endocytosis. These results are the first to demonstrate sequential SR (s-SR) imaging using direct stochastic reconstruction microscopy (dSTORM), and this method for sequential imaging can be applied to any superresolution technique. PMID:25860558

Raman Imaging in Cell Membranes, Lipid-Rich Organelles, and Lipid Bilayers.

PubMed

Syed, Aleem; Smith, Emily A

2017-06-12

Raman-based optical imaging is a promising analytical tool for noninvasive, label-free chemical imaging of lipid bilayers and cellular membranes. Imaging using spontaneous Raman scattering suffers from a low intensity that hinders its use in some cellular applications. However, developments in coherent Raman imaging, surface-enhanced Raman imaging, and tip-enhanced Raman imaging have enabled video-rate imaging, excellent detection limits, and nanometer spatial resolution, respectively. After a brief introduction to these commonly used Raman imaging techniques for cell membrane studies, this review discusses selected applications of these modalities for chemical imaging of membrane proteins and lipids. Finally, recent developments in chemical tags for Raman imaging and their applications in the analysis of selected cell membrane components are summarized. Ongoing developments toward improving the temporal and spatial resolution of Raman imaging and small-molecule tags with strong Raman scattering cross sections continue to expand the utility of Raman imaging for diverse cell membrane studies.

The Enceladus Atlas

NASA Image and Video Library

2010-05-13

This map sheet covers a 15-series image set covering the entire surface of Enceladus. The map data was acquired by NASA Cassini imaging experiment. Individual images can be viewed via the Photojournal.

Texture Analysis of Chaotic Coupled Map Lattices Based Image Encryption Algorithm

NASA Astrophysics Data System (ADS)

Khan, Majid; Shah, Tariq; Batool, Syeda Iram

2014-09-01

As of late, data security is key in different enclosures like web correspondence, media frameworks, therapeutic imaging, telemedicine and military correspondence. In any case, a large portion of them confronted with a few issues, for example, the absence of heartiness and security. In this letter, in the wake of exploring the fundamental purposes of the chaotic trigonometric maps and the coupled map lattices, we have presented the algorithm of chaos-based image encryption based on coupled map lattices. The proposed mechanism diminishes intermittent impact of the ergodic dynamical systems in the chaos-based image encryption. To assess the security of the encoded image of this scheme, the association of two nearby pixels and composition peculiarities were performed. This algorithm tries to minimize the problems arises in image encryption.

Deep embedding convolutional neural network for synthesizing CT image from T1-Weighted MR image.

PubMed

Xiang, Lei; Wang, Qian; Nie, Dong; Zhang, Lichi; Jin, Xiyao; Qiao, Yu; Shen, Dinggang

2018-07-01

Recently, more and more attention is drawn to the field of medical image synthesis across modalities. Among them, the synthesis of computed tomography (CT) image from T1-weighted magnetic resonance (MR) image is of great importance, although the mapping between them is highly complex due to large gaps of appearances of the two modalities. In this work, we aim to tackle this MR-to-CT synthesis task by a novel deep embedding convolutional neural network (DECNN). Specifically, we generate the feature maps from MR images, and then transform these feature maps forward through convolutional layers in the network. We can further compute a tentative CT synthesis from the midway of the flow of feature maps, and then embed this tentative CT synthesis result back to the feature maps. This embedding operation results in better feature maps, which are further transformed forward in DECNN. After repeating this embedding procedure for several times in the network, we can eventually synthesize a final CT image in the end of the DECNN. We have validated our proposed method on both brain and prostate imaging datasets, by also comparing with the state-of-the-art methods. Experimental results suggest that our DECNN (with repeated embedding operations) demonstrates its superior performances, in terms of both the perceptive quality of the synthesized CT image and the run-time cost for synthesizing a CT image. Copyright © 2018. Published by Elsevier B.V.

Physical limits to autofluorescence signals in vivo recordings in the rat olfactory bulb: a Monte Carlo study

NASA Astrophysics Data System (ADS)

L'Heureux, B.; Gurden, H.; Pinot, L.; Mastrippolito, R.; Lefebvre, F.; Lanièce, P.; Pain, F.

2007-07-01

Understanding the cellular mechanisms of energy supply to neurons following physiological activation is still challenging and has strong implications to the interpretation of clinical functional images based on metabolic signals such as Blood Oxygen Level Dependent Magnetic Resonance Imaging or 18F-Fluorodexoy-Glucose Positron Emission Tomography. Intrinsic Optical Signal Imaging provides with high spatio temporal resolution in vivo imaging in the anaesthetized rat. In that context, intrinsic signals are mainly related to changes in the optical absorption of haemoglobin depending on its oxygenation state. This technique has been validated for imaging of the rat olfactory bulb, providing with maps of the actived olfactory glomeruli, the functional modules involved in the first step of olfactory coding. A complementary approach would be autofluorescence imaging relying on the fluorescence properties of endogenous Flavin Adenine Dinucleotide (FAD) or Nicotinamide Adenine Dinucleotide (NADH) both involved in intracellular metabolic pathways. The purpose of the present study was to investigate the feasibility of in vivo autofluorescence imaging in the rat olfactory bulb. We performed standard Monte Carlo simulations of photons scattering and absorption at the excitation and emission wavelengths of FAD and NADH fluorescence. Characterization of the fluorescence distribution in the glomerulus, effect of hemoglobin absorption at the excitation and absorption wavelengths as well as the effect of the blurring due to photon scattering and the depth of focus of the optical apparatus have been studied. Finally, optimal experimental parameters are proposed to achieve in vivo validation of the technique in the rat olfactory bulb.

Earlier Detection of Tumor Treatment Response Using Magnetic Resonance Diffusion Imaging with Oscillating Gradients

PubMed Central

Colvin, Daniel C.; Loveless, Mary E.; Does, Mark D.; Yue, Zou; Yankeelov, Thomas E.; Gore, John C.

2011-01-01

An improved method for detecting early changes in tumors in response to treatment, based on a modification of diffusion-weighted magnetic resonance imaging, has been demonstrated in an animal model. Early detection of therapeutic response in tumors is important both clinically and in pre-clinical assessments of novel treatments. Non-invasive imaging methods that can detect and assess tumor response early in the course of treatment, and before frank changes in tumor morphology are evident, are of considerable interest as potential biomarkers of treatment efficacy. Diffusion-weighted magnetic resonance imaging is sensitive to changes in water diffusion rates in tissues that result from structural variations in the local cellular environment, but conventional methods mainly reflect changes in tissue cellularity and do not convey information specific to micro-structural variations at sub-cellular scales. We implemented a modified imaging technique using oscillating gradients of the magnetic field for evaluating water diffusion rates over very short spatial scales that are more specific for detecting changes in intracellular structure that may precede changes in cellularity. Results from a study of orthotopic 9L gliomas in rat brains indicate that this method can detect changes as early as 24 hours following treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), when conventional approaches do not find significant effects. These studies suggest that diffusion imaging using oscillating gradients may be used to obtain an earlier indication of treatment efficacy than previous magnetic resonance imaging methods. PMID:21190804

MO-DE-206-01: Cellular Metabolism of FDG

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pratx, G.

In this symposium jointly sponsored by the World Molecular Imaging Society (WMIS) and the AAPM, luminary speakers on imaging metabolism will discuss three impactful topics. The first presentation on Cellular Metabolism of FDG will be given by Guillem Pratx (Stanford). This presentation will detail new work on looking at how the most common molecular imaging agent, fluoro-deoxy-glucose is metabolized at a cellular level. This will be followed by a talk on an improved approach to whole-body PET imaging by Simon Cherry (UC Davis). Simon’s work on a new whole-body PET imaging system promises to have dramatic improvement in our abilitymore » to detect and characterize cancer using PET. Finally, Jim Bankson (MD Anderson) will discuss extremely sophisticated approaches to quantifying hyperpolarized-13-C pyruvate metabolism using MR imaging. This technology promises to compliment the exquisite sensitivity of PET with an ability to measure not just uptake, but tumor metabolism. Learning Objectives: Understand the metabolism of FDG at a cellular level. Appreciate the engineering related to a novel new high-sensitivity whole-body PET imaging system. Understand the process of hyperpolarization, how pyruvate relates to metabolism and how advanced modeling can be used to better quantify this data. G. Pratx, Funding: 5R01CA186275, 1R21CA193001, and Damon Runyon Cancer Foundation. S. Cherry, National Institutes of Health; University of California, Davis; Siemens Medical SolutionsJ. Bankson, GE Healthcare; NCI P30-CA016672; CPRIT PR140021-P5.« less

MO-DE-206-02: Cellular Metabolism of FDG

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cherry, S.

In this symposium jointly sponsored by the World Molecular Imaging Society (WMIS) and the AAPM, luminary speakers on imaging metabolism will discuss three impactful topics. The first presentation on Cellular Metabolism of FDG will be given by Guillem Pratx (Stanford). This presentation will detail new work on looking at how the most common molecular imaging agent, fluoro-deoxy-glucose is metabolized at a cellular level. This will be followed by a talk on an improved approach to whole-body PET imaging by Simon Cherry (UC Davis). Simon’s work on a new whole-body PET imaging system promises to have dramatic improvement in our abilitymore » to detect and characterize cancer using PET. Finally, Jim Bankson (MD Anderson) will discuss extremely sophisticated approaches to quantifying hyperpolarized-13-C pyruvate metabolism using MR imaging. This technology promises to compliment the exquisite sensitivity of PET with an ability to measure not just uptake, but tumor metabolism. Learning Objectives: Understand the metabolism of FDG at a cellular level. Appreciate the engineering related to a novel new high-sensitivity whole-body PET imaging system. Understand the process of hyperpolarization, how pyruvate relates to metabolism and how advanced modeling can be used to better quantify this data. G. Pratx, Funding: 5R01CA186275, 1R21CA193001, and Damon Runyon Cancer Foundation. S. Cherry, National Institutes of Health; University of California, Davis; Siemens Medical SolutionsJ. Bankson, GE Healthcare; NCI P30-CA016672; CPRIT PR140021-P5.« less

Coherent X-Ray Diffraction Imaging of Chloroplasts from Cyanidioschyzon merolae by Using X-Ray Free Electron Laser.

PubMed

Takayama, Yuki; Inui, Yayoi; Sekiguchi, Yuki; Kobayashi, Amane; Oroguchi, Tomotaka; Yamamoto, Masaki; Matsunaga, Sachihiro; Nakasako, Masayoshi

2015-07-01

Coherent X-ray diffraction imaging (CXDI) is a lens-less technique for visualizing the structures of non-crystalline particles with the dimensions of submicrometer to micrometer at a resolution of several tens of nanometers. We conducted cryogenic CXDI experiments at 66 K to visualize the internal structures of frozen-hydrated chloroplasts of Cyanidioschyzon merolae using X-ray free electron laser (XFEL) as a coherent X-ray source. Chloroplast dispersed specimen disks at a number density of 7/(10×10 µm(2)) were flash-cooled with liquid ethane without staining, sectioning or chemical labeling. Chloroplasts are destroyed at atomic level immediately after the diffraction by XFEL pulses. Thus, diffraction patterns with a good signal-to-noise ratio from single chloroplasts were selected from many diffraction patterns collected through scanning specimen disks to provide fresh specimens into the irradiation area. The electron density maps of single chloroplasts projected along the direction of the incident X-ray beam were reconstructed by using the iterative phase-retrieval method and multivariate analyses. The electron density map at a resolution of 70 nm appeared as a C-shape. In addition, the fluorescence image of proteins stained with Flamingo™ dye also appeared as a C-shape as did the autofluorescence from Chl. The similar images suggest that the thylakoid membranes with an abundance of proteins distribute along the outer membranes of chloroplasts. To confirm the present results statistically, a number of projection structures must be accumulated through high-throughput data collection in the near future. Based on the results, we discuss the feasibility of XFEL-CXDI experiments in the structural analyses of cellular organelles. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging

PubMed Central

Quinto-Su, Pedro A.; Lai, Hsuan-Hong; Yoon, Helen H.; Sims, Christopher E.; Allbritton, Nancy L.; Venugopalan, Vasan

2008-01-01

We use time-resolved imaging to examine the lysis dynamics of non-adherent BAF-3 cells within a microfluidic channel produced by the delivery of single highly-focused 540 ps duration laser pulses at λ = 532 nm. Time-resolved bright-field images reveal that the delivery of the pulsed laser microbeam results in the formation of a laser-induced plasma followed by shock wave emission and cavitation bubble formation. The confinement offered by the microfluidic channel constrains substantially the cavitation bubble expansion and results in significant deformation of the PDMS channel walls. To examine the cell lysis and dispersal of the cellular contents, we acquire time-resolved fluorescence images of the process in which the cells were loaded with a fluorescent dye. These fluorescence images reveal cell lysis to occur on the nanosecond to microsecond time scale by the plasma formation and cavitation bubble dynamics. Moreover, the time-resolved fluorescence images show that while the cellular contents are dispersed by the expansion of the laser-induced cavitation bubble, the flow associated with the bubble collapse subsequently re-localizes the cellular contents to a small region. This capacity of pulsed laser microbeam irradiation to achieve rapid cell lysis in microfluidic channels with minimal dilution of the cellular contents has important implications for their use in lab-on-a-chip applications. PMID:18305858

Images of the World: Mental Maps of U.S. Military Officers

DTIC Science & Technology

1992-05-01

only displayed the geographic characteristics of the countries, (the direction, distance, and size; all relative to the U.S.) but the maps also indicate...individual’s mental map is based upon learned facts and exposure to impressionable images. Facts and images of the world constitute the foundation from... characteristics of the world are mostly static, images people have of places are dynamic and fluid. Graphically, this is the main difference between

X-ray fluorescence microscopy reveals the role of selenium in spermatogenesis

PubMed Central

Kehr, Sebastian; Malinouski, Mikalai; Finney, Lydia; Vogt, Stefan; Labunskyy, Vyacheslav M.; Kasaikina, Marina V.; Carlson, Bradley A.; Zhou, You; Hatfield, Dolph L.; Gladyshev, Vadim N.

2009-01-01

Selenium (Se) is a trace element with important roles in human health. Several selenoproteins have essential functions in development. However, the cellular and tissue distribution of Se remains largely unknown because of the lack of analytical techniques that image this element with sufficient sensitivity and resolution. Herein, we report that X-ray fluorescence microscopy (XFM) can be used to visualize and quantify the tissue, cellular and subcellular topography of Se. We applied this technique to characterize the role of Se in spermatogenesis and identified a dramatic Se enrichment specifically in late spermatids, a pattern that was not seen in any other elemental maps. This enrichment was due to elevated levels of the mitochondrial form of glutathione peroxidase 4 and was fully dependent on the supplies of Se by Selenoprotein P. High-resolution scans revealed that Se concentrated near the lumen side of elongating spermatids, where structural components of sperm are formed. During spermatogenesis, maximal Se associated with decreased phosphorus, whereas Zn did not change. In sperm, Se was primarily in the midpiece and co-localized with Cu and Fe. XFM allowed quantification of Se in the midpiece (0.8 fg) and head (0.14 fg) of individual sperm cells, revealing the ability of sperm cells to handle the amounts of this element well above its toxic levels. Overall, the use of XFM allowed visualization of tissue and cellular Se and provided important insights in the role of this and other trace elements in spermatogenesis. PMID:19379757

36 CFR 1194.22 - Web-based intranet and internet information and applications.

Code of Federal Regulations, 2012 CFR

2012-07-01

... active region of a server-side image map. (f) Client-side image maps shall be provided instead of server-side image maps except where the regions cannot be defined with an available geometric shape. (g) Row...) Frames shall be titled with text that facilitates frame identification and navigation. (j) Pages shall be...

36 CFR 1194.22 - Web-based intranet and internet information and applications.

Code of Federal Regulations, 2014 CFR

2014-07-01

... active region of a server-side image map. (f) Client-side image maps shall be provided instead of server-side image maps except where the regions cannot be defined with an available geometric shape. (g) Row...) Frames shall be titled with text that facilitates frame identification and navigation. (j) Pages shall be...

36 CFR § 1194.22 - Web-based intranet and internet information and applications.

Code of Federal Regulations, 2013 CFR

2013-07-01

... active region of a server-side image map. (f) Client-side image maps shall be provided instead of server-side image maps except where the regions cannot be defined with an available geometric shape. (g) Row...) Frames shall be titled with text that facilitates frame identification and navigation. (j) Pages shall be...

Integrating polarimetric synthetic aperture radar and imaging spectrometry for wildland fuel mapping in southern California

Treesearch

P.E. Dennison; D.A. Roberts; J. Regelbrugge; S.L. Ustin

2000-01-01

Polarimetric synthetic aperture radar (SAR) and imaging spectrometry exemplify advanced technologies for mapping wildland fuels in chaparral ecosystems. In this study, we explore the potential of integrating polarimetric SAR and imaging spectrometry for mapping wildland fuels. P-band SAR and ratios containing P-band polarizations are sensitive to variations in stand...

A chaotic cryptosystem for images based on Henon and Arnold cat map.

PubMed

Soleymani, Ali; Nordin, Md Jan; Sundararajan, Elankovan

2014-01-01

The rapid evolution of imaging and communication technologies has transformed images into a widespread data type. Different types of data, such as personal medical information, official correspondence, or governmental and military documents, are saved and transmitted in the form of images over public networks. Hence, a fast and secure cryptosystem is needed for high-resolution images. In this paper, a novel encryption scheme is presented for securing images based on Arnold cat and Henon chaotic maps. The scheme uses Arnold cat map for bit- and pixel-level permutations on plain and secret images, while Henon map creates secret images and specific parameters for the permutations. Both the encryption and decryption processes are explained, formulated, and graphically presented. The results of security analysis of five different images demonstrate the strength of the proposed cryptosystem against statistical, brute force and differential attacks. The evaluated running time for both encryption and decryption processes guarantee that the cryptosystem can work effectively in real-time applications.