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Bioluminescence Imaging  

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Bioluminescence refers to the process of visible light emission in living organisms. Bioluminescence imaging is a powerful methodology that has been developed over the last decade as a tool for molecular imaging of small laboratory animals, enabling the study of ongoing biological processes in vivo. This form of optical imaging is low cost and noninvasive and facilitates real-time analysis of disease processes at the molecular level in living organisms. In this article, we provide a brief int...

Sadikot, Ruxana T.; Blackwell, Timothy S.

2005-01-01

2

Bioanalytical Applications of Real-Time ATP Imaging Via Bioluminescence  

Energy Technology Data Exchange (ETDEWEB)

The research discussed within involves the development of novel applications of real-time imaging of adenosine 5'-triphosphate (ATP). ATP was detected via bioluminescence and the firefly luciferase-catalyzed reaction of ATP and luciferin. The use of a microscope and an imaging detector allowed for spatially resolved quantitation of ATP release. Employing this method, applications in both biological and chemical systems were developed. First, the mechanism by which the compound 48/80 induces release of ATP from human umbilical vein endothelial cells (HUVECs) was investigated. Numerous enzyme activators and inhibitors were utilized to probe the second messenger systems involved in release. Compound 48/80 activated a G{sub q}-type protein to initiate ATP release from HUVECs. Ca{sup 2+} imaging along with ATP imaging revealed that activation of phospholipase C and induction of intracellular Ca{sup 2+} signaling were necessary for release of ATP. Furthermore, activation of protein kinase C inhibited the activity of phospholipase C and thus decreased the magnitude of ATP release. This novel release mechanism was compared to the existing theories of extracellular release of ATP. Bioluminescence imaging was also employed to examine the role of ATP in the field of neuroscience. The central nervous system (CNS) was dissected from the freshwater snail Lymnaea stagnalis. Electrophysiological experiments demonstrated that the neurons of the Lymnaea were not damaged by any of the components of the imaging solution. ATP was continuously released by the ganglia of the CNS for over eight hours and varied from ganglion to ganglion and within individual ganglia. Addition of the neurotransmitters K{sup +} and serotonin increased release of ATP in certain regions of the Lymnaea CNS. Finally, the ATP imaging technique was investigated for the study of drug release systems. MCM-41-type mesoporous nanospheres were loaded with ATP and end-capped with mercaptoethanol functionalized CdS monocrystals. Aggregates of nanospheres were bathed in imaging solution, and ATP bioluminescence was monitored to investigated the release kinetics of the nanosphere drug delivery systems. Addition of disulfide bond-cleaving molecules induced uncapping of the nanospheres and subsequently, the release of ATP. Increasing the concentration of the uncapping molecule decreased the temporal maximum and increased the magnitude of release of encapsulated ATP from the nanospheres. Furthermore, the release kinetics from the nanospheres varied with the size of the particle aggregates.

Jason Alan Gruenhagen

2003-12-12

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Timing of Imaging after D-Luciferin Injection Affects the Longitudinal Assessment of Tumor Growth Using In Vivo Bioluminescence Imaging  

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The peak signal or the signal at a predetermined, fixed time point after D-luciferin injection may be used for the quantitative analysis of in vivo bioluminescence imaging. We repeatedly performed sequential bioluminescence imaging after subcutaneous injection of D-luciferin in mice bearing subcutaneous tumors. The peak time in each measurement became shorter early after cell inoculation, presumably due to gradual establishment of intratumoral vasculature, and reached a plateau of about 10 mi...

2010-01-01

4

Bioluminescence microscopy using a short focal-length imaging lens.  

Science.gov (United States)

Bioluminescence from cells is so dim that bioluminescence microscopy is performed using an ultra low-light imaging camera. Although the image sensor of such cameras has been greatly improved over time, such improvements have not been made commercially available for microscopes until now. Here, we customized the optical system of a microscope for bioluminescence imaging. As a result, bioluminescence images of cells could be captured with a conventional objective lens and colour imaging camera. As bioluminescence microscopy requires no excitation light, it lacks the photo-toxicity associated with fluorescence imaging and permits the long-term, nonlethal observation of living cells. Thus, bioluminescence microscopy would be a powerful tool in cellular biology that complements fluorescence microscopy. PMID:24386879

Ogoh, K; Akiyoshi, R; May-Maw-Thet; Sugiyama, T; Dosaka, S; Hatta-Ohashi, Y; Suzuki, H

2014-03-01

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Measuring prions by bioluminescence imaging  

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Prions are infectious proteins that cause fatal neurodegenerative diseases. Because astrocytic gliosis marked by the deposition of fibrils composed of GFAP is a prominent feature of prion disease, we asked whether GFAP might be used as a surrogate marker for prions. To interrogate this posit, we inoculated prions into transgenic (Tg) mice expressing luciferase (luc) under the GFAP gene (Gfap) promoter, denoted Tg(Gfap-luc) mice. Weekly noninvasive, bioluminescence imaging (BLI) detected an in...

Tamgu?ney, Gu?ltekin; Francis, Kevin P.; Giles, Kurt; Lemus, Azucena; Dearmond, Stephen J.; Prusiner, Stanley B.

2009-01-01

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Bioluminescent imaging of Trypanosoma cruzi infection in Rhodnius prolixus  

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Full Text Available Abstract Background Usually the analysis of the various developmental stages of Trypanosoma cruzi in the experimentally infected vertebrate and invertebrate hosts is based on the morphological observations of tissue fragments from animals and insects. The development of techniques that allow the imaging of animals infected with parasites expressing luciferase open up possibilities to follow the fate of bioluminescent parasites in infected vectors. Methods D-luciferin (60 ?g was injected into the hemocoel of the whole insect before bioluminescence acquisition. In dissected insects, the whole gut was incubated with D-luciferin in PBS (300 ?g/ml for ex vivo bioluminescence acquisition in the IVIS® Imaging System, Xenogen. Results Herein, we describe the results obtained with the luciferase gene integrated into the genome of the Dm28c clone of T. cruzi, and the use of these parasites to follow, in real time, the infection of the insect vector Rhodnius prolixus, by a non- invasive method. The insects were evaluated by in vivo bioluminescent imaging on the feeding day, and on the 7 th, 14 th, 21 st and 28 th days after feeding. To corroborate the bioluminescent imaging made in vivo, and investigate the digestive tract region, the insects were dissected. The bioluminescence emitted was proportional to the number of protozoans in regions of the gut. The same digestive tracts were also macerated to count the parasites in distinct morphological stages with an optical microscope, and for bioluminescence acquisition in a microplate using the IVIS® Imaging System. A positive correlation of parasite numbers and bioluminescence in the microplate was obtained. Conclusions This is the first report of bioluminescent imaging in Rhodnius prolixus infected with trypomastigotes of the Dm28c-luc stable strain, expressing firefly luciferase. In spite of the distribution limitations of the substrate (D-luciferin in the insect body, longitudinal evaluation of infected insects by bioluminescent imaging is a valuable tool. Bioluminescent imaging of the digestive tract infected with Dm28c-luc is highly sensitive and accurate method to track the fate of the parasite in the vector, in the crop, intestine and rectum. This methodology is useful to gain a better understanding of the parasite – insect vector interactions.

Henriques Cristina

2012-09-01

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In vivo dual substrate bioluminescent imaging.  

Science.gov (United States)

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate since the advent of in vivo bioluminescent imaging technologies. Indeed, the ability to locate and quantify tumor growth longitudinally in a single cohort of animals to completion of the study as opposed to sacrificing individual groups of animals at specific assay times has revolutionized how researchers investigate breast cancer metastasis. Unfortunately, current methodologies preclude the real-time assessment of critical changes that transpire in cell signaling systems as breast cancer cells (i) evolve within primary tumors, (ii) disseminate throughout the body, and (iii) reinitiate proliferative programs at sites of a metastatic lesion. However, recent advancements in bioluminescent imaging now make it possible to simultaneously quantify specific spatiotemporal changes in gene expression as a function of tumor development and metastatic progression via the use of dual substrate luminescence reactions. To do so, researchers take advantage for two light-producing luciferase enzymes isolated from the firefly (Photinus pyralis) and sea pansy (Renilla reniformis), both of which react to mutually exclusive substrates that previously facilitated their wide-spread use in in vitro cell-based reporter gene assays. Here we demonstrate the in vivo utility of these two enzymes such that one luminescence reaction specifically marks the size and location of a developing tumor, while the second luminescent reaction serves as a means to visualize the activation status of specific signaling systems during distinct stages of tumor and metastasis development. Thus, the objectives of this study are two-fold. First, we will describe the steps necessary to construct dual bioluminescent reporter cell lines, as well as those needed to facilitate their use in visualizing the spatiotemporal regulation of gene expression during specific steps of the metastatic cascade. Using the 4T1 model of breast cancer metastasis, we show that the in vivo activity of a synthetic Smad Binding Element (SBE) promoter was decreased dramatically in pulmonary metastasis as compared to that measured in the primary tumor. Recently, breast cancer metastasis was shown to be regulated by changes within the primary tumor microenvironment and reactive stroma, including those occurring in fibroblasts and infiltrating immune cells. Thus, our second objective will be to demonstrate the utility of dual bioluminescent techniques in monitoring the growth and localization of two unique cell populations harbored within a single animal during breast cancer growth and metastasis. PMID:22006228

Wendt, Michael K; Molter, Joseph; Flask, Christopher A; Schiemann, William P

2011-01-01

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Bioluminescent system for dynamic imaging of cell and animal behavior  

Energy Technology Data Exchange (ETDEWEB)

Highlights: Black-Right-Pointing-Pointer We combined a yellow variant of GFP and firefly luciferase to make ffLuc-cp156. Black-Right-Pointing-Pointer ffLuc-cp156 showed improved photon yield in cultured cells and transgenic mice. Black-Right-Pointing-Pointer ffLuc-cp156 enabled video-rate bioluminescence imaging of freely-moving animals. Black-Right-Pointing-Pointer ffLuc-cp156 mice enabled tracking real-time drug delivery in conscious animals. -- Abstract: The current utility of bioluminescence imaging is constrained by a low photon yield that limits temporal sensitivity. Here, we describe an imaging method that uses a chemiluminescent/fluorescent protein, ffLuc-cp156, which consists of a yellow variant of Aequorea GFP and firefly luciferase. We report an improvement in photon yield by over three orders of magnitude over current bioluminescent systems. We imaged cellular movement at high resolution including neuronal growth cones and microglial cell protrusions. Transgenic ffLuc-cp156 mice enabled video-rate bioluminescence imaging of freely moving animals, which may provide a reliable assay for drug distribution in behaving animals for pre-clinical studies.

Hara-Miyauchi, Chikako [Department of Physiology, Keio University School of Medicine, Tokyo 160-8582 (Japan); Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Saitama 351-0198 (Japan); Department of Biophysics and Biochemistry, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Tokyo 113-8510 (Japan); Tsuji, Osahiko [Department of Physiology, Keio University School of Medicine, Tokyo 160-8582 (Japan); Department of Orthopedic Surgery, Keio University School of Medicine, Tokyo 160-8582 (Japan); Hanyu, Aki [Division of Biochemistry, The Cancer Institute of the Japanese Foundation for Cancer Research, Tokyo 135-8550 (Japan); Okada, Seiji [Department of Advanced Medical Initiatives, Faculty of Medical Sciences, Kyushu University, Fukuoka 812-8582 (Japan); Yasuda, Akimasa [Department of Physiology, Keio University School of Medicine, Tokyo 160-8582 (Japan); Department of Orthopedic Surgery, Keio University School of Medicine, Tokyo 160-8582 (Japan); Fukano, Takashi [Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Saitama 351-0198 (Japan); Akazawa, Chihiro [Department of Biophysics and Biochemistry, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Tokyo 113-8510 (Japan); Nakamura, Masaya [Department of Orthopedic Surgery, Keio University School of Medicine, Tokyo 160-8582 (Japan); Imamura, Takeshi [Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Toon, Ehime 791-0295 (Japan); Core Research for Evolutional Science and Technology, The Japan Science and Technology Corporation, Tokyo 135-8550 (Japan); Matsuzaki, Yumi [Department of Physiology, Keio University School of Medicine, Tokyo 160-8582 (Japan); Okano, Hirotaka James, E-mail: hjokano@jikei.ac.jp [Department of Physiology, Keio University School of Medicine, Tokyo 160-8582 (Japan); Division of Regenerative Medicine Jikei University School of Medicine, Tokyo 150-8461 (Japan); and others

2012-03-09

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Bioluminescent system for dynamic imaging of cell and animal behavior  

International Nuclear Information System (INIS)

Highlights: ? We combined a yellow variant of GFP and firefly luciferase to make ffLuc-cp156. ? ffLuc-cp156 showed improved photon yield in cultured cells and transgenic mice. ? ffLuc-cp156 enabled video-rate bioluminescence imaging of freely-moving animals. ? ffLuc-cp156 mice enabled tracking real-time drug delivery in conscious animals. -- Abstract: The current utility of bioluminescence imaging is constrained by a low photon yield that limits temporal sensitivity. Here, we describe an imaging method that uses a chemiluminescent/fluorescent protein, ffLuc-cp156, which consists of a yellow variant of Aequorea GFP and firefly luciferase. We report an improvement in photon yield by over three orders of magnitude over current bioluminescent systems. We imaged cellular movement at high resolution including neuronal growth cones and microglial cell protrusions. Transgenic ffLuc-cp156 mice enabled video-rate bioluminescence imaging of freely moving animals, which may provide a reliable assay for drug distribution in behaving animals for pre-clinical studies.

2012-03-09

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Dynamic bioluminescence imaging for quantitative tumour burden assessment using IV or IP administration of d-luciferin: effect on intensity, time kinetics and repeatability of photon emission  

International Nuclear Information System (INIS)

In vivo bioluminescence imaging (BLI) is a promising technique for non-invasive tumour imaging. d-luciferin can be administrated intraperitonealy or intravenously. This will influence its availability and, therefore, the bioluminescent signal. The aim of this study is to compare the repeatability of BLI measurement after IV versus IP administration of d-luciferin and assess the correlation between photon emission and histological cell count both in vitro and in vivo. Fluc-positive R1M cells were subcutaneously inoculated in nu/nu mice. Dynamic BLI was performed after IV or IP administration of d-luciferin. Maximal photon emission (PEmax) was calculated. For repeatability assessment, every acquisition was repeated after 4 h and analysed using Bland-Altman method. A second group of animals was serially imaged, alternating IV and IP administration up to 21 days. When mice were killed, PEmax after IV administration was correlated with histological cell number. The coefficients of repeatability were 80.2% (IV) versus 95.0% (IP). Time-to-peak is shorter, and its variance lower for IV (p max was 5.6 times higher for IV. A trend was observed towards lower photon emission per cell in larger tumours. IV administration offers better repeatability and better sensitivity when compared to IP. In larger tumours, multiple factors may contribute to underestimation of tumour burden. It might, therefore, be beneficial to test novel therapeutics on small tumours to enable an accurate evaluation of tumour burden. (orig.)

2008-05-01

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Bioluminescence imaging of myeloperoxidase activity in vivo  

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The myeloperoxidase (MPO) system of activated phagocytes is central to normal host defense mechanisms, and dysregulated MPO contributes to the pathogenesis of inflammatory disease states ranging from atherosclerosis to cancer. Here we show that upon systemic administration, the small molecule luminol enables noninvasive bioluminescence imaging (BLI) of MPO activity in vivo. Luminol-BLI allowed quantitative longitudinal monitoring of MPO activity in animal models of acute dermatitis, mixed all...

Gross, Shimon; Gammon, Seth T.; Moss, Britney L.; Rauch, Daniel; Harding, John; Heinecke, Jay W.; Ratner, Lee; Piwnica-worms, David

2009-01-01

12

Dynamic bioluminescence imaging for quantitative tumour burden assessment using IV or IP administration of d-luciferin: effect on intensity, time kinetics and repeatability of photon emission  

Energy Technology Data Exchange (ETDEWEB)

In vivo bioluminescence imaging (BLI) is a promising technique for non-invasive tumour imaging. d-luciferin can be administrated intraperitonealy or intravenously. This will influence its availability and, therefore, the bioluminescent signal. The aim of this study is to compare the repeatability of BLI measurement after IV versus IP administration of d-luciferin and assess the correlation between photon emission and histological cell count both in vitro and in vivo. Fluc-positive R1M cells were subcutaneously inoculated in nu/nu mice. Dynamic BLI was performed after IV or IP administration of d-luciferin. Maximal photon emission (PE{sub max}) was calculated. For repeatability assessment, every acquisition was repeated after 4 h and analysed using Bland-Altman method. A second group of animals was serially imaged, alternating IV and IP administration up to 21 days. When mice were killed, PE{sub max} after IV administration was correlated with histological cell number. The coefficients of repeatability were 80.2% (IV) versus 95.0% (IP). Time-to-peak is shorter, and its variance lower for IV (p < 0.0001). PE{sub max} was 5.6 times higher for IV. A trend was observed towards lower photon emission per cell in larger tumours. IV administration offers better repeatability and better sensitivity when compared to IP. In larger tumours, multiple factors may contribute to underestimation of tumour burden. It might, therefore, be beneficial to test novel therapeutics on small tumours to enable an accurate evaluation of tumour burden. (orig.)

Keyaerts, Marleen; Vanhove, Chris; Caveliers, Vicky; Bossuyt, Axel; Lahoutte, Tony [Vrije Universiteit Brussel (VUB), In Vivo Cellular and Molecular Imaging (ICMI) Laboratory, Brussels (Belgium); University Hospital Brussels (UZ-Brussel), Department of Nuclear Medicine, Brussels (Belgium); Verschueren, Jacob [University of Antwerp, Bio-Imaging lab, Department of Biomedical Sciences, Antwerp (Belgium); Bos, Tomas J. [Vrije Universiteit Brussel (VUB), Department of Haematology and Immunology, Brussels (Belgium); Tchouate-Gainkam, Lea O.; Peleman, Cindy [Vrije Universiteit Brussel (VUB), In Vivo Cellular and Molecular Imaging (ICMI) Laboratory, Brussels (Belgium); Breckpot, Karine [Vrije Universiteit Brussel (VUB), Laboratory of Molecular and Cellular Therapy, Department of Physiology and Immunology, Brussels (Belgium)

2008-05-15

13

Filtering and deconvolution for bioluminescence imaging of small animals  

International Nuclear Information System (INIS)

This thesis is devoted to analysis of bioluminescence images applied to the small animal. This kind of imaging modality is used in cancerology studies. Nevertheless, some problems are related to the diffusion and the absorption of the tissues of the light of internal bioluminescent sources. In addition, system noise and the cosmic rays noise are present. This influences the quality of the images and makes it difficult to analyze. The purpose of this thesis is to overcome these disturbing effects. We first have proposed an image formation model for the bioluminescence images. The processing chain is constituted by a filtering stage followed by a deconvolution stage. We have proposed a new median filter to suppress the random value impulsive noise which corrupts the acquired images; this filter represents the first block of the proposed chain. For the deconvolution stage, we have performed a comparative study of various deconvolution algorithms. It allowed us to choose a blind deconvolution algorithm initialized with the estimated point spread function of the acquisition system. At first, we have validated our global approach by comparing our obtained results with the ground truth. Through various clinical tests, we have shown that the processing chain allows a significant improvement of the spatial resolution and a better distinction of very close tumor sources, what represents considerable contribution for the users of bioluminescence images. (author)

2010-01-01

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Computer-aided photometric analysis of dynamic digital bioluminescent images  

Science.gov (United States)

The paper deals with photometric and morphologic analysis of bioluminescent images obtained by registration of light radiated directly from some plant objects. Registration of images obtained from ultra-weak light sources by the single photon counting (SPC) technique is the subject of this work. The radiation is registered by use of a 16-bit charge coupled device (CCD) camera "Night Owl" together with WinLight EG&G Berthold software. Additional application-specific software has been developed in order to deal with objects that are changing during the exposition time. Advantages of the elaborated set of easy configurable tools named FCT for a computer-aided photometric and morphologic analysis of numerous series of quantitatively imperfect chemiluminescent images are described. Instructions are given how to use these tools and exemplified with several algorithms for the transformation of images library. Using the proposed FCT set, automatic photometric and morphologic analysis of the information hidden within series of chemiluminescent images reflecting defensive processes in poinsettia (Euphorbia pulcherrima Willd) leaves affected by a pathogenic fungus Botrytis cinerea is revealed.

Gorski, Zbigniew; Bembnista, T.; Floryszak-Wieczorek, J.; Domanski, Marek; Slawinski, Janusz

2003-04-01

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Triple bioluminescence imaging for in vivo monitoring of cellular processes.  

Science.gov (United States)

Bioluminescence imaging (BLI) has shown to be crucial for monitoring in vivo biological processes. So far, only dual bioluminescence imaging using firefly (Fluc) and Renilla or Gaussia (Gluc) luciferase has been achieved due to the lack of availability of other efficiently expressed luciferases using different substrates. Here, we characterized a codon-optimized luciferase from Vargula hilgendorfii (Vluc) as a reporter for mammalian gene expression. We showed that Vluc can be multiplexed with Gluc and Fluc for sequential imaging of three distinct cellular phenomena in the same biological system using vargulin, coelenterazine, and D-luciferin substrates, respectively. We applied this triple imaging system to monitor the effect of soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL) delivered using an adeno-associated viral vector (AAV) on brain tumors in mice. Vluc imaging showed efficient sTRAIL gene delivery to the brain, while Fluc imaging revealed a robust antiglioma therapy. Further, nuclear factor-?B (NF-?B) activation in response to sTRAIL binding to glioma cells death receptors was monitored by Gluc imaging. This work is the first demonstration of trimodal in vivo bioluminescence imaging and will have a broad applicability in many different fields including immunology, oncology, virology, and neuroscience.Molecular Therapy - Nucleic Acids (2013) 2, e99; doi:10.1038/mtna.2013.25; published online 18 June 2013. PMID:23778500

Maguire, Casey A; Bovenberg, M Sarah; Crommentuijn, Matheus Hw; Niers, Johanna M; Kerami, Mariam; Teng, Jian; Sena-Esteves, Miguel; Badr, Christian E; Tannous, Bakhos A

2013-01-01

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A synthetic luciferin improves bioluminescence imaging in live mice.  

Science.gov (United States)

Firefly luciferase is the most widely used optical reporter for noninvasive bioluminescence imaging (BLI) in rodents. BLI relies on the ability of the injected luciferase substrate D-luciferin to access luciferase-expressing cells and tissues within the animal. Here we show that injection of mice with a synthetic luciferin, CycLuc1, improves BLI with existing luciferase reporters and enables imaging in the brain that could not be achieved with D-luciferin. PMID:24509630

Evans, Melanie S; Chaurette, Joanna P; Adams, Spencer T; Reddy, Gadarla R; Paley, Miranda A; Aronin, Neil; Prescher, Jennifer A; Miller, Stephen C

2014-04-01

17

Autonomously bioluminescent mammalian cells for continuous and real-time monitoring of cytotoxicity.  

Science.gov (United States)

Mammalian cell-based in vitro assays have been widely employed as alternatives to animal testing for toxicological studies but have been limited due to the high monetary and time costs of parallel sample preparation that are necessitated due to the destructive nature of firefly luciferase-based screening methods. This video describes the utilization of autonomously bioluminescent mammalian cells, which do not require the destructive addition of a luciferin substrate, as an inexpensive and facile method for monitoring the cytotoxic effects of a compound of interest. Mammalian cells stably expressing the full bacterial bioluminescence (luxCDABEfrp) gene cassette autonomously produce an optical signal that peaks at 490 nm without the addition of an expensive and possibly interfering luciferin substrate, excitation by an external energy source, or destruction of the sample that is traditionally performed during optical imaging procedures. This independence from external stimulation places the burden for maintaining the bioluminescent reaction solely on the cell, meaning that the resultant signal is only detected during active metabolism. This characteristic makes the lux-expressing cell line an excellent candidate for use as a biosentinel against cytotoxic effects because changes in bioluminescent production are indicative of adverse effects on cellular growth and metabolism. Similarly, the autonomous nature and lack of required sample destruction permits repeated imaging of the same sample in real-time throughout the period of toxicant exposure and can be performed across multiple samples using existing imaging equipment in an automated fashion. PMID:24193545

Xu, Tingting; Close, Dan M; Webb, James D; Ripp, Steven A; Sayler, Gary S

2013-01-01

18

Hyperspectral and multispectral bioluminescence optical tomography for small animal imaging  

Energy Technology Data Exchange (ETDEWEB)

For bioluminescence imaging studies in small animals, it is important to be able to accurately localize the three-dimensional (3D) distribution of the underlying bioluminescent source. The spectrum of light produced by the source that escapes the subject varies with the depth of the emission source because of the wavelength-dependence of the optical properties of tissue. Consequently, multispectral or hyperspectral data acquisition should help in the 3D localization of deep sources. In this paper, we describe a framework for fully 3D bioluminescence tomographic image acquisition and reconstruction that exploits spectral information. We describe regularized tomographic reconstruction techniques that use semi-infinite slab or FEM-based diffusion approximations of photon transport through turbid media. Singular value decomposition analysis was used for data dimensionality reduction and to illustrate the advantage of using hyperspectral rather than achromatic data. Simulation studies in an atlas-mouse geometry indicated that sub-millimeter resolution may be attainable given accurate knowledge of the optical properties of the animal. A fixed arrangement of mirrors and a single CCD camera were used for simultaneous acquisition of multispectral imaging data over most of the surface of the animal. Phantom studies conducted using this system demonstrated our ability to accurately localize deep point-like sources and show that a resolution of 1.5 to 2.2 mm for depths up to 6 mm can be achieved. We also include an in vivo study of a mouse with a brain tumour expressing firefly luciferase. Co-registration of the reconstructed 3D bioluminescent image with magnetic resonance images indicated good anatomical localization of the tumour.

Chaudhari, Abhijit J [Signal and Image Processing Institute, Department of Electrical Engineering-Systems, University of Southern California, Los Angeles, CA 90089 (United States); Darvas, Felix [Signal and Image Processing Institute, Department of Electrical Engineering-Systems, University of Southern California, Los Angeles, CA 90089 (United States); Bading, James R [Department of Radiology, University of Southern California, Los Angeles, CA 90033 (United States); Moats, Rex A [Department of Radiology, University of Southern California, Los Angeles, CA 90033 (United States); Conti, Peter S [Department of Radiology, University of Southern California, Los Angeles, CA 90033 (United States); Smith, Desmond J [Department of Molecular and Medical Pharmacology, UCLA School of Medicine, Los Angeles, CA 90095 (United States); Cherry, Simon R [Department of Biomedical Engineering, University of California-Davis, Davis, CA 95616 (United States); Leahy, Richard M [Signal and Image Processing Institute, Department of Electrical Engineering-Systems, University of Southern California, Los Angeles, CA 90089 (United States)

2005-12-07

19

Hyperspectral and multispectral bioluminescence optical tomography for small animal imaging  

International Nuclear Information System (INIS)

For bioluminescence imaging studies in small animals, it is important to be able to accurately localize the three-dimensional (3D) distribution of the underlying bioluminescent source. The spectrum of light produced by the source that escapes the subject varies with the depth of the emission source because of the wavelength-dependence of the optical properties of tissue. Consequently, multispectral or hyperspectral data acquisition should help in the 3D localization of deep sources. In this paper, we describe a framework for fully 3D bioluminescence tomographic image acquisition and reconstruction that exploits spectral information. We describe regularized tomographic reconstruction techniques that use semi-infinite slab or FEM-based diffusion approximations of photon transport through turbid media. Singular value decomposition analysis was used for data dimensionality reduction and to illustrate the advantage of using hyperspectral rather than achromatic data. Simulation studies in an atlas-mouse geometry indicated that sub-millimeter resolution may be attainable given accurate knowledge of the optical properties of the animal. A fixed arrangement of mirrors and a single CCD camera were used for simultaneous acquisition of multispectral imaging data over most of the surface of the animal. Phantom studies conducted using this system demonstrated our ability to accurately localize deep point-like sources and show that a resolution of 1.5 to 2.2 mm for depths up to 6 mm can be achieved. We also include an in vivo study of a mouse with a brain tumour expressing firefly luciferase. Co-registration of the reconstructed 3D bioluminescent image with magnetic resonance images indicated good anatomical localization of the tumour

2005-12-07

20

Bioluminescence Imaging for Assessment and Normalization in Transfected Cell Arrays  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transfected cell arrays (TCAs) represent a high-throughput technique to correlate gene expression with functional cell responses. Despite advances in TCAs, improvements are needed for the widespread application of this technology. We have developed a TCA that combines a two-plasmid system and dual-bioluminescence imaging to quantitatively normalize for variability in transfection and increase sensitivity. The two-plasmids consist of: (i) normalization plasmid present within each spot, and (ii...

Pannier, Angela K.; Ariazi, Eric A.; Bellis, Abigail D.; Bengali, Zain; Jordan, V. Craig; Shea, Lonnie D.

2007-01-01

 
 
 
 
21

Enhanced in vivo bioluminescence imaging using liposomal luciferin delivery system  

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To provide a continuous and prolonged delivery of the substrate D-luciferin for bioluminescence imaging in vivo, luciferin was encapsulated into liposomes using either the pH-gradient or acetate-gradient method. Under optimum loading conditions, 0.17 mg luciferin was loaded per mg of lipid with 90–95% encapsulation efficiency, where active loading was 6 to 18-fold higher than obtained with passive loading. Liposomal luciferin in a long-circulating formulation had good shelf stability, with ...

2010-01-01

22

Expedient synthesis of electronically modified luciferins for bioluminescence imaging  

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Bioluminescence imaging with luciferase enzymes requires access to light-emitting, small molecule luciferins. Here, we describe a rapid method to synthesize d-luciferin, the substrate for firefly luciferase (Fluc), along with a novel set of electronically modified analogs. Our procedure utilizes a relatively rare, but synthetically useful dithiazolium reagent to generate heteroaromatic scaffolds in a divergent fashion. Two of the luciferin analogs produced with this approach emit light with F...

2012-01-01

23

A human brainstem glioma xenograft model enabled for bioluminescence imaging  

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Despite the use of radiation and chemotherapy, the prognosis for children with diffuse brainstem gliomas is extremely poor. There is a need for relevant brainstem tumor models that can be used to test new therapeutic agents and delivery systems in pre-clinical studies. We report the development of a brainstem-tumor model in rats and the application of bioluminescence imaging (BLI) for monitoring tumor growth and response to therapy as part of this model. Luciferase-modified human glioblastoma...

Hashizume, Rintaro; Ozawa, Tomoko; Dinca, Eduard B.; Banerjee, Anuradha; Prados, Michael D.; James, Charles D.; Gupta, Nalin

2010-01-01

24

Bioluminescence Imaging of Stem Cell-Based Therapeutics for Vascular Regeneration  

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Stem cell-based therapeutics show promise for treatment of vascular diseases. However, the survival of the cells after in vivo injection into diseased tissues remains a concern. In the advent of non-invasive optical imaging techniques such as bioluminescence imaging (BLI), cell localization and survival can be easily monitored over time. This approach has recently been applied towards monitoring stem cell treatments for vascular regeneration of the coronary or peripheral arteries. In this rev...

Huang, Ngan F.; Okogbaa, Janet; Babakhanyan, Anna; Cooke, John P.

2012-01-01

25

Bioluminescence imaging for assessment and normalization in transfected cell arrays.  

Science.gov (United States)

Transfected cell arrays (TCAs) represent a high-throughput technique to correlate gene expression with functional cell responses. Despite advances in TCAs, improvements are needed for the widespread application of this technology. We have developed a TCA that combines a two-plasmid system and dual-bioluminescence imaging to quantitatively normalize for variability in transfection and increase sensitivity. The two-plasmids consist of: (i) normalization plasmid present within each spot, and (ii) functional plasmid that varies between spots, responsible for the functional endpoint of the array. Bioluminescence imaging of dual-luciferase reporters (renilla, firefly luciferase) provides sensitive and quantitative detection of cellular response, with minimal post-transfection processing. The array was applied to quantify estrogen receptor alpha (ERalpha) activity in MCF-7 breast cancer cells. A plasmid containing an ERalpha-regulated promoter directing firefly luciferase expression was mixed with a normalization plasmid, complexed with cationic lipids and deposited into an array. ER induction mimicked results obtained through traditional assays methods, with estrogen inducing luciferase expression 10-fold over the antiestrogen fulvestrant or vehicle. Furthermore, the array captured a dose response to estrogen, demonstrating the sensitivity of bioluminescence quantification. This system provides a tool for basic science research, with potential application for the development of patient specific therapies. PMID:17486653

Pannier, Angela K; Ariazi, Eric A; Bellis, Abigail D; Bengali, Zain; Jordan, V Craig; Shea, Lonnie D

2007-10-01

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Bioluminescence imaging of A? deposition in bigenic mouse models of Alzheimer's disease  

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Transgenic (Tg) mouse models of Alzheimer's disease have served as valuable tools for investigating pathogenic mechanisms related to A? accumulation. However, assessing disease status in these animals has required time-consuming behavioral assessments or postmortem neuropathological analysis. Here, we report a method for tracking the progression of A? accumulation in vivo using bioluminescence imaging (BLI) on two lines of Tg mice, which express luciferase (luc) under control of the Gfap pr...

Watts, Joel C.; Giles, Kurt; Grillo, Sunny K.; Lemus, Azucena; Dearmond, Stephen J.; Prusiner, Stanley B.

2011-01-01

27

Video-rate bioluminescence imaging of protein secretion from a living cell.  

Science.gov (United States)

We have established a method of video-rate bioluminescence imaging to visualize exocytotic protein secretion from a single living cell using Gaussia luciferase (GLase) as a reporter protein. The luminescence signals produced by the enzymatic reaction of secreted GLase (luciferase) and coelenterazine (luciferin) are detected with an electron-multiplying charge-coupled device camera. An exocytotic event of protein secretion can be visualized using the protein fused to GLase with a time resolution of 30-500 ms. Signal analyses of the bioluminescence video images reveal a number of exocytotic sites, the frequency of exocytotic events, and the amount of secreted protein on a whole cell. Furthermore, the method can distinguish between secreted and cell surface-bound proteins. Our method is a direct approach to investigate the secretion and localization of proteins on the whole surface of living cells. PMID:24166369

Suzuki, Takahiro; Inouye, Satoshi

2014-01-01

28

Assessing laser-tissue damage with bioluminescent imaging.  

Science.gov (United States)

Effective medical laser procedures are achieved by selecting laser parameters that minimize undesirable tissue damage. Traditionally, human subjects, animal models, and monolayer cell cultures have been used to study wound healing, tissue damage, and cellular effects of laser radiation. Each of these models has significant limitations, and consequently, a novel skin model is needed. To this end, a highly reproducible human skin model that enables noninvasive and longitudinal studies of gene expression was sought. In this study, we present an organotypic raft model (engineered skin) used in combination with bioluminescent imaging (BLI) techniques. The efficacy of the raft model was validated and characterized by investigating the role of heat shock protein 70 (hsp70) as a sensitive marker of thermal damage. The raft model consists of human cells incorporated into an extracellular matrix. The raft cultures were transfected with an adenovirus containing a murine hsp70 promoter driving transcription of luciferase. The model enables quantitative analysis of spatiotemporal expression of proteins using BLI. Thermal stress was induced on the raft cultures by means of a constant temperature water bath or with a carbon dioxide (CO2) laser (lambda=10.6 microm, 0.679 to 2.262 Wcm2, cw, unfocused Gaussian beam, omegaL=4.5 mm, 1 min exposure). The bioluminescence was monitored noninvasively with an IVIS 100 Bioluminescent Imaging System. BLI indicated that peak hsp70 expression occurs 4 to 12 h after exposure to thermal stress. A minimum irradiance of 0.679 Wcm2 activated the hsp70 response, and a higher irradiance of 2.262 Wcm2 was associated with a severe reduction in hsp70 response due to tissue ablation. Reverse transcription polymerase chain reaction demonstrated that hsp70 mRNA levels increased with prolonged heating exposures. Enzyme-linked immunosorbent protein assays confirmed that luciferase was an accurate surrogate for hsp70 intracellular protein levels. Hematoxylin and eosin stains verified the presence of the thermally denatured tissue regions. Immunohistochemical analyses confirmed that maximal hsp70 expression occurred at a depth of 150 microm. Bioluminescent microscopy was employed to corroborate these findings. These results indicate that quantitative BLI in engineered tissue equivalents provides a powerful model that enables sequential gene expression studies. Such a model can be used as a high throughput screening platform for laser-tissue interaction studies. PMID:16965142

Wilmink, Gerald J; Opalenik, Susan R; Beckham, Joshua T; Davidson, Jeffrey M; Jansen, E Duco

2006-01-01

29

Assessing laser-tissue damage with bioluminescent imaging  

Science.gov (United States)

Effective medical laser procedures are achieved by selecting laser parameters that minimize undesirable tissue damage. Traditionally, human subjects, animal models, and monolayer cell cultures have been used to study wound healing, tissue damage, and cellular effects of laser radiation. Each of these models has significant limitations, and consequently, a novel skin model is needed. To this end, a highly reproducible human skin model that enables noninvasive and longitudinal studies of gene expression was sought. In this study, we present an organotypic raft model (engineered skin) used in combination with bioluminescent imaging (BLI) techniques. The efficacy of the raft model was validated and characterized by investigating the role of heat shock protein 70 (hsp70) as a sensitive marker of thermal damage. The raft model consists of human cells incorporated into an extracellular matrix. The raft cultures were transfected with an adenovirus containing a murine hsp70 promoter driving transcription of luciferase. The model enables quantitative analysis of spatiotemporal expression of proteins using BLI. Thermal stress was induced on the raft cultures by means of a constant temperature water bath or with a carbon dioxide (CO2) laser (?=10.6 µm, 0.679 to 2.262 W/cm2, cw, unfocused Gaussian beam, ?L=4.5 mm, 1 min exposure). The bioluminescence was monitored noninvasively with an IVIS 100 Bioluminescent Imaging System. BLI indicated that peak hsp70 expression occurs 4 to 12 h after exposure to thermal stress. A minimum irradiance of 0.679 W/cm2 activated the hsp70 response, and a higher irradiance of 2.262 W/cm2 was associated with a severe reduction in hsp70 response due to tissue ablation. Reverse transcription polymerase chain reaction demonstrated that hsp70 mRNA levels increased with prolonged heating exposures. Enzyme-linked immunosorbent protein assays confirmed that luciferase was an accurate surrogate for hsp70 intracellular protein levels. Hematoxylin and eosin stains verified the presence of the thermally denatured tissue regions. Immunohistochemical analyses confirmed that maximal hsp70 expression occurred at a depth of 150 µm. Bioluminescent microscopy was employed to corroborate these findings. These results indicate that quantitative BLI in engineered tissue equivalents provides a powerful model that enables sequential gene expression studies. Such a model can be used as a high throughput screening platform for laser-tissue interaction studies.

Wilmink, Gerald J.; Opalenik, Susan R.; Beckham, Josh T.; Davidson, Jeffrey M.; Jansen, Eric D.

2006-07-01

30

DEVELOPMENT OF A DUAL MODALITY TOMOGRAPHIC IMAGING SYSTEM FOR BIOLUMINESCENCE AND PET  

Energy Technology Data Exchange (ETDEWEB)

The goal of this proposal was to develop a new hybrid imaging modality capable to simultaneously image optical bioluminescence signals, as well as radionuclide emissions from the annihilation of positrons originating from molecular imaging probes in preclinical mouse models. This new technology enables the simultaneous in-vivo measurements of both emissions that could be produced from a single or a combination of two different biomarkers. It also facilitates establishing the physical limitations of bioluminescence imaging, its tomographic and spectral image reconstruction potential and the quantification of bioluminescence signals.

CHATZIIOANNOU, ARION

2011-12-21

31

In Vivo Bioluminescence Imaging of Tumor Cells Using Optimized Firefly Luciferase luc2  

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Full Text Available The present study was aimed to establish a tumor cell line stably expressing luciferase luc2, and to develop the technique to observe primary tumor nodes and metastases using in vivo bioluminescence imaging. Materials and Methods. In this research we used pLuc2-N plasmid, lentiviral vector pLVT-1, Colo 26 cell line and BALB/c mice to generate new bioluminescent tumor model. Bioluminescence imaging in vitro ? in vivo was carried out on IVIS-Spectrum system (Caliper Life Sciences, USA. Primary tumor model was created by subcutaneous injection of 500 000 Colo 26-luc2 cells. Model of metastases was generated by i.v. injection of 75 000 Colo 26-luc2 cells. Histological analysis was performed to verify the results of the imaging. Results. We created the lentiviral vector containing luc2 gene using molecular cloning. Then Colo 26-luc2 tumor cell line was generated. We assessed the sensitivity of luc2-based bioluminescence imaging. The intensity of bioluminescent signal in vitro averaged about 5000 photon/s per cell, in vivo — 250 photon/sec per cell. In vivo monitoring of Colo 26-luc2 primary tumor and metastases was demonstrated. The results of bioluminescence imaging correlated with histological analysis data. Conclusion. The present work shows the possibility of bioluminescent system based on optimized luciferase luc2 for in vivo noninvasive high-sensitive whole-body imaging of tumors.

N.V. Klementyeva

2013-08-01

32

Bioluminescence Imaging of Stem Cell-Based Therapeutics for Vascular Regeneration  

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Full Text Available Stem cell-based therapeutics show promise for treatment of vascular diseases. However, the survival of the cells after in vivo injection into diseased tissues remains a concern. In the advent of non-invasive optical imaging techniques such as bioluminescence imaging (BLI, cell localization and survival can be easily monitored over time. This approach has recently been applied towards monitoring stem cell treatments for vascular regeneration of the coronary or peripheral arteries. In this review, we will describe the application of BLI for tracking transplanted stem cells and associating their viability with therapeutic efficacy, in preclinical disease models of vascular disease.

Ngan F. Huang, Janet Okogbaa, Anna Babakhanyan, John P. Cooke

2012-01-01

33

Real-Time Bioluminescent Tracking of Cellular Population Dynamics  

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Cellular population dynamics are routinely monitored across many diverse fields for a variety of purposes. In general, these dynamics are assayed either through the direct counting of cellular aliquots followed by extrapolation to the total population size, or through the monitoring of signal intensity from any number of externally stimulated reporter proteins. While both viable methods, here we describe a novel technique that allows for the automated, non-destructive tracking of cellular population dynamics in real-time. This method, which relies on the detection of a continuous bioluminescent signal produced through expression of the bacterial luciferase gene cassette, provides a low cost, low time-intensive means for generating additional data compared to alternative methods.

Close, Dan [University of Tennessee, Knoxville (UTK); Sayler, Gary Steven [ORNL; Xu, Tingting [ORNL; Ripp, Steven Anthony [ORNL

2014-01-01

34

Multimodal imaging of orthotopic hepatocellular carcinoma using small animal PET, bioluminescence and contrast enhanced CT imaging  

Energy Technology Data Exchange (ETDEWEB)

Molecular imaging with small-animal PET and bioluminescence imaging has been used as an important tool in cancer research. One of the disadvantages of these imaging modalities is the lack of anatomic information. To obtain fusion images with both molecular and anatomical information, small-animal PET and bioluminescence images fused with contrast enhance CT image in orthotopic hepatocellular carcinoma (HCC) model. We retrovially transfected dual gene (HSV1-tk and firefly luciferase) to morris hepatoma cells. The expression of HSV1-tk and luciferase was checked by optical imager and in vitro radiolabeled FIAU uptake, respectively and also checked by RT-PCR analysis. MCA-TL cells (5X10{sup 5}/ 0.05 ml) mixed with matrigel (1: 10) injected into left lobe of liver in nude mice. {sup 124}I-FIAU-PET, bioluminescence and contrast enhanced CT images were obtained in the orthotopic HCC model and digital whole body autoradiography (DWBA) was performed. Small animal PET image was obtained at 2 h post injection of {sup 124}I-FIAU and contrast enhanced CT image was obtained at 3 h post injection of Fenestra LC (0.3 ml). MCA-TL cells showed more specific {sup 124}I-FIAU uptake and higher luminescent activity than parental cells. The orthotopic HCC was detected by {sup 124}I-FIAU PET, contrast enhanced CT, and BLI and confirmed by DWBA. Registered image in orthotopic HCC t models showed a good correlation of images from both PET and CT. Contrast enhanced CT image delineated margin of HCC. Multimodal imaging with {sup 124}I-FIAU PET, bioluminescence and contrast enhanced CT allows a precise and improved detection of tumor in orthotopic hepatocellular carcinoma model. Multimodal imaging is potentially useful for monitoring progression of hepatic metastasis and for the evaluation of cancer treatments.

Lee, T. S.; Woo, S. G.; Jeong, J. H.; Woo, K. S.; Jeong, E. S.; Kang, J. H.; Cheon, G. J.; Choi, C. W.; Lim, S. M. [Korea Institute of Radioligical and Medical Sciences, Seoul (Korea, Republic of)

2007-07-01

35

Multimodal imaging of orthotopic hepatocellular carcinoma using small animal PET, bioluminescence and contrast enhanced CT imaging  

International Nuclear Information System (INIS)

Molecular imaging with small-animal PET and bioluminescence imaging has been used as an important tool in cancer research. One of the disadvantages of these imaging modalities is the lack of anatomic information. To obtain fusion images with both molecular and anatomical information, small-animal PET and bioluminescence images fused with contrast enhance CT image in orthotopic hepatocellular carcinoma (HCC) model. We retrovially transfected dual gene (HSV1-tk and firefly luciferase) to morris hepatoma cells. The expression of HSV1-tk and luciferase was checked by optical imager and in vitro radiolabeled FIAU uptake, respectively and also checked by RT-PCR analysis. MCA-TL cells (5X105/ 0.05 ml) mixed with matrigel (1: 10) injected into left lobe of liver in nude mice. 124I-FIAU-PET, bioluminescence and contrast enhanced CT images were obtained in the orthotopic HCC model and digital whole body autoradiography (DWBA) was performed. Small animal PET image was obtained at 2 h post injection of 124I-FIAU and contrast enhanced CT image was obtained at 3 h post injection of Fenestra LC (0.3 ml). MCA-TL cells showed more specific 124I-FIAU uptake and higher luminescent activity than parental cells. The orthotopic HCC was detected by 124I-FIAU PET, contrast enhanced CT, and BLI and confirmed by DWBA. Registered image in orthotopic HCC t models showed a good correlation of images from both PET and CT. Contrast enhanced CT image delineated margin of HCC. Multimodal imaging with 124I-FIAU PET, bioluminescence and contrast enhanced CT allows a precise and improved detection of tumor in orthotopic hepatocellular carcinoma model. Multimodal imaging is potentially useful for monitoring progression of hepatic metastasis and for the evaluation of cancer treatments

2007-10-26

36

Use of a highly sensitive two-dimensional luminescence imaging system to monitor endogenous bioluminescence in plant leaves  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background All living organisms emit spontaneous low-level bioluminescence, which can be increased in response to stress. Methods for imaging this ultra-weak luminescence have previously been limited by the sensitivity of the detection systems used. Results We developed a novel configuration of a cooled charge-coupled device (CCD for 2-dimensional imaging of light emission from biological material. In this study, we imaged photon emission from plant leaves. The equipment allowed short integration times for image acquisition, providing high resolution spatial and temporal information on bioluminescence. We were able to carry out time course imaging of both delayed chlorophyll fluorescence from whole leaves, and of low level wound-induced luminescence that we showed to be localised to sites of tissue damage. We found that wound-induced luminescence was chlorophyll-dependent and was enhanced at higher temperatures. Conclusions The data gathered on plant bioluminescence illustrate that the equipment described here represents an improvement in 2-dimensional luminescence imaging technology. Using this system, we identify chlorophyll as the origin of wound-induced luminescence from leaves.

Flor-Henry Michel

2004-11-01

37

Compartmentalization of algal bioluminescence: autofluorescence of bioluminescent particles in the dinoflagellate Gonyaulax as studied with image-intensified video microscopy and flow cytometry  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Compartmentalization of specialized functions to discrete locales is a fundamental theme of eucaryotic organization in cells. We report here that bioluminescence of the dinoflagellate alga Gonyaulax originates in vivo from discrete subcellular loci that are intrinsically fluorescent. We demonstrate this localization by comparing the loci of fluorescence and bioluminescence as visualized by image-intensified video microscopy. These fluorescent particles appeared to be the same as the previousl...

1985-01-01

38

Functional imaging of interleukin 1 beta expression in inflammatory process using bioluminescence imaging in transgenic mice  

Science.gov (United States)

Background Interleukin 1 beta (IL-1?) plays an important role in a number of chronic and acute inflammatory diseases. To understand the role of IL-1? in disease processes and develop an in vivo screening system for anti-inflammatory drugs, a transgenic mouse line was generated which incorporated the transgene firefly luciferase gene driven by a 4.5-kb fragment of the human IL-1? gene promoter. Luciferase gene expression was monitored in live mice under anesthesia using bioluminescence imaging in a number of inflammatory disease models. Results In a LPS-induced sepsis model, dramatic increase in luciferase activity was observed in the mice. This transgene induction was time dependent and correlated with an increase of endogenous IL-1? mRNA and pro-IL-1? protein levels in the mice. In a zymosan-induced arthritis model and an oxazolone-induced skin hypersensitivity reaction model, luciferase expression was locally induced in the zymosan injected knee joint and in the ear with oxazolone application, respectively. Dexamethasone suppressed the expression of luciferase gene both in the acute sepsis model and in the acute arthritis model. Conclusion Our data suggest that the transgenic mice model could be used to study transcriptional regulation of the IL-1? gene expression in the inflammatory process and evaluation the effect of anti-inflammatory drug in vivo.

Li, Limei; Fei, Zhaoliang; Ren, Jianke; Sun, Ruilin; Liu, Zhihui; Sheng, Zhejin; Wang, Long; Sun, Xia; Yu, Jun; Wang, Zhugang; Fei, Jian

2008-01-01

39

Functional imaging of interleukin 1 beta expression in inflammatory process using bioluminescence imaging in transgenic mice  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Interleukin 1 beta (IL-1? plays an important role in a number of chronic and acute inflammatory diseases. To understand the role of IL-1? in disease processes and develop an in vivo screening system for anti-inflammatory drugs, a transgenic mouse line was generated which incorporated the transgene firefly luciferase gene driven by a 4.5-kb fragment of the human IL-1? gene promoter. Luciferase gene expression was monitored in live mice under anesthesia using bioluminescence imaging in a number of inflammatory disease models. Results In a LPS-induced sepsis model, dramatic increase in luciferase activity was observed in the mice. This transgene induction was time dependent and correlated with an increase of endogenous IL-1? mRNA and pro-IL-1? protein levels in the mice. In a zymosan-induced arthritis model and an oxazolone-induced skin hypersensitivity reaction model, luciferase expression was locally induced in the zymosan injected knee joint and in the ear with oxazolone application, respectively. Dexamethasone suppressed the expression of luciferase gene both in the acute sepsis model and in the acute arthritis model. Conclusion Our data suggest that the transgenic mice model could be used to study transcriptional regulation of the IL-1? gene expression in the inflammatory process and evaluation the effect of anti-inflammatory drug in vivo.

Liu Zhihui

2008-08-01

40

In vitro influence of hypoxia on bioluminescence imaging in brain tumor cells  

Science.gov (United States)

Bioluminescence Imaging (BLI) has been employed as an imaging modality to identify and characterize fundamental processes related to cancer development and response at cellular and molecular levels. This technique is based on the reaction of luciferin with oxygen in the presence of luciferase and ATP. A major concern in this technique is that tumors are generally hypoxic, either constitutively and/or as a result of treatment, therefore the oxygen available for the bioluminescence reaction could possibly be reduced to limiting levels, and thus leading to underestimation of the actual number of luciferase-labeled cells during in vivo procedures. In this report, we present the initial in vitro results of the oxygen dependence of the bioluminescence signal in rat gliosarcoma 9L cells tagged with the luciferase gene (9L luc cells). Bioluminescence photon emission from cells exposed to different oxygen tensions was detected by a sensitive CCD camera upon exposure to luciferin. The results showed that bioluminescence signal decreased at administered pO II levels below about 5%, falling by approximately 50% at 0.2% pO II. Additional experiments showed that changes in BLI was due to the cell inability to maintain normal levels of ATP during the hypoxic period reducing the ATP concentration to limiting levels for BLI.

Moriyama, Eduardo H.; Jarvi, Mark; Niedre, Mark; Mocanu, Joseph D.; Moriyama, Yumi; Li, Buhong; Lilge, Lothar; Wilson, Brian C.

2007-03-01

 
 
 
 
41

Bioluminescence imaging to monitor the prolongation of stem cell survival by pharmaceutical intervention  

Energy Technology Data Exchange (ETDEWEB)

The rapid donor cell death and rejection owing to humoral and cellular immune reactions are a basic limitation encountered in stem cell therapy for treatment of cardiovascular disease. We investigated the potential for longitudinal bioluminescence imaging to monitor the survival of transplanted stem cells prolonged by immunosuppressive agents. Embryonic rat H9c2 cardio myoblasts were transfected with adenovirus containing luciferase reporter gene (Ad-CMV-Fluc) in different MOI (1,10,100) and various cell doses (1x10{sup 5} - 5x10{sup 6})followed by injection in the thigh muscle of nude mice (n=6 per group), Other mice (n = 18) were undergone transient immunosuppression provided by either Cyclosporine (5mg/kg) or Tacrolimus (1mg/kg) or Dexamethasone (4mg/kg) beginning 3 days prior to and continuing to 2 weeks after transplantation. Optical bioluminescent imaging was then daily carried out using cooled CCD camera (Xenogen) Viral transfection at MOI 100 and the 5x10{sup 6} cell dose implantation resulted in optimal transgene efficiency. Mice received immunosuppressive agents displayed long-term in vivo reporter gene expression for a time course of 14 days. Tacrolimus (Prograf) and Cyclosporine successfully suppressed the transplanted cell loss in animals, that obviously observed until day 8 as compared to Dexamethasone-treated and non-treated mice (day 1: 1.00E+08 (Prograf), 9.47E+07 (Cys), 5.25E+07 (Dex), and 1.25E+07 p/s/cm{sup 2}/sr (control); day 8: 3.27E+05 (Prograf), 1.02E+05 (Cys), 6.17E+04 (Dex) and 2.73E+04 p/s/cm{sup 2}/sr (control)) and continued expressing bioluminescence until day 13 ( 6.42E+05 (Prograf), 4.99E+05 (Cys), and 4.10E+04 p/s/cm{sup 2}/sr. Induction of immune tolerance using pharmaceutical agents during cardio myoblast transplantation improved long-term donor cell survival in murine muscles. Optical imaging technique is capable of being used for tracking implanted stem cells in myocardium of living subjects over time.

Le, Uyenchi N.; Min, Jung Joon; Moon, Sung Min; Ahn, Young Keun; Kim, Yong Sook; Joo, Soo Yeon; Hong, Moon Hwa; Jeong, Myung Ho; Song, Ho Cheon; Bom, Hee Seung [Chonnam National University Medical School, Gwangju (Korea, Republic of)

2005-07-01

42

Bioluminescence imaging to monitor the prolongation of stem cell survival by pharmaceutical intervention  

International Nuclear Information System (INIS)

The rapid donor cell death and rejection owing to humoral and cellular immune reactions are a basic limitation encountered in stem cell therapy for treatment of cardiovascular disease. We investigated the potential for longitudinal bioluminescence imaging to monitor the survival of transplanted stem cells prolonged by immunosuppressive agents. Embryonic rat H9c2 cardio myoblasts were transfected with adenovirus containing luciferase reporter gene (Ad-CMV-Fluc) in different MOI (1,10,100) and various cell doses (1x105 - 5x106)followed by injection in the thigh muscle of nude mice (n=6 per group), Other mice (n = 18) were undergone transient immunosuppression provided by either Cyclosporine (5mg/kg) or Tacrolimus (1mg/kg) or Dexamethasone (4mg/kg) beginning 3 days prior to and continuing to 2 weeks after transplantation. Optical bioluminescent imaging was then daily carried out using cooled CCD camera (Xenogen) Viral transfection at MOI 100 and the 5x106 cell dose implantation resulted in optimal transgene efficiency. Mice received immunosuppressive agents displayed long-term in vivo reporter gene expression for a time course of 14 days. Tacrolimus (Prograf) and Cyclosporine successfully suppressed the transplanted cell loss in animals, that obviously observed until day 8 as compared to Dexamethasone-treated and non-treated mice (day 1: 1.00E+08 (Prograf), 9.47E+07 (Cys), 5.25E+07 (Dex), and 1.25E+07 p/s/cm2/sr (control); day 8: 3.27E+05 (Prograf), 1.02E+05 (Cys), 6.17E+04 (Dex) and 2.73E+04 p/s/cm2/sr (control)) and continued expressing bioluminescence until day 13 ( 6.42E+05 (Prograf), 4.99E+05 (Cys), and 4.10E+04 p/s/cm2/sr. Induction of immune tolerance using pharmaceutical agents during cardio myoblast transplantation improved long-term donor cell survival in murine muscles. Optical imaging technique is capable of being used for tracking implanted stem cells in myocardium of living subjects over time

2005-11-18

43

Evaluation of biolistic gene transfer methods in vivo using non-invasive bioluminescent imaging techniques  

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Full Text Available Abstract Background Gene therapy continues to hold great potential for treating many different types of disease and dysfunction. Safe and efficient techniques for gene transfer and expression in vivo are needed to enable gene therapeutic strategies to be effective in patients. Currently, the most commonly used methods employ replication-defective viral vectors for gene transfer, while physical gene transfer methods such as biolistic-mediated ("gene-gun" delivery to target tissues have not been as extensively explored. In the present study, we evaluated the efficacy of biolistic gene transfer techniques in vivo using non-invasive bioluminescent imaging (BLI methods. Results Plasmid DNA carrying the firefly luciferase (LUC reporter gene under the control of the human Cytomegalovirus (CMV promoter/enhancer was transfected into mouse skin and liver using biolistic methods. The plasmids were coupled to gold microspheres (1 ?m diameter using different DNA Loading Ratios (DLRs, and "shot" into target tissues using a helium-driven gene gun. The optimal DLR was found to be in the range of 4-10. Bioluminescence was measured using an In Vivo Imaging System (IVIS-50 at various time-points following transfer. Biolistic gene transfer to mouse skin produced peak reporter gene expression one day after transfer. Expression remained detectable through four days, but declined to undetectable levels by six days following gene transfer. Maximum depth of tissue penetration following biolistic transfer to abdominal skin was 200-300 ?m. Similarly, biolistic gene transfer to mouse liver in vivo also produced peak early expression followed by a decline over time. In contrast to skin, however, liver expression of the reporter gene was relatively stable 4-8 days post-biolistic gene transfer, and remained detectable for nearly two weeks. Conclusions The use of bioluminescence imaging techniques enabled efficient evaluation of reporter gene expression in vivo. Our results demonstrate that different tissues show different expression kinetics following gene transfer of the same reporter plasmid to different mouse tissues in vivo. We evaluated superficial (skin and abdominal organ (liver targets, and found that reporter gene expression peaked within the first two days post-transfer in each case, but declined most rapidly in the skin (3-4 days compared to liver (10-14 days. This information is essential for designing effective gene therapy strategies in different target tissues.

Daniell Henry

2011-06-01

44

Development of Optical Molecular Imaging System for the Acquisition of Bioluminescence Signals from Small Animals  

International Nuclear Information System (INIS)

Optical imaging is providing great advance and improvement in genetic and molecular imaging of animals and humans. Optical imaging system consists of optical imaging devices, which carry out major function for monitoring, tracing, and imaging in most of molecular in-vivo researches. In bio-luminescent imaging, small animals containing luciferase gene locally irradiate light, and emitted photons transmitted through skin of the small animals are imaged by using a high sensitive charged coupled device (CCD) camera. In this paper, we introduced optical imaging system for the image acquisition of bio-luminescent signals emitted from small animals. In the system, Nikon lens and four LED light sources were mounted at the inside of a dark box. A cooled CCD camera equipped with a control module was used. We tested the performance of the optical imaging system using effendorf tube and light emitting bacteria which injected intravenously into CT26 tumor bearing nude mouse. The performance of implemented optical imaging system for bio-luminescence imaging was demonstrated and the feasibility of the system in small animal imaging application was proved. We anticipate this system could be a useful tool for the molecular imaging of small animals adaptable for various experimental conditions in future

2009-08-01

45

Bioluminescence imaging of chondrocytes in rabbits by intraarticular injection of D-luciferin  

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Luciferase is one of the most commonly used reporter enzymes in the field of in vivo optical imaging. D-luciferin, the substrate for firefly luciferase has very high cost that allows this kind of experiment limited to small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in the articular capsule for bioluminescence imaging in rabbits. Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase (Fluc). Chondrocytes expressing Fluc were injected or implanted in the left knee joint. The rabbits underwent optical imaging studies after local injection of D-luciferin at 1, 5, 7, 9 days after cellular administration. We sought whether optimal imaging signals was could be by a cooled CCD camera after local injection of D-luciferin. Imaging signal was not observed from the left knee joint after intraperitoneal injection of D-luciferin (15 mg/kg), whereas it was observed after intraarticular injection. Photon intensity from the left knee joint of rabbits was compared between cell injected and implanted groups after intraarticular injection of D-luciferin. During the period of imaging studies, photon intensity of the cell implanted group was 5-10 times higher than that of the cell injected group. We successfully imaged chondrocytes expressing Fluc after intraarticular injection of D-luciferin. This technique may be further applied to develop new drugs for knee joint disease.

Moon, Sung Min; Min, Jung Joon; Kim, Sung Mi; Bom, Hee Seung [Chonnam National University Medical School, Gwangju (Korea, Republic of); Oh, Suk Jung; Kang, Han Saem; Kim, Kwang Yoon [ECOBIO INC., Gwangju (Korea, Republic of); Kim, Young Ho [College of Natural Science, Chosun University, Gwangju (Korea, Republic of)

2007-02-15

46

Bioluminescence imaging of chondrocytes in rabbits by intraarticular injection of D-luciferin  

International Nuclear Information System (INIS)

Luciferase is one of the most commonly used reporter enzymes in the field of in vivo optical imaging. D-luciferin, the substrate for firefly luciferase has very high cost that allows this kind of experiment limited to small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in the articular capsule for bioluminescence imaging in rabbits. Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase (Fluc). Chondrocytes expressing Fluc were injected or implanted in the left knee joint. The rabbits underwent optical imaging studies after local injection of D-luciferin at 1, 5, 7, 9 days after cellular administration. We sought whether optimal imaging signals was could be by a cooled CCD camera after local injection of D-luciferin. Imaging signal was not observed from the left knee joint after intraperitoneal injection of D-luciferin (15 mg/kg), whereas it was observed after intraarticular injection. Photon intensity from the left knee joint of rabbits was compared between cell injected and implanted groups after intraarticular injection of D-luciferin. During the period of imaging studies, photon intensity of the cell implanted group was 5-10 times higher than that of the cell injected group. We successfully imaged chondrocytes expressing Fluc after intraarticular injection of D-luciferin. This technique may be further applied to develop new drugs for knee joint disease

2007-02-01

47

Uptake kinetics and biodistribution of 14C-d-luciferin - a radiolabeled substrate for the firefly luciferase catalyzed bioluminescence reaction: impact on bioluminescence based reporter gene imaging  

International Nuclear Information System (INIS)

Firefly luciferase catalyzes the oxidative decarboxylation of d-luciferin to oxyluciferin in the presence of cofactors, producing bioluminescence. This reaction is used in optical bioluminescence-based molecular imaging approaches to detect the expression of the firefly luciferase reporter gene. Biokinetics and distribution of the substrate most likely have a significant impact on levels of light signal and therefore need to be investigated. Benzene ring 14C(U)-labeled d-luciferin was utilized. Cell uptake and efflux assays, murine biodistribution, autoradiography and CCD-camera based optical bioluminescence imaging were carried out to examine the in vitro and in vivo characteristics of the tracer in cell culture and in living mice respectively. Radiolabeled and unlabeled d-luciferin revealed comparable levels of light emission when incubated with equivalent amounts of the firefly luciferase enzyme. Cell uptake assays in pCMV-luciferase-transfected cells showed slow trapping of the tracer and relatively low uptake values (up to 22.9-fold higher in firefly luciferase gene-transfected vs. nontransfected cells, p=0.0002). Biodistribution studies in living mice after tail-vein injection of 14C-d-luciferin demonstrated inhomogeneous tracer distribution with early predominant high radioactivity levels in kidneys (10.6% injected dose [ID]/g) and liver (11.9% ID/g), followed at later time points by the bladder (up to 81.3% ID/g) and small intestine (6.5% ID/g), reflecting the elimination routes of the tracer. Kinetics and uptake levels profoundly differed when using alternate injection routes (intravenous versus intraperitoneal). No clear trapping of 14C-d-luciferin in firefly luciferase-expressing tissues could be observed in vivo. The data obtained with 14C-d-luciferin provide insights into the dynamics of d-luciferin cell uptake, intracellular accumulation, and efflux. Results of the biodistribution and autoradiographic studies should be useful for optimizing and adapting optical imaging protocols to specific experimental settings when utilizing the firefly luciferase and d-luciferin system. (orig.)

2008-12-01

48

Uptake kinetics and biodistribution of {sup 14}C-d-luciferin - a radiolabeled substrate for the firefly luciferase catalyzed bioluminescence reaction: impact on bioluminescence based reporter gene imaging  

Energy Technology Data Exchange (ETDEWEB)

Firefly luciferase catalyzes the oxidative decarboxylation of d-luciferin to oxyluciferin in the presence of cofactors, producing bioluminescence. This reaction is used in optical bioluminescence-based molecular imaging approaches to detect the expression of the firefly luciferase reporter gene. Biokinetics and distribution of the substrate most likely have a significant impact on levels of light signal and therefore need to be investigated. Benzene ring {sup 14}C(U)-labeled d-luciferin was utilized. Cell uptake and efflux assays, murine biodistribution, autoradiography and CCD-camera based optical bioluminescence imaging were carried out to examine the in vitro and in vivo characteristics of the tracer in cell culture and in living mice respectively. Radiolabeled and unlabeled d-luciferin revealed comparable levels of light emission when incubated with equivalent amounts of the firefly luciferase enzyme. Cell uptake assays in pCMV-luciferase-transfected cells showed slow trapping of the tracer and relatively low uptake values (up to 22.9-fold higher in firefly luciferase gene-transfected vs. nontransfected cells, p=0.0002). Biodistribution studies in living mice after tail-vein injection of {sup 14}C-d-luciferin demonstrated inhomogeneous tracer distribution with early predominant high radioactivity levels in kidneys (10.6% injected dose [ID]/g) and liver (11.9% ID/g), followed at later time points by the bladder (up to 81.3% ID/g) and small intestine (6.5% ID/g), reflecting the elimination routes of the tracer. Kinetics and uptake levels profoundly differed when using alternate injection routes (intravenous versus intraperitoneal). No clear trapping of {sup 14}C-d-luciferin in firefly luciferase-expressing tissues could be observed in vivo. The data obtained with {sup 14}C-d-luciferin provide insights into the dynamics of d-luciferin cell uptake, intracellular accumulation, and efflux. Results of the biodistribution and autoradiographic studies should be useful for optimizing and adapting optical imaging protocols to specific experimental settings when utilizing the firefly luciferase and d-luciferin system. (orig.)

Berger, Frank [Ludwig-Maximilians University Munich (Germany). Department of Clinical Radiology; Paulmurugan, Ramasamy; Gambhir, Sanjiv Sam [Molecular Imaging Program at Stanford, Departments of Radiology and Bioengineering, Stanford, CA (United States); Bhaumik, Srabani [GE Global Research, Niskayuna, NY (United States)

2008-12-15

49

Bioluminescence imaging of DNA synthetic phase of cell cycle in living animals.  

Science.gov (United States)

Bioluminescence reporter proteins have been widely used in the development of tools for monitoring biological events in living cells. Currently, some assays like flow cytometry analysis are available for studying DNA synthetic phase (S-phase) targeted anti-cancer drug activity in vitro; however, techniques for imaging of in vivo models remain limited. Cyclin A2 is known to promote S-phase entry in mammals. Its expression levels are low during G1-phase, but they increase at the onset of S-phase. Cyclin A2 is degraded during prometaphase by ubiquitin-dependent, proteasome-mediated proteolysis. In this study, we have developed a cyclin A2-luciferase (CYCA-Luc) fusion protein targeted for ubiquitin-proteasome dependent degradation, and have evaluated its utility in screening S-phase targeted anti-cancer drugs. Similar to endogenous cyclin A2, CYCA-Luc accumulates during S-phase and is degraded during G2/M-phase. Using Cdc20 siRNA we have demonstrated that Cdc20 can mediate CYCA-Luc degradation. Moreover, using noninvasive bioluminescent imaging, we demonstrated accumulation of CYCA-Luc in response to 10-hydroxycamptothecin (HCPT), an S-phase targeted anti-cancer drug, in human tumor cells in vivo and in vitro. Our results indicate that a CYCA-Luc fusion reporter system can be used to monitor S-phase of cell cycle, and evaluate pharmacological activity of anti-cancer drug HCPT in real time in vitro and in vivo, and is likely to provide an important tool for screening such drugs. PMID:23301056

Chen, Zhi-Hong; Zhao, Rui-Jun; Li, Rong-Hui; Guo, Cui-Ping; Zhang, Guo-Jun

2013-01-01

50

Bioluminescence Imaging of Reporter Mice for Studies of Infection and Inflammation  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In vivo bioluminescence imaging offers the opportunity to study biological processes in living animals, and the study of viral infections and host immune responses can be enhanced substantially through this imaging modality. For most studies of viral pathogenesis and effects of anti-viral therapies, investigators have used recombinant viruses engineered to express a luciferase enzyme. This strategy requires stable insertion of an imaging reporter gene into the viral genome, which is not feasi...

2010-01-01

51

Bioluminescent imaging: a critical tool in pre-clinical oncology research.  

LENUS (Irish Health Repository)

Bioluminescent imaging (BLI) is a non-invasive imaging modality widely used in the field of pre-clinical oncology research. Imaging of small animal tumour models using BLI involves the generation of light by luciferase-expressing cells in the animal following administration of substrate. This light may be imaged using an external detector. The technique allows a variety of tumour-associated properties to be visualized dynamically in living models. The increasing use of BLI as a small-animal imaging modality has led to advances in the development of xenogeneic, orthotopic, and genetically engineered animal models expressing luciferase genes. This review aims to provide insight into the principles of BLI and its applications in cancer research. Many studies to assess tumour growth and development, as well as efficacy of candidate therapeutics, have been performed using BLI. More recently, advances have also been made using bioluminescent imaging in studies of protein-protein interactions, genetic screening, cell-cycle regulators, and spontaneous cancer development. Such novel studies highlight the versatility and potential of bioluminescent imaging in future oncological research.

O'Neill, Karen

2010-02-01

52

Comparison of in vivo optical systems for bioluminescence and fluorescence imaging.  

Science.gov (United States)

In vivo optical imaging has become a popular tool in animal laboratories. Currently, many in vivo optical imaging systems are available on the market, which often makes it difficult for research groups to decide which system fits their needs best. In this work we compared different commercially available systems, which can measure both bioluminescent and fluorescent light. The systems were tested for their bioluminescent and fluorescent sensitivity both in vitro and in vivo. The IVIS Lumina II was found to be most sensitive for bioluminescence imaging, with the Photon Imager a close second. Contrary, the Kodak system was, in vitro, the most sensitive system for fluorescence imaging. In vivo, the fluorescence sensitivity of the systems was similar. Finally, we examined the added value of spectral unmixing algorithms for in vivo optical imaging and demonstrated that spectral unmixing resulted in at least a doubling of the in vivo sensitivity. Additionally, spectral unmixing also enabled separate imaging of dyes with overlapping spectra which were, without spectral unmixing, not distinguishable. PMID:23579930

Cool, Steven K; Breyne, Koen; Meyer, Evelyne; De Smedt, Stefaan C; Sanders, Niek N

2013-09-01

53

Non-invasive visualisation of the development of peritoneal carcinomatosis and tumour regression after {sup 213}Bi-radioimmunotherapy using bioluminescence imaging  

Energy Technology Data Exchange (ETDEWEB)

Non-invasive imaging of tumour development remains a challenge, especially for tumours in the intraperitoneal cavity. Therefore, the aim of this study was the visualisation of both the development of peritoneal carcinomatosis and tumour regression after radioimmunotherapy with tumour-specific {sup 213}Bi-Immunoconjugates, via in vivo bioluminescence imaging of firefly luciferase-transfected cells. Human diffuse-type gastric cancer cells expressing mutant d9-E-cadherin were stably transfected with firefly luciferase (HSC45-M2-luc). For bioluminescence imaging, nude mice were inoculated intraperitoneally with 1 x 10{sup 7} HSC45-M2-luc cells. On days 4 and 8 after tumour cell inoculation, imaging was performed following D-luciferin injection using a cooled CCD camera with an image intensifier unit. For therapy, mice were injected with 2.7 MBq {sup 213}Bi-d9MAb targeting d9-E-cadherin on day 8 after tumour cell inoculation. Bioluminescence images were taken every 4 days to monitor tumour development. After i.p. inoculation of HSC45-M2-luc cells into nude mice, development as well as localisation of peritoneal carcinomatosis could be visualised using bioluminescence imaging. Following {sup 213}Bi-d9MAb therapy on day 8 after intraperitoneal inoculation of HSC45-M2-luc cells, small tumour nodules were totally eliminated and larger nodules showed a clear reduction in size on day 12 after tumour cell inoculation. Subsequently a recurrence of tumour mass was observed, starting from the remaining tumour spots. By measuring the mean grey level intensity, tumour development over time could be demonstrated. Non-invasive bioluminescence imaging permits visualisation of the development of peritoneal carcinomatosis, localisation of tumour in the intraperitoneal cavity and evaluation of therapeutic success after {sup 213}Bi-d9MAb treatment. (orig.)

Buchhorn, H. Matthias; Seidl, Christof; Beck, Roswitha; Schwaiger, Markus [Technische Universitaet Muenchen, Department of Nuclear Medicine, Munich (Germany); Saur, Dieter [Technische Universitaet Muenchen, Department of Internal Medicine 2, Munich (Germany); Apostolidis, Christos; Morgenstern, Alfred [Institute for Transuranium Elements, European Commission, Joint Research Centre, Karlsruhe (Germany); Senekowitsch-Schmidtke, Reingard [Technische Universitaet Muenchen, Department of Nuclear Medicine, Munich (Germany); Technische Universitaet Muenchen, Nuklearmedizinische Klinik, Klinikum r. d. Isar, Muenchen (Germany)

2007-06-15

54

Non-invasive visualisation of the development of peritoneal carcinomatosis and tumour regression after 213Bi-radioimmunotherapy using bioluminescence imaging  

International Nuclear Information System (INIS)

Non-invasive imaging of tumour development remains a challenge, especially for tumours in the intraperitoneal cavity. Therefore, the aim of this study was the visualisation of both the development of peritoneal carcinomatosis and tumour regression after radioimmunotherapy with tumour-specific 213Bi-Immunoconjugates, via in vivo bioluminescence imaging of firefly luciferase-transfected cells. Human diffuse-type gastric cancer cells expressing mutant d9-E-cadherin were stably transfected with firefly luciferase (HSC45-M2-luc). For bioluminescence imaging, nude mice were inoculated intraperitoneally with 1 x 107 HSC45-M2-luc cells. On days 4 and 8 after tumour cell inoculation, imaging was performed following D-luciferin injection using a cooled CCD camera with an image intensifier unit. For therapy, mice were injected with 2.7 MBq 213Bi-d9MAb targeting d9-E-cadherin on day 8 after tumour cell inoculation. Bioluminescence images were taken every 4 days to monitor tumour development. After i.p. inoculation of HSC45-M2-luc cells into nude mice, development as well as localisation of peritoneal carcinomatosis could be visualised using bioluminescence imaging. Following 213Bi-d9MAb therapy on day 8 after intraperitoneal inoculation of HSC45-M2-luc cells, small tumour nodules were totally eliminated and larger nodules showed a clear reduction in size on day 12 after tumour cell inoculation. Subsequently a recurrence of tumour mass was observed, starting from the remaining tumour spots. By measuring the mean grey level intensity, tumour development over time could be demonstrated. Non-invasive bioluminescence imaging permits visualisation of the development of peritoneal carcinomatosis, localisation of tumour in the intraperitoneal cavity and evaluation of therapeutic success after 213Bi-d9MAb treatment. (orig.)

2007-06-01

55

In Vivo Bioluminescence Imaging of Cell Differentiation in Biomaterials: A Platform for Scaffold Development  

Science.gov (United States)

In vivo testing is a mandatory last step in scaffold development. Agile longitudinal noninvasive real-time monitoring of stem cell behavior in biomaterials implanted in live animals should facilitate the development of scaffolds for tissue engineering. We report on a noninvasive bioluminescence imaging (BLI) procedure for simultaneous monitoring of changes in the expression of multiple genes to evaluate scaffold performance in vivo. Adipose tissue-derived stromal mensenchymal cells were dually labeled with Renilla red fluorescent protein and firefly green fluorescent protein chimeric reporters regulated by cytomegalovirus and tissue-specific promoters, respectively. Labeled cells were induced to differentiate in vitro and in vivo, by seeding in demineralized bone matrices (DBMs) and monitored by BLI. Imaging results were validated by RT-polymerase chain reaction and histological procedures. The proposed approach improves molecular imaging and measurement of changes in gene expression of cells implanted in live animals. This procedure, applicable to the simultaneous analysis of multiple genes from cells seeded in DBMs, should facilitate engineering of scaffolds for tissue repair.

Bago, Juli R.; Aguilar, Elisabeth; Alieva, Maria; Soler-Botija, Carolina; Vila, Olaia F.; Claros, Silvia; Andrades, Jose A.; Becerra, Jose; Rubio, Nuria

2013-01-01

56

Bioluminescence imaging to track bacterial dissemination of Yersinia pestis using different routes of infection in mice  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Plague is caused by Yersinia pestis, a bacterium that disseminates inside of the host at remarkably high rates. Plague bacilli disrupt normal immune responses in the host allowing for systematic spread that is fatal if left untreated. How Y. pestis disseminates from the site of infection to deeper tissues is unknown. Dissemination studies for plague are typically performed in mice by determining the bacterial burden in specific organs at various time points. To follow bacterial dissemination during plague infections in mice we tested the possibility of using bioluminescence imaging (BLI, an alternative non-invasive approach. Fully virulent Y. pestis was transformed with a plasmid containing the luxCDABE genes, making it able to produce light; this lux-expressing strain was used to infect mice by subcutaneous, intradermal or intranasal inoculation. Results We successfully obtained images from infected animals and were able to follow bacterial dissemination over time for each of the three different routes of inoculation. We also compared the radiance signal from animals infected with a wild type strain and a ?caf1?psaA mutant that we previously showed to be attenuated in colonization of the lymph node and systemic dissemination. Radiance signals from mice infected with the wild type strain were larger than values obtained from mice infected with the mutant strain (linear regression of normalized values, P? Conclusions We demonstrate that BLI is useful for monitoring dissemination from multiple inoculation sites, and for characterization of mutants with defects in colonization or dissemination.

Gonzalez Rodrigo J

2012-07-01

57

Stably Luminescent Staphylococcus aureus Clinical Strains for Use in Bioluminescent Imaging  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In vivo bioluminescent imaging permits the visualization of bacteria in live animals, allowing researchers to monitor, both temporally and spatially, the progression of infection in each animal. We sought to engineer stably luminescent clinical strains of Staphylococcus aureus, with the goal of using such strains in mouse models. The gram-positive shuttle vector pMAD was used as the backbone for an integration plasmid. A chloramphenicol resistance gene, a modified lux operon from Photorhabdus...

Plaut, Roger D.; Mocca, Christopher P.; Prabhakara, Ranjani; Merkel, Tod J.; Stibitz, Scott

2013-01-01

58

Identification of Inhibitors of ABCG2 by a Bioluminescence Imaging-based High-throughput Assay  

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ABCG2 is a member of the ATP-binding cassette (ABC) family of transporters, the overexpression of which is associated with tumor resistance to a variety of chemotherapeutic agents. Accordingly, combining ABCG2 inhibitor(s) with chemotherapy has the potential to improve treatment outcome. To search for clinically useful ABCG2 inhibitors, a bioluminescence imaging (BLI)-based assay was developed to allow high-throughput compound screening. This assay exploits our finding that D-luciferin, the s...

2009-01-01

59

Compartment-specific bioluminescence imaging platform for the high-throughput evaluation of antitumor immune function  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Conventional assays evaluating antitumor activity of immune effector cells have limitations that preclude their high-throughput application. We adapted the recently developed Compartment-Specific Bioluminescence Imaging (CS-BLI) technique to perform high-throughput quantification of innate antitumor activity and to show how pharmacologic agents (eg, lenalidomide, pomalidomide, bortezomib, and dexamethasone) and autologous BM stromal cells modulate that activity. CS-BLI–based screening allow...

Mcmillin, Douglas W.; Delmore, Jake; Negri, Joseph M.; Vanneman, Matthew; Koyama, Shohei; Schlossman, Robert L.; Munshi, Nikhil C.; Laubach, Jacob; Richardson, Paul G.; Dranoff, Glenn; Anderson, Kenneth C.; Mitsiades, Constantine S.

2012-01-01

60

Bioluminescence imaging in a medium-sized animal by local injection of d-luciferin  

Energy Technology Data Exchange (ETDEWEB)

Luciferase is one of the most commonly used reporter enzymes in the field of molecular imaging. D-luciferin is known as the substrate for luciferase enzyme and its cost is very expensive. Therefore, the bioluminescence molecular imaging study has been allowed in small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in articular capsule for bioluminescence imaging in rabbits. Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase. And then was performed different method of chondrocyte cell injection and transplantation into the knee of rabbits. The rabbits underwent imaging by cooled CCD camera after local injection of D-luciferin (3mg) into experimental knee joint as well as contralateral normal knee joint on days 1, 5, 7, 9. We sought whether optimal imaging signal was acquired by using cooled CCD camera after local injection of D-luciferin. We successfully visualized injected or transplanted cells in knee joint by local injection of D-luciferin. Total photon flux (7.86E+08 p/s/cm{sup 2}/sr) from the knee joint transplanted with cells approximately increased 10-fold more than (9.43E+07p/s/cm{sup 2}/sr) that from injected knee joints until 7 day. Imaging signal was observed in transplanted joints until day 9 after surgery while signal from injected knee was observed by day 7 after injection. We successfully carried out bioluminescence imaging study with medium sized animal by local injection of small amount of D-luciferin. Survival of chondrocytes were prolonged when surgically transplanted in joints than when directly injected in joint space.

Moon, Sung Min; Min, Jung Joon; Kim, Sung Mi; Bom, Hee Seung [Chonnam National University Medical School, Gwangju (Korea, Republic of); Oh, Suk Jung; Kang, Han Saem; Kim, Kwang Yoon [ECOBIO INC., Gwangju (Korea, Republic of); Kim, Young Ho [Chosun University, Gwangju (Korea, Republic of)

2005-07-01

 
 
 
 
61

Bioluminescence imaging in a medium-sized animal by local injection of d-luciferin  

International Nuclear Information System (INIS)

Luciferase is one of the most commonly used reporter enzymes in the field of molecular imaging. D-luciferin is known as the substrate for luciferase enzyme and its cost is very expensive. Therefore, the bioluminescence molecular imaging study has been allowed in small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in articular capsule for bioluminescence imaging in rabbits. Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase. And then was performed different method of chondrocyte cell injection and transplantation into the knee of rabbits. The rabbits underwent imaging by cooled CCD camera after local injection of D-luciferin (3mg) into experimental knee joint as well as contralateral normal knee joint on days 1, 5, 7, 9. We sought whether optimal imaging signal was acquired by using cooled CCD camera after local injection of D-luciferin. We successfully visualized injected or transplanted cells in knee joint by local injection of D-luciferin. Total photon flux (7.86E+08 p/s/cm2/sr) from the knee joint transplanted with cells approximately increased 10-fold more than (9.43E+07p/s/cm2/sr) that from injected knee joints until 7 day. Imaging signal was observed in transplanted joints until day 9 after surgery while signal from injected knee was observed by day 7 after injection. We successfully carried out bioluminescence imaging study with medium sized animal by local injection of small amount of D-luciferin. Survival of chondrocytes were prolonged when surgically transplanted in joints than when directly injected in joint space

2005-11-18

62

Development of a Novel Preclinical Pancreatic Cancer Research Model: Bioluminescence Image-Guided Focal Irradiation and Tumor Monitoring of Orthotopic Xenografts1  

Digital Repository Infrastructure Vision for European Research (DRIVER)

PURPOSE: We report on a novel preclinical pancreatic cancer research model that uses bioluminescence imaging (BLI)-guided irradiation of orthotopic xenograft tumors, sparing of surrounding normal tissues, and quantitative, noninvasive longitudinal assessment of treatment response. MATERIALS AND METHODS: Luciferase-expressing MiaPaCa-2 pancreatic carcinoma cells were orthotopically injected in nude mice. BLI was compared to pathologic tumor volume, and photon emission was assessed over time. B...

2012-01-01

63

Tomographic bioluminescence imaging by use of a combined optical-PET (OPET) system: a computer simulation feasibility study  

International Nuclear Information System (INIS)

The feasibility and limits in performing tomographic bioluminescence imaging with a combined optical-PET (OPET) system were explored by simulating its image formation process. A micro-MRI based virtual mouse phantom was assigned appropriate tissue optical properties to each of its segmented internal organs at wavelengths spanning the emission spectrum of the firefly luciferase at 37 deg. C. The TOAST finite-element code was employed to simulate the diffuse transport of photons emitted from bioluminescence sources in the mouse. OPET measurements were simulated for single-point, two-point and distributed bioluminescence sources located in different organs such as the liver, the kidneys and the gut. An expectation maximization code was employed to recover the intensity and location of these simulated sources. It was found that spectrally resolved measurements were necessary in order to perform tomographic bioluminescence imaging. The true location of emission sources could be recovered if the mouse background optical properties were known a priori. The assumption of a homogeneous optical property background proved inadequate for describing photon transport in optically heterogeneous tissues and led to inaccurate source localization in the reconstructed images. The simulation results pointed out specific methodological challenges that need to be addressed before a practical implementation of OPET-based bioluminescence tomography is achieved

2005-09-07

64

Tomographic bioluminescence imaging by use of a combined optical-PET (OPET) system: a computer simulation feasibility study  

Energy Technology Data Exchange (ETDEWEB)

The feasibility and limits in performing tomographic bioluminescence imaging with a combined optical-PET (OPET) system were explored by simulating its image formation process. A micro-MRI based virtual mouse phantom was assigned appropriate tissue optical properties to each of its segmented internal organs at wavelengths spanning the emission spectrum of the firefly luciferase at 37 deg. C. The TOAST finite-element code was employed to simulate the diffuse transport of photons emitted from bioluminescence sources in the mouse. OPET measurements were simulated for single-point, two-point and distributed bioluminescence sources located in different organs such as the liver, the kidneys and the gut. An expectation maximization code was employed to recover the intensity and location of these simulated sources. It was found that spectrally resolved measurements were necessary in order to perform tomographic bioluminescence imaging. The true location of emission sources could be recovered if the mouse background optical properties were known a priori. The assumption of a homogeneous optical property background proved inadequate for describing photon transport in optically heterogeneous tissues and led to inaccurate source localization in the reconstructed images. The simulation results pointed out specific methodological challenges that need to be addressed before a practical implementation of OPET-based bioluminescence tomography is achieved.

Alexandrakis, George [David Geffen School of Medicine at UCLA, Crump Institute for Molecular Imaging, University of California, 700 Westwood Plaza, Los Angeles, CA 90095 (United States); Rannou, Fernando R [Departamento de Ingenieria Informatica, Universidad de Santiago de Chile (USACH), Av. Ecuador 3659, Santiago (Chile); Chatziioannou, Arion F [David Geffen School of Medicine at UCLA, Crump Institute for Molecular Imaging, University of California, 700 Westwood Plaza, Los Angeles, CA 90095 (United States)

2005-09-07

65

Bioluminescence imaging of Clavibacter michiganensis subsp. michiganensis infection of tomato seeds and plants.  

Science.gov (United States)

Clavibacter michiganensis subsp. michiganensis is a Gram-positive bacterium that causes wilting and cankers, leading to severe economic losses in commercial tomato production worldwide. The disease is transmitted from infected seeds to seedlings and mechanically from plant to plant during seedling production, grafting, pruning, and harvesting. Because of the lack of tools for genetic manipulation, very little is known regarding the mechanisms of seed and seedling infection and movement of C. michiganensis subsp. michiganensis in grafted plants, two focal points for application of bacterial canker control measures in tomato. To facilitate studies on the C. michiganensis subsp. michiganensis movement in tomato seed and grafted plants, we isolated a bioluminescent C. michiganensis subsp. michiganensis strain using the modified Tn1409 containing a promoterless lux reporter. A total of 19 bioluminescent C. michiganensis subsp. michiganensis mutants were obtained. All mutants tested induced a hypersensitive response in Mirabilis jalapa and caused wilting of tomato plants. Real-time colonization studies of germinating seeds using a virulent, stable, constitutively bioluminescent strain, BL-Cmm17, showed that C. michiganensis subsp. michiganensis aggregated on hypocotyls and cotyledons at an early stage of germination. In grafted seedlings in which either the rootstock or scion was exposed to BL-Cmm17 via a contaminated grafting knife, bacteria were translocated in both directions from the graft union at higher inoculum doses. These results emphasize the use of bioluminescent C. michiganensis subsp. michiganensis to help better elucidate the C. michiganensis subsp. michiganensis-tomato plant interactions. Further, we demonstrated the broader applicability of this tool by successful transformation of C. michiganensis subsp. nebraskensis with Tn1409::lux. Thus, our approach would be highly useful to understand the pathogenesis of diseases caused by other subspecies of the agriculturally important C. michiganensis. PMID:20400561

Xu, Xiulan; Miller, Sally A; Baysal-Gurel, Fulya; Gartemann, Karl-Heinz; Eichenlaub, Rudolf; Rajashekara, Gireesh

2010-06-01

66

Bioluminescence Imaging of Toxoplasma gondii Infection in Living Mice Reveals Dramatic Differences between Strains  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We examined the in vivo growth, dissemination, and reactivation of strains of the protozoan parasite Toxoplasma gondii using a bioluminescence-based imaging system. Two T. gondii strains, one with a highly virulent disease phenotype in mice (S23) and the other with a 1,000-fold-lower virulence phenotype (S22), were engineered to stably express the light-emitting protein luciferase. One clone of each wild-type strain was isolated, and the two clones (S23-luc7 and S22-luc2) were found to expres...

Saeij, Jeroen P. J.; Boyle, Jon P.; Grigg, Michael E.; Arrizabalaga, Gustavo; Boothroyd, John C.

2005-01-01

67

In Vivo Bioluminescent Imaging (BLI: Noninvasive Visualization and Interrogation of Biological Processes in Living Animals  

Directory of Open Access Journals (Sweden)

Full Text Available In vivo bioluminescent imaging (BLI is increasingly being utilized as a method for modern biological research. This process, which involves the noninvasive interrogation of living animals using light emitted from luciferase-expressing bioreporter cells, has been applied to study a wide range of biomolecular functions such as gene function, drug discovery and development, cellular trafficking, protein-protein interactions, and especially tumorigenesis, cancer treatment, and disease progression. This article will review the various bioreporter/biosensor integrations of BLI and discuss how BLI is being applied towards a new visual understanding of biological processes within the living organism.

Steven Ripp

2010-12-01

68

Functional Imaging of Rel Expression in Inflammatory Processes Using Bioluminescence Imaging System in Transgenic Mice  

Science.gov (United States)

c-Rel plays important roles in many inflammatory diseases. Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs. In this paper, a transgenic mouse line, B6-Tg(c-Rel-luc)Mlit, which incorporated the transgene firefly luciferase driven by a 14.5-kb fragment containing mouse c-Rel gene Rel promoter, was generated to monitor Rel expression in vivo. Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model. The luciferase expression in transgenic mice was comparable to the endogenous Rel expression and could be suppressed by administration of anti-inflammatory drug dexamethasone or aspirin. These results indicate that the B6-Tg(c-Rel-luc)Mlit mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

Yang, Xingyu; Jing, Hua; Zhao, Kai; Sun, Ruilin; Liu, Zhenze; Ying, Yue; Ci, Lei; Kuang, Ying; Huang, Fang; Wang, Zhugang; Fei, Jian

2013-01-01

69

A biocompatible in vivo ligation reaction and its application for noninvasive bioluminescent imaging of protease activity in living mice.  

Science.gov (United States)

The discovery of biocompatible reactions had a tremendous impact on chemical biology, allowing the study of numerous biological processes directly in complex systems. However, despite the fact that multiple biocompatible reactions have been developed in the past decade, very few work well in living mice. Here we report that D-cysteine and 2-cyanobenzothiazoles can selectively react with each other in vivo to generate a luciferin substrate for firefly luciferase. The success of this "split luciferin" ligation reaction has important implications for both in vivo imaging and biocompatible labeling strategies. First, the production of a luciferin substrate can be visualized in a live mouse by bioluminescence imaging (BLI) and furthermore allows interrogation of targeted tissues using a "caged" luciferin approach. We therefore applied this reaction to the real-time noninvasive imaging of apoptosis associated with caspase 3/7. Caspase-dependent release of free D-cysteine from the caspase 3/7 peptide substrate Asp-Glu-Val-Asp-D-Cys (DEVD-(D-Cys)) allowed selective reaction with 6-amino-2-cyanobenzothiazole (NH(2)-CBT) in vivo to form 6-amino-D-luciferin with subsequent light emission from luciferase. Importantly, this strategy was found to be superior to the commercially available DEVD-aminoluciferin substrate for imaging of caspase 3/7 activity. Moreover, the split luciferin approach enables the modular construction of bioluminogenic sensors, where either or both reaction partners could be caged to report on multiple biological events. Lastly, the luciferin ligation reaction is 3 orders of magnitude faster than Staudinger ligation, suggesting further applications for both bioluminescence and specific molecular targeting in vivo. PMID:23463944

Godinat, Aurélien; Park, Hyo Min; Miller, Stephen C; Cheng, Ke; Hanahan, Douglas; Sanman, Laura E; Bogyo, Matthew; Yu, Allen; Nikitin, Gennady F; Stahl, Andreas; Dubikovskaya, Elena A

2013-05-17

70

Bioluminescence imaging allows measuring CD8 T cell function in the liver.  

Science.gov (United States)

In vivo evaluation of CD8 T cell effector (cytotoxic T lymphocyte [CTL]) function in peripheral organs such as the liver is currently not possible but would greatly improve our understanding of local immune regulation, because simple determination of antigen-specific CTL numbers does not predict the outcome of immune responses. In particular, measurement of alanine aminotransferase serum levels is not sensitive enough to detect T cell immunity against low numbers of target hepatocytes. We developed a procedure that detects virus-specific effector function of CTLs in the liver after simultaneous adenoviral transfer of reporter and immune target genes into hepatocytes, followed by bioluminescence imaging of reporter genes. Bioluminescence imaging enabled detection of as few as 10,000 infected hepatocytes in vivo, and even more importantly, quantification of antiviral effector function of as few as 50,000 CTLs. Conclusion: Our results provide evidence that low numbers of antigen-specific CTLs are sufficient to control viral gene expression and eliminate viral infection from hepatocytes. The experimental system established here is a highly sensitive method to simultaneously detect viral infection of hepatocytes and to quantify antiviral CTL function in the liver in vivo and will help in characterizing principles of hepatic immune regulation. PMID:20373369

Stabenow, Dirk; Frings, Marianne; Trück, Christina; Gärtner, Katja; Förster, Irmgard; Kurts, Christian; Tüting, Thomas; Odenthal, Margarete; Dienes, Hans-Peter; Cederbrant, Karin; Protzer, Ulrike; Knolle, Percy A

2010-04-01

71

Multimodality Imaging of Tumor Xenografts and Metastases in Mice with Combined Small-Animal PET, Small-Animal CT, and Bioluminescence Imaging  

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Recent developments have established molecular imaging of mouse models with small-animal PET and bioluminescence imaging (BLI) as an important tool in cancer research. One of the disadvantages of these imaging modalities is the lack of anatomic information. We combined small-animal PET and BLI technology with small-animal CT to obtain fusion images with both molecular and anatomic information.

Deroose, Christophe M.; De, Abhijit; Loening, Andreas M.; Chow, Patrick L.; Ray, Pritha; Chatziioannou, Arion F.; Gambhir, Sanjiv S.

2007-01-01

72

Bioluminescent imaging of Trypanosoma cruzi infection in Rhodnius prolixus  

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Abstract Background Usually the analysis of the various developmental stages of Trypanosoma cruzi in the experimentally infected vertebrate and invertebrate hosts is based on the morphological observations of tissue fragments from animals and insects. The development of techniques that allow the imaging of animals infected with parasites expressing luciferase open up possibilities to follow the fate of bioluminescent parasites in infected vectors. Methods ...

2012-01-01

73

Registration of planar bioluminescence to magnetic resonance and x-ray computed tomography images as a platform for the development of bioluminescence tomography reconstruction algorithms  

Science.gov (United States)

The procedures we propose make possible the mapping of two-dimensional (2-D) bioluminescence image (BLI) data onto a skin surface derived from a three-dimensional (3-D) anatomical modality [magnetic resonance (MR) or computed tomography (CT)] dataset. This mapping allows anatomical information to be incorporated into bioluminescence tomography (BLT) reconstruction procedures and, when applied using sources visible to both optical and anatomical modalities, can be used to evaluate the accuracy of those reconstructions. Our procedures, based on immobilization of the animal and a priori determined fixed projective transforms, should be more robust and accurate than previously described efforts, which rely on a poorly constrained retrospectively determined warping of the 3-D anatomical information. Experiments conducted to measure the accuracy of the proposed registration procedure found it to have a mean error of 0.36+/-0.23 mm. Additional experiments highlight some of the confounds that are often overlooked in the BLT reconstruction process, and for two of these confounds, simple corrections are proposed.

Beattie, Bradley J.; Klose, Alexander D.; Le, Carl H.; Longo, Valerie A.; Dobrenkov, Konstantine; Vider, Jelena; Koutcher, Jason A.; Blasberg, Ronald G.

2009-03-01

74

Bioluminescent Imaging of HPV-Positive Oral Tumor Growth and Its Response to Image-Guided Radiotherapy.  

Science.gov (United States)

The treatment paradigms for head and neck squamous cell cancer (HNSCC) are changing due to the emergence of human papillomavirus (HPV)-associated tumors possessing distinct molecular profiles and responses to therapy. Although patients with HNSCCs are often treated with radiotherapy, preclinical models are limited by the ability to deliver precise radiation to orthotopic tumors and to monitor treatment responses accordingly. To better model this clinical scenario, we developed a novel autochthonous HPV-positive oral tumor model to track responses to small molecules and image-guided radiation. We used a tamoxifen-regulated Cre recombinase system to conditionally express the HPV oncogenes E6 and E7 as well as a luciferase reporter (iHPV-Luc) in the epithelial cells of transgenic mice. In the presence of activated Cre recombinase, luciferase activity, and by proxy, HPV oncogenes were induced to 11-fold higher levels. In triple transgenic mice containing the iHPV-Luc, K14-CreER(tam), and LSL-Kras transgenes, tamoxifen treatment resulted in oral tumor development with increased bioluminescent activity within 6 days that reached a maximum of 74.8-fold higher bioluminescence compared with uninduced mice. Oral tumors expressed p16 and MCM7, two biomarkers associated with HPV-positive tumors. After treatment with rapamycin or image-guided radiotherapy, tumors regressed and possessed decreased bioluminescence. Thus, this novel system enables us to rapidly visualize HPV-positive tumor growth to model existing and new interventions using clinically relevant drugs and radiotherapy techniques. Cancer Res; 74(7); 2073-81. ©2014 AACR. PMID:24525739

Zhong, Rong; Pytynia, Matt; Pelizzari, Charles; Spiotto, Michael

2014-04-01

75

Assessing the effect of EPO on tumor oxygenation and radioresponsiveness via in vivo bioluminescence imaging  

International Nuclear Information System (INIS)

Evaluating tumor kill by volume measurement lacks sensitivity while in vivo-in vitro and histological assays are unsuitable for serial measurements. In vivo bioluminescence imaging (BI) nondestructively measures the number of metabolically active cells containing luciferase (LUC) over time. In this paper, the effect of erythropoietin (EPO) on tumor oxygenation and radioresponsivenessis is studied using BI and conventional methods. Murine adenocarcinoma cells, transfected with the LUC gene, were placed in the flank of BALB/C mice. EPO 1 u/g or saline was injected sc tiw for two weeks, starting the day of transplant. Mice then underwent irradiation (XRT) or pO2 measurement with an optical probe. In BI, mice were injected with luciferin and total photon flux (TPF) measured with a CCD camera. In vitro, cells were plated, irradiated and incubated at 37 deg C. Initial hematocrit was 47% (n=119) vs. 61% in EPO-treated mice (n=23, p2 (6.4 vs. 4.7 mm Hg, p=0.04) than controls. For 1-3x7 Gy, TPF was stable for 2 days after the start of XRT, then fell precipitously. Two weeks post XRT, TPF was 10-5 the initial value and a nidus of LUC activity persisted for months in most tumors. Tumor volume decreased only 1-2 orders of magnitude. For 3x7 Gy, tumor regrew in 1/11 EPO-TM and controls (p=NS.) For 1x7 Gy, tumors regrew in 4/6 EPO-TM and 2/4 controls (p=NS). TPF did not increase with tumor regrowth. Recurrent tumors exhibited lower median pO2 (2.1 mm Hg, p=.003) and higher hypoxic fraction than controls. A clonogenic assay yielded D10 = 3.7 Gy with all colonies expressing LUC. The TPF of 0-Gy treated wells rose significantly over incubation, while that of wells treated to 10 Gy was unchanged. Though EPO improved tumor oxygenation, no effect on XRT-mediated cell kill was seen. BI measured tumor killing in vivo over a broad dynamic range. The results suggest that cell killing in vivo is a multistep process, amplified by humoral factors

2003-08-17

76

Self-illuminating in vivo lymphatic imaging using a bioluminescence resonance energy transfer quantum dot nano-particle  

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Autofluorescence arising from normal tissues can compromise the sensitivity and specificity of in vivo fluorescence imaging by lowering the target-to-background signal ratio. Since bioluminescence resonance energy transfer quantum dot (BRET-QDot) nano-particles can self-illuminate in near-infrared in the presence of the substrate, coelenterazine, without irradiating excitation lights, imaging using BRET-QDots does not produce any autofluorescence. In this study, we applied this BRET-QDot nano...

Kosaka, Nobuyuki; Mitsunaga, Makoto; Bhattacharyya, Sukanta; Miller, Steven C.; Choyke, Peter L.; Kobayashi, Hisataka

2011-01-01

77

Sensitive in situ monitoring of a recombinant bioluminescent Yersinia enterocolitica reporter mutant in real time on Camembert cheese.  

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Bioluminescent mutants of Yersinia enterocolitica were generated by transposon mutagenesis using a promoterless, complete lux operon (luxCDABE) derived from Photorhabdus luminescens, and their production of light in the cheese environment was monitored. Mutant B94, which had the lux cassette inserted into an open reading frame of unknown function was used for direct monitoring of Y. enterocolitica cells on cheeses stored at 10 degrees C by quantifying bioluminescence using a photon-counting, intensified charge-coupled device camera. The detection limit on cheese was 200 CFU/cm(2). Bioluminescence of the reporter mutant was significantly regulated by its environment (NaCl, temperature, and cheese), as well as by growth phase, via the promoter the lux operon had acquired upon transposition. At low temperatures, mutant B94 did not exhibit the often-reported decrease of photon emission in older cells. It was not necessary to include either antibiotics or aldehyde in the food matrix in order to gain quantitative, reproducible bioluminescence data. As far as we know, this is the first time a pathogen has been monitored in situ, in real time, in a "real-product" status, and at a low temperature. PMID:12406772

Maoz, Ariel; Mayr, Ralf; Bresolin, Geraldine; Neuhaus, Klaus; Francis, Kevin P; Scherer, Siegfried

2002-11-01

78

Fast iterative image reconstruction methods for fully 3D multispectral bioluminescence tomography  

International Nuclear Information System (INIS)

We investigate fast iterative image reconstruction methods for fully 3D multispectral bioluminescence tomography for applications in small animal imaging. Our forward model uses a diffusion approximation for optically inhomogeneous tissue, which we solve using a finite element method (FEM). We examine two approaches to incorporating the forward model into the solution of the inverse problem. In a conventional direct calculation approach one computes the full forward model by repeated solution of the FEM problem, once for each potential source location. We describe an alternative on-the-fly approach where one does not explicitly solve for the full forward model. Instead, the solution to the forward problem is included implicitly in the formulation of the inverse problem, and the FEM problem is solved at each iteration for the current image estimate. We evaluate the convergence speeds of several representative iterative algorithms. We compare the computation cost of those two approaches, concluding that the on-the-fly approach can lead to substantial reductions in total cost when combined with a rapidly converging iterative algorithm

2008-07-21

79

Bioluminescence imaging of invasive intracranial xenografts: implications for translational research and targeted therapeutics of brain tumors.  

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Despite decades of study, the etiology of brain cancer remains elusive. However, extensive molecular characterization of primary brain tumors has been accomplished, outlining recurrent features that are proving useful for devising targeted therapies. There are far too few patients available for comparing the efficacy of therapeutic combinations, especially when variations in dosing, frequency, and sequencing are taken into account. Consequently, there is a substantial need for increasing preclinical testing throughput using clinically relevant models. We review luminescent optical imaging for its potential in facilitating in vivo assessment of intracranial tumor growth and response to therapy in rodent orthotopic xenograft models of primary brain malignancies. We review the rationale behind the need of an in vivo model, why orthotopic tumor models displaying an invasive phenotype may be a superior choice when compared to flank-implanted tumors, and what advantages may be drawn from the use of modified cells, suitable for sequential monitoring by in vivo optical imaging. Studies show that luminescent signal correlates highly both with tumor burden and Kaplan-Meier survival curves of rodents bearing intracranial xenografts. We conclude that bioluminescent imaging is a highly sensitive technique for assessment of tumor burden, response to therapy, tumor recurrence, and behavior to salvage therapy, making it a superior option for longitudinal monitoring in intracranial rodent models of primary brain tumors. PMID:20652720

Dinca, Eduard B; Voicu, Ramona V; Ciurea, Alexandru V

2010-10-01

80

Evaluating reporter genes of different luciferases for optimized in vivo bioluminescence imaging of transplanted neural stem cells in the brain  

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Bioluminescence imaging (BLI) has become the method of choice for optical tracking of cells in small laboratory animals. However, the use of luciferases from different species, depending on different substrates and emitting at distinct wavelengths, has not been optimized for sensitive neuroimaging. In order to identify the most suitable luciferase, this quantitative study compared the luciferases Luc2, CBG99, PpyRE9 and hRluc. Human embryonic kidney (HEK-293) cells and mouse neural stem cells...

2013-01-01

 
 
 
 
81

Effect of optical property estimation accuracy on tomographic bioluminescence imaging: simulation of a combined optical–PET (OPET) system  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Inevitable discrepancies between the mouse tissue optical properties assumed by an experimenter and the actual physiological values may affect the tomographic localization of bioluminescent sources. In a previous work, the simplifying assumption of optically homogeneous tissues led to inaccurate localization of deep sources. Improved results may be obtained if a mouse anatomical map is provided by a high-resolution imaging modality and optical properties are assigned to segmented tissues. In ...

Alexandrakis, George; Rannou, Fernando R.; Chatziioannou, Arion F.

2006-01-01

82

Making the brain glow: in vivo bioluminescence imaging to study neurodegeneration.  

Science.gov (United States)

Bioluminescence imaging (BLI) takes advantage of the light-emitting properties of luciferase enzymes, which produce light upon oxidizing a substrate (i.e., D-luciferin) in the presence of molecular oxygen and energy. Photons emitted from living tissues can be detected and quantified by a highly sensitive charge-coupled device camera, enabling the investigator to noninvasively analyze the dynamics of biomolecular reactions in a variety of living model organisms such as transgenic mice. BLI has been used extensively in cancer research, cell transplantation, and for monitoring of infectious diseases, but only recently experimental models have been designed to study processes and pathways in neurological disorders such as Alzheimer disease, Parkinson disease, or amyotrophic lateral sclerosis. In this review, we highlight recent applications of BLI in neuroscience, including transgene expression in the brain, longitudinal studies of neuroinflammatory responses to neurodegeneration and injury, and in vivo imaging studies of neurogenesis and mitochondrial toxicity. Finally, we highlight some new developments of BLI compounds and luciferase substrates with promising potential for in vivo studies of neurological dysfunctions. PMID:23192390

Hochgräfe, Katja; Mandelkow, Eva-Maria

2013-06-01

83

Bioluminescence in the Sea  

Science.gov (United States)

Bioluminescence spans all oceanic dimensions and has evolved many times—from bacteria to fish—to powerfully influence behavioral and ecosystem dynamics. New methods and technology have brought great advances in understanding of the molecular basis of bioluminescence, its physiological control, and its significance in marine communities. Novel tools derived from understanding the chemistry of natural light-producing molecules have led to countless valuable applications, culminating recently in a related Nobel Prize. Marine organisms utilize bioluminescence for vital functions ranging from defense to reproduction. To understand these interactions and the distributions of luminous organisms, new instruments and platforms allow observations on individual to oceanographic scales. This review explores recent advances, including the chemical and molecular, phylogenetic and functional, community and oceanographic aspects of bioluminescence.

Haddock, Steven H. D.; Moline, Mark A.; Case, James F.

2010-01-01

84

Evaluation of monkeypox virus infection of prairie dogs (Cynomys ludovicianus) using in vivo bioluminescent imaging  

Science.gov (United States)

Monkeypox (MPX) is a re-emerging zoonotic disease that is endemic in Central and West Africa, where it can cause a smallpox-like disease in humans. Despite many epidemiologic and field investigations of MPX, no definitive reservoir species has been identified. Using recombinant viruses expressing the firefly luciferase (luc) gene, we previously demonstrated the suitability of in vivo bioluminescent imaging (BLI) to study the pathogenesis of MPX in animal models. Here, we evaluated BLI as a novel approach for tracking MPX virus infection in black-tailed prairie dogs (Cynomys ludovicianus). Prairie dogs were affected during a multistate outbreak of MPX in the US in 2003 and have since been used as an animal model of this disease. Our BLI results were compared with PCR and virus isolation from tissues collected postmortem. Virus was easily detected and quantified in skin and superficial tissues by BLI before and during clinical phases, as well as in subclinical secondary cases, but was not reliably detected in deep tissues such as the lung. Although there are limitations to viral detection in larger wild rodent species, BLI can enhance the use of prairie dogs as an animal model of MPX and can be used for the study of infection, disease progression, and transmission in potential wild rodent reservoirs.

Falendysz, Elizabeth A.; Londońo-Navas, Angela M.; Meteyer, Carol U.; Pussini, Nicola; Lopera, Juan G.; Osorio, Jorge E.; Rocke, Tonie E.

2014-01-01

85

Real-time monitoring of bioaerosols via cell-lysis by air ion and ATP bioluminescence detection.  

Science.gov (United States)

In this study, we introduce a methodology for disrupting cell membranes with air ions coupled with ATP bioluminescence detection for real-time monitoring of bioaerosol concentrations. A carbon fiber ionizer was used to extract ATP from bacterial cells for generating ATP bioluminescence. Our methodology was tested using Staphylococcus epidermidis and Escherichia coli, which were aerosolized with an atomizer, and then indoor bioaerosols were also used for testing the methodology. Bioaerosol concentrations were estimated without culturing which requires several days for colony formation. Correlation equations were obtained for results acquired using our methodology (Relative Luminescent Unit (RLU)/m(3)) and a culture-based (Colony Forming Unit (CFU)/m(3)) method; CFU/m(3)=1.8 × measured RLU/m(3) for S. epidermidis and E. coli, and CFU/m(3)=1.1 × measured RLU/m(3) for indoor bioaerosols under the experimental conditions. Our methodology is an affordable solution for rapidly monitoring bioaerosols due to rapid detection time (cell-lysis time: 3 min; bioluminescence detection time: <1 min) and easy operation. PMID:24080217

Park, Chul Woo; Park, Ji-Woon; Lee, Sung Hwa; Hwang, Jungho

2014-02-15

86

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model  

Science.gov (United States)

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1? (HIF-1?) to drive transcription of various genes. Therefore, we have used a HIF-1? reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1? bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1? binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1? bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

Saha, Debabrata; Dunn, Henry; Zhou, Heling; Harada, Hiroshi; Hiraoka, Masahiro; Mason, Ralph P.; Zhao, Dawen

2011-01-01

87

In vivo cell tracking by bioluminescence imaging after transplantation of bioengineered cell sheets to the knee joint.  

Science.gov (United States)

In our previous studies, we have demonstrated effective regeneration of cartilage through the creation and application of layered cell sheets that combine both chondrocytes and synovial cells. In this study, we were able to demonstrate that cells derived from cell sheets can survive for long periods after transplantation into rat knee joints having osteochondral defects. We established a method for generating cell sheets from firefly luciferase-expressing chondrocytes obtained from transgenic Lewis rats, and carried out allogenic transplantation of these cell sheets into wild-type Lewis rats. We then administered luciferin and monitored the survival of the transplanted cells by using bioluminescence imaging (BLI). Our data showed that the transplanted cells survived and could be detected for more than 21 months, which was longer than expected. Furthermore, the BLI data showed that the transplanted cells remained in the knee joint and did not migrate to other parts of the body, thus confirming the safety of the cell sheets. In this study, we monitored the duration of survival of cell sheets composed of only chondrocytes, only synovial cells, or both chondrocytes and synovial cells, and found that all three types of cell sheets survived for an extended period of time. PMID:24360579

Takaku, Yuko; Murai, Kunihiko; Ukai, Taku; Ito, Satoshi; Kokubo, Mami; Satoh, Masaaki; Kobayashi, Eiji; Yamato, Masayuki; Okano, Teruo; Takeuchi, Mamoru; Mochida, Joji; Sato, Masato

2014-02-01

88

Rat model of metastatic breast cancer monitored by MRI at 3 tesla and bioluminescence imaging with histological correlation  

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Full Text Available Abstract Background Establishing a large rodent model of brain metastasis that can be monitored using clinically relevant magnetic resonance imaging (MRI techniques is challenging. Non-invasive imaging of brain metastasis in mice usually requires high field strength MR units and long imaging acquisition times. Using the brain seeking MDA-MB-231BR transfected with luciferase gene, a metastatic breast cancer brain tumor model was investigated in the nude rat. Serial MRI and bioluminescence imaging (BLI was performed and findings were correlated with histology. Results demonstrated the utility of multimodality imaging in identifying unexpected sights of metastasis and monitoring the progression of disease in the nude rat. Methods Brain seeking breast cancer cells MDA-MB-231BR transfected with firefly luciferase (231BRL were labeled with ferumoxides-protamine sulfate (FEPro and 1-3 × 106 cells were intracardiac (IC injected. MRI and BLI were performed up to 4 weeks to monitor the early breast cancer cell infiltration into the brain and formation of metastases. Rats were euthanized at different time points and the imaging findings were correlated with histological analysis to validate the presence of metastases in tissues. Results Early metastasis of the FEPro labeled 231BRL were demonstrated onT2*-weighted MRI and BLI within 1 week post IC injection of cells. Micro-metastatic tumors were detected in the brain on T2-weighted MRI as early as 2 weeks post-injection in greater than 85% of rats. Unexpected skeletal metastases from the 231BRL cells were demonstrated and validated by multimodal imaging. Brain metastases were clearly visible on T2 weighted MRI by 3-4 weeks post infusion of 231BRL cells, however BLI did not demonstrate photon flux activity originating from the brain in all animals due to scattering of the photons from tumors. Conclusion A model of metastatic breast cancer in the nude rat was successfully developed and evaluated using multimodal imaging including MRI and BLI providing the ability to study the temporal and spatial distribution of metastases in the brain and skeleton.

Klaunberg Brenda

2009-10-01

89

Novel mouse mammary cell lines for in vivo bioluminescence imaging (BLI of bone metastasis  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Tumor cell lines that can be tracked in vivo during tumorigenesis and metastasis provide vital tools for studying the specific cellular mechanisms that mediate these processes as well as investigating therapeutic targets to inhibit them. The goal of this study was to engineer imageable mouse mammary tumor cell lines with discrete propensities to metastasize to bone in vivo. Two novel luciferase expressing cell lines were developed and characterized for use in the study of breast cancer metastasis to bone in a syngeneic mouse model. Results The 4 T1.2 luc3 and 66c14 luc2 cell lines were shown to have high levels of bioluminescence intensity in vitro and in vivo after orthotopic injection into mouse mammary fat pads. The 4 T1.2 luc3 cell line was found to closely model the sites of metastases seen in human patients including lung, liver, and bone. Specifically, 4 T1.2 luc3 cells demonstrated a high incidence of metastasis to spine, with an ex-vivo BLI intensity three orders of magnitude above the commercially available 4 T1 luc2 cells. 66c14 luc2 cells also demonstrated metastasis to spine, which was lower than that of 4 T1.2 luc3 cells but higher than 4 T1 luc2 cells, in addition to previously unreported metastases in the liver. High osteolytic activity of the 4 T1.2 luc3 cells in vivo in the bone microenvironment was also detected. Conclusions The engineered 4 T1.2 luc3 and 66c14 luc2 cell lines described in this study are valuable tools for studying the cellular events moderating the metastasis of breast tumor cells to bone.

Bolin Celeste

2012-04-01

90

Bioluminescent imaging of drug efflux at the blood-brain barrier mediated by the transporter ABCG2.  

Science.gov (United States)

ATP-binding cassette (ABC) transporters are a group of transmembrane proteins that maintain chemical homeostasis through efflux of compounds out of organelles and cells. Among other functions, ABC transporters play a key role in protecting the brain parenchyma by efflux of xenobiotics from capillary endothelial cells at the blood-brain barrier (BBB). They also prevent the entry of therapeutic drugs at the BBB, thereby limiting their efficacy. One of the key transporters playing this role is ABCG2. Although other ABC transporters can be studied through various imaging modalities, no specific probe exists for imaging ABCG2 function in vivo. Here we show that D-luciferin, the endogenous substrate of firefly luciferase, is a specific substrate for ABCG2. We hypothesized that ABCG2 function at the BBB could be evaluated by using bioluminescence imaging in transgenic mice expressing firefly luciferase in the brain. Bioluminescence signal in the brain of mice increased with coadministration of the ABCG2 inhibitors Ko143, gefitinib, and nilotinib, but not an ABCB1 inhibitor. This method for imaging ABCG2 function at the BBB will facilitate understanding of the function and pharmacokinetic inhibition of this transporter. PMID:24297888

Bakhsheshian, Joshua; Wei, Bih-Rong; Chang, Ki-Eun; Shukla, Suneet; Ambudkar, Suresh V; Simpson, R Mark; Gottesman, Michael M; Hall, Matthew D

2013-12-17

91

Establishment of cell strains stably-transfected with luciferase gene mediated by retrovirus and their detection with bioluminescence imaging system  

Directory of Open Access Journals (Sweden)

Full Text Available Objective ?To establish cell strains stably transfected with Luciferase gene (Luc2, which was mediated by retrovirus, and explore the relationship between the number of Luc2-positive cells and light flux from bioluminescence imaging system by experiments in vitro and in vivo. Methods ?We co-transfected pMX-Luc2 plasmid and pMD.G plasmid into 293T gag-pol cells to get retrovirus expressing Luc2 gene. Stable Luc2 positive cell lines were generated and screened by transduction of Retro-Luc2 in mouse colon cancer cell line CT26, human non-small cell lung cancer cell line NCI-H446, human colon cancer cell line HT-29, human ovarian carcinoma cell line SKOV3 and human hepatocellular carcinoma cell line SMMC-7721, all of them were identified by bioluminescence imaging system. Different numbers of SKOV3-Luc2 cells ranging from 10 to 10000 were plated onto culture dishes. Two xenograft models of ovarian cancer were reproduced by subcutaneous injection of 200?l SKOV3-Luc2 cell suspension with different concentrations (1×107, 5×106, 2.5×106, 1×106, 5×105, 2.5×105, 1×105 and 5×104/ml into 16 sites on the back of 4 nude mice, or intravenous injection of 1×106 or 3 ×106 SKOV3-Luc2 cells into the tail vein. Light flux value of SKOV3-Luc2 cells in dishes and in mice was measured by bioluminescence imaging system. Results ?Retro-Luc2 was constructed successfully and expressed Luc2 stably in transduced CT26, NCI-H446, HT-29, SKOV3 and SMMC-7721 cell lines. Light flux was correlated in a linear manner with the number of Luc2-positive cells in dishes and in mice (R2=0.944, ?=0.972; R2=0.991, ?=0.996; R2=0.351, ?=0.628; P < 0.01. Conclusion ?Luc2-positive cell lines could be established rapidly and accurately by infecting tumor cells with retrovirus expressing Luc2. The number of Luc2 positive cells is significantly related in a linear manner to light flux from bioluminescence imaging system in vitro and in vivo.

Hai-juan WANG

2012-05-01

92

Effect of optical property estimation accuracy on tomographic bioluminescence imaging: simulation of a combined optical-PET (OPET) system  

Energy Technology Data Exchange (ETDEWEB)

Inevitable discrepancies between the mouse tissue optical properties assumed by an experimenter and the actual physiological values may affect the tomographic localization of bioluminescent sources. In a previous work, the simplifying assumption of optically homogeneous tissues led to inaccurate localization of deep sources. Improved results may be obtained if a mouse anatomical map is provided by a high-resolution imaging modality and optical properties are assigned to segmented tissues. In this work, the feasibility of this approach was explored by simulating the effect of different magnitude optical property errors on the image formation process of a combined optical-PET system. Some comparisons were made with corresponding simulations using higher spatial resolution data that are typically attainable by CCD cameras. In addition, simulation results provided insights on some of the experimental conditions that could lead to poor localization of bioluminescent sources. They also provided a rough guide on how accurately tissue optical properties need to be known in order to achieve correct localization of point sources with increasing tissue depth under low background noise conditions.

Alexandrakis, George [Crump Institute for Molecular Imaging, David Geffen School of Medicine at UCLA, University of California, 700 Westwood Plaza, Los Angeles, CA 90095 (United States); Rannou, Fernando R [Departamento de Ingenieria Informatica, Universidad de Santiago de Chile (USACH), Av. Ecuador 3659, Santiago (Chile); Chatziioannou, Arion F [Crump Institute for Molecular Imaging, David Geffen School of Medicine at UCLA, University of California, 700 Westwood Plaza, Los Angeles, CA 90095 (United States)

2006-04-21

93

Effect of optical property estimation accuracy on tomographic bioluminescence imaging: simulation of a combined optical-PET (OPET) system  

International Nuclear Information System (INIS)

Inevitable discrepancies between the mouse tissue optical properties assumed by an experimenter and the actual physiological values may affect the tomographic localization of bioluminescent sources. In a previous work, the simplifying assumption of optically homogeneous tissues led to inaccurate localization of deep sources. Improved results may be obtained if a mouse anatomical map is provided by a high-resolution imaging modality and optical properties are assigned to segmented tissues. In this work, the feasibility of this approach was explored by simulating the effect of different magnitude optical property errors on the image formation process of a combined optical-PET system. Some comparisons were made with corresponding simulations using higher spatial resolution data that are typically attainable by CCD cameras. In addition, simulation results provided insights on some of the experimental conditions that could lead to poor localization of bioluminescent sources. They also provided a rough guide on how accurately tissue optical properties need to be known in order to achieve correct localization of point sources with increasing tissue depth under low background noise conditions

2006-04-21

94

Application of Bioluminescence Imaging to the Prediction of Lethality in Vaccinia Virus-Infected Mice? †  

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To find an alternative endpoint for the efficacy of antismallpox treatments, bioluminescence was measured in live BALB/c mice following lethal challenge with a recombinant WR vaccinia virus expressing luciferase. Intravenous vaccinia immunoglobulin treatments were used to confer protection on a proportion of animals. Using known lethality outcomes in 200 animals and total fluxes recorded daily in live animals, we performed univariate receiver operating characteristic (ROC) curve analysis to a...

Zaitseva, Marina; Kapnick, Senta M.; Scott, John; King, Lisa R.; Manischewitz, Jody; Sirota, Lev; Kodihalli, Shantha; Golding, Hana

2009-01-01

95

Destabilized bioluminescent proteins  

Energy Technology Data Exchange (ETDEWEB)

Purified nucleic acids, vectors and cells containing a gene cassette encoding at least one modified bioluminescent protein, wherein the modification includes the addition of a peptide sequence. The duration of bioluminescence emitted by the modified bioluminescent protein is shorter than the duration of bioluminescence emitted by an unmodified form of the bioluminescent protein.

Allen, Michael S. (Knoxville, TN); Rakesh, Gupta (New Delhi, IN); Gary, Sayler S. (Blaine, TN)

2007-07-31

96

A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague.  

Science.gov (United States)

The gold standard in microbiology for monitoring bacterial dissemination in infected animals has always been viable plate counts. This method, despite being quantitative, requires sacrificing the infected animals. Recently, however, an alternative method of in vivo imaging of bioluminescent bacteria (IVIBB) for monitoring microbial dissemination within the host has been employed. Yersinia pestis is a Gram-negative bacterium capable of causing bubonic, septicemic, and pneumonic plague. In this study, we compared the conventional counting of bacterial colony forming units (cfu) in the various infected tissues to IVIBB in monitoring Y. pestis dissemination in a mouse model of pneumonic plague. By using a transposon mutagenesis system harboring the luciferase (luc) gene, we screened approximately 4000 clones and obtained a fully virulent, luc-positive Y. pestis CO92 (Y. pestis-luc2) reporter strain in which transposition occurred within the largest pMT1 plasmid which possesses murine toxin and capsular antigen encoding genes. The aforementioned reporter strain and the wild-type CO92 exhibited similar growth curves, formed capsule based on immunofluorescence microscopy and flow cytometry, and had a similar LD(50). Intranasal infection of mice with 15 LD(50) of CO92-luc2 resulted in animal mortality by 72 h, and an increasing number of bioluminescent bacteria were observed in various mouse organs over a 24-72 h period when whole animals were imaged. However, following levofloxacin treatment (10 mg/kg/day) for 6 days 24 h post infection, no luminescence was observed after 72 h of infection, indicating that the tested antimicrobial killed bacteria preventing their detection in host peripheral tissues. Overall, we demonstrated that IVIBB is an effective and non-invasive way of monitoring bacterial dissemination in animals following pneumonic plague having strong correlation with cfu, and our reporter CO92-luc2 strain can be employed as a useful tool to monitor the efficacy of antimicrobial countermeasures in real time. PMID:23063826

Sha, Jian; Rosenzweig, Jason A; Kirtley, Michelle L; van Lier, Christina J; Fitts, Eric C; Kozlova, Elena V; Erova, Tatiana E; Tiner, Bethany L; Chopra, Ashok K

2013-02-01

97

Firefly luciferase inhibitor-conjugated Peptide quenches bioluminescence: a versatile tool for real time monitoring cellular uptake of biomolecules.  

Science.gov (United States)

In this paper, novel firefly luciferase-specific inhibitor compounds (FLICs) are evaluated as potential tools for cellular trafficking of transporter conjugates. As a proof-of-concept, we designed FLICs that were suitable for solid phase peptide synthesis and could be covalently conjugated to peptides via an amide bond. The spacer between inhibitor and peptide was optimized to gain efficient inhibition of recombinant firefly luciferase (FLuc) without compromising the activity of the model peptides. The hypothesis of using FLICs as tools for cellular trafficking studies was ensured with U87Fluc glioblastoma cells expressing firefly luciferase. Results show that cell penetrating peptide (penetratin) FLIC conjugate 9 inhibited FLuc penetrated cells efficiently (IC50 = 1.6 ?M) and inhibited bioluminescence, without affecting the viability of the cells. Based on these results, peptide-FLIC conjugates can be used for the analysis of cellular uptake of biomolecules in a new way that can at the same time overcome some downsides seen with other methods. Thus, FLICs can be considered as versatile tools that broaden the plethora of methods that take advantage of the bioluminescence phenomena. PMID:24341748

Poutiainen, Pekka K; Rönkkö, Teemu; Hinkkanen, Ari E; Palvimo, Jorma J; Närvänen, Ale; Turhanen, Petri; Laatikainen, Reino; Weisell, Janne; Pulkkinen, Juha T

2014-01-15

98

A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging  

Energy Technology Data Exchange (ETDEWEB)

Two genetic reporter systems were developed for multimodality reporter gene imaging of different molecular-genetic processes using fluorescence, bioluminescence (BLI), and nuclear imaging techniques. The eGFP cDNA was fused at the N-terminus with HSV1-tk cDNA bearing a nuclear export signal from MAPKK (NES-HSV1-tk) or with truncation at the N-terminus of the first 45 amino acids ({delta}45HSV1-tk) and with firefly luciferase at the C-terminus. A single fusion protein with three functional subunits is formed following transcription and translation from a single open reading frame. The NES-TGL (NES-TGL) or {delta}45HSV1-tk/GFP/luciferase ({delta}45-TGL) triple-fusion gene cDNAs were cloned into a MoMLV-based retrovirus, which was used for transduction of U87 human glioma cells. The integrity, fluorescence, bioluminescence, and enzymatic activity of the TGL reporter proteins were assessed in vitro. The predicted molecular weight of the fusion proteins (130 kDa) was confirmed by western blot. The U87-NES-TGL and U87-{delta}45-TGL cells had cytoplasmic green fluorescence. The in vitro BLI was 7- and 13-fold higher in U87-NES-TGL and U87-{delta}45-TGL cells compared to nontransduced control cells. The Ki of {sup 14}C-FIAU was 0.49{+-}0.02, 0.51{+-}0.03, and 0.003{+-}0.001 ml/min/g in U87-NES-TGL, U87-{delta}45-TGL, and wild-type U87 cells, respectively. Multimodality in vivo imaging studies were performed in nu/nu mice bearing multiple s.c. xenografts established from U87-NES-TGL, U87-{delta}45-TGL, and wild-type U87 cells. BLI was performed after administration of d-luciferin (150 mg/kg i.v.). Gamma camera or PET imaging was conducted at 2 h after i.v. administration of [{sup 131}I]FIAU (7.4 MBq/animal) or [{sup 124}I]FIAU (7.4 MBq/animal), respectively. Whole-body fluorescence imaging was performed in parallel with the BLI and radiotracer imaging studies. In vivo BLI and gamma camera imaging showed specific localization of luminescence and radioactivity to the TGL transduced xenografts with background levels of activity in the wild-type xenografts. Tissue sampling yielded values of 0.47%{+-}0.08%, 0.86%{+-}0.06%, and 0.03%{+-}0.01%dose/g [{sup 131}I]FIAU in U87-NES-TGL, U87-{delta}45-TGL, and U87 xenografts, respectively. The TGL triple-fusion reporter gene preserves the functional activity of its subunits and is very effective for multimodality imaging. It provides for the seamless transition from fluorescence microscopy and FACS to whole-body bioluminescence imaging, to nuclear (PET, SPET, gamma camera) imaging, and back to in situ fluorescence image analysis. (orig.)

Ponomarev, Vladimir; Vider, Jelena; Shavrin, Aleksander; Ageyeva, Ludmila; Tourkova, Vilia [Department of Radiology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, NY 10021, New York (United States); Doubrovin, Michael; Serganova, Inna; Beresten, Tatiana; Ivanova, Anna; Blasberg, Ronald [Department of Neurology, Memorial Sloan-Kettering Cancer Center, New York (United States); Balatoni, Julius [Radiochemistry/Cyclotron Core Facility, Memorial Sloan-Kettering Cancer Center, New York (United States); Bornmann, William [Organic Chemistry Synthesis Core Facility, Memorial Sloan-Kettering Cancer Center, New York (United States); Gelovani Tjuvajev, Juri [Department of Radiology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, NY 10021, New York (United States); MD Anderson Cancer Center, 1515 Holcombe Road, Box 0057, TX 77030, Houston (United States)

2004-05-01

99

A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging  

International Nuclear Information System (INIS)

Two genetic reporter systems were developed for multimodality reporter gene imaging of different molecular-genetic processes using fluorescence, bioluminescence (BLI), and nuclear imaging techniques. The eGFP cDNA was fused at the N-terminus with HSV1-tk cDNA bearing a nuclear export signal from MAPKK (NES-HSV1-tk) or with truncation at the N-terminus of the first 45 amino acids (?45HSV1-tk) and with firefly luciferase at the C-terminus. A single fusion protein with three functional subunits is formed following transcription and translation from a single open reading frame. The NES-TGL (NES-TGL) or ?45HSV1-tk/GFP/luciferase (?45-TGL) triple-fusion gene cDNAs were cloned into a MoMLV-based retrovirus, which was used for transduction of U87 human glioma cells. The integrity, fluorescence, bioluminescence, and enzymatic activity of the TGL reporter proteins were assessed in vitro. The predicted molecular weight of the fusion proteins (130 kDa) was confirmed by western blot. The U87-NES-TGL and U87-?45-TGL cells had cytoplasmic green fluorescence. The in vitro BLI was 7- and 13-fold higher in U87-NES-TGL and U87-?45-TGL cells compared to nontransduced control cells. The Ki of 14C-FIAU was 0.49±0.02, 0.51±0.03, and 0.003±0.001 ml/min/g in U87-NES-TGL, U87-?45-TGL, and wild-type U87 cells, respectively. Multimodality in vivo imaging studies were performed in nu/nu mice bearing multiple s.c. xenografts established from U87-NES-TGL, U87-?45-TGL, and wild-type U87 cells. BLI was performed after administration of d-luciferin (150 mg/kg i.v.). Gamma camera or PET imaging was conducted at 2 h after i.v. administration of [131I]FIAU (7.4 MBq/animal) or [124I]FIAU (7.4 MBq/animal), respectively. Whole-body fluorescence imaging was performed in parallel with the BLI and radiotracer imaging studies. In vivo BLI and gamma camera imaging showed specific localization of luminescence and radioactivity to the TGL transduced xenografts with background levels of activity in the wild-type xenografts. Tissue sampling yielded values of 0.47%±0.08%, 0.86%±0.06%, and 0.03%±0.01%dose/g [131I]FIAU in U87-NES-TGL, U87-?45-TGL, and U87 xenografts, respectively. The TGL triple-fusion reporter gene preserves the functional activity of its subunits and is very effective for multimodality imaging. It provides for the seamless transition from fluorescence microscopy and FACS to whole-body bioluminescence imaging, to nuclear (PET, SPET, gamma camera) imaging, and back to in situ fluorescence image analysis. (orig.)

2004-05-01

100

Bioluminescence imaging as a marker for cellular Hsp70 response to thermal laser injury  

Science.gov (United States)

Assessment of laser tissue damage is not complete without an investigation into the cellular effects that are induced. In the past, tissue damage was quantified by such macroscopically visual results as tissue mass removal, carbonization, and melting. In this research, we used heat shock protein (hsp70) transcription, to track cellular response to laser injury. A stable cell line was generated containing the luciferase reporter gene attached to the heat shock protein (Hsp70) promoter. After thermal injury with a Holmium:YAG pulsed laser (wavelength= 2.1 ?m, pulsetime = 250 ?s, 30 pulses, 3 Hz), luciferase is produced upon hsp70 activation and emits bioluminescence at 563 nm. The luminescence was quantified with a liquid nitrogen cooled CCD camera. A minimum pulse energy (65 mJ/pulse, 2.0 mJ/mm2) was needed to activate the hsp70 response and a higher energy (103 mJ/pulse, 3.2 mJ/mm2) was associated with a reduction in hsp70 response. Bioluminescence levels correlated well to actual hsp70 protein concentrations as determined by ELISA assay. Photon counts were normalized to the percentage of live cells by means of a flow cytometry cell viability assay. The hsp70 response followed an Arrhenius relationship in nature when constant temperature water bath and constant area laser experiments were carried out.

Beckham, Joshua T.; Baran, Jennifer A.; Mackanos, Mark A.; Crooke, Cornelia; Takahashi, Takamune; O'Connell-Rodwell, Caitlin E.; Contag, Christopher H.; Jansen, E. Duco

2003-07-01

 
 
 
 
101

In vivo bioluminescence imaging of Escherichia coli O104:H4 and role of aerobactin during colonization of a mouse model of infection  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background A major outbreak of bloody diarrhea associated with Shiga toxin-producing Escherichia coli O104:H4 occurred early in 2011, to which an unusual number of hemolytic uremic syndrome cases were linked. Due to limited information regarding pathogenesis and/or virulence properties of this particular serotype, we investigated the contribution of the aerobactin iron transport system during in vitro and in vivo conditions. Results A bioluminescent reporter construct was used to perform real-time monitoring of E. coli O104:H4 in a mouse model of infection. We verified that our reporter strain maintained characteristics and growth kinetics that were similar to those of the wild-type E. coli strain. We found that the intestinal cecum of ICR (CD-1 mice was colonized by O104:H4, with bacteria persisting for up to 7?days after intragastric inoculation. MALDI-TOF analysis of heat-extracted proteins was performed to identify putative surface-exposed virulence determinants. A protein with a high similarity to the aerobactin iron receptor was identified and further demonstrated to be up-regulated in E. coli O104:H4 when grown on MacConkey agar or during iron-depleted conditions. Because the aerobactin iron acquisition system is a key virulence factor in Enterobacteriaceae, an isogenic aerobactin receptor (iutA mutant was created and its intestinal fitness assessed in the murine model. We demonstrated that the aerobactin mutant was out-competed by the wild-type E. coli O104:H4 during in vivo competition experiments, and the mutant was unable to persist in the cecum. Conclusion Our findings demonstrate that bioluminescent imaging is a useful tool to monitor E. coli O104:H4 colonization properties, and the murine model can become a rapid way to evaluate bacterial factors associated with fitness and/or colonization during E. coli O104:H4 infections.

Torres Alfredo G

2012-06-01

102

Synthetic strategies for controlling inter- and intramolecular interactions: Applications in single-molecule fluorescence imaging, bioluminescence imaging, and palladium catalysis  

Science.gov (United States)

The field of synthetic organic chemistry has reached such maturity that, with sufficient effort and resources, the synthesis of virtually any small molecule which exhibits reasonable stability at room temperature can be realized. While representing a monumental achievement for the field, the ability to exert precise control over molecular structure is just a means to an end, and it is frequently the responsibility of the synthetic chemist to determine which molecules should actually be synthesized. For better or worse, there exists no competitive free market in academia for new molecules, and as a result, the decision of which compounds should be synthesized is seldom driven by the forces of supply and demand; rather, it is guided by the synthetic chemist's interest in an anticipated structure-function relationship or in the properties of a previously unstudied class of molecules. As a consequence, there exists a pervasive need for chemists with synthetic expertise in fields (e.g., molecular imaging) and subdisciplines of chemistry (e.g., physical chemistry) in which the identification of promising synthetic targets dramatically outpaces the synthetic output in that field or subdiscipline, and ample opportunities are available for synthetic chemists who choose to pursue such cross-disciplinary research. This thesis describes synthetic efforts that leverage these opportunities to realize applications in biological imaging and in palladium catalysis. In Part I, the synthesis and characterization of three novel luminophores and their imaging applications are discussed. The first is a molecular beacon that utilizes a fluorophorefluorophore pair which exhibits H-dimer quenching in the closed conformation. This probe offers several advantages over conventional fluorophore-quencher molecular beacons in the detection of oligonucleotides, both in bulk and at the single-molecule level. Secondly, a fluorescent, Cy3-Cy5 covalent heterodimer is reported, which on account of the proximity of the Cy3 and Cy5 fluorophores, behaves as an optical photoswitch in the presence of a thiol reagent. This unique property was employed to achieve sub-diffraction-limited imaging of the stalks of Caulobacter crescentus cells with 30-nm resolution using STORM (stochastic optical reconstruction microscopy). Lastly, the synthesis of the first selenium analogue of firefly luciferin is described, and this analogue is shown to be a competent substrate for firefly luciferase (fLuc). Remarkably, it exhibits red-shifted bioluminescence emission relative to the native sulfur analogue. The in vivo performance of the selenium and sulfur analogues in imaging are compared by tail-vein injection into nude mice bearing subcutaneous tumor xenografts of a human breast cancer cell line that was stably transduced to express fLuc. Part II of this thesis begins by addressing design considerations in the development of palladium catalysts that effect oxidative transformations under mild conditions (i.e., 1 atm air, room temperature) using molecular oxygen as the terminal oxidant. A newly synthesized cationic palladium complex, [(2,9-dimethylphenanthroline)Pd(OAc)]2[OTf]2, is shown to catalyze aerobic alcohol oxidation under such conditions with an unprecedented initial turnover frequency, but the presence of partially reduced oxygen species results in competitive ligand oxidation with concomitant decrease in catalyst activity. To remedy this, oxidatively resistant ligands, which are essential for the development of next-generation, high-turnover-frequency palladium catalysts that utilize oxygen as a terminal oxidant, have been prepared and effectively employed. In addition, the first general palladium-catalyzed route to the carbonylation of diols is reported. In this system, carbon monoxide (1 atm) serves the carbonyl source, (2,9-dimethylphenanthroline)Pd(OAc) 2 acts as the catalyst, and N-chlorosuccinimide and iodosobenzene are the oxidants for 1,2- and 1,3-diols, respectively. This thesis illustrates the power of synthetic organic chemistry to exert precise control ove

Conley, Nicholas R.

103

In vivo bioluminescence imaging validation of a human biopsy-derived orthotopic mouse model of glioblastoma multiforme.  

Science.gov (United States)

Glioblastoma multiforme (GBM), the most aggressive brain malignancy, is characterized by extensive cellular proliferation, angiogenesis, and single-cell infiltration into the brain. We have previously shown that a xenograft model based on serial xenotransplantation of human biopsy spheroids in immunodeficient rodents maintains the genotype and phenotype of the original patient tumor. The present work further extends this model for optical assessment of tumor engraftment and growth using bioluminescence imaging (BLI). A method for successful lentiviral transduction of the firefly luciferase gene into multicellular spheroids was developed and implemented to generate optically active patient tumor cells. Luciferase-expressing spheroids were injected into the brains of immunodeficient mice. BLI photon counts and tumor volumes from magnetic resonance imaging (MRI) were correlated. Luciferase-expressing tumors recapitulated the histopathologic hallmarks of human GBMs and showed proliferation rates and microvessel density counts similar to those of wild-type xenografts. Moreover, we detected widespread invasion of luciferase-positive tumor cells in the mouse brains. Herein we describe a novel optically active model of GBM that closely mimics human pathology with respect to invasion, angiogenesis, and proliferation indices. The model may thus be routinely used for the assessment of novel anti-GBM therapeutic approaches implementing well-established and cost-effective optical imaging strategies. PMID:23490442

Jarzabek, Monika A; Huszthy, Peter C; Skaftnesmo, Kai O; McCormack, Emmet; Dicker, Patrick; Prehn, Jochen H M; Bjerkvig, Rolf; Byrne, Annette T

2013-05-01

104

Imaging tumor angiogenesis in breast cancer experimental lung metastasis with positron emission tomography, near-infrared fluorescence, and bioluminescence.  

Science.gov (United States)

The goal of this study was to develop a molecular imaging agent that can allow for both positron emission tomography (PET) and near-infrared fluorescence (NIRF) imaging of CD105 expression in metastatic breast cancer. TRC105, a chimeric anti-CD105 monoclonal antibody, was labeled with both a NIRF dye (i.e., IRDye 800CW) and (64)Cu to yield (64)Cu-NOTA-TRC105-800CW. Flow cytometry analysis revealed no difference in CD105 binding affinity/specificity between TRC105 and NOTA-TRC105-800CW. Serial bioluminescence imaging (BLI) was carried out to non-invasively monitor the lung tumor burden in BALB/c mice, after intravenous injection of firefly luciferase-transfected 4T1 (i.e., fLuc-4T1) murine breast cancer cells to establish the experimental lung metastasis model. Serial PET imaging revealed that fLuc-4T1 lung tumor uptake of (64)Cu-NOTA-TRC105-800CW was 11.9 ± 1.2, 13.9 ± 3.9, and 13.4 ± 2.1 %ID/g at 4, 24, and 48 h post-injection respectively (n = 3). Biodistribution studies, blocking fLuc-4T1 lung tumor uptake with excess TRC105, control experiments with (64)Cu-NOTA-cetuximab-800CW (which served as an isotype-matched control), ex vivo BLI/PET/NIRF imaging, autoradiography, and histology all confirmed CD105 specificity of (64)Cu-NOTA-TRC105-800CW. Successful PET/NIRF imaging of tumor angiogenesis (i.e., CD105 expression) in the breast cancer experimental lung metastasis model warrants further investigation and clinical translation of dual-labeled TRC105-based agents, which can potentially enable early detection of small metastases and image-guided surgery for tumor removal. PMID:23471463

Zhang, Yin; Hong, Hao; Nayak, Tapas R; Valdovinos, Hector F; Myklejord, Duane V; Theuer, Charles P; Barnhart, Todd E; Cai, Weibo

2013-07-01

105

Time courses and time-resolved spectra of firefly bioluminescence initiated by two methods of ATP injection and photolysis of caged ATP.  

Science.gov (United States)

The time-dependent characteristics of firefly bioluminescence initiated by manual injection of adenosine triphosphate (ATP) into buffer solution containing luciferin (Ln), luciferase (Luc) and Mg(2+) were measured with a resolution of 10 ms, and compared with those obtained by photolysis of caged ATP. The time course depends on pH; both rise and decay rates decrease when pH is lowered from 7.8 to 6.8. In contrast, the parameter ? in the kinetic formula related to diffusion of ATP is almost independent of pH. The pH dependence of the time course of bioluminescence can be explained by the same pH tendency as the rate of ATP binding at the active site of Luc. The time-resolved spectra can be decomposed into two Gaussian components with maxima at 2.2 and 2.0 eV. At pH 7.8, the band at 2.2 eV is more intense than that at 2.0 eV for all three concentration conditions. At lower pH, the band at 2.2 eV becomes weaker than that at 2.0 eV. The intensity ratio of the 2.0 and 2.2 eV bands is constant for duration time of 600 s for both injection and photolysis experiments, and the above conclusions are unaffected by the concentration ratio [Ln]/[Luc]. PMID:23875889

Yanagisawa, Yuki; Kageyama, Takeshi; Wada, Naohisa; Tanaka, Masatoshi; Ohno, Shin-Ya

2013-01-01

106

Glioblastoma Therapy with Cytotoxic Mesenchymal Stromal Cells Optimized by Bioluminescence Imaging of Tumor and Therapeutic Cell Response  

Science.gov (United States)

Genetically modified adipose tissue derived mesenchymal stromal cells (hAMSCs) with tumor homing capacity have been proposed for localized therapy of chemo- and radiotherapy resistant glioblastomas. We demonstrate an effective procedure to optimize glioblastoma therapy based on the use of genetically modified hAMSCs and in vivo non invasive monitoring of tumor and therapeutic cells. Glioblastoma U87 cells expressing Photinus pyralis luciferase (Pluc) were implanted in combination with hAMSCs expressing a trifunctional Renilla reniformis luciferase-red fluorescent protein-thymidine kinase reporter in the brains of SCID mice that were subsequently treated with ganciclovir (GCV). The resulting optimized therapy was effective and monitoring of tumor cells by bioluminescence imaging (BLI) showed that after 49 days GCV treatment reduced significantly the hAMSC treated tumors; by a factor of 104 relative to controls. Using a Pluc reporter regulated by an endothelial specific promoter and in vivo BLI to image hAMSC differentiation we gained insight on the therapeutic mechanism. Implanted hAMSCs homed to tumor vessels, where they differentiated to endothelial cells. We propose that the tumor killing efficiency of genetically modified hAMSCs results from their association with the tumor vascular system and should be useful vehicles to deliver localized therapy to glioblastoma surgical borders following tumor resection.

Alieva, Maria; Bago, Juli R.; Aguilar, Elisabet; Soler-Botija, Carolina; Vila, Olaia F.; Molet, Joan; Gambhir, Sanjiv S.; Rubio, Nuria; Blanco, Jeronimo

2012-01-01

107

Spectral Unmixing of Multicolored Bioluminescence Emitted from Heterogeneous Biological Sources  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A wide variety of bioluminescent luciferase proteins are available for use in transcriptional or biochemical reporter assays. However, spectral overlap normally prevents them from being monitored simultaneously. To address this problem, a Java plug-in for ImageJ was written to deconvolute bioluminescent images composed of signals from multiple luciferases. The methodology was validated by testing the program with both simulated and real luciferase images. Bioluminescent images were acquired u...

2006-01-01

108

Sensitive In Situ Monitoring of a Recombinant Bioluminescent Yersinia enterocolitica Reporter Mutant in Real Time on Camembert Cheese  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Bioluminescent mutants of Yersinia enterocolitica were generated by transposon mutagenesis using a promoterless, complete lux operon (luxCDABE) derived from Photorhabdus luminescens, and their production of light in the cheese environment was monitored. Mutant B94, which had the lux cassette inserted into an open reading frame of unknown function was used for direct monitoring of Y. enterocolitica cells on cheeses stored at 10°C by quantifying bioluminescence using a photon-counting, intensi...

Maoz, Ariel; Mayr, Ralf; Bresolin, Geraldine; Neuhaus, Klaus; Francis, Kevin P.; Scherer, Siegfried

2002-01-01

109

Comparison of red-shifted firefly luciferase Ppy RE9 and conventional Luc2 as bioluminescence imaging reporter genes for in vivo imaging of stem cells  

Science.gov (United States)

One critical issue for noninvasive imaging of transplanted bioluminescent cells is the large amount of light absorption in tissue when emission wavelengths below 600 nm are used. Luciferase with a red-shifted spectrum can potentially bypass this limitation. We assessed and compared a mutant of firefly luciferase (Ppy RE9, PRE9) against the yellow luciferase luc2 gene for use in cell transplantation studies. C17.2 neural stem cells expressing PRE9-Venus and luc2-Venus were sorted by flow cytometry and assessed for bioluminescence in vitro in culture and in vivo after transplantation into the brain of immunodeficient Rag2-/- mice. We found that the luminescence from PRE9 was stable, with a peak emission at 620 nm, shifted to the red compared to that of luc2. The emission peak for PRE9 was pH-independent, in contrast to luc2, and much less affected by tissue absorbance compared to that of luc2. However, the total emitted light radiance from PRE9 was substantially lower than that of luc2, both in vitro and in vivo. We conclude that PRE9 has favorable properties as compared to luc2 in terms of pH independence, red-shifted spectrum, tissue light penetration, and signal quantification, justifying further optimization of protein expression and enzymatic activity.

Liang, Yajie; Walczak, Piotr; Bulte, Jeff W. M.

2012-01-01

110

Bioluminescence imaging of mitochondrial Ca2+ dynamics in soma and neurites of individual adult mouse sympathetic neurons.  

Science.gov (United States)

Changes in the cytosolic Ca(2+) concentration ([Ca(2+)](c)) are essential for triggering neurotransmitter release from presynaptic nerve terminals. Calcium-induced Ca(2+) release (CICR) from the endoplasmic reticulum (ER) may amplify the [Ca(2+)](c) signals and facilitate neurotransmitter release in sympathetic neurons. In adrenal chromaffin cells, functional triads are formed by voltage-operated Ca(2+) channels (VOCCs), CICR sites and mitochondria. In fact, mitochondria take up most of the Ca(2+) load entering the cells and are essential for shaping [Ca(2+)](c) signals and exocytosis. Here we have investigated the existence of such functional triads in sympathetic neurons. The mitochondrial Ca(2+) concentration ([Ca(2+)](m)) in soma and neurites of individual mouse superior cervical ganglion (SCG) neurons was monitored by bioluminescence imaging of targeted aequorins. In soma, Ca(2+) entry through VOCCs evoked rapid, near millimolar [Ca(2+)](m) increases in a subpopulation of mitochondria containing about 40% of the aequorin. Caffeine evoked a similar [Ca(2+)](m) increase in a mitochondrial pool containing about 30% of the aequorin and overlapping with the VOCC-sensitive pool. These observations suggest the existence of functional triads similar to the ones described in chromaffin cells. In neurites, mitochondria were able to buffer [Ca(2+)](c) increases resulting from activation of VOCCs but not those mediated by caffeine-induced Ca(2+) release from the ER. The weaker Ca(2+) buffering by mitochondria in neurites could contribute to facilitate Ca(2+)-induced exocytosis at the presynaptic sites. PMID:17234693

Núńez, Lucía; Senovilla, Laura; Sanz-Blasco, Sara; Chamero, Pablo; Alonso, María T; Villalobos, Carlos; García-Sancho, Javier

2007-04-15

111

Bio-luminescent imaging and characterization of organ-specific metastasis of human cancer in NOD/SCID mice  

Science.gov (United States)

Many clinical evidences demonstrate that the sites of distant metastasis are not random and certain malignant tumors show a tendency to develop metastases in specific organs (e.g., brain, liver, and lungs). However, an appropriate animal model to characterize the metastatic nature of transplantable human cancer cell lines has not been reported well. Recent advances in bio-luminescent imaging (BLI) technologies have facilitated the quantitative analysis of various cellular processes in vivo. To visualize the fate of tumor progression in the living mice, we are constructing a luciferaseexpressing human cancer cell library (including melanoma, colon, breast, and prostate cancer). Herein we demonstrate that the BLI technology in couple with a fine ultrasonic guidance realizes cancer cell-type dependent metastasis to the specific organs. For example, some melanoma cell lines showed frequent metastasis to brain, lungs, and lymph nodes in the mouse model. Notably, reflecting the clinical features of melanoma, breast, and prostate cancer, some of the cell lines showed preferential metastasis to the brain. Moreover, these cellular resources for BLI allow a high throughput screening for potential anti-cancer drugs. Thus, this BLI-mediated additional strategy with the luciferase-expressing cancer cell resources should promote many translational studies for human cancer therapy.

Chun, Nicole A. L.; Murakami, Takashi

2010-02-01

112

Real-time analysis of agonist-induced activation of protease-activated receptor 1/Galphai1 protein complex measured by bioluminescence resonance energy transfer in living cells.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

G protein-coupled receptors transmit extracellular signals into the cells by activating heterotrimeric G proteins, a process that is often followed by receptor desensitization. Monitoring such a process in real time and in living cells will help better understand how G protein activation occurs. Energy transfer-based approaches [fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET)] were recently shown to be powerful methods to monitor the G protei...

Ayoub, Mohammed A.; Maurel, Damien; Binet, Virginie; Fink, Michel; Pre?zeau, Laurent; Ansanay, Herve?; Pin, Jean-philippe

2007-01-01

113

In vivo bioluminescent imaging of irradiated orthotopic pancreatic cancer xenografts in nonobese diabetic-severe combined immunodeficient mice: a novel method for targeting and assaying efficacy of ionizing radiation.  

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Adenocarcinoma of the pancreas is a lethal malignancy, and better models to study tumor behavior in vivo are needed for the development ofmore effective therapeutics. Ionizing radiation is a treatment modality that is commonly used in the clinical setting, in particular, for locally confined disease; however, good model systems to study the effect of ionizing radiation in orthotopic tumors have not been established. In an attempt to create clinically relevant models for studying treatments directed against pancreatic cancer, we have defined a methodology to measure the effect of varying doses of radiation in established human pancreatic cancer orthotopic xenografts using two different pancreatic cancer cell lines (Panc-1 and BXPC3) infected with a lentiviral vector expressing CMV promoter-driven luciferase to allow bioluminescence imaging of live animals in real time. Quantifiable photon emission from luciferase signaling in vivo correlated well with actual tumor growth. Bioluminescence imaging of the established pancreatic xenografts was used to direct delivery of radiation to the orthotopic tumors and minimize off-target adverse effects. Growth delay was observed with schedules in the range of 7.5 Gy in five fractions to 10 Gy in four fractions, whereas doses 3 Gy or higher produced toxic adverse effects. In conclusion, we describe a model in which the effects of ionizing radiation, alone or in combination with other therapeutics, in orthotopic xenografts, can be studied. PMID:20563256

Lee, Cheong J; Spalding, Aaron C; Ben-Josef, Edgar; Wang, Lidong; Simeone, Diane M

2010-01-01

114

Evaluating reporter genes of different luciferases for optimized in vivo bioluminescence imaging of transplanted neural stem cells in the brain.  

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Bioluminescence imaging (BLI) has become the method of choice for optical tracking of cells in small laboratory animals. However, the use of luciferases from different species, depending on different substrates and emitting at distinct wavelengths, has not been optimized for sensitive neuroimaging. In order to identify the most suitable luciferase, this quantitative study compared the luciferases Luc2, CBG99, PpyRE9 and hRluc. Human embryonic kidney (HEK-293) cells and mouse neural stem cells were transduced by lentiviral vector-mediated transfer to express one of the four luciferases, together with copGFP. A T2A peptide linker promoted stoichiometric expression between both imaging reporters and the comparison of cell populations upon flow cytometry. Cell dilution series were used to determine highest BLI sensitivity in vitro for Luc2. However, Coelenterazine h-dependent hRluc signals clearly exceeded d-luciferin-dependent BLI in vitro. For the quantitative in vivo analysis, cells were transplanted into mouse brain and BLI was performed including the recording of emission kinetics and spectral characteristics. Differences in light kinetics were observed for d-luciferin vs Coelenterazine h. The emission spectra of Luc2 and PpyRE9 remained almost unchanged, while the emission spectrum of CBG99 became biphasic. Most importantly, photon emission decreased in the order of Luc2, CBG99, PpyRE9 to hRluc. The feasibility of combining different luciferases for dual color and dual substrate neuroimaging was tested and discussed. This investigation provides the first complete quantitative comparison of different luciferases expressed by neural stem cells. It results in a clear recommendation of Luc2 as the best luciferase selection for in vivo neuroimaging. PMID:24375906

Mezzanotte, Laura; Aswendt, Markus; Tennstaedt, Annette; Hoeben, Rob; Hoehn, Mathias; Löwik, Clemens

2013-01-01

115

Real-time luminescence imaging of cellular ATP release.  

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Extracellular ATP and other purines are ubiquitous mediators of local intercellular signaling within the body. While the last two decades have witnessed enormous progress in uncovering and characterizing purinergic receptors and extracellular enzymes controlling purinergic signals, our understanding of the initiating step in this cascade, i.e., ATP release, is still obscure. Imaging of extracellular ATP by luciferin-luciferase bioluminescence offers the advantage of studying ATP release and distribution dynamics in real time. However, low-light signal generated by bioluminescence reactions remains the major obstacle to imaging such rapid processes, imposing substantial constraints on its spatial and temporal resolution. We have developed an improved microscopy system for real-time ATP imaging, which detects ATP-dependent luciferin-luciferase luminescence at ?10 frames/s, sufficient to follow rapid ATP release with sensitivity of ?10 nM and dynamic range up to 100 ?M. In addition, simultaneous differential interference contrast cell images are acquired with infra-red optics. Our imaging method: (1) identifies ATP-releasing cells or sites, (2) determines absolute ATP concentration and its spreading manner at release sites, and (3) permits analysis of ATP release kinetics from single cells. We provide instrumental details of our approach and give several examples of ATP-release imaging at cellular and tissue levels, to illustrate its potential utility. PMID:23973809

Furuya, Kishio; Sokabe, Masahiro; Grygorczyk, Ryszard

2014-03-15

116

Boosting Bioluminescence Neuroimaging: An Optimized Protocol for Brain Studies  

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Bioluminescence imaging is widely used for optical cell tracking approaches. However, reliable and quantitative bioluminescence of transplanted cells in the brain is highly challenging. In this study we established a new bioluminescence imaging protocol dedicated for neuroimaging, which increases sensitivity especially for noninvasive tracking of brain cell grafts. Different D-Luciferin concentrations (15, 150, 300 and 750 mg/kg), injection routes (iv, ip, sc), types of anesthesia (Isoflurane...

2013-01-01

117

GMO detection using a bioluminescent real time reporter (BART of loop mediated isothermal amplification (LAMP suitable for field use  

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Full Text Available Abstract Background There is an increasing need for quantitative technologies suitable for molecular detection in a variety of settings for applications including food traceability and monitoring of genetically modified (GM crops and their products through the food processing chain. Conventional molecular diagnostics utilising real-time polymerase chain reaction (RT-PCR and fluorescence-based determination of amplification require temperature cycling and relatively complex optics. In contrast, isothermal amplification coupled to a bioluminescent output produced in real-time (BART occurs at a constant temperature and only requires a simple light detection and integration device. Results Loop mediated isothermal amplification (LAMP shows robustness to sample-derived inhibitors. Here we show the applicability of coupled LAMP and BART reactions (LAMP-BART for determination of genetically modified (GM maize target DNA at low levels of contamination (0.1-5.0% GM using certified reference material, and compare this to RT-PCR. Results show that conventional DNA extraction methods developed for PCR may not be optimal for LAMP-BART quantification. Additionally, we demonstrate that LAMP is more tolerant to plant sample-derived inhibitors, and show this can be exploited to develop rapid extraction techniques suitable for simple field-based qualitative tests for GM status determination. We also assess the effect of total DNA assay load on LAMP-BART quantitation. Conclusions LAMP-BART is an effective and sensitive technique for GM detection with significant potential for quantification even at low levels of contamination and in samples derived from crops such as maize with a large genome size. The resilience of LAMP-BART to acidic polysaccharides makes it well suited to rapid sample preparation techniques and hence to both high throughput laboratory settings and to portable GM detection applications. The impact of the plant sample matrix and genome loading within a reaction must be controlled to ensure quantification at low target concentrations.

Kiddle Guy

2012-04-01

118

zebraflash transgenic lines for in vivo bioluminescence imaging of stem cells and regeneration in adult zebrafish  

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The zebrafish has become a standard model system for stem cell and tissue regeneration research, based on powerful genetics, high tissue regenerative capacity and low maintenance costs. Yet, these studies can be challenged by current limitations of tissue visualization techniques in adult animals. Here we describe new imaging methodology and present several ubiquitous and tissue-specific luciferase-based transgenic lines, which we have termed zebraflash, that facilitate the assessment of regeneration and engraftment in freely moving adult zebrafish. We show that luciferase-based live imaging reliably estimates muscle quantity in an internal organ, the heart, and can longitudinally follow cardiac regeneration in individual animals after major injury. Furthermore, luciferase-based detection enables visualization and quantification of engraftment in live recipients of transplanted hematopoietic stem cell progeny, with advantages in sensitivity and gross spatial resolution over fluorescence detection. Our findings present a versatile resource for monitoring and dissecting vertebrate stem cell and regeneration biology.

Chen, Chen-Hui; Durand, Ellen; Wang, Jinhu; Zon, Leonard I.; Poss, Kenneth D.

2013-01-01

119

zebraflash transgenic lines for in vivo bioluminescence imaging of stem cells and regeneration in adult zebrafish.  

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The zebrafish has become a standard model system for stem cell and tissue regeneration research, based on powerful genetics, high tissue regenerative capacity and low maintenance costs. Yet, these studies can be challenged by current limitations of tissue visualization techniques in adult animals. Here we describe new imaging methodology and present several ubiquitous and tissue-specific luciferase-based transgenic lines, which we have termed zebraflash, that facilitate the assessment of regeneration and engraftment in freely moving adult zebrafish. We show that luciferase-based live imaging reliably estimates muscle quantity in an internal organ, the heart, and can longitudinally follow cardiac regeneration in individual animals after major injury. Furthermore, luciferase-based detection enables visualization and quantification of engraftment in live recipients of transplanted hematopoietic stem cell progeny, with advantages in sensitivity and gross spatial resolution over fluorescence detection. Our findings present a versatile resource for monitoring and dissecting vertebrate stem cell and regeneration biology. PMID:24198277

Chen, Chen-Hui; Durand, Ellen; Wang, Jinhu; Zon, Leonard I; Poss, Kenneth D

2013-12-01

120

Influence of MSA on Cell Growth and Spontaneousn Metastasis of L9981-Luc Lung Cancer Transplanted Model in Nude Mice by Bioluminescence Imaging  

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Full Text Available Background and objective Methylseleninic acid (MSA is an artificially developed selenium compound. It has been proven that MSA could inhibit growth and metastasis on many tumor cells. This study investigated whether MSA has an impact on the growth and metastasis of L9981-Luc lung cancer transplanted model in nude mice or not. Methods A transplantated tumor model was established in nude mice. Fifteen nude mice were randomly divided into three groups: the control group treated with normal saline (0.2 mL/d, the MSA group treated with MSA solution (0.2 mL, and the cisplatin (DDP group injected intraperitoneally with DDP (4 mg/kg/w. Inhibition of MSA on tumor growth and tumor metastasis was observed using the IVIS Imaging System 200 Series. Results A significant difference was obserced in the primary tumor bioluminescence among the three groups (P=0.002 on 21 days post-inoculation. Primary tumor bioluminescence in the DDP group (P=0.001 and in the MSA group (P=0.031 was significantly lower than that in the control group (P=0.001. No significant difference in the metastasis bioluminescence of the thoracic area was indicated among the three groups (P>0.05. Conclusion MSA can inhibit the growth of planted tumor of transgenic lung cancer cell lines L9981-Luc in nude mice. MSA may also suppress the distant metastasis of the transplanted tumor of transgenic lung cancer cell lines L9981-Luc in nude mice.

Yuanrong REN

2013-02-01

 
 
 
 
121

Fungi bioluminescence revisited.  

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A review of the research conducted during the past 30 years on the distribution, taxonomy, phylogeny, ecology, physiology and bioluminescence mechanisms of luminescent fungi is presented. We recognize 64 species of bioluminescent fungi belonging to at least three distinct evolutionary lineages, termed Omphalotus, Armillaria and mycenoid. An accounting of their currently accepted names, distributions, citations reporting luminescence and whether their mycelium and/or basidiomes emit light are provided. We address the physiological and ecological aspects of fungal bioluminescence and provide data on the mechanisms responsible for bioluminescence in the fungi. PMID:18264584

Desjardin, Dennis E; Oliveira, Anderson G; Stevani, Cassius V

2008-02-01

122

Formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium  

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Full Text Available Abstract Background The bioluminescent enzyme firefly luciferase (Luc or variants of green fluorescent protein (GFP in transformed cells can be effectively used to reveal molecular and cellular features of neoplasia in vivo. Tumor cell growth and regression in response to various therapies can be evaluated by using bioluminescent imaging. In bioluminescent imaging, light propagates in highly scattering tissue, and the diffusion approximation is sufficiently accurate to predict the imaging signal around the biological tissue. The numerical solutions to the diffusion equation take large amounts of computational time, and the studies for its analytic solutions have attracted more attention in biomedical engineering applications. Methods Biological tissue is a turbid medium that both scatters and absorbs photons. An accurate model for the propagation of photons through tissue can be adopted from transport theory, and its diffusion approximation is applied to predict the imaging signal around the biological tissue. The solution to the diffusion equation is formulated by the convolution between its Green's function and source term. The formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium can be obtained to accelerate the forward simulation of bioluminescent phenomena. Results The closed form solutions have been derived for the time-dependent diffusion equation and the steady-state diffusion equation with solid and hollow spherical sources in a homogeneous medium, respectively. Meanwhile, the relationship between solutions with a solid sphere source and ones with a surface sphere source is obtained. Conclusion We have formulated solutions for the diffusion equation with solid and hollow spherical sources in an infinite homogeneous medium. These solutions have been verified by Monte Carlo simulation for use in biomedical optical imaging studies. The closed form solution is highly accurate and more computationally efficient in biomedical engineering applications. By using our analytic solutions for spherical sources, we can better predict bioluminescent signals and better understand both the potential for, and the limitations of, bioluminescent tomography in an idealized case. The formulas are particularly valuable for furthering the development of bioluminescent tomography.

Wang Lihong V

2004-05-01

123

Luciferase expression and bioluminescence does not affect tumor cell growth in vitro or in vivo  

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Full Text Available Abstract Live animal imaging is becoming an increasingly common technique for accurate and quantitative assessment of tumor burden over time. Bioluminescence imaging systems rely on a bioluminescent signal from tumor cells, typically generated from expression of the firefly luciferase gene. However, previous reports have suggested that either a high level of luciferase or the resultant light reaction produced upon addition of D-luciferin substrate can have a negative influence on tumor cell growth. To address this issue, we designed an expression vector that allows simultaneous fluorescence and luminescence imaging. Using fluorescence activated cell sorting (FACS, we generated clonal cell populations from a human breast cancer (MCF-7 and a mouse melanoma (B16-F10 cell line that stably expressed different levels of luciferase. We then compared the growth capabilities of these clones in vitro by MTT proliferation assay and in vivo by bioluminescence imaging of tumor growth in live mice. Surprisingly, we found that neither the amount of luciferase nor biophotonic activity was sufficient to inhibit tumor cell growth, in vitro or in vivo. These results suggest that luciferase toxicity is not a necessary consideration when designing bioluminescence experiments, and therefore our approach can be used to rapidly generate high levels of luciferase expression for sensitive imaging experiments.

Rasko John EJ

2010-11-01

124

Enhanced healing of diabetic wounds by topical administration of adipose tissue-derived stromal cells overexpressing stromal-derived factor-1: biodistribution and engraftment analysis by bioluminescent imaging.  

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Chronic ulcers represent a major health problem in diabetic patients resulting in pain and discomfort. Conventional therapy does not guarantee adequate wound repair. In diabetes, impaired healing is partly due to poor endothelial progenitor cells mobilisation and homing, with altered levels of the chemokine stromal-derived factor-1 (SDF-1) at the wound site. Adipose tissue-associated stromal cells (AT-SCs) can provide an accessible source of progenitor cells secreting proangiogenic factors and differentiating into endothelial-like cells. We demonstrated that topical administration of AT-SCs genetically modified ex vivo to overexpress SDF-1, promotes wound healing into diabetic mice. In particular, by in vivo bioluminescent imaging analysis, we monitored biodistribution and survival after transplantation of luciferase-expressing cells. In conclusion, this study indicates the therapeutic potential of AT-SCs administration in wound healing, through cell differentiation, enhanced cellular recruitment at the wound site, and paracrine effects associated with local growth-factors production. PMID:21234108

Di Rocco, Giuliana; Gentile, Antonietta; Antonini, Annalisa; Ceradini, Francesca; Wu, Joseph C; Capogrossi, Maurizio C; Toietta, Gabriele

2010-01-01

125

Enhanced Healing of Diabetic Wounds by Topical Administration of Adipose Tissue-Derived Stromal Cells Overexpressing Stromal-Derived Factor-1: Biodistribution and Engraftment Analysis by Bioluminescent Imaging  

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Chronic ulcers represent a major health problem in diabetic patients resulting in pain and discomfort. Conventional therapy does not guarantee adequate wound repair. In diabetes, impaired healing is partly due to poor endothelial progenitor cells mobilisation and homing, with altered levels of the chemokine stromal-derived factor-1 (SDF-1) at the wound site. Adipose tissue-associated stromal cells (AT-SCs) can provide an accessible source of progenitor cells secreting proangiogenic factors and differentiating into endothelial-like cells. We demonstrated that topical administration of AT-SCs genetically modified ex vivo to overexpress SDF-1, promotes wound healing into diabetic mice. In particular, by in vivo bioluminescent imaging analysis, we monitored biodistribution and survival after transplantation of luciferase-expressing cells. In conclusion, this study indicates the therapeutic potential of AT-SCs administration in wound healing, through cell differentiation, enhanced cellular recruitment at the wound site, and paracrine effects associated with local growth-factors production.

Di Rocco, Giuliana; Gentile, Antonietta; Antonini, Annalisa; Ceradini, Francesca; Wu, Joseph C.; Capogrossi, Maurizio C.; Toietta, Gabriele

2011-01-01

126

Multimodality imaging of breast cancer experimental lung metastasis with bioluminescence and a monoclonal antibody dual-labeled with 89Zr and IRDye 800CW.  

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Metastatic breast cancer is incurable. The goal of this study was to develop a positron emission tomography (PET)/near-infrared fluorescent (NIRF) probe for imaging CD105 expression in breast cancer experimental lung metastasis. TRC105, a chimeric anti-CD105 antibody, was dual-labeled with a NIRF dye (IRDye 800CW) and (89)Zr to yield (89)Zr-Df-TRC105-800CW. Luciferase-transfected 4T1 murine breast cancer cells were injected intravenously into female mice to establish the tumor model. Bioluminescence imaging (BLI) was carried out to noninvasively monitor the lung tumor burden. PET imaging revealed that 4T1 lung tumor uptake of (89)Zr-Df-TRC105-800CW was 8.7 ± 1.4, 10.9 ± 0.5, and 9.7 ± 1.1% ID/g at 4, 24, and 48 h postinjection (n = 4), with excellent tumor contrast. Biodistribution studies, blocking, control studies with (89)Zr-Df-cetuximab-800CW, ex vivo BLI/PET/NIRF imaging, and histology all confirmed CD105 specificity of the tracer. Broad clinical potential of TRC105-based agents was shown in many tumor types, which also enabled early detection of small metastasis and intraoperative guidance for tumor removal. PMID:22784250

Hong, Hao; Zhang, Yin; Severin, Gregory W; Yang, Yunan; Engle, Jonathan W; Niu, Gang; Nickles, Robert J; Chen, Xiaoyuan; Leigh, Bryan R; Barnhart, Todd E; Cai, Weibo

2012-08-01

127

Validation of method for enhanced production of red-shifted bioluminescent photons in vivo  

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Bioluminescence Imaging (BLI) is an increasingly useful and applicable technique that allows for the non-invasive observation of biological events in intact living organisms, ranging from single cells to small rodents. Though the photon production occurs within the host, significant exposure times can be necessary due to the low photon flux compared to fluorescence imaging. The optical absorption spectrum of haemoglobin strongly overlaps most bioluminescent emission spectra, greatly attenuating the total detectable photons in animal models. We have developed and validated a technique that is able to red-shift the bioluminescent photons to the more desirable optical region of > 650 nm, a region of minimal absorbance by hemoglobin. This red-shift occurs by using bioluminescence as an internal light source capable of exciting a fluorophore, such as a fluorescent protein or a quantum dot, that emits in the red. Interestingly, in the absence of an absorber, this excitation can occur over substantial distances (microns to centimeters), far exceeding distances associated to, and thereby precluding, resonance energy transfer phenomena. We show this novel technique yields a substantial increase in the number of red photons for in vitro and ex vivo conditions, suggesting eventually utility for in vivo studies on, for example, intact living mice.

Dragavon, Joe; Blazquez, Samantha; Rogers, Kelly L.; Samson, Chelsea; Tournebize, Régis; Shorte, Spencer

2011-02-01

128

Thoughts on the diversity of convergent evolution of bioluminescence on earth  

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The widespread independent evolution of analogous bioluminescent systems is one of the most impressive and diverse examples of convergent evolution on earth. There are roughly 30 extant bioluminescent systems that have evolved independently on Earth, with each system likely having unique enzymes responsible for catalysing the bioluminescent reaction. Bioluminescence is a chemical reaction involving a luciferin molecule and a luciferase or photoprotein that results in the emission of light. Some independent systems utilize the same luciferin, such as the use of tetrapyrrolic compounds by krill and dinoflagellates, and the wide use of coelenterazine by marine organisms, while the enzymes involved are unique. One common thread among all the different bioluminescent systems is the requirement of molecular oxygen. Bioluminescence is found in most forms of life, especially marine organisms. Bioluminescence in known to benefit the organism by: attraction, repulsion, communication, camouflage, and illumination. The marine ecosystem is significantly affected by bioluminescence, the only light found in the pelagic zone and below is from bioluminescent organisms. Transgenic bioluminescent organisms have revolutionized molecular research, medicine and the biotechnology industry. The use of bioluminescence in studying molecular pathways and disease allows for non-invasive and real-time analysis. Bioluminescence-based assays have been developed for several analytes by coupling luminescence to many enzyme-catalysed reactions.

Waldenmaier, Hans E.; Oliveira, Anderson G.; Stevani, Cassius V.

2012-10-01

129

Use of Bioluminescence to Study Reactive Solute Transport and Biofilm Growth and Activity in Porous Media  

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Using a meso-scale porous media flat plate reactor we utilized a naturally bioluminescent biofilm (V. fischeri) and tracer studies to obtain information on the interactions between biofilms and reactive flow in porous media. The growth and development of the V. fischeri biofilm in a porous media geometry was studied using digital time lapse images of the bioluminescent signal given off by the developing biofilm. The effect of biofilm development on porous media hydrodynamics was examined using dye tracer studies and image analysis. The natural bioluminescence of the V. fischeri allowed real-time, in-situ study of biofilm development in porous media, without destruction of the biofilm. Dye studies and image analysis enabled the study of effects of biofilm accumulation on porous media hydraulics, with comparisons to plug flow and completely mixed systems with varying degrees of biofilm accumulation. The hydraulic conductivity of the porous media/biofilm system was continuously monitored showing a 1 to 4 order of magnitude decrease in hydraulic conductivity as a function of biofilm thickness and accumulation. The real-time nature of the study permitted us to visualize dynamic flow channel formation within the biofilm/porous media system. In addition, the sensitivity of the V. fischeri biofilm to dissolved oxygen allowed us to capture real-time images of reactive transport within the system. Using bioluminescent imaging, the location of active biomass, as well as the relative degree of biological activity, could be visualized and monitored over time. This work is the first meso-scale visualization of the interactions between biofilm and flow in porous media.

Sharp, R. R.; Gerlach, R.; Al, C. B.

2004-12-01

130

Formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium  

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Abstract Background The bioluminescent enzyme firefly luciferase (Luc) or variants of green fluorescent protein (GFP) in transformed cells can be effectively used to reveal molecular and cellular features of neoplasia in vivo. Tumor cell growth and regression in response to various therapies can be evaluated by using bioluminescent imaging. In bioluminescent imaging, light propagates in highly scattering tissue, and the diffusion approximation is sufficiently accurat...

Cong Wenxiang; Wang Lihong V; Wang Ge

2004-01-01

131

Dual-wavelength imaging of tumor progression by activatable and targeting near-infrared fluorescent probes in a bioluminescent breast cancer model.  

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Bioluminescence imaging (BLI) has shown its appeal as a sensitive technique for in vivo whole body optical imaging. However, the development of injectable tumor-specific near-infrared fluorescent (NIRF) probes makes fluorescence imaging (FLI) a promising alternative to BLI in situations where BLI cannot be used or is unwanted (e.g., spontaneous transgenic tumor models, or syngeneic mice to study immune effects).In this study, we addressed the questions whether it is possible to detect tumor progression using FLI with appropriate sensitivity and how FLI correlates with BLI measurements. In addition, we explored the possibility to simultaneously detect multiple tumor characteristics by dual-wavelength FLI (~700 and ~800 nm) in combination with spectral unmixing. Using a luciferase-expressing 4T1-luc2 mouse breast cancer model and combinations of activatable and targeting NIRF probes, we showed that the activatable NIRF probes (ProSense680 and MMPSense680) and the targeting NIRF probes (IRDye 800CW 2-DG and IRDye 800CW EGF) were either activated by or bound to 4T1-luc2 cells. In vivo, we implanted 4T1-luc2 cells orthotopically in nude mice and were able to follow tumor progression longitudinally both by BLI and dual-wavelength FLI. We were able to reveal different probe signals within the tumor, which co-localized with immuno-staining. Moreover, we observed a linear correlation between the internal BLI signals and the FLI signals obtained from the NIRF probes. Finally, we could detect pulmonary metastases both by BLI and FLI and confirmed their presence histologically.Taken together, these data suggest that dual-wavelength FLI is a feasible approach to simultaneously detect different features of one tumor and to follow tumor progression with appropriate specificity and sensitivity. This study may open up new perspectives for the detection of tumors and metastases in various experimental models and could also have clinical applications, such as image-guided surgery. PMID:22348134

Xie, Bang-Wen; Mol, Isabel M; Keereweer, Stijn; van Beek, Ermond R; Que, Ivo; Snoeks, Thomas J A; Chan, Alan; Kaijzel, Eric L; Löwik, Clemens W G M

2012-01-01

132

Bioluminescence lights the way to food safety  

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The food industry is increasingly adopting food safety and quality management systems that are more proactive and preventive than those used in the past which have tended to rely on end product testing and visual inspection. The regulatory agencies in many countries are promoting one such management tool, Hazard Analysis Critical Control Point (HACCP), as a way to achieve a safer food supply and as a basis for harmonization of trading standards. Verification that the process is safe must involve microbiological testing but the results need not be generated in real-time. Of all the rapid microbiological tests currently available, the only ones that come close to offering real-time results are bioluminescence-based methods. Recent developments in application of bioluminescence for food safety issues are presented in the paper. These include the use of genetically engineered microorganisms with bioluminescent and fluorescent phenotypes as a real time indicator of physiological state and survival of food-borne pathogens in food and food processing environments as well as novel bioluminescent-based methods for rapid detection of pathogens in food and environmental samples. Advantages and pitfalls of the methods are discussed.

Brovko, Lubov Y.; Griffiths, Mansel W.

2003-07-01

133

Circadian regulation of bioluminescence in Gonyaulax involves translational control.  

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A 10-fold circadian variation in the amount of luciferin binding protein (LBP) in the marine dinoflagellate Gonyaulax polyedra is reported. This protein binds and stabilizes luciferin, the bioluminescence substrate. In early night phase, when bioluminescence is increasing and LBP levels are rising in the cell, pulse labeling experiments show that LBP is being rapidly synthesized in vivo. At other times, the rate of LBP synthesis is at least 50 times lower, while the rate of synthesis of most ...

1989-01-01

134

Bioluminescence imaging of therapy response does not correlate with FDG-PET response in a mouse model of Burkitt lymphoma  

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Since the development and evaluation of novel anti-cancer therapies require molecular insight in the disease state, both FDG-PET and BLI imaging were evaluated in a Burkitt B-cell lymphoma xenograft model treated with cyclophosphamide or temsirolimus. Daudi xenograft mice were treated with either cyclophosphamide or temsirolimus and imaged with BLI and FDG-PET on d0 (before treatment), d2, d4, d7, d9 and d14 following the start of therapy. Besides tumor volume changes, therapy response was as...

Saint-hubert, Marijke; Devos, Ellen; Ibrahimi, Abdelilah; Debyser, Zeger; Mortelmans, Luc; Mottaghy, Felix M.

2012-01-01

135

Bioluminescence-based visualization of CD4 T cell dynamics using a T lineage-specific luciferase transgenic model1  

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Full Text Available Abstract Background Rapid clonal expansion of T cells occurs in response to antigenic challenges. The kinetics of the T cell response has previously been described using tissue-based studies performed at defined time points. Luciferase bioluminescence has recently been utilized for non-invasive analysis of in vivo biologic processes in real-time. Results We have created a novel transgenic mouse model (T-Lux using a human CD2 mini-gene to direct luciferase expression specifically to the T cell compartment. T-Lux T cells demonstrated normal homing patterns within the intact mouse and following adoptive transfer. Bioluminescent signal correlated with T cell numbers in the whole body images as well as within specific organ regions of interest. Following transfer into lymphopenic (RAG2-/- recipients, homeostatic proliferation of T-Lux T cells was visualized using bioluminescent imaging. Real-time bioluminescent analysis of CD4+ T cell antigen-specific responses enabled real-time comparison of the kinetics and magnitude of clonal expansion and contraction in the inductive lymph node and tissue site of antigen injection. T cell expansion was dose-dependent despite the presence of supraphysiologic numbers of OVA-specific OT-II transgenic TCR T-Lux T cells. CD4+ T cells subsequently underwent a rapid (3–4 day contraction phase in the draining lymph node, with a delayed contraction in the antigen delivery site, with bioluminescent signal diminished below initial levels, representing TCR clonal frequency control. Conclusion The T-Lux mouse provides a novel, efficient model for tracking in vivo aspects of the CD4+ T cell response to antigen, providing an attractive approach for studies directed at immunotherapy or vaccine design.

Zinn Kurt R

2009-08-01

136

GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use  

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Abstract Background There is an increasing need for quantitative technologies suitable for molecular detection in a variety of settings for applications including food traceability and monitoring of genetically modified (GM) crops and their products through the food processing chain. Conventional molecular diagnostics utilising real-time polymerase chain reaction (RT-PCR) and fluorescence-based determination of amplification require temperature cycling and relatively complex ...

Kiddle Guy; Hardinge Patrick; Buttigieg Neil; Gandelman Olga; Pereira Clint; McElgunn Cathal J; Rizzoli Manuela; Jackson Rebecca; Appleton Nigel; Moore Cathy; Tisi Laurence C; Ah, Murray James

2012-01-01

137

Bioluminescent response of individual dinoflagellate cells to hydrodynamic stress measured with millisecond resolution in a microfluidic device.  

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Dinoflagellate bioluminescence serves as a model system for examining mechanosensing by suspended motile unicellular organisms. The response latency, i.e. the delay time between the mechanical stimulus and luminescent response, provides information about the mechanotransduction and signaling process, and must be accurately known for dinoflagellate bioluminescence to be used as a flow visualization tool. This study used a novel microfluidic device to measure the response latency of a large number of individual dinoflagellates with a resolution of a few milliseconds. Suspended cells of several dinoflagellate species approximately 35 microm in diameter were directed through a 200 microm deep channel to a barrier with a 15 microm clearance impassable to the cells. Bioluminescence was stimulated when cells encountered the barrier and experienced an abrupt increase in hydrodynamic drag, and was imaged using high numerical aperture optics and a high-speed low-light video system. The average response latency for Lingulodinium polyedrum strain HJ was 15 ms (N>300 cells) at the three highest flow rates tested, with a minimum latency of 12 ms. Cells produced multiple flashes with an interval as short as 5 ms between individual flashes, suggesting that repeat stimulation involved a subset of the entire intracellular signaling pathway. The mean response latency for the dinoflagellates Pyrodinium bahamense, Alexandrium monilatum and older and newer isolates of L. polyedrum ranged from 15 to 22 ms, similar to the latencies previously determined for larger dinoflagellates with different morphologies, possibly reflecting optimization of dinoflagellate bioluminescence as a rapid anti-predation behavior. PMID:18723546

Latz, Michael I; Bovard, Michelle; VanDelinder, Virginia; Segre, Enrico; Rohr, Jim; Groisman, Alex

2008-09-01

138

Bathyphotometer bioluminescence potential measurements: A framework for characterizing flow agitators and predicting flow-stimulated bioluminescence intensity  

Science.gov (United States)

Bathyphotometer measurements of bioluminescence are used as a proxy for the abundance of luminescent organisms for studying population dynamics; the interaction of luminescent organisms with physical, chemical, and biological oceanographic processes; and spatial complexity especially in coastal areas. However, the usefulness of bioluminescence measurements has been limited by the inability to compare results from different bathyphotometer designs, or even the same bathyphotometer operating at different volume flow rates. The primary objective of this study was to compare measurements of stimulated bioluminescence of four species of cultured dinoflagellates, the most common source of bioluminescence in coastal waters, using two different bathyphotometer flow agitators as a function of bathyphotometer volume flow rate and dinoflagellate concentration. For both the NOSC and BIOLITE flow agitators and each species of dinoflagellate tested, there was a critical volume flow rate, above which average bioluminescence intensity, designated as bathyphotometer bioluminescence potential (BBP), remained relatively constant and scaled directly with dinoflagellate cell concentration. At supra-critical volume flow rates, the ratio of BIOLITE to NOSC BBP was nearly constant for the same species studied, but varied between species. The spatial pattern and residence time of flash trajectories within the NOSC flow agitator indicated the presence of dominant secondary recirculating flows, where most of the bioluminescence was detected. A secondary objective (appearing in the Appendix) was to study the feasibility of using NOSC BBP to scale flow-stimulated bioluminescence intensity across similar flow fields, where the contributing composition of luminescent species remained the same. Fully developed turbulent pipe flow was chosen because it is hydrodynamically well characterized. Average bioluminescence intensity in a 2.54-cm i.d. pipe was highly correlated with wall shear stress and BBP. This correlation, when further scaled by pipe diameter, effectively predicted bioluminescence intensity in fully developed turbulent flow in a 0.83-cm i.d. pipe. Determining similar correlations between other bathyphotometer flow agitators and flow fields will allow bioluminescence potential measurements to become a more powerful tool for the oceanographic community.

Latz, Michael I.; Rohr, Jim

2013-07-01

139

Phage-amplified bioluminescent bioreporters for the detection of foodborne pathogens  

Science.gov (United States)

The objective of this investigation is to develop a bioluminescent bioreporter system for the detection and monitoring of pathogenic microbial species. Current detection methodologies typically rely on time-consuming sample pre-enrichment steps to elevate pathogen concentrations to detectable levels or DNA based polymerase chain reaction (PCR) techniques that require extensive user training and expensive instrumentation. Detection utilizing bioluminescent bioreporter organisms, however, can provide a simple and rapid means of monitoring foodborne pathogens. Bioluminescent bioreporters are engineered to produce light in response to specific environmental inducers. The light signal is then measured with photodetector devices to generate a quantitative assessment of inducer concentration. The immediate goal of this research effort is to integrate key quorum sensing signal transduction elements into pathogen specific bacteriophages. Upon infection of a unique pathogenic species by the bacteriophages, quorum sensing signals will be generated that will subsequently stimulate bioluminescence in neighboring bioluminescent bioreporter cells. Utilizing both bacteriophages and bioluminescent bioreporters, we realize exceptional pathogen specificity while attaining enhanced bioluminescence production. This integrative approach will lead to rapid pathogen identification without requisite sample pre-enrichment. Additionally, since the bioluminescent response is completely intrinsic to the bioreporter organism, no user interventions are required for generating light signals; the protocol requires only addition of the food sample with the bacteriophage/bioluminescent bioreporter system. Measurement of light responses can be achieved using high-throughput microtiter plate readers, hand-held photomultiplier units, or microchip luminometers.

Ripp, Steven; Young, Jacque C.; Ozen, Aysu; Jegier, Patricia; Johnson, Courtney; Daumer, Kathleen; Garland, Jay; Sayler, Gary S.

2004-06-01

140

Bioluminescence tomography with Gaussian prior  

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Parameterizing the bioluminescent source globally in Gaussians provides several advantages over voxel representation in bioluminescence tomography. It is mathematically unique to recover Gaussians [Med. Phys. 31(8), 2289 (2004)] and practically sufficient to approximate various shapes by Gaussians in diffusive medium. The computational burden is significantly reduced since much fewer unknowns are required. Besides, there are physiological evidences that the source can be modeled by Gaussians....

Gao, Hao; Zhao, Hongkai; Cong, Wenxiang; Wang, Ge

2010-01-01

 
 
 
 
141

Early Time Points Perfusion Imaging  

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The aim was to investigate the feasibility of making relative cerebral blood flow (rCBF) maps from MR images acquired with short TR by measuring the initial arrival amount of Gd-DTPA evaluated within a time window before any contrast agent has a chance to leave the tissue. We named this rCBF measurement technique utilizing the early data points of the Gd-DTPA bolus the “early time points” method (ET), based on the hypothesis that early time point signals were proportional to rCBF. Simulat...

2011-01-01

142

Sustained accurate recording of intracellular acidification in living tissues with a photo-controllable bioluminescent protein.  

Science.gov (United States)

Regulation of an intracellular acidic environment plays a pivotal role in biological processes and functions. However, spatiotemporal analysis of the acidification in complex tissues of living subjects persists as an important challenge. We developed a photo-inactivatable bioluminescent indicator, based on a combination of luciferase-fragment complementation and a photoreaction of a light, oxygen, and voltage domain from Avena sativa Phototropin1 (LOV2), to visualize temporally dynamic acidification in living tissue samples. Bioluminescence of the indicator diminished upon light irradiation and it recovered gradually in the dark state thereafter. The recovery rate was remarkably sensitive to pH changes but unsusceptible to fluctuation of luciferin or ATP concentrations. Bioluminescence imaging, taken as an index of the recovery rates, enabled long-time recording of acidification in apoptotic and autophagous processes in a cell population and an ischemic condition in living mice. This technology using the indicator is widely applicable to sense organelle-specific acidic changes in target biological tissues. PMID:23690604

Hattori, Mitsuru; Haga, Sanae; Takakura, Hideo; Ozaki, Michitaka; Ozawa, Takeaki

2013-06-01

143

Early time points perfusion imaging.  

Science.gov (United States)

The aim was to investigate the feasibility of making relative cerebral blood flow (rCBF) maps from MR images acquired with short TR by measuring the initial arrival amount of Gd-DTPA evaluated within a time window before any contrast agent has a chance to leave the tissue. We named this rCBF measurement technique utilizing the early data points of the Gd-DTPA bolus the "early time points" method (ET), based on the hypothesis that early time point signals were proportional to rCBF. Simulation data were used successfully to examine the ideal behavior of ET while monkey's MRI results offered encouraging support to the utility of ET for rCBF calculation. A better brain coverage for ET could be obtained by applying the Simultaneous Echo Refocusing (SER) EPI technique. A recipe to run ET was presented, with attention paid to the noise problem around the time of arrival (TOA) of the contrast agent. PMID:20851196

Kwong, Kenneth K; Reese, Timothy G; Nelissen, Koen; Wu, Ona; Chan, Suk-Tak; Benner, Thomas; Mandeville, Joseph B; Foley, Mary; Vanduffel, Wim; Chesler, David A

2011-01-15

144

Real-time in vivo imaging of p16Ink4a reveals cross talk with p53  

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Expression of the p16Ink4a tumor suppressor gene, a sensor of oncogenic stress, is up-regulated by a variety of potentially oncogenic stimuli in cultured primary cells. However, because p16Ink4a expression is also induced by tissue culture stress, physiological mechanisms regulating p16Ink4a expression remain unclear. To eliminate any potential problems arising from tissue culture–imposed stress, we used bioluminescence imaging for noninvasive and real-time analysis of p16Ink4a expression u...

Yamakoshi, Kimi; Takahashi, Akiko; Hirota, Fumiko; Nakayama, Rika; Ishimaru, Naozumi; Kubo, Yoshiaki; Mann, David J.; Ohmura, Masako; Hirao, Atsushi; Saya, Hideyuki; Arase, Seiji; Hayashi, Yoshio; Nakao, Kazuki; Matsumoto, Mitsuru; Ohtani, Naoko

2009-01-01

145

Real-time metabolic imaging  

Science.gov (United States)

The endogenous substance pyruvate is of major importance to maintain energy homeostasis in the cells and provides a window to several important metabolic processes essential to cell survival. Cell viability is therefore reflected in the metabolism of pyruvate. NMR spectroscopy has until now been the only noninvasive method to gain insight into the fate of pyruvate in the body, but the low NMR sensitivity even at high field strength has only allowed information about steady-state conditions. The medically relevant information about the distribution, localization, and metabolic rate of the substance during the first minute after the injection has not been obtainable. Use of a hyperpolarization technique has enabled 10-15% polarization of 13C1 in up to a 0.3 M pyruvate solution. i.v. injection of the solution into rats and pigs allows imaging of the distribution of pyruvate and mapping of its major metabolites lactate and alanine within a time frame of 10 s. Real-time molecular imaging with MRI has become a reality. 13C | dynamic nuclear polarization | hyperpolarized | MRI | spectroscopy

Golman, Klaes; in 't Zandt, René; Thaning, Mikkel

2006-07-01

146

Effect of irradiation on bioluminescence spectrum of microbial ATP  

International Nuclear Information System (INIS)

The effect of irradiation on bioluminescence spectrum of dehydrated cabbage microbial ATP was studied. The results showed that the spectral bandwidth of ATP standard was from 490 to 640 nm and the peak wavelength was at 563 nm. The spectral bandwidths of irradiated dehydrated cabbage microbial ATP and CK did not change. Peak wavelengths of dehydrated cabbage irradiated at different dosages were not significantly different from that of CK. The peaks of bioluminescence spectrum of irradiated samples were higher than that of CK, which may be because of the increasing concentration of ATP, and this effect would be kept for quite a long time after irradiation. (authors)

2010-12-01

147

A multithread based new sparse matrix method in bioluminescence tomography  

Science.gov (United States)

Among many molecular imaging modalities, bioluminescence tomography (BLT) stands out as an effective approach for in vivo imaging because of its noninvasive molecular and cellular level detection ability, high sensitivity and low cost in comparison with other imaging technologies. However, there exists the case that large scale problem with large number of points and elements in the structure of mesh standing for the small animal or phantom. And the large scale problem's system matrix generated by the diffuse approximation (DA) model using finite element method (FEM) is large. So there wouldn't be enough random access memory (RAM) for the program and the related inverse problem couldn't be solved. Considering the sparse property of the BLT system matrix, we've developed a new sparse matrix (ZSM) to overcome the problem. And the related algorithms have all been speeded up by multi-thread technologies. Then the inverse problem is solved by Tikhonov regularization method in adaptive finite element (AFE) framework. Finally, the performance of this method is tested on a heterogeneous phantom and the boundary data is obtained through Monte Carlo simulation. During the process of solving the forward model, the ZSM can save more processing time and memory space than the usual way, such as those not using sparse matrix and those using Triples or Cross Linked sparse matrix. Numerical experiments have shown when more CPU cores are used, the processing speed is increased. By incorporating ZSM, BLT can be applied to large scale problems with large system matrix.

Zhang, Bo; Tian, Jie; Liu, Dan; Sun, Li; Yang, Xin; Han, Dong

2010-03-01

148

Expression of a Humanized Viral 2A-Mediated lux Operon Efficiently Generates Autonomous Bioluminescence in Human Cells  

Science.gov (United States)

Background Expression of autonomous bioluminescence from human cells was previously reported to be impossible, suggesting that all bioluminescent-based mammalian reporter systems must therefore require application of a potentially influential chemical substrate. While this was disproven when the bacterial luciferase (lux) cassette was demonstrated to function in a human cell, its expression required multiple genetic constructs, was functional in only a single cell type, and generated a significantly reduced signal compared to substrate-requiring systems. Here we investigate the use of a humanized, viral 2A-linked lux genetic architecture for the efficient introduction of an autobioluminescent phenotype across a variety of human cell lines. Methodology/Principal Findings The lux cassette was codon optimized and assembled into a synthetic human expression operon using viral 2A elements as linker regions. Human kidney, breast cancer, and colorectal cancer cell lines were both transiently and stably transfected with the humanized operon and the resulting autobioluminescent phenotype was evaluated using common imaging instrumentation. Autobioluminescent cells were screened for cytotoxic effects resulting from lux expression and their utility as bioreporters was evaluated through the demonstration of repeated monitoring of single populations over a prolonged period using both a modified E-SCREEN assay for estrogen detection and a classical cytotoxic compound detection assay for the antibiotic Zeocin. Furthermore, the use of self-directed bioluminescent initiation in response to target detection was assessed to determine its amenability towards deployment as fully autonomous sensors. In all cases, bioluminescent measurements were supported with traditional genetic and transcriptomic evaluations. Conclusions/Significance Our results demonstrate that the viral 2A-linked, humanized lux genetic architecture successfully produced autobioluminescent phenotypes in all cell lines tested without the induction of cytotoxicity. This autobioluminescent phenotype allowed for repeated interrogation of populations and self-directed control of bioluminescent activation with detection limits and EC50 values similar to traditional reporter systems, making the autobioluminescent cells amenable to automated monitoring and significantly reducing the time and cost required to perform bioluminescent workflows.

Xu, Tingting; Ripp, Steven; Sayler, Gary S.; Close, Dan M.

2014-01-01

149

Generation of topically transgenic rats by in utero electroporation and in vivo bioluminescence screening.  

Science.gov (United States)

In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the use of IUE for investigating behavior or neuropathology in the adult brain has been limited by insufficient methods for monitoring of IUE transfection success by non-invasive techniques in postnatal animals. For the present study, E16 rats were used for IUE. After intraventricular injection of the nucleic acids into the embryos, positioning of the tweezer electrodes was critical for targeting either the developing cortex or the hippocampus. Ventricular co-injection and electroporation of a luciferase gene allowed monitoring of the transfected cells postnatally after intraperitoneal luciferin injection in the anesthetized live P7 pup by in vivo bioluminescence, using an IVIS Spectrum device with 3D quantification software. Area definition by bioluminescence could clearly differentiate between cortical and hippocampal electroporations and detect a signal longitudinally over time up to 5 weeks after birth. This imaging technique allowed us to select pups with a sufficient number of transfected cells assumed necessary for triggering biological effects and, subsequently, to perform behavioral investigations at 3 month of age. As an example, this study demonstrates that IUE with the human full length DISC1 gene into the rat cortex led to amphetamine hypersensitivity. Co-transfected GFP could be detected in neurons by post mortem fluorescence microscopy in cryosections indicating gene expression present at ?6 months after birth. We conclude that postnatal bioluminescence imaging allows evaluating the success of transient transfections with IUE in rats. Investigations on the influence of topical gene manipulations during neurodevelopment on the adult brain and its connectivity are greatly facilitated. For many scientific questions, this technique can supplement or even replace the use of transgenic rats and provide a novel technology for behavioral neuroscience. PMID:24084570

Vomund, Sandra; Sapir, Tamar; Reiner, Orly; Silva, Maria A de Souza; Korth, Carsten

2013-01-01

150

Automatic Segmentation Framework of Building Anatomical Mouse Model for Bioluminescence Tomography  

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Full Text Available Bioluminescence tomography is known as a highly ill-posed inverse problem. To improve the reconstruction performance by introducing anatomical structures as a priori knowledge, an automatic segmentation framework has been proposed in this paper to extract the mouse whole-body organs and tissues, which enables to build up a heterogeneous mouse model for reconstruction of bioluminescence tomography. Finally, an in vivo mouse experiment has been conducted to evaluate this framework by using an X-ray computed tomography system and a multi-view bioluminescence imaging system. The findings suggest that the proposed method can realize fast automatic segmentation of mouse anatomical structures, ultimately enhancing the reconstruction performance of bioluminescence tomography.

Abdullah Alali

2013-09-01

151

Differential Evolution Approach for Regularized Bioluminescence Tomography  

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Bioluminescence tomography (BLT) is an inverse source problem that localizes and quantifies bioluminescent probe distribution in 3D. The generic BLT model is ill-posed, leading to non-unique solutions and aberrant reconstruction in the presence of measurement noise and optical parameter mismatches. In this paper, we introduce the knowledge of the number of bioluminescence sources to stabilize the BLT problem. Based on this regularized BLT model, we develop a differential evolution-based recon...

Cong, Alexander; Cong, Wenxiang; Lu, Yujie; Santago, Pete; Chatziiouannou, Arion; Wang, Ge

2010-01-01

152

Improved bioluminescent assay of somatic cell counts in raw milk.  

Science.gov (United States)

Somatic cell count (SCC) in milk is considered to be a valuable indicator of cow mastitis. For assessment of SCC in milk, the bioluminescent assay based on determination of ATP from somatic cells ([ATPsom]) in milk was proposed earlier. However, this assay is still not widely used in practice owing to lower reliability compared with conventional methods such as direct microscopy and flow cytometry. We revised the bioluminescent SCC assay and developed a simple protocol based on determination of the total non-bacterial ATP concentration in milk. It was shown that the novel ATP-releasing agent Neonol-10 (oxy-ethylated iso-nonyl phenol) has superior performance providing 100% lysis of somatic cells while not disrupting bacterial cells of milk at a concentration of 1.5% w/w. There was high correlation (R2=0.99) between measured bioluminescence and SCC as measured by direct microscopy. The observed detection limit of the bioluminescent milk SCC assay was as low as 900 cell/ml, time of analysis was 2-3 min per sample. The proposed method has high potential for on-site mastitis diagnostics. PMID:18513459

Frundzhyan, Valery G; Parkhomenko, Inna M; Brovko, Lubov Y; Ugarova, Natalia N

2008-08-01

153

Mathematical Study and Numerical Simulation of Multispectral Bioluminescence Tomography  

Directory of Open Access Journals (Sweden)

Full Text Available Multispectral bioluminescence tomography (BLT attracts increasingly more attention in the area of optical molecular imaging. In this paper, we analyze the properties of the solutions to the regularized and discretized multispectral BLT problems. First, we show the solution existence, uniqueness, and its continuous dependence on the data. Then, we introduce stable numerical schemes and derive error estimates for numerical solutions. We report some numerical results to illustrate the performance of the numerical methods on the quality of multispectral BLT reconstruction.

Ge Wang

2006-12-01

154

Preclinical evaluation of destruxin B as a novel Wnt signaling target suppressing proliferation and metastasis of colorectal cancer using non-invasive bioluminescence imaging  

International Nuclear Information System (INIS)

In continuation to our studies toward the identification of direct anti-cancer targets, here we showed that destruxin B (DB) from Metarhizium anisopliae suppressed the proliferation and induced cell cycle arrest in human colorectal cancer (CRC) HT29, SW480 and HCT116 cells. Additionally, DB induced apoptosis in HT29 cells by decreased expression level of anti-apoptotic proteins Bcl-2 and Bcl-xL while increased pro-apoptotic Bax. On the other hand, DB attenuated Wnt-signaling by downregulation of ?-catenin, Tcf4 and ?-catenin/Tcf4 transcriptional activity, concomitantly with decreased expression of ?-catenin target genes cyclin D1, c-myc and survivin. Furthermore, DB affected the migratory and invasive ability of HT29 cells through suppressed MMPs-2 and -9 enzymatic activities. We also found that DB targeted the MAPK and/or PI3K/Akt pathway by reduced expression of Akt, IKK-?, JNK, NF-?B, c-Jun and c-Fos while increased that of I?B?. Finally, we demonstrated that DB inhibited tumorigenesis in HT29 xenograft mice using non-invasive bioluminescence technique. Consistently, tumor samples from DB-treated mice demonstrated suppressed expression of ?-catenin, cyclin D1, survivin, and endothelial marker CD31 while increased caspase-3 expression. Collectively, our data supports DB as an inhibitor of Wnt/?-catenin/Tcf signaling pathway that may be beneficial in the CRC management. Highlights: ? Destruxin B (DB) inhibited colorectal cancer cells growth and induced apoptosis. ? MAPK and/or PI3K/Akt cascade cooperates in DB induced apoptosis. ? DB affected the migratory and invasive ability of HT29 cells through MMP-9. ? DB attenuated Wnt-signaling components ?-catenin, Tcf4. ? DB attenuated cyclin D1, c-myc, survivin and tumorigenesis in HT29 xenograft mice.

2012-05-15

155

Zero-scan-time vascular MR imaging  

International Nuclear Information System (INIS)

This paper describes a method that integrates vascular imaging with a routine high-resolution MR imaging examination in the same acquisition. All studies are performed after injection of Gd-DTPA using gradient recalled acquisition in a steady state with RF spoiling. Half-echo processing is used to reduce the echo time to 5 msec. Vascular images were extracted from the routinely acquired imaging examinations in 15 patients. High-quality basic images optimized for gadolinium-enhanced routine diagnostic MR imaging also provide suitable vascular enhancement for MR angiography in all cases

1990-11-25

156

Spectrally resolved bioluminescence tomography with the third-order simplified spherical harmonics approximation  

Energy Technology Data Exchange (ETDEWEB)

Bioluminescence imaging has been extensively applied to in vivo small animal imaging. Quantitative three-dimensional bioluminescent source information obtained by using bioluminescence tomography can directly and much more accurately reflect biological changes as opposed to planar bioluminescence imaging. Preliminary simulated and experimental reconstruction results demonstrate the feasibility and promise of bioluminescence tomography. However, the use of multiple approximations, particularly the diffusion approximation theory, affects the quality of in vivo small animal-based image reconstructions. In the development of new reconstruction algorithms, high-order approximation models of the radiative transfer equation and spectrally resolved data introduce new challenges to the reconstruction algorithm and speed. In this paper, an SP{sub 3}-based (the third-order simplified spherical harmonics approximation) spectrally resolved reconstruction algorithm is proposed. The simple linear relationship between the unknown source distribution and the spectrally resolved data is established in this algorithm. A parallel version of this algorithm is realized, making BLT reconstruction feasible for the whole body of small animals especially for fine spatial domain discretization. In simulation validations, the proposed algorithm shows improved reconstruction quality compared with diffusion approximation-based methods when high absorption, superficial sources and detection modes are considered. In addition, comparisons between fine and coarse mesh-based BLT reconstructions show the effects of numerical errors in reconstruction image quality. Finally, BLT reconstructions using in vivo mouse experiments further demonstrate the potential and effectiveness of the SP{sub 3}-based reconstruction algorithm.

Lu Yujie; Douraghy, Ali; Stout, David; Herschman, Harvey; Chatziioannou, Arion F [Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Machado, Hidevaldo B [Department of Biological Chemistry, Molecular Biology Institute, University of California, Los Angeles, CA 90095 (United States); Tian Jie [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, PO Box 2728, Beijing 100190 (China)], E-mail: archatziioann@mednet.ucla.edu, E-mail: hherschman@mednet.ucla.edu

2009-11-07

157

Spectrally resolved bioluminescence tomography with the third-order simplified spherical harmonics approximation  

International Nuclear Information System (INIS)

Bioluminescence imaging has been extensively applied to in vivo small animal imaging. Quantitative three-dimensional bioluminescent source information obtained by using bioluminescence tomography can directly and much more accurately reflect biological changes as opposed to planar bioluminescence imaging. Preliminary simulated and experimental reconstruction results demonstrate the feasibility and promise of bioluminescence tomography. However, the use of multiple approximations, particularly the diffusion approximation theory, affects the quality of in vivo small animal-based image reconstructions. In the development of new reconstruction algorithms, high-order approximation models of the radiative transfer equation and spectrally resolved data introduce new challenges to the reconstruction algorithm and speed. In this paper, an SP3-based (the third-order simplified spherical harmonics approximation) spectrally resolved reconstruction algorithm is proposed. The simple linear relationship between the unknown source distribution and the spectrally resolved data is established in this algorithm. A parallel version of this algorithm is realized, making BLT reconstruction feasible for the whole body of small animals especially for fine spatial domain discretization. In simulation validations, the proposed algorithm shows improved reconstruction quality compared with diffusion approximation-based methods when high absorption, superficial sources and detection modes are considered. In addition, comparisons between fine and coarse mesh-based BLT reconstructions show the effects of numerical errors in reconstruction image quality. Finally, BLT reconstructions using in vivo mouse experiments further demonstrate the potential and effectiveness of the SP3-based reconstruction algorithm.

2009-11-07

158

Real-time snapshot hyperspectral imaging endoscope  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Hyperspectral imaging has tremendous potential to detect important molecular biomarkers of early cancer based on their unique spectral signatures. Several drawbacks have limited its use for in vivo screening applications: most notably the poor temporal and spatial resolution, high expense, and low optical throughput of existing hyperspectral imagers. We present the development of a new real-time hyperspectral endoscope (called the image mapping spectroscopy endoscope) based on an image mappin...

Kester, Robert T.; Bedard, Noah; Gao, Liang; Tkaczyk, Tomasz S.

2011-01-01

159

Pharmacokinetic Modeling of Tumor Bioluminescence Implicates Efflux, and Not Influx, as the Bigger Hurdle in Cancer Drug Therapy  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In vivo bioluminescence imaging is a powerful tool for assessing tumor burden and quantifying therapeutic response in xenograft models. However this technique exhibits significant variability as a consequence of differences in substrate administration, as well as the tumor size, type, and location. Here we present a novel pharmacokinetic (PK) approach that utilizes bioluminescence image data. The sample data are taken from mice implanted with a melanoma tumor cell line that was transfected to...

Sim, Hoon; Bibee, Kristin; Wickline, Samuel A.; Sept, David

2011-01-01

160

Sparse Approximation of Computational Time Reversal Imaging  

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Computational time reversal imaging can be used to locate the position of multiple scatterers in a known background medium. Here, we discuss a sparse approximation method for computational time-reversal imaging. The method is formulated entirely in the frequency domain, and besides imaging it can also be used for denoising, and to determine the magnitude of the scattering coefficients in the presence of moderate noise levels.

Andrecut, M.

2009-01-01

 
 
 
 
161

Colonization of tomato seedlings by bioluminescent Clavibacter michiganensis subsp. michiganensis under different humidity regimes.  

Science.gov (United States)

Tomato bacterial canker, caused by Clavibacter michiganensis subsp. michiganensis, is transmitted by infected or infested seed and mechanically from plant to plant. Wounds occurring during seedling production and crop maintenance facilitate the dissemination of the pathogen. However, the effects of environmental factors on C. michiganensis subsp. michiganensis translocation and growth as an endophyte have not been fully elucidated. A virulent, stable, constitutively bioluminescent C. michiganensis subsp. michiganensis strain BL-Cmm 17 coupled with an in vivo imaging system allowed visualization of the C. michiganensis subsp. michiganensis colonization process in tomato seedlings in real time. The dynamics of bacterial infection in seedlings through wounds were compared under low (45%) and high (83%) relative humidity. Bacteria multiplied rapidly in cotyledon petioles remaining after clip inoculation and moved in the stem toward both root and shoot. Luminescent signals were also observed in tomato seedling roots over time, and root development was reduced in inoculated plants maintained under both humidity regimes. Wilting was more severe in seedlings under high-humidity regimes. A strong positive correlation between light intensity and bacterial population in planta suggests that bioluminescent C. michiganensis subsp. michiganensis strains will be useful in evaluating the efficacy of bactericides and host resistance. PMID:21936661

Xu, Xiulan; Rajashekara, Gireesh; Paul, Pierce A; Miller, Sally A

2012-02-01

162

Facile synthesis of gold-silver alloy nanoparticles for application in metal enhanced bioluminescence.  

Science.gov (United States)

In the present study we explored metal enhanced bioluminescence in luciferase enzymes for the first time. For this purpose a simple and reproducible one pot synthesis of gold-silver alloy nanoparticles was developed. By changing the molar ratio of tri-sodium citrate and silver nitrate we could synthesize spherical Au-Ag colloids of sizes ranging from 10 to 50 nm with a wide range of localized surface plasmon resonance (LSPR) peaks (450-550 nm). The optical tunability of the Au-Ag colloids enabled their effective use in enhancement of bioluminescence in a luminescent bacterium Photobacterium leiognathi and in luciferase enzyme systems from fireflies and bacteria. Enhancement of bioluminescence was 250% for bacterial cells, 95% for bacterial luciferase and 52% for firefly luciferase enzyme. The enhancement may be a result of energy transfer or plasmon induced enhancement. Such an increase can lead to higher sensitivity in detection of bioluminescent signals with potential applications in bio-analysis. PMID:24865663

Abhijith, K S; Sharma, Richa; Ranjan, Rajeev; Thakur, M S

2014-07-18

163

Hydrodynamic stimulation of dinoflagellate bioluminescence: a computational and experimental study.  

Science.gov (United States)

Dinoflagellate bioluminescence provides a near-instantaneous reporter of cell response to flow. Although both fluid shear stress and acceleration are thought to be stimulatory, previous studies have used flow fields dominated by shear. In the present study, computational and experimental approaches were used to assess the relative contributions to bioluminescence stimulation of shear stress and acceleration in a laminar converging nozzle. This flow field is characterized by separate regions of pronounced acceleration away from the walls, and shear along the wall. Bioluminescence of the dinoflagellates Lingulodinium polyedrum and Ceratocorys horrida, chosen because of their previously characterized different flow sensitivities, was imaged with a low-light video system. Numerical simulations were used to calculate the position of stimulated cells and the levels of acceleration and shear stress at these positions. Cells were stimulated at the nozzle throat within the wall boundary layer where, for that downstream position, shear stress was relatively high and acceleration relatively low. Cells of C. horrida were always stimulated significantly higher in the flow field than cells of L. polyedrum and at lower flow rates, consistent with their greater flow sensitivity. For both species, shear stress levels at the position of stimulated cells were similar to but slightly greater than previously determined response thresholds using independent flow fields. L. polyedrum did not respond in conditions where acceleration was as high as 20 g. These results indicate that shear stress, rather than acceleration, was the stimulatory component of flow. Thus, even in conditions of high acceleration, dinoflagellate bioluminescence is an effective marker of shear stress. PMID:15107447

Latz, Michael I; Juhl, Andrew R; Ahmed, Abdel M; Elghobashi, Said E; Rohr, Jim

2004-05-01

164

Bioluminescent orthotopic model of pancreatic cancer progression.  

Science.gov (United States)

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis. PMID:23852391

Chai, Ming G; Kim-Fuchs, Corina; Angst, Eliane; Sloan, Erica K

2013-01-01

165

Time Dimension in Consumers’ Image Construction Processes: Introducing Image Heritage and Image-in-use  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This paper focuses on the time dimension in consumers’ image construction processes. Two new concepts are introduced to cover past consumer experiences about the company – image heritage, and the present image construction process - image-in-use. Image heritage and image-in-use captures the dynamic, relational, social, and contextual features of corporate image construction processes. Qualitative data from a retailing context were collected and analysed following a grounded theory approac...

Rindell, Anne

2010-01-01

166

Coherent interferometric imaging, time gating and beamforming  

International Nuclear Information System (INIS)

Coherent interferometric imaging is based on the backpropagation of local spacetime cross-correlations of array data and was introduced in order to improve images when the medium between the array and the object to be imaged is inhomogeneous and unknown (Borcea et al 2005 Inverse Problems 21 1419). Although this method has been shown to be effective and is well founded theoretically, the coherent interferometric imaging function is computationally expensive and therefore difficult to use. In this paper, we show that this function is equivalent to a windowed beamformer energy function, that is, a quadratic function that involves only time gating and time delaying signals in emission and in reception. In this form the coherent interferometric imaging can be implemented efficiently both in hardware and software, that is, at a computational cost that is comparable to the usual beamforming and migration imaging methods. We also revisit the trade-off between enhanced image stability and loss of resolution in coherent interferometry from the point of view of its equivalence to a windowed beamformer energy imaging function

2011-06-01

167

Real-time multiple image manipulations  

International Nuclear Information System (INIS)

There are many situations in which it is desired to manipulate two or more images under real-time operator control. The authors have investigated a number of such cases in order to determine their value and applicability in clinical medicine and laboratory research. Several examples are presented in detail. The DICOM-8 video image computer system was used due to its capability of storing two 512 x 512 x 8 bit images and operating on them, and/or an incoming video frame, with any of a number of real time operations including addition, subtraction, inversion, averaging, logical AND, NAND, OR, NOR, NOT, XOR and XNOR, as well as combinations of these. Some applications involve manipulations of or among the stored images. In others, a stored image is used as a mask or template for positioning or adjusting a second image to be grabbed via a video camera. The accuracy of radiotherapy treatment is verified by comparing port films with the original radiographic planning film, which is previously digitized and stored. Moving the port film on the light box while viewing the real-time subtraction image allows for adjustments of zoom, translation and rotation, together with contrast and edge enhancement

1984-01-01

168

Can compression reduce forensic image time?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Creating a forensic copy (image) of a hard disk drive is one of the fundamental tasks a computer forensic analyst must perform. Time is often critical, and there is a need to consider a trade-off between a number of factors to achieve best results. This paper reports the results from an exploratory study into the impact of using disk drive compression on the time needed to image (and verify) a hard disk drive. It was found that time reduction may be achieved once the trade-off of contributing...

2011-01-01

169

Analytical Applications of Bioluminescence and Chemiluminescence  

Science.gov (United States)

Bioluminescence and chemiluminescence studies were used to measure the amount of adenosine triphosphate and therefore the amount of energy available. Firefly luciferase - luciferin enzyme system was emphasized. Photometer designs are also considered.

Chappelle, E. W. (editor); Picciolo, G. L. (editor)

1975-01-01

170

Doubling time of liver metastase images  

International Nuclear Information System (INIS)

For our study, where clinical and scintigraphic observation seldom lasts more than two years and where measurable metastases always exceed 1 cm"3, the exponential model was adopted and our results were all calculated with GERSTENBERG's formula which gives an apparent doubling time. The liver metastases were measured on the scintigraphic image obtained, a more or less sharply limited blank which makes for a first difficulty of judgement. Two magnascanner V type PICKER 5-inch crystal scintigraphs were used, giving three images simultaneously by a transcriber made up of a stylus and a light spot built into the detection system. The isotope used is colloidal gold ("1"9"8Au) phagocytized by the Kuepfferian reticulo-endothelial system. The doubling time for liver metastase scintigraphic images calculated for fifteen patients having undergone one or more isotopic checks after a first metastase image was discovered range from 10 to 103 days

1975-01-01

171

Time-encoded imaging of energetic radiation  

Science.gov (United States)

Time-encoded imaging (TEI) is a new approach to directional detection of energetic radiation that produces images by inducing a time-dependent modulation of detected particles. TEI-based detectors use single-scatter events and have a low channel count, reducing complexity and cost while maintaining high efficiency with respect to other radiation imaging techniques such as double-scatter or coded aperture imaging. The scalability of TEI systems makes them a very promising detector class for weak source detection. Extension of the technique to high-resolution imaging is also under study. With a prototype time-encoding detector, we demonstrated detection of a neutron source at 60 m with neutron output equivalent to an IAEA significant quantity of WGPu. We have since designed and built a full-scale detector based on the time-encoding concept. We will present results from characterization of very large liquid scintillator cells, including pulse shape discrimination, as well as from studies of the detector system performance in weak source detection scenarios.

Brennan, James; Brubaker, Erik; Gerling, Mark; Marleau, Peter; Nowack, Aaron; Schuster, Patricia

2013-09-01

172

Real-time imaging detectors for portal imaging  

International Nuclear Information System (INIS)

This paper reviews the status of real-time imaging systems which are used in radiation-therapy for radiotherapy localization and verification. Imaging systems under review include (1) metal-fluorescent screens, optically coupled to video cameras, (2) metal-phosphor screen in direct contact with two-dimensional photo-diode array (flat panel detector), (3) two-dimensional liquid ionization chamber and (5) linear diode arrays. These systems permit frequent verification during the treatment and have been shown to be very useful. Unfortunately the image quality achieved, while impressive considering the short time the devices have been on the market, is significantly inferior to that which is available form the metal/film combination (port film). The spatial resolution is about 1 lp/mm at 10% MTF, the Detective Quantum Efficiency (DQE) is less than 1% at 0 lp/mm and less than 0.1% at 1 lp/mm. It is also noted that these systems have not reached their ultimate limits of performance yet. The levels of x-ray fluence in the radiotherapy beam should allow a significant increase in the image quality. Nevertheless, the digital imaging systems available now are superior to analog film based systems because they provide a separation between the important functions of detection and display. They can provide almost instant image processing to optimize the information to be presented to the human observer. 100 refs., 11 figs., 4 tabs

1993-07-11

173

A fast dynamic linked library based mixed-language programming technology for the trust region method in bioluminescence tomography  

Science.gov (United States)

Bioluminescence tomography (BLT) is a novel optical molecular imaging (MI) modality. It can reconstruct the inner bioluminescent light source distribution, according to the surface light distribution. The trust region method (TRM) can overcome the ill-posedness of BLT for its regularization property. As there exists a "TRUST" function that can solve the trust region subproblem in Matlab and Matlab's powerful matrix operation ability suited for TRM, the TRM is implemented in Matlab. Then the Matlab code of TRM is transformed into a dynamic linked library (DDL) and mixed together with the C++ code of the adaptive finite element (AFE) framework, using the mixed-language programming technology (MLPT). There are two main advantages of the MLPT. The first is taking advantages of all the participated programming languages. The second is time efficient. The usual way of transferring data between programmes written in different programming languages is to write the data first into files that are stored in the hard discs in one programme, and then read the files from another programme. Besides wasting time on writing and reading, it is difficult to keep the precision of the data. The DLL based MLPT can eliminate the need of installing code compilers in the platform running the software. Furthermore, in DLL, the code is implemented in C/C++ with high time efficiency, while the code in Matlab remains relatively low time efficiency. Finally, a numerical experiment is carried out to show MLPT's usage in the source reconstruction procedure of BLT, using the MLPT based on DLL.

Zhang, Bo; Tian, Jie; Yang, Xin; Qin, Chenghu; Han, Dong; Ma, Xibo

2011-03-01

174

Time-Resolved Imaging Of Transient Plasma  

International Nuclear Information System (INIS)

Pulsed capillary discharge is a compact device that is used to perform fast electrical discharge that is used to produce transient plasma. In this work, a more economical imaging technique is developed in order to study the dynamics of the plasma that is formed in a capillary tube. The imaging system consists of two main devices, a four-frame gated micro-channel plate and a Nikon Coolpix5000 camera. The time-resolved imaging that we have performed in order to study the dynamics of the plasma that is formed in a 10 mm long and 1 mm diameter low pressure capillary tube is reported. The images obtained portrayed that the plasma is heated up when the magnitude of the current is around the maximum and cools down when the current magnitude is around the minimum.

2009-07-07

175

Small animal fluorescence and bioluminescence tomography: a review of approaches, algorithms and technology update  

Science.gov (United States)

Emerging fluorescence and bioluminescence tomography approaches have several common, yet several distinct features from established emission tomographies of PET and SPECT. Although both nuclear and optical imaging modalities involve counting of photons, nuclear imaging techniques collect the emitted high energy (100-511 keV) photons after radioactive decay of radionuclides while optical techniques count low-energy (1.5-4.1 eV) photons that are scattered and absorbed by tissues requiring models of light transport for quantitative image reconstruction. Fluorescence imaging has been recently translated into clinic demonstrating high sensitivity, modest tissue penetration depth, and fast, millisecond image acquisition times. As a consequence, the promise of quantitative optical tomography as a complement of small animal PET and SPECT remains high. In this review, we summarize the different instrumentation, methodological approaches and schema for inverse image reconstructions for optical tomography, including luminescence and fluorescence modalities, and comment on limitations and key technological advances needed for further discovery research and translation.

Darne, Chinmay; Lu, Yujie; Sevick-Muraca, Eva M.

2014-01-01

176

Real-time image and video processing  

CERN Document Server

This book presents an overview of the guidelines and strategies for transitioning an image or video processing algorithm from a research environment into a real-time constrained environment. Such guidelines and strategies are scattered in the literature of various disciplines including image processing, computer engineering, and software engineering, and thus have not previously appeared in one place. By bringing these strategies into one place, the book is intended to serve the greater community of researchers, practicing engineers, industrial professionals, who are interested in taking an im

Kehtarnavaz, Nasser

2006-01-01

177

Real-Time Imaging of Quantum Entanglement  

CERN Document Server

Quantum Entanglement - correlations between at least two systems that are stronger than classically explainable - is widely regarded as one of the most prominent features of quantum mechanics and quantum information science. Although, the creation of entanglement between two systems has become possible in laboratories, it has been out of the grasp of one of the most natural ways to investigate nature: direct visual observation. Here we show that modern imaging technology, namely a triggered intensified charge coupled device (ICCD) camera, is fast and sensitive enough to image in real-time the influence of the measurement of one photon on its entangled partner. To demonstrate the non-classicality of the measurements quantitatively from the registered intensity we develop a novel method to statistically analyze the image and precisely quantify the number of photons within a certain region. In addition, we show the high flexibility of our experimental setup in creating any desired spatial-mode entanglement, even...

Fickler, Robert; Lapkiewicz, Radek; Ramelow, Sven; Zeilinger, Anton

2013-01-01

178

Immobilized Bioluminescent Reagents in Flow Injection Analysis.  

Science.gov (United States)

Available from UMI in association with The British Library. Bioluminescent reactions exhibits two important characteristics from an analytical viewpoint; they are selective and highly sensitive. Furthermore, bioluminescent emissions are easily measured with a simple flow-through detector based on a photomultiplier tube and the rapid and reproducible mixing of sample and expensive reagent is best achieved by a flow injection manifold. The two most important bioluminescent systems are the enzyme (luciferase)/substrate (luciferin) combinations extracted from fireflies (Photinus pyralis) and marine bacteria (Virio harveyi) which requires ATP and NAD(P)H respectively as cofactors. Reactions that generate or consume these cofactors can also be coupled to the bioluminescent reaction to provide assays for a wide range of clinically important species. A flow injection manifold for the study of bioluminescent reactions is described, as are procedures for the extraction, purification and immobilization of firefly and bacterial luciferase and oxidoreductase. Results are presented for the determination of ATP using firefly system and the determination of other enzymes and substrates participating in ATP-converting reactions e.g. creatine kinase, ATP-sulphurylase, pyruvate kinase, creatine phosphate, pyrophosphate and phophoenolypyruvate. Similarly results are presented for the determination of NAD(P)H, FMN, FMNH_2 and several dehydrogenases which produce NAD(P)H and their substrates, e.g. alcohol, L-lactate, L-malate, L-glutamate, Glucose-6-phosphate and primary bile acid.

Nabi, Abdul

179

Shear-stress dependence of dinoflagellate bioluminescence.  

Science.gov (United States)

Fluid flow stimulates bioluminescence in dinoflagellates. However, many aspects of the cellular mechanotransduction are incompletely known. The objective of our study was to formally test the hypothesis that flow-stimulated dinoflagellate bioluminescence is dependent on shear stress, signifying that organisms are responding to the applied fluid force. The dinoflagellate Lingulodinium polyedrum was exposed to steady shear using simple Couette flow in which fluid viscosity was manipulated to alter shear stress. At a constant shear rate, a higher shear stress due to increased viscosity increased both bioluminescence intensity and decay rate, supporting our hypothesis that bioluminescence is shear-stress dependent. Although the flow response of non-marine attached cells is known to be mediated through shear stress, our results indicate that suspended cells such as dinoflagellates also sense and respond to shear stress. Shear-stress dependence of flow-stimulated bioluminescence in dinoflagellates is consistent with mechanical stimulation due to direct predator handling in the context of predator-prey interactions. PMID:17565113

Maldonado, Elisa M; Latz, Michael I

2007-06-01

180

Understanding Bioluminescence in Dinoflagellates—How Far Have We Come?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Some dinoflagellates possess the remarkable genetic, biochemical, and cellular machinery to produce bioluminescence. Bioluminescent species appear to be ubiquitous in surface waters globally and include numerous cosmopolitan and harmful taxa. Nevertheless, bioluminescence remains an enigmatic topic in biology, particularly with regard to the organisms’ lifestyle. In this paper, we review the literature on the cellular mechanisms, molecular evolution, diversity, and ecology of bioluminescenc...

2013-01-01

 
 
 
 
181

Bioluminescent bioreporter assays for targeted detection of chemical and biological agents  

Science.gov (United States)

Bioluminescent bioreporters carrying the bacterial lux gene cassette have been well established for the sensing and monitoring of select chemical agents. Their ability to generate target specific visible light signals with no requirement for extraneous additions of substrate or other hands-on manipulations affords a real-time, repetitive assaying technique that is remarkable in its simplicity and accuracy. Although the predominant application of lux-based bioluminescent bioreporters has been towards chemical compound detection, novel genetic engineering schemes are yielding a variety of new bioreporter systems that extend the lux sensing mechanism beyond mere analyte discrimination. For example, the unique specificity of bacteriophage (bacterial viruses) has been exploited in lux bioluminescent assays for specific identification of foodborne bacterial pathogens such as Escherichia coli O157:H7. With the concurrent ability to interface bioluminescent bioreporter assays onto integrated circuit microluminometers (BBICs; bioluminescent bioreporter integrated circuits), the potential exists for the development of sentinel microchips that can function as environmental monitors for multiplexed recognition of chemical and biological agents in air, food, and water. The size and portability of BBIC biosensors may ultimately provide a deployable, interactive network sensing technology adaptable towards chem/bio defense.

Ripp, Steven; Jegier, Pat; Johnson, Courtney; Moser, Scott; Islam, Syed; Sayler, Gary

2008-05-01

182

Interactive Real-time Magnetic Resonance Imaging  

DEFF Research Database (Denmark)

Real-time acquisition, reconstruction and interactively changing the slice position using magnetic resonance imaging (MRI) have been possible for years. However, the current clinical use of interactive real-time MRI is limited due to an inherent low spatial and temporal resolution. This PhD project seeks to implement and assess existing reconstruction algorithms using multi-processors of modern graphics cards and many-core computer processors and to cover some of the potential clinical applications which might benefit from using an interactive real-time MRI system. First an off-line, but interactive, slice alignment tool was used to support the notion that 3D blood flow quantification in the heart possesses the ability to obtain curves and volumes which are not statistical different from standard 2D flow. Secondly, the feasibility of an interactive real-time MRI system was exploited with regard to optimal sampling strategy for detecting motion in four different anatomies on two different MRI scanner brands. A fully implemented interactive real-time MRI system was exploited in a group of healthy fetuses and proved its eligibility as an alternative diagnostic tool for fetal imaging. Finally, the system was used for 3D motion tracking of the liver, and its use was proposed for future integrations of MRI scanners and linear accelerators in the field of radiotherapy treatment.

Brix, Lau

2013-01-01

183

Towards real-time image quality assessment  

Science.gov (United States)

We introduce a real-time implementation and evaluation of a new fast accurate full reference based image quality metric. The popular general image quality metric known as the Structural Similarity Index Metric (SSIM) has been shown to be an effective, efficient and useful, finding many practical and theoretical applications. Recently the authors have proposed an enhanced version of the SSIM algorithm known as the Rotated Gaussian Discrimination Metric (RGDM). This approach uses a Gaussian-like discrimination function to evaluate local contrast and luminance. RGDM was inspired by an exploration of local statistical parameter variations in relation to variation of Mean Opinion Score (MOS) for a range of particular distortion types. In this paper we out-line the salient features of the derivation of RGDM and show how analyses of local statistics of distortion type necessitate variation in discrimination function width. Results on the LIVE image database show tight banding of RGDM metric value when plotted against mean opinion score indicating the usefulness of this metric. We then explore a number of strategies for algorithmic speed-up including the application of Integral Images for patch based computation optimisation, cost reduction for the evaluation of the discrimination function and general loop unrolling. We also employ fast Single Instruction Multiple Data (SIMD) intrinsics and explore data parallel decomposition on a multi-core Intel Processor.

Geary, Bobby; Grecos, Christos

2011-02-01

184

Time Variant Change Analysis in Satellite Images  

Directory of Open Access Journals (Sweden)

Full Text Available This paper describes the time variant changes in satellite images using Self Organizing Feature Map (SOFM technique associated with Artificial Neural Network. In this paper, we take a satellite image and find the time variant changes using above technique with the help of MATLAB. This paper reviews remotely sensed data analysis with neural networks. First, we present an overview of the main concepts underlying Artificial Neural Networks (ANNs, including the main architectures and learning algorithms. Then, the main tasks that involve ANNs in remote sensing are described. We first make a brief introduction to models of networks, for then describing in general terms Artificial Neural Networks (ANNs. As an application, we explain the back propagation algorithm, since it is widely used and many other algorithms are derived from it. There are two techniques that are used for classification in pattern recognition such as Supervised Classification and Unsupervised Classification. In supervised learning technique the network knows about the target and it has to change accordingly to get the desired output corresponding to the presented input sample data. Most of the previous work has already been done on supervised classification. In this study we are going to present the classification of satellite images using unsupervised classification method of ANN.

Rachita Sharma

2013-05-01

185

Improving medical imaging report turnaround times.  

Science.gov (United States)

Southern Ohio Medical Center (SOMC), a 232-bed community-based teaching hospital, is equipped with state-of-the-art equipment such as 2 16-slice computed tomography (CT) scanners, 3 MR scanners, 3 ultrasound scanners, 2 digital mammography units, and 3 nuclear medicine cameras. One hundred twenty-six employees--ranging from support personnel to technologists along with 7 board-certified radiologists--staff the medical imaging department. Procedure volume is approximately 164,000 per year and is performed in all American College of Radiology (ACR)-accredited modalities. Filmless since 1998, SOMC's medical imaging department has resulted in productivity gains to the estimated 164,000 procudures for fiscal year 2005. The catalyst for the department is a robust picture archiving and communication system (PACS). Working with the radiologists, staff, and transcription services, turnaround time was reduced to from 13 hours to 9 hours from exam start to report sign off. Additional technology intervention was essential to further decrease report turnaround time. SOMC served as a beta site for a radiology information system (RIS). The new RIS has allowed the medical imaging department to move from a paper department to a "pseudo paperless" department. Orders, history sheets, consents, and other forms are scanned into the RIS for staff and radiologist use. Requisitions are no longer printed, and staff has access to review workstations to ensure that patients are called back into the department for procedures. This new workflow has also reduced paper traffic within the department. The last piece of the technology puzzle to improve report turnaround time was voice recognition technology. From its implementation, voice recognition enhanced the RIS technology. All of the radiologists began to use the product as soon as it was available. They perform all the editing and corrections either by voice command or by manual typing. The medical imaging department has noted that voice command corrections and editing are more efficient for the radiologist. The overall impact on decreased radiology report turnaround times is not only seen in medical imaging, but also has a global affect within the hospital. SOMC plans to realize a reduction length of patient stays, and a faster process for plotting the course of patient treatment, e.g., faster visits from emergency department (ED) physicians to patients. PMID:15794377

Marquez, Luis O

2005-01-01

186

Papyrus imaging with terahertz time domain spectroscopy  

Science.gov (United States)

Terahertz time domain spectroscopic imaging (THz-TDSI) is a non-ionizing, non-contact and non-destructive measurement technique that has been recently utilized to study cultural heritage artifacts. We will present this technique and the results of non-contact measurements of papyrus texts, including images of hidden papyri. Inks for modern papyrus specimens were prepared using the historical binder, Arabic gum, and two common pigments used to write ancient texts, carbon black and red ochre. The samples were scanned in reflection at normal incidence with a pulse with a spectral range between 0.1 and 1.5 THz. Temporal analysis of the signals provides the depths of the layers, and their frequency spectra give information about the inks.

Labaune, J.; Jackson, J. B.; Pagčs-Camagna, S.; Duling, I. N.; Menu, M.; Mourou, G. A.

2010-09-01

187

A Multi-Camera System for Bioluminescence Tomography in Preclinical Oncology Research  

Directory of Open Access Journals (Sweden)

Full Text Available Bioluminescent imaging (BLI of cells expressing luciferase is a valuable noninvasive technique for investigating molecular events and tumor dynamics in the living animal. Current usage is often limited to planar imaging, but tomographic imaging can enhance the usefulness of this technique in quantitative biomedical studies by allowing accurate determination of tumor size and attribution of the emitted light to a specific organ or tissue. Bioluminescence tomography based on a single camera with source rotation or mirrors to provide additional views has previously been reported. We report here in vivo studies using a novel approach with multiple rotating cameras that, when combined with image reconstruction software, provides the desired representation of point source metastases and other small lesions. Comparison with MRI validated the ability to detect lung tumor colonization in mouse lung.

Ralph P. Mason

2013-07-01

188

Observed and modeled bio-optical, bioluminescent, and physical properties during a coastal upwelling event in Monterey Bay, California  

Science.gov (United States)

During spring and summer time, coastal upwelling influences circulation and ecosystem dynamics of the Monterey Bay, California, which is recognized as a National Marine Sanctuary. Observations of physical, bio-optical properties (including bioluminescence) together with results from dynamical biochemical and bioluminescence models are used to interpret the development of the upwelling event during August 2003 in Monterey Bay, California. Observations and the biochemical model show the development of a phytoplankton bloom in the southern portion of Monterey Bay. Model results show an increase of nutrients in the southern portion of the bay, where nutrient-rich water masses are brought in by the southward flow and cyclonic circulation inside the bay. This increase in nutrients together with the sluggish circulation in the southern portion of the bay provides favorable conditions for phytoplankton growth. Our observations and models suggest that with the development of upwelling the offshore water masses with the subsurface layer of bioluminescent zooplankton were replaced by water masses advected from the northern coast of the bay with a relatively high presence of mostly nonbioluminescent phytoplankton. Inshore observations from autonomous underwater vehicles (AUVs) show consistent coincidence of chlorophyll, backscatter, and bioluminescence maxima during upwelling development. Offshore AUV observations (taken at the entrance to the bay) show a deeper bioluminescence maximum below the surface layers of high chlorophyll and backscatter values during the earlier stages of upwelling development. Later, the observed deep offshore bioluminescence maximum disappeared and became a shallower and much weaker signal, coinciding with high chlorophyll and backscatter values offshore. Based on the biochemical and bioluminescence models, a methodology for estimating the nighttime water-leaving radiance due to stimulated bioluminescence is demonstrated and evaluated.

Shulman, Igor; Moline, Mark A.; Penta, Bradley; Anderson, Stephanie; Oliver, Matthew; Haddock, Steven H. D.

2011-01-01

189

Phase Time and Envelope Time in Time-Distance Analysis and Acoustic Imaging  

Science.gov (United States)

Time-distance analysis and acoustic imaging are two related techniques to probe the local properties of solar interior. In this study, we discuss the relation of phase time and envelope time between the two techniques. The location of the envelope peak of the cross correlation function in time-distance analysis is identified as the travel time of the wave packet formed by modes with the same w/l. The phase time of the cross correlation function provides information of the phase change accumulated along the wave path, including the phase change at the boundaries of the mode cavity. The acoustic signals constructed with the technique of acoustic imaging contain both phase and intensity information. The phase of constructed signals can be studied by computing the cross correlation function between time series constructed with ingoing and outgoing waves. In this study, we use the data taken with the Taiwan Oscillation Network (TON) instrument and the Michelson Doppler Imager (MDI) instrument. The analysis is carried out for the quiet Sun. We use the relation of envelope time versus distance measured in time-distance analyses to construct the acoustic signals in acoustic imaging analyses. The phase time of the cross correlation function of constructed ingoing and outgoing time series is twice the difference between the phase time and envelope time in time-distance analyses as predicted. The envelope peak of the cross correlation function between constructed ingoing and outgoing time series is located at zero time as predicted for results of one-bounce at 3 mHz for all four data sets and two-bounce at 3 mHz for two TON data sets. But it is different from zero for other cases. The cause of the deviation of the envelope peak from zero is not known.

Chou, Dean-Yi; Duvall, Thomas L.; Sun, Ming-Tsung; Chang, Hsiang-Kuang; Jimenez, Antonio; Rabello-Soares, Maria Cristina; Ai, Guoxiang; Wang, Gwo-Ping; Goode Philip; Marquette, William; Ehgamberdiev, Shuhrat; Landenkov, Oleg

1999-01-01

190

Practical application of bioluminescence enzyme immunoassay using enhancer for firefly luciferin-luciferase bioluminescence.  

Science.gov (United States)

Firefly luciferin-luciferase bioluminescence is known for its high quantum yield (41.0 ± 7.4%). Given this high quantum yield, application of this bioluminescence is expected to be useful in the field of clinical diagnostics. The kinetic profile of this bioluminescence exhibits an instant rise (<1 s) and a rapid decay in light emission (decreased to 42% after 5 s). In this study, we applied four enhancers including coenzyme A, inosine5'-triphosphate sodium salt, sodium tripolyphosphate and potassium pyrophosphate to prolong light emission. When these enhancers were used, luminescence was only decreased to 89, 83, 87 and 82% after 5 s, respectively. These materials modified the kinetic profile of bioluminescence so that the luminescence is more suitable for clinical application. It becomes more suitable because they enable highly sensitive integration and simplification of a device by separating luminescence measurements from dispensing of reagents. Using these enhancers, we then developed a bioluminescent enzyme immunoassay (BLEIA) for hepatitis B virus surface antigen (HBsAg) that employed firefly luciferase as a labeling enzyme. We compared the results obtained from the HBsAg BLEIA method with the conventional chemiluminescent enzyme immunoassay method, and found a satisfactory correlation (r=0.984, n=118). PMID:21681909

Minekawa, Takayuki; Ohkuma, Hiroshi; Abe, Katsushi; Maekawa, Hiroaki; Arakawa, Hidetoshi

2011-01-01

191

Reliability of a bioluminescence ATP assay for detection of bacteria.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The reliability of bioluminescence assays which employ the luciferin-luciferase ATP-dependent reaction to evaluate bacterial counts was studied, both in vitro and on urine specimens. Bioluminescence and cultural results for the most common urinary tract pathogens were analyzed. Furthermore, the influence of the culture medium, of the assaying method, and of the phase of growth on bioluminescence readings was studied. Results show that Proteus, Providencia, and Morganella strains are not corre...

1992-01-01

192

Cloning and characterization of new bioluminescent proteins  

Science.gov (United States)

Over the past two years Prolume has undertaken a comprehensive program to clone luciferases and associated 'green fluorescent proteins' (GFPs) from marine animals that use coelenterazine as the luciferin. To data we have cloned several bioluminescent proteins, including two novel copepod luciferases and two anthozoan GFPs. These four proteins have sequences that differ greatly form previously cloned analogous proteins; the sequence diversity apparently is due to independent evolutionary origins and unusual evolutionary constraints. Thus coelenterazine-based bioluminescent systems may also manifest a variety of useful properties. We discuss form this taxonomic perspective the initial biochemical and spectral characterization of our cloned proteins. Emphasis is placed on the anthozoan luciferase-GFP systems, whose efficient resonance energy transfer has elicited much current interest.

Szent-Gyorgyi, Christopher; Ballou, Byron T.; Dagnal, Erich; Bryan, Bruce

1999-07-01

193

Bioluminescence enhancement through an added washing protocol enabling a greater sensitivity to carbofuran toxicity.  

Science.gov (United States)

The effects of carbofuran toxicity on a genetically modified bacterial strain E. coli DPD2794 were enhanced using a new bioluminescent protocol which consisted of three consecutive steps: incubation, washing and luminescence reading. Specifically, in the first step, several concentrations of carbofuran aqueous solutions were incubated with different bacterial suspensions at recorded optical densities for different lengths of time. Thereafter, the resulting bacterial/toxicant mixtures were centrifuged and the aged cellular supernatant replaced with fresh medium. In the final step, the carbofuran- induced bioluminescence to the exposed E. coli DPD2794 bacteria was shown to provide a faster and higher intensity when recorded at a higher temperature at30°C which is not usually used in the literature. It was found that the incubation time and the replacement of aged cellular medium were essential factors to distinguish different concentrations of carbofuran in the bioluminescent assays. From our results, the optimum incubation time for a "light ON" bioluminescence detection of the effect of carbofuran was 6h. Thanks to the replacement of the aged cellular medium, a group of additional peaks starting around 30min were observed and we used the corresponding areas under the curve (AUC) at different contents of carbofuran to produce the calibration curve. Based on the new protocol, a carbofuran concentration of 0.5pg/mL can be easily determined in a microtiter plate bioluminescent assay, while a non-wash protocol provides an unexplainable order of curve evolutionswhich does not allow the user to determine the concentration. PMID:23867093

Jia, Kun; Eltzov, Evgeni; Marks, Robert S; Ionescu, Rodica E

2013-10-01

194

Novel tools for use in bioluminescence resonance energy transfer (BRET) assays.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Recent advances in imaging assays based on bioluminescence resonance energy transfer (BRET) have made it possible to study protein/protein interactions in living cells under physiological conditions. Here we describe protocols for these assays including relevant positive and negative controls, and we also show how they can be combined with protein complementation assays such as bimolecular fluorescence complementation (BiFC) to study three- and four-partner interactions. We also describe a BR...

Baragli, Alessandra

2009-01-01

195

A Monte-Carlo-Based Network Method for Source Positioning in Bioluminescence Tomography  

Directory of Open Access Journals (Sweden)

Full Text Available We present an approach based on the improved Levenberg Marquardt (LM algorithm of backpropagation (BP neural network to estimate the light source position in bioluminescent imaging. For solving the forward problem, the table-based random sampling algorithm (TBRS, a fast Monte Carlo simulation method we developed before, is employed here. Result shows that BP is an effective method to position the light source.

Zhun Xu

2007-10-01

196

Image timing and detector performance of a matrix ion-chamber electronic portal imaging device  

International Nuclear Information System (INIS)

The Oncology Centre of Auckland Hospital recently purchased a Varian PortalVisionTM electronic portal imaging device (EPID). Image acquisition times, input-output characteristics and contrast-detail curves of this matrix liquid ion-chamber EPID have been measured to examine the variation in imaging performance with acquisition mode. The variation in detector performance with acquisition mode has been examined. The HV cycle time can be increased to improve image quality. Consideration should be given to the acquisition mode and HV cycle time used when imaging to ensure adequate imaging performance with reasonable imaging time. (author)

1995-11-20

197

Rapid detection of bacteria in green tea using a novel pretreatment method in a bioluminescence assay.  

Science.gov (United States)

Tea is one of the most popular beverages consumed in the world, and green tea has become a popular beverage in Western as well as Asian countries. A novel pretreatment method for a commercial bioluminescence assay to detect bacteria in green tea was developed and evaluated in this study. Pretreatment buffers with pH levels ranging from 6.0 to 9.0 were selected from MES (morpholineethanesulfonic acid), HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), or Tricine buffers. To evaluate the effect of pretreatment and the performance of the assay, serially diluted cultures of Enterobacter cloacae, Escherichia coli, Bacillus subtilis, and Staphylococcus aureus were tested. The improved methods, which consisted of a pretreatment of the sample in alkaline buffer, significantly decreased the background bioluminescence intensity of green tea samples when compared with the conventional method. Pretreatment with alkaline buffers with pH levels ranging from 8.0 to 9.0 increased the bioluminescence intensities of cultures of E. cloacae and S. aureus. Strong log-linear relationships between the bioluminescence intensities and plate counts emerged for the tested strains. Furthermore, the microbial detection limit was 15 CFU in 500 ml of bottled green tea after an 8-h incubation at 35°C and an assay time of 1 h. The results showed that contaminated samples could be detected within 1 h of operation using our improved bioluminescence assay. This method could be used to test for contamination during the manufacturing process as well as for statistical sampling for quality control. PMID:24853516

Shinozaki, Yohei; Harada, Yasuhiro

2014-06-01

198

Iterative reconstruction for bioluminescence tomography with total variation regularization  

Science.gov (United States)

Bioluminescence tomography(BLT) is an instrumental molecular imaging modality designed for the 3D location and quantification of bioluminescent sources distribution in vivo. In our context, the diffusion approximation(DA) to radiative transfer equation(RTE) is utilized to model the forward process of light propagation. Mathematically, the solution uniqueness does not hold for DA-based BLT which is an inverse source problem of partial differential equations and hence is highly ill-posed. In the current work, we concentrate on a general regularization framework for BLT with Bregman distance as data fidelity and total variation(TV) as regularization. Two specializations of the Bregman distance, the least squares(LS) distance and Kullback-Leibler(KL) divergence, which correspond to the Gaussian and Poisson environments respectively, are demonstrated and the resulting regularization problems are denoted as LS+TV and KL+TV. Based on the constrained Landweber(CL) scheme and expectation maximization(EM) algorithm for BLT, iterative algorithms for the LS+TV and KL+TV problems in the context of BLT are developed, which are denoted as CL-TV and EM-TV respectively. They are both essentially gradient-based algorithms alternatingly performing the standard CL or EM iteration step and the TV correction step which requires the solution of a weighted ROF model. Chambolle's duality-based approach is adapted and extended to solving the weighted ROF subproblem. Numerical experiments for a 3D heterogeneous mouse phantom are carried out and preliminary results are reported to verify and evaluate the proposed algorithms. It is found that for piecewise-constant sources both CL-TV and EM-TV outperform the conventional CL and EM algorithms for BLT.

Jin, Wenma; He, Yonghong

2012-12-01

199

Characterization of bioluminescent derivatives of assimilable organic carbon test bacteria.  

Science.gov (United States)

The assimilable organic carbon (AOC) test is a standardized measure of the bacterial growth potential of treated water. We describe the design and initial development of an AOC assay that uses bioluminescent derivatives of AOC test bacteria. Our assay is based on the observation that bioluminescence peaks at full cell yield just prior to the onset of the stationary phase during growth in a water sample. Pseudomonas fluorescens P-17 and Spirillum sp. strain NOX bacteria were mutagenized with luxCDABE operon fusion and inducible transposons and were selected on minimal medium. Independent mutants were screened for high luminescence activity and predicted AOC assay sensitivity. All mutants tested were able to grow in tap water under AOC assay conditions. Strains P-17 I5 (with p-aminosalicylate inducer) and NOX I3 were chosen for use in the bioluminescence AOC test. Peak bioluminescence and plate count AOC were linearly related for both test bacteria, though data suggest that the P-17 bioluminescence assay requires more consistent luminescence monitoring. Bioluminescence results were obtained 2 or 3 days postinoculation, compared with 5 days for the ATP luminescence AOC assay and 8 days for the plate count assay. Plate count AOC assay results for nonmutant and bioluminescent bacteria from 36 water samples showed insignificant differences, indicating that the luminescent bacteria retained a full range of AOC measurement capability. This bioluminescence method is amenable to automation with a microplate format with programmable reagent injection. PMID:14766564

Haddix, Pryce L; Shaw, Nancy J; LeChevallier, Mark W

2004-02-01

200

Time-delay compensation for stabilization imaging system  

Science.gov (United States)

The spatial resolution of imaging systems for airborne and space-borne remote sensing are often limited by image degradation resulting from mechanical vibrations of platforms during image exposure. A straightforward way to overcome this problem is to actively stabilize the optical axis or drive the focal plane synchronous to the motion image during exposure. Thus stabilization imaging system usually consists of digital image motion estimation and micromechanical compensation. The performance of such kind of visual servo system is closely related to precision of motion estimation and time delay. Large time delay results in larger phase delay between motion estimation and micromechanical compensation, and leads to larger uncompensated residual motion and limited bandwidth. The paper analyzes the time delay caused by image acquisition period and introduces a time delay compensation method based on SVM (Support Vector Machine) motion prediction. The main idea to cancel the time delay is to predict the current image motion from delayed measurements. A support vector machine based method is designed to predict the image motion. A prototype of stabilization imaging system has been implemented in the lab. To analyze the influences of time delay on system performance and to verify the proposed time delay cancelation method, comparative experiments over various frequencies of vibration are taken. The experimental results show that, the accuracy of motion compensation and the bandwidth of the system can be significantly improved with time delay cancelation.

Chen, Yueting; Xu, Zhihai; Li, Qi; Feng, Huajun

2014-05-01

 
 
 
 
201

Dinoflagellate bioluminescence in response to mechanical stimuli in water flows  

Directory of Open Access Journals (Sweden)

Full Text Available Bioluminescence of plankton organisms induced by water movements has long been observed and is still under investigations because of its great complexity. In particular, the exact mechanism occurring at the level of the cell has not been yet fully understood. This work is devoted to the study of the bioluminescence of the dinoflagellates plankton species Pyrocystis noctiluca in response to mechanical stimuli generated by water flows. Several experiments were performed with different types of flows in a Couette shearing apparatus. All of them converge to the conclusion that stationary homogeneous laminar shear does not trigger massive bioluminescence, but that acceleration and shear are both necessary to stimulate together an intense bioluminescence response. The distribution of the experimental bioluminescence thresholds is finally calculated from the light emission response for the Pyrocystis noctiluca species.

A. S. Cussatlegras

2005-01-01

202

Real time neutron radiography using a Lixi neutron imaging device  

International Nuclear Information System (INIS)

A real time neutron radiography system has been developed at the University of Michigan Phoenix Memorial Laboratory (PML) and has recently been used to test the imaging capabilities of a neutron imaging device developed by Lixi, Inc. of Downers Grove, Ill. This device uses an input phosphor that is high in gadolinium to generate a light image which is then sent through an intensifier stage to provide images that can be viewed by eye, video camera, or standard 35 mm camera. It was determined that this device provides images of much higher resolution and sensitivity than those obtained with the imaging system currently being used at PML. Using computerized image enhancement techniques, the images obtained with the Lixi neutron imaging device can then be further enhanced or processed to obtain quantitative information on the object being imaged. (orig.)

1986-01-15

203

Cerenkov imaging - a new modality for molecular imaging.  

Science.gov (United States)

Cerenkov luminescence imaging (CLI) is an emerging hybrid modality that utilizes the light emission from many commonly used medical isotopes. Cerenkov radiation (CR) is produced when charged particles travel through a dielectric medium faster than the speed of light in that medium. First described in detail nearly 100 years ago, CR has only recently applied for biomedical imaging purposes. The modality is of considerable interest as it enables the use of widespread luminescence imaging equipment to visualize clinical diagnostic (all PET radioisotopes) and many therapeutic radionuclides. The amount of light detected in CLI applications is significantly lower than other that in other optical imaging techniques such as bioluminescence and fluorescence. However, significant advantages include the use of approved radiotracers and lack of an incident light source, resulting in high signal to background ratios. As well, multiple subjects may be imaged concurrently (up to 5 in common bioluminescent equipment), conferring both cost and time benefits. This review summarizes the field of Cerenkov luminescence imaging to date. Applications of CLI discussed include intraoperative radionuclide-guided surgery, monitoring of therapeutic efficacy, tomographic optical imaging capabilities, and the ability to perform multiplexed imaging using fluorophores excited by the Cerenkov radiation. While technical challenges still exist, Cerenkov imaging has materialized as an important molecular imaging modality. PMID:23133811

Thorek, Daniel Lj; Robertson, Robbie; Bacchus, Wassifa A; Hahn, Jaeseung; Rothberg, Julie; Beattie, Bradley J; Grimm, Jan

2012-01-01

204

Bioluminescent bacteria: lux genes as environmental biosensors Bactérias bioluminescentes: os genes lux como biosensores ambientais  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Bioluminescent bacteria are widespread in natural environments. Over the years, many researchers have been studying the physiology, biochemistry and genetic control of bacterial bioluminescence. These discoveries have revolutionized the area of Environmental Microbiology through the use of luminescent genes as biosensors for environmental studies. This paper will review the chronology of scientific discoveries on bacterial bioluminescence and the current applications of bioluminescence in env...

2003-01-01

205

Image detection in real time based on fuzzy fractal theory.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Real time image detection is still a challenge in research. Several methods have been used, but all can be divide in two approaches: the first is based on image field estimation in this case the quality of image is depending on the estimation method. The second is based on electrons collection, the particularity of this approach is that more the collection time is longer, better will be the quality of image. In both of these approaches, the global image should be obtained by assembling the mo...

Abraham Berthe, Kya; Yan, Yan; Dembe?le?, Sounkalo

2006-01-01

206

Local statistical analysis of real-time NRG images  

International Nuclear Information System (INIS)

An analysis of real-time neutron images, taken with the cyclotron-based neutron radiography system of Sumitomo Heavy Industries, Ltd., which uses a set of neutron-photon converter, LiF + ZnS (Ag) scintillation screen, and a high sensitive SIT television camera, is described. The fluctuation of image intensity at a constant neutron flux domain is well represented by a Poisson distribution and ripple noise appearing in the real-time image can be attributed to the deficiency of neutron flux. The image integration technique, therefore, would be effective for improvement of the statistical quality of neutron image. Some digital image processing techniques based on statistical method would be effective for a ruffled image like real-time neutron radiography. (author)

1986-01-01

207

Digital Image Processing: An Overview of Computational Time Requirement.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Image processing is a growing field covering a wide range of techniques for the manipulation of digital images [1].Many image processing tasks can be characterized as being computationally intensive. One reason for this is the vast amount of data that requires processing, more than seven million pixels per second for typical image sources. To keep up with these data rates and demanding computations in real-time, the processing engine must provide specialized data paths, application-specific o...

2013-01-01

208

The design of time resolved intensified CCD imaging system  

International Nuclear Information System (INIS)

Coupled the CCD with an image intensifier, the intensified CCD imaging system has such advantages as higher signal gain, higher signal-to-noise ratio (SNR) and higher dynamic range. Time resolution can be achieved by controlling the shutter time of the image intensifier with an electronic pulse. The structures and set-up of the intensified CCD imaging system, and the performance parameters in field use are detailed. The object plane spatial resolution of 5 lp/mm is achieved when enlargement rate is 1. The dynamic range of this system is 38.7, and the synchronous precision is less than 2 ns, the time resolution is 5 ns. (authors)

2010-08-01

209

Volumetric Real-Time Imaging Using a CMUT Ring Array  

Science.gov (United States)

A ring array provides a very suitable geometry for forward-looking volumetric intracardiac and intravascular ultrasound imaging. We fabricated an annular 64-element capacitive micromachined ultrasonic transducer (CMUT) array featuring a 10-MHz operating frequency and a 1.27-mm outer radius. A custom software suite was developed to run on a PC-based imaging system for real-time imaging using this device. This paper presents simulated and experimental imaging results for the described CMUT ring array. Three different imaging methods—flash, classic phased array (CPA), and synthetic phased array (SPA)—were used in the study. For SPA imaging, two techniques to improve the image quality—Hadamard coding and aperture weighting—were also applied. The results show that SPA with Hadamard coding and aperture weighting is a good option for ring-array imaging. Compared with CPA, it achieves better image resolution and comparable signal-to-noise ratio at a much faster image acquisition rate. Using this method, a fast frame rate of up to 463 volumes per second is achievable if limited only by the ultrasound time of flight; with the described system we reconstructed three cross-sectional images in real-time at 10 frames per second, which was limited by the computation time in synthetic beamforming.

Choe, Jung Woo; Oralkan, Omer; Nikoozadeh, Amin; Gencel, Mustafa; Stephens, Douglas N.; O'Donnell, Matthew; Sahn, David J.; Khuri-Yakub, Butrus T.

2012-01-01

210

Volumetric real-time imaging using a CMUT ring array.  

Science.gov (United States)

A ring array provides a very suitable geometry for forward-looking volumetric intracardiac and intravascular ultrasound imaging. We fabricated an annular 64-element capacitive micromachined ultrasonic transducer (CMUT) array featuring a 10-MHz operating frequency and a 1.27-mm outer radius. A custom software suite was developed to run on a PC-based imaging system for real-time imaging using this device. This paper presents simulated and experimental imaging results for the described CMUT ring array. Three different imaging methods--flash, classic phased array (CPA), and synthetic phased array (SPA)--were used in the study. For SPA imaging, two techniques to improve the image quality--Hadamard coding and aperture weighting--were also applied. The results show that SPA with Hadamard coding and aperture weighting is a good option for ring-array imaging. Compared with CPA, it achieves better image resolution and comparable signal-to-noise ratio at a much faster image acquisition rate. Using this method, a fast frame rate of up to 463 volumes per second is achievable if limited only by the ultrasound time of flight; with the described system we reconstructed three cross-sectional images in real-time at 10 frames per second, which was limited by the computation time in synthetic beamforming. PMID:22718870

Choe, Jung Woo; Oralkan, Ömer; Nikoozadeh, Amin; Gencel, Mustafa; Stephens, Douglas N; O'Donnell, Matthew; Sahn, David J; Khuri-Yakub, Butrus T

2012-06-01

211

UWGSP7: a real-time optical imaging workstation  

Science.gov (United States)

With the development of UWGSP7, the University of Washington Image Computing Systems Laboratory has a real-time workstation for continuous-wave (cw) optical reflectance imaging. Recent discoveries in optical science and imaging research have suggested potential practical use of the technology as a medical imaging modality and identified the need for a machine to support these applications in real time. The UWGSP7 system was developed to provide researchers with a high-performance, versatile tool for use in optical imaging experiments with the eventual goal of bringing the technology into clinical use. One of several major applications of cw optical reflectance imaging is tumor imaging which uses a light-absorbing dye that preferentially sequesters in tumor tissue. This property could be used to locate tumors and to identify tumor margins intraoperatively. Cw optical reflectance imaging consists of illumination of a target with a band-limited light source and monitoring the light transmitted by or reflected from the target. While continuously illuminating the target, a control image is acquired and stored. A dye is injected into a subject and a sequence of data images are acquired and processed. The data images are aligned with the control image and then subtracted to obtain a signal representing the change in optical reflectance over time. This signal can be enhanced by digital image processing and displayed in pseudo-color. This type of emerging imaging technique requires a computer system that is versatile and adaptable. The UWGSP7 utilizes a VESA local bus PC as a host computer running the Windows NT operating system and includes ICSL developed add-on boards for image acquisition and processing. The image acquisition board is used to digitize and format the analog signal from the input device into digital frames and to the average frames into images. To accommodate different input devices, the camera interface circuitry is designed in a small mezzanine board that supports the RS-170 standard. The image acquisition board is connected to the image- processing board using a direct connect port which provides a 66 Mbytes/s channel independent of the system bus. The image processing board utilizes the Texas Instruments TMS320C80 Multimedia Video Processor chip. This chip is capable of 2 billion operations per second providing the UWGSP7 with the capability to perform real-time image processing functions like median filtering, convolution and contrast enhancement. This processing power allows interactive analysis of the experiments as compared to current practice of off-line processing and analysis. Due to its flexibility and programmability, the UWGSP7 can be adapted into various research needs in intraoperative optical imaging.

Bush, John E.; Kim, Yongmin; Pennington, Stan D.; Alleman, Andrew P.

1995-04-01

212

Timing and Operating Mode Design for Time-Gated Fluorescence Lifetime Imaging Microscopy  

Science.gov (United States)

Steady-state fluorence imaging and time-resolved fluorescence imaging are two important areas in fluorescence imaging research. Fluorescence lifetime imaging is an absolute measurement method which is independent of excitation laser intensity, fluorophore concentration, and photobleaching compared to fluorescence intensity imaging techniques. Time-gated fluorescence lifetime imaging microscopy (FLIM) can provide high resolution and high imaging frame during mature FLIM methods. An abstract time-gated FLIM model was given, and important temporal parameters are shown as well. Aiming at different applications of steady and transient fluorescence processes, two different operation modes, timing and lifetime computing algorithm are designed. High resolution and high frame can be achieved by one-excitation one-sampling mode and least square algorithm for steady imaging applications. Correspondingly, one-excitation two-sampling mode and rapid lifetime determination algorithm contribute to transient fluorescence situations.

Wang, Xinwei; Zhou, Yan; Liu, Yuliang

2013-01-01

213

Photo-real rendering of bioluminescence and iridescence in creatures from the abyss  

Science.gov (United States)

The generation of photo-real renderings of bioluminescence is developed for creatures from the abyss. Bioluminescence results from a chemical reaction with examples found in deep-sea marine environments including: algae, copepods, jellyfish, squid, and fish. In bioluminescence, the excitation energy is supplied by a chemical reaction, not by a source of light. The greatest transparency window in seawater is in the blue region of the visible spectrum. From small creatures like single-cell algae, to large species of siphonophore Praya dubia (40m), luminescent phenomena can be produced by mechanical excitement from disturbances of objects passing by. Deep sea fish, like the Pacific Black Dragonfish are covered with photophores along the upper and lower surfaces which emits light when disturbed. Other animals like small squids have several different types of light organs oscillating at different rates. Custom shaders and material phenomena incorporate indirect lighting like: global illumination, final gathering, ambient occlusion and subsurface scattering to provide photo real images. Species like the Hydomedusae jellyfish, produce colors that are also generated by iridescence of thin tissues. The modeling and rendering of these tissues requires thin film multilayer stacks. These phenomena are simulated by semi-rigid body dynamics in a procedural animation environment. These techniques have been applied to develop spectral rendering of scenes outside the normal visible window in typical computer animation render engines.

Prusten, Mark

2008-08-01

214

An ebCMOS camera system for marine bioluminescence observation: The LuSEApher prototype  

International Nuclear Information System (INIS)

The ebCMOS camera, called LuSEApher, is a marine bioluminescence recorder device adapted to extreme low light level. This prototype is based on the skeleton of the LUSIPHER camera system originally developed for fluorescence imaging. It has been installed at 2500 m depth off the Mediterranean shore on the site of the ANTARES neutrino telescope. The LuSEApher camera is mounted on the Instrumented Interface Module connected to the ANTARES network for environmental science purposes (European Seas Observatory Network). The LuSEApher is a self-triggered photo detection system with photon counting ability. The presentation of the device is given and its performances such as the single photon reconstruction, noise performances and trigger strategy are presented. The first recorded movies of bioluminescence are analyzed. To our knowledge, those types of events have never been obtained with such a sensitivity and such a frame rate. We believe that this camera concept could open a new window on bioluminescence studies in the deep sea.

2012-12-11

215

Design of real-time remote sensing image compression system  

Science.gov (United States)

This paper focuses on the issue of CCD remote sensing image compression. Compared with other images, CCD remote sensing image data is characterized with high speed and high quantized bits. A high speed CCD image compression system is proposed based on ADV212 chip. The system is mainly composed of three devices: FPGA, SRAM and ADV212. In this system, SRAM plays the role of data buffer, ADV212 focuses on data compression and the FPGA is used for image storage and interface bus control. Finally, a system platform is designed to test the performance of compression. Test results show that the proposed scheme can satisfy the real-time processing requirement and there is no obvious difference between the sourced image and the compressed image in respect of image quality.

Wu, Wenbo; Lei, Ning; Wang, Kun; Wang, Qingyuan; Li, Tao

2013-08-01

216

Real-time extended dynamic range imaging in shearography  

International Nuclear Information System (INIS)

Extended dynamic range (EDR) imaging is a postprocessing technique commonly associated with photography. Multiple images of a scene are recorded by the camera using different shutter settings and are merged into a single higher dynamic range image. Speckle interferometry and holography techniques require a well-modulated intensity signal to extract the phase information, and of these techniques shearography is most sensitive to different object surface reflectivities as it uses self-referencing from a sheared image. In this paper the authors demonstrate real-time EDR imaging in shearography and present experimental results from a difficult surface reflectivity sample: a wooden panel painting containing gold and dark earth color paint

2008-10-20

217

Is bioluminescent turbulence an example of self-organized critically?  

Science.gov (United States)

When mechanically stimulated by flow, some marine cells react by flashing blue light or bioluminescing. The flash response give a relatively non-invasive measure of turbulent behavior which lends itself readily and quantitatively to statistical models of fluid structure. We report substantial agreement between a series of experiments performed using stimulated light from the bioluminescence of seawater flowing through a glass pipe and theoretical predictions based on models proposed for describing a self-organized critical phenomenon. In this way, bioluminescent turbulent flow can be taken as a natural and potentially robust example connecting long-held scaling problems in turbulence to recent thinking on fractals and self-organized critically.

Noever, David; Cronise, Raymond

1994-06-01

218

Construction of a bioluminescent reporter strain to detect polychlorinated biphenyls  

Energy Technology Data Exchange (ETDEWEB)

A bioluminescent reporter strain, Ralstonia eutropha ENV307 (pUTK60), was constructed for the detection of polychlorinated biphenyls by inserting the biphenyl promoter upstream of the bioluminescence genes. In the presence of a nonionic surfactant, which enhances the solubility of chlorinated biphenyls, bioluminescence was induced three- to fourfold over background by biphenyl, monochlorinated biphenyls, and Aroclor 1242. The minimum detection limits for these compounds ranged from 0.15 mg/liter for 4-chlorobiphenyl to 1.5 mg/liter for Aroclor 1242.

Layton, A.C.; Muccini, M.; Ghosh, M.M.; Sayler, G.S. [Univ. of Tennessee, Knoxville, TN (United States)

1998-12-01

219

Real-time image deblurring by optoelectronic hybrid processing.  

Science.gov (United States)

An efficient approach is presented to restore a motion-blurred image in real time by optoelectronic hybrid processing, by which an image motion vector can be effectively detected and an accurate point spread function is constructed rapidly. A simple Wiener filter algorithm is employed to restore the blurred image. It greatly alleviates the complexity of the restoration algorithm. The proposed approach can apply to arbitrary motion-blurred image restoration. An optoelectronic hybrid joint transform correlation is constructed to verify the efficiency. The experimental results show that the proposed method has distinct advantages of preferable effect and good real time. PMID:22108875

Qian, Yixian; Hu, Fangrong; Cheng, Xiaowei; Jin, Weimin

2011-11-20

220

Noninvasive Imaging Technologies Reveal Edema Toxin as a Key Virulence Factor in Anthrax  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Powerful noninvasive imaging technologies enable real-time tracking of pathogen-host interactions in vivo, giving access to previously elusive events. We visualized the interactions between wild-type Bacillus anthracis and its host during a spore infection through bioluminescence imaging coupled with histology. We show that edema toxin plays a central role in virulence in guinea pigs and during inhalational infection in mice. Edema toxin (ET), but not lethal toxin (LT), markedly modified the ...

Dumetz, Fabien; Jouvion, Gre?gory; Khun, Huot; Glomski, Ian Justin; Corre, Jean-philippe; Rougeaux, Cle?mence; Tang, Wei-jen; Mock, Miche?le; Huerre, Michel; Goossens, Pierre Louis

2011-01-01

 
 
 
 
221

Real-time digital x-ray subtraction imaging  

International Nuclear Information System (INIS)

The invention provides a method of producing visible difference images derived from an X-ray image of an anatomical subject, comprising the steps of directing X-rays through the anatomical subject for producing an image, converting the image into television fields comprising trains of on-going video signals, digitally storing and integrating the on-going video signals over a time interval corresponding to several successive television fields and thereby producing stored and integrated video signals, recovering the video signals from storage and producing integrated video signals, producing video difference signals by performing a subtraction between the integrated video signals and the on-going video signals outside the time interval, and converting the difference signals into visible television difference images representing on-going changes in the X-ray image

1978-05-16

222

Digital Image Processing: An Overview of Computational Time Requirement.  

Directory of Open Access Journals (Sweden)

Full Text Available Image processing is a growing field covering a wide range of techniques for the manipulation of digital images [1].Many image processing tasks can be characterized as being computationally intensive. One reason for this is the vast amount of data that requires processing, more than seven million pixels per second for typical image sources. To keep up with these data rates and demanding computations in real-time, the processing engine must provide specialized data paths, application-specific operators, creative data management, and careful sequencing and pipelining. The paper is confined to the major drawback of digital image processing which is the computational time required and is also said to be the computational power of a digital image processing system.

Ahaiwe J

2013-08-01

223

Real-Time Image Composition with Correction of Drift Distortion  

CERN Multimedia

In this article, a new scanning electron microscopy (SEM) image composition technique is described, which can significantly reduce drift-distortion related image corruptions. Drift-distortion commonly causes blur and distortions in the SEM images. Such corruption ordinarily appears when conventional image-acquisition methods, i.e. "slow scan" and "fast scan", are applied. The corruption is often very significant; it may render images unusable for metrology applications, especially, when sub-nanometer accuracy is required. The described correction technique works with a large number of quickly taken frames, which are properly aligned and then composed into a single image. Such image contains much less noise than the individual frames, whilst the blur and deformation is minimized. Also, this technique provides useful information about changes of the sample position in time, which may be well used to investigate the drift properties of the instrument.

Cizmar, Petr; Postek, Michael T

2009-01-01

224

Volumetric Real-Time Imaging Using a CMUT Ring Array  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A ring array provides a very suitable geometry for forward-looking volumetric intracardiac and intravascular ultrasound imaging. We fabricated an annular 64-element capacitive micromachined ultrasonic transducer (CMUT) array featuring a 10-MHz operating frequency and a 1.27-mm outer radius. A custom software suite was developed to run on a PC-based imaging system for real-time imaging using this device.

Choe, Jung Woo; Oralkan, O?mer; Nikoozadeh, Amin; Gencel, Mustafa; Stephens, Douglas N.; O’donnell, Matthew; Sahn, David J.; Khuri-yakub, Butrus T.

2012-01-01

225

Preserving time structures while denoising a dynamical image  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In restoration or denoising of a movie, the classical procedures often do not take into account the full information provided by the movie. These pro- cedures are either applied spatially "image per image" or locally on some neighborhood combining both closeness in one image and closeness in time. The local information is then combined homogeneously in order to realize the treatment. There is one type of movie where both approaches fail to provide a relevant treatment. Such a movie, c...

Rozenholc, Yves; Reiss, Markus

2012-01-01

226

Mechanism of Energy Conversion and Transfer in Bioluminescence. Final Report.  

Science.gov (United States)

Bioluminescence in the sea pansy, Renilla reniformis, a marine anthozoan coelenterate, is a complex process involving the participation of three proteins specific to anthozoan coelenterate-type systems. These are: (1) the luciferin binding protein, (2) th...

M. J. Cormier

1979-01-01

227

Real-time neutron coded aperture imaging: A technique for nondestructive three-dimensional imaging  

Energy Technology Data Exchange (ETDEWEB)

Neutron Coded Aperture Imaging is a nondestructive imaging technique that utilizes neutrons scattered from an object through specially designed apertures. Coded Aperture Imaging is an alternative technique to Computed Tomography for three-dimensional imaging. Coded Aperture Imaging has the advantage that all of the three-dimensional information is contained in a single image, whereas Computed Tomography requires several images or projections. This technique has been implemented by other using photographic film as an image recording medium and optical reconstruction or decoding of the images. In this work, the possibility of using a real-time neutron video camera to record the images, followed by digital decoding methodology has been investigated. Because only a small fraction of the neutrons incident on the object are scattered to the neutron camera, a new neutron beamport facility, with a larger neutron flux (7.3 x 10[sup 7] n/cm[sup 2]/s) than the previous facility was constructed. A real-time imaging system has been assembled that also allows for image integration directly on the video tube for low flux applications. A technique for aperture fabrication has been developed. This technique utilizes vapor deposition of neutron absorbing material (Gadolinium) onto a substrate followed by photolithographic etching of the coded aperture pattern. Various method for digital decoding have been examined including a novel use of the Wiener filter for direct image decoding. These methods have been successfully implemented on simulated images to insure that they function properly. Algorithms for decoding multiple planes to exploit the three-dimensional information have been developed and tested. In addition, a method to remove the contributions due to out-of-focus planes from a given reconstructed or decoded image was developed. Coded images were successfully recorded using the real-time imaging system with integration times of approximately 5 minutes.

Gibbs, K.M.

1992-01-01

228

Real time magneto-optical imaging of vortices in superconductors  

CERN Document Server

We demonstrate here real-time imaging of individual vortices in a NbSe2 single crystal using polarized light microscopy. A new high-sensitivity magneto-optical (MO) imaging system enables observation of the static vortex lattice as well as single vortex motion at low flux densities.

Goa, P E; Baziljevich, M; Ilyashenko, E I; Gammel, P L; Johansen, T H; Goa, Paal Erik; Hauglin, Harald; Baziljevich, Michael; Il'yashenko, Eugene; Gammel, Peter L.; Johansen, Tom H.

2001-01-01

229

Photographic documentation in real time radiographic imaging procedures and other inspection procedures using video imaging techniques  

International Nuclear Information System (INIS)

With an automatic daylight video imaging system, based on the use of wet processing type of film, it is possible to produce permanent documents of real time video images with almost the same convenience as video tape recording while maintaining all the advantages of display flexibility and image quality offered by photographic documents

1985-01-01

230

Monitoring Neural Activity with Bioluminescence during Natural Behavior  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Existing techniques for monitoring neural activity in awake, freely behaving vertebrates are invasive and difficult to target to genetically identified neurons. Here we describe the use of bioluminescence to non-invasively monitor the activity of genetically specified neurons in freely behaving zebrafish. Transgenic fish expressing the Ca2+-sensitive photoprotein GFP-apoAequorin (GA) in most neurons generated large and fast bioluminescent signals related to neural activity, neuroluminescence,...

2010-01-01

231

Monitoring Neural Activity with Bioluminescence during Natural Behavior  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Existing techniques for monitoring neural activity in awake, freely behaving vertebrates are invasive and difficult to target to genetically identified neurons. We used bioluminescence to non-invasively monitor the activity of genetically specified neurons in freely behaving zebrafish. Transgenic fish with the Ca\\(^{2+}\\)-sensitive photoprotein green fluorescent protein (GFP)-Aequorin in most neurons generated large and fast bioluminescent signals that were related to neural activity, neurolu...

2010-01-01

232

Characterization of Bioluminescent Derivatives of Assimilable Organic Carbon Test Bacteria  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The assimilable organic carbon (AOC) test is a standardized measure of the bacterial growth potential of treated water. We describe the design and initial development of an AOC assay that uses bioluminescent derivatives of AOC test bacteria. Our assay is based on the observation that bioluminescence peaks at full cell yield just prior to the onset of the stationary phase during growth in a water sample. Pseudomonas fluorescens P-17 and Spirillum sp. strain NOX bacteria were mutagenized with l...

Haddix, Pryce L.; Shaw, Nancy J.; Lechevallier, Mark W.

2004-01-01

233

Rapid time-gated polarimetric Stokes imaging using photoelastic modulators.  

Science.gov (United States)

We report a rapid time-gated full Stokes imaging approach without mechanically moving parts, which is well-suited for biomedical applications, using two photoelastic modulators (PEMs). A charge-coupled device (CCD) with microsecond time-gating capability was used to acquire the images. To synchronize the CCD with the PEMs, thus gaining signal-to-noise ratio advantage, a field programmable gate array was employed. After calibration, an evolutionary algorithm was used to select four time points from which the full Stokes vector can be recovered. Using the images taken by the camera at these four times (in ~80 ms), the images of the full Stokes vectors of different incident polarization states were accurately derived. PMID:24104631

Alali, Sanaz; Yang, Tianyu; Vitkin, I Alex

2013-08-15

234

Analysis of time-varying psoriasis lesion image patterns  

DEFF Research Database (Denmark)

The multivariate alteration detection transform is applied to pairs of within and between time varying registered psoriasis image patterns. Color band contribution to the variates explaining maximal change is analyzed.

Maletti, Gabriela Mariel; Ersbøll, Bjarne Kjær

2004-01-01

235

Imaging regional cerebral vascular transit time  

International Nuclear Information System (INIS)

Regional variations in cerebral capillary transit time (CTT) could affect the accuracy of models used for measurements of cerebral blood flow and metabolism based on positron emission tomography (PET). The authors of this paper developed an autoradiographic method for mapping CTT by simultaneously measuring local cerebral flow (LCBF) and local cerebral capillary blood volume (LCBV). Rats were given intravenous injections of mixtures of 1,850 MBq of Tc-99 m DTPA for (LCBV) and 1,850 kBq of C-14 iodoantipyrine (for LCBF). At the end of the infusions, the rats were killed and their brains were sectioned for autoradiography. To sets of autoradiograms were produced, each predominantly representing exposure from one of the tracers

1990-11-25

236

Real-time movie image enhancement in NMR  

International Nuclear Information System (INIS)

Clinical NMR motion picture (movie) images can now be produced routinely in real-time by ultra-high-speed echo-planar imaging (EPI). The single-shot image quality depends on both pixel resolution and signal-to-noise ratio (S/N), both factors being intertradeable. If image S/N is sacrificed rather than resolution, it is shown that S/N may be greatly enhanced subsequently without vitiating spatial resolution or foregoing real motional effects when the object motion is periodic. This is achieved by a Fourier filtering process. Experimental results are presented which demonstrate the technique for a normal functioning heart. (author)

1986-01-01

237

Imager Evaluation of Diabetic Retinopathy at the Time of Imaging in a Telemedicine Program  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Objective: To evaluate the ability of certified retinal imagers to identify presence versus absence of sight-threatening diabetic retinopathy (stDR) (moderate nonproliferative diabetic retinopathy or worse or diabetic macular edema) at the time of retinal imaging in a telemedicine program. Research Design and Methods: Diabetic patients in a primary care setting or specialty diabetes clinic received Joslin Vision Network protocol retinal imaging as part of their care. Trained nonphysician imag...

2012-01-01

238

Deep-Sea Bioluminescence Blooms after Dense Water Formation at the Ocean Surface  

Science.gov (United States)

The deep ocean is the largest and least known ecosystem on Earth. It hosts numerous pelagic organisms, most of which are able to emit light. Here we present a unique data set consisting of a 2.5-year long record of light emission by deep-sea pelagic organisms, measured from December 2007 to June 2010 at the ANTARES underwater neutrino telescope in the deep NW Mediterranean Sea, jointly with synchronous hydrological records. This is the longest continuous time-series of deep-sea bioluminescence ever recorded. Our record reveals several weeks long, seasonal bioluminescence blooms with light intensity up to two orders of magnitude higher than background values, which correlate to changes in the properties of deep waters. Such changes are triggered by the winter cooling and evaporation experienced by the upper ocean layer in the Gulf of Lion that leads to the formation and subsequent sinking of dense water through a process known as “open-sea convection”. It episodically renews the deep water of the study area and conveys fresh organic matter that fuels the deep ecosystems. Luminous bacteria most likely are the main contributors to the observed deep-sea bioluminescence blooms. Our observations demonstrate a consistent and rapid connection between deep open-sea convection and bathypelagic biological activity, as expressed by bioluminescence. In a setting where dense water formation events are likely to decline under global warming scenarios enhancing ocean stratification, in situ observatories become essential as environmental sentinels for the monitoring and understanding of deep-sea ecosystem shifts.

Tamburini, Christian; Canals, Miquel; Durrieu de Madron, Xavier; Houpert, Loic; Lefevre, Dominique; Martini, Severine; D'Ortenzio, Fabrizio; Robert, Anne; Testor, Pierre; Aguilar, Juan Antonio; Samarai, Imen Al; Albert, Arnaud; Andre, Michel; Anghinolfi, Marco; Anton, Gisela; Anvar, Shebli; Ardid, Miguel; Jesus, Ana Carolina Assis; Astraatmadja, Tri L.; Aubert, Jean-Jacques; Baret, Bruny; Basa, Stephane; Bertin, Vincent; Biagi, Simone; Bigi, Armando; Bigongiari, Ciro; Bogazzi, Claudio; Bou-Cabo, Manuel; Bouhou, Boutayeb; Bouwhuis, Mieke C.; Brunner, Jurgen; Busto, Jose; Camarena, Francisco; Capone, Antonio; Carloganu, Christina; Carminati, Giada; Carr, John; Cecchini, Stefano; Charif, Ziad; Charvis, Philippe; Chiarusi, Tommaso; Circella, Marco; Coniglione, Rosa; Costantini, Heide; Coyle, Paschal; Curtil, Christian; Decowski, Patrick; Dekeyser, Ivan; Deschamps, Anne; Donzaud, Corinne; Dornic, Damien; Dorosti, Hasankiadeh Q.; Drouhin, Doriane; Eberl, Thomas; Emanuele, Umberto; Ernenwein, Jean-Pierre; Escoffier, Stephanie; Fermani, Paolo; Ferri, Marcelino; Flaminio, Vincenzo; Folger, Florian; Fritsch, Ulf; Fuda, Jean-Luc; Galata, Salvatore; Gay, Pascal; Giacomelli, Giorgio; Giordano, Valentina; Gomez-Gonzalez, Juan-Pablo; Graf, Kay; Guillard, Goulven; Halladjian, Garadeb; Hallewell, Gregory; van Haren, Hans; Hartman, Joris; Heijboer, Aart J.; Hello, Yann; Hernandez-Rey, Juan Jose; Herold, Bjoern; Hossl, Jurgen; Hsu, Ching-Cheng; de Jong, Marteen; Kadler, Matthias; Kalekin, Oleg; Kappes, Alexander; Katz, Uli; Kavatsyuk, Oksana; Kooijman, Paul; Kopper, Claudio; Kouchner, Antoine; Kreykenbohm, Ingo; Kulikovskiy, Vladimir; Lahmann, Robert; Lamare, Patrick; Larosa, Giuseppina; Lattuada, Dario; Lim, Gordon; Presti, Domenico Lo; Loehner, Herbert; Loucatos, Sotiris; Mangano, Salvatore; Marcelin, Michel; Margiotta, Annarita; Martinez-Mora, Juan Antonio; Meli, Athina; Montaruli, Teresa; Motz, Holger; Neff, Max; Nezri, Emma nuel; Palioselitis, Dimitris; Pavalas, Gabriela E.; Payet, Kevin; Payre, Patrice; Petrovic, Jelena; Piattelli, Paolo; Picot-Clemente, Nicolas; Popa, Vlad; Pradier, Thierry; Presani, Eleonora; Racca, Chantal; Reed, Corey; Riccobene, Giorgio; Richardt, Carsten; Richter, Roland; Riviere, Colas; Roensch, Kathrin; Rostovtsev, Andrei; Ruiz-Rivas, Joaquin; Rujoiu, Marius; Russo, Valerio G.; Salesa, Francisco; Sanchez-Losa, Augustin; Sapienza, Piera; Schock, Friederike; Schuller, Jean-Pierre; Schussler, Fabian; Shanidze, Rezo; Simeone, Francesco; Spies, Andreas; Spurio, Maurizio; Steijger, Jos J. M.; Stolarczyk, Thierry; Taiuti, Mauro G. F.; Toscano, Simona; Vallage, Bertrand; Van Elewyck, Veronique; Vannoni, Giulia; Vecchi, Manuela; Vernin, Pascal; Wijnker, Guus; Wilms, Jorn; de Wolf, Els; Yepes, Harold; Zaborov, Dmitry; De Dios Zornoza, Juan; Zuniga, Juan

2013-01-01

239

Kinetics of bacterial bioluminescence and the fluorescent transient.  

Science.gov (United States)

The addition of FMNH(2), to Vibrio harveyi luciferase at 2°C in the presence of tetradecanal results in the formation of a highly fluorescent transient species with a spectral distribution indistinguishable from that of the bioluminescence. The bioluminescence reaches maximum intensity in 1.5 s and decays in a complex manner with exponential components of 10(-1) s(-1) , 7 x 10(-3)S(-1). and 7 x10(4)s(-1). The fluorescent transient rises exponentially at 7 x 10(-2)s(-1) and decays at 3 x 10 (4)s(-1) . The slowest bioluminescence component. comprising the bulk of the bioluminescence. decays at twice the rate of the fluorescent transient under all variations of reaction conditions: concentration of reactants.temperature 2 - 20°C. and aldehyde chain length - decana1, dodecanal and tetradecanal. The activation energy for both the slowest bioluminescence decay and the transient fluorescence decay is 80 kJ-mol(-1). An energy transfer scheme is proposed to explain the results where two distinct chemically energized species utilize the fluorescent transient as emitter for the slower bioluminescences, and for the faster process a fluorophore present in the protein preparation. Kinetic observations suggest that typical preparations of V. harveyi luciferase comprise 15% active protein. PMID:23479819

Matheso, I B; Lee, J

1983-08-01

240

Salience detection in time-evolving image sequences  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This paper presents a new approach to detect salient regions in an image. A number of units is used to respond to the most salient regions, adapting their response to the size and range of colors present in each region. The system can be directly used on a sequence of images, continuously adapting its output with time. A quality estimation of each unit allows selecting the most relevant regions present in the image at any time. The output of this process can be used as a visual attention mech...

Celaya Llover, Enric; Jime?nez Schlegl, Pablo

2003-01-01

 
 
 
 
241

Time-of-flight flow imaging using NMR remote detection  

Energy Technology Data Exchange (ETDEWEB)

A time-of-flight imaging technique is introduced to visualize fluid flow and dispersion through porous media using NMR. As the fluid flows through a sample, the nuclear spin magnetization is modulated by RF pulses and magnetic field gradients to encode the spatial coordinates of the fluid. When the fluid leaves the sample, its magnetization is recorded by a second RF coil. This scheme not only facilitates a time-dependent imaging of fluid flow, it also allows a separate optimization of encoding and detection subsystems to enhance overall sensitivity. The technique is demonstrated by imaging gas flow through a porous rock.

Granwehr, Josef; Harel, Elad; Han, Song-I; Garcia, Sandra; Pines,Alex; Sen, Pabitra N.; Song, Yi-Qiao

2005-05-05

242

Predictive modelling of cardiac real-time 2D images  

DEFF Research Database (Denmark)

Prediction of the respiratory and beating motion of the heart has several useful applications in cardiovascular MRI. Despite its usefulness, there is currently no available technique that establishes a predictive model of both cardiac and respiratory 3D motion. The main bottleneck is the inherent slow nature of MRI that prevents obtaining real-time 3D images of the heart with both high spatial and temporal resolution. In this work, we present a novel technique that predicts 2D real-time images for any cardiac and respiratory motion state. By combining several 2D acquisitions, the method predicts 3D images of the heart, including through-plane motion.

Fuglø, Dan; B.W. Larsson, Henrik

2009-01-01

243

Time-resolved fast-neutron imaging with a pulse-counting image intensifier  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A new imaging method that combines high-efficiency fast-neutron detection with sub-ns time resolution is presented. This is achieved by exploiting the high neutron detection efficiency of a thick scintillator and the fast timing capability and flexibility of light-pulse detection with a dedicated image intensifier. The neutron converter is a plastic scintillator slab or, alternatively, a scintillating fibre screen. The scintillator is optically coupled to a pulse counting image intensifier wh...

Dangendorf, Volker

2007-01-01

244

Focus-drift correction in time-lapse confocal imaging.  

Science.gov (United States)

In long-term time-lapse imaging of living cells, where recording takes several minutes or longer, a drift of focus may be significant. Focus-drift is due to the slippage in the microscope focus mechanism and/or the thermal gradients in the microscope. Software and hardware solutions may be introduced to correct for the focus-drift. Some autofocus techniques measure position of the specimen by sensing the light or sound reflected from a well-defined surface, such as the microscope slide. An autofocusing approach, where a focus measure is computed for images acquired at different objective positions is less appropriate in confocal microscopy, since more than one section is in focus. To correct for the focal-drift in long-term time-lapse confocal imaging, we acquired an image stack of the specimen periodically. The software calculated Pearson's correlation coefficient between each image in the z-stack and the reference image in the stack, which was selected at the beginning of the experiment. The maximal correlation coefficient of pixel intensities was taken to identify the image, which corresponded to the focal plane of the reference image. To test our approach, we used confocal images of living rat lactotroph cells, which discharged preloaded green fluorescent probe from a single secretory granule. Simultaneously, an extracellularly applied FM 4-64 red fluorescent probe loaded the discharging vesicle and the plasma membrane. We show that our approach is appropriate to correct for focal-drift in long term time-lapse imaging and analysis of living cells. PMID:16154944

Kreft, Marko; Stenovec, Matjaz; Zorec, Robert

2005-06-01

245

[A technology of real-time image compression for convex grating imaging spectrometer].  

Science.gov (United States)

The huge amount of convex grating imaging spectrometer image data brings much pressure to data transmission and storage, so the image must be compressed in real time. Firstly, the image characteristics were analyzed according to the imaging principle, and the compression approach to removing spatial correlation and spectral correlation was achieved; Secondly, the compression algorithms were analyzed and the 3-D compression scheme of one-order linear compression in spectral dimension and JPEG2000 compression in spatial dimension was proposed. Finally, a real-time compression system based on FPGA and ADV212 was designed, in which FPGA was used for logic control and implementation of prediction algorithm, and ADV212 was used for JPEG2000 compression. The analysis result shows that the system has the ability of lossless and lossy compression, enabling real-time image compression. PMID:22715801

Liu, Yang-chuan; Bayanheshig; Cui, Ji-cheng; Tang, Yu-guo

2012-04-01

246

Effect of exposure time and image resolution on fractal dimension  

International Nuclear Information System (INIS)

To evaluate the effect of exposure time and image resolution on fractal dimension calculations for determining the optimal range of these two variances. Thirty-one radiographs of the mandibular angle area of sixteen human dry mandibles were taken at different exposure times (0.01, 0.08, 0.16, 0.25, 0.40, 0.64, and 0.80 s). Each radiograph was digitized at 1200 dpi, 8 bit, 256 gray level using a film scanner. We selected an Region of Interest (ROI) that corresponded to the same region as in each radiograph, but the resolution of ROI was degraded to 1000, 800, 600, 500, 400, 300, 200, and 100 dpi. The fractal dimension was calculated by using the tile-counting method for each image, and the calculated values were then compared statistically. As the exposure time and the image resolution increased, the mean value of the fractal dimension decreased, except the case where exposure time was set at 0.01 seconds (alpha = 0.05). The exposure time and image resolution affected the fractal dimension by interaction (p<0.001). When the exposure time was set to either 0.64 seconds or 0.80 seconds, the resulting fractal dimensions were lower, irrespective of image resolution, than at shorter exposure times (alpha = 0.05). The optimal range for exposure time and resolution was determined to be 0.08-0.40 seconds and from 400-1000 dpi, respectively. Adequate exposure time and image resolution is essential for acquiring the fractal dimension using tile-counting method for evaluation of the mandible.

2002-06-01

247

Time-reverse imaging for detection of landmines  

Science.gov (United States)

Time Reversal is based on the fact that most physical laws of nature are invariant for time reversal, i.e., when time t is replaced by -t, most physical laws remain unchanged. Physically this means that by time reversing, a particle will retrace its original path or trajectory. Based on this fact, systems were built which receive reflections or scattering from targets. If this reflected data is recorded, time reversed and launched into the medium again, it will focus back on the targets. This is the basis for experimental time reversal. Time reverse imaging is somewhat different in the sense that scattering from targets are recorded on the sensors, but then back propagated numerically. Narrow-band or single frequency MUSIC based time-reverse imaging algorithms have been proposed in literature for point-like targets. When this algorithm is applied to scattering from an extended target, such as a landmine, the image has good cross-range resolution, but rather poor range resolution. We propose the use of 2-D MUSIC-based algorithm to improve the near-field range resolution, which can then be used in conjunction with single frequency MUSIC to produce a final high-resolution image. A FDTD elastic-wave simulation is used to verify the results using mines and mine-like targets embedded in a heterogenous soil.

Alam, Mubashir; McClellan, James H.; Norville, Pelham D.; Scott, Waymond R., Jr.

2004-09-01

248

Parallelisable method for real-time aerial images matching  

Science.gov (United States)

This paper proposes an algorithm for approximate affine plane matching of aerial images. The concept of a projective transform associated with partial Hausdorff distance is used in order to reduce the computational burden, and meet the real-time constraints. Potential algorithm parallelization for the SPMD computers is proposed as well. The running time evaluation evaluation of a sequential algorithm is given.

Pissaloux, Edwige; Bonnin, Patrick J.; Durbin, F.; Bezencenet, Gilles

1995-08-01

249

Mathematical methods in time series analysis and digital image processing  

CERN Document Server

The aim of this volume is to bring together research directions in theoretical signal and imaging processing developed rather independently in electrical engineering, theoretical physics, mathematics and the computer sciences. In particular, mathematically justified algorithms and methods, the mathematical analysis of these algorithms, and methods as well as the investigation of connections between methods from time series analysis and image processing are reviewed. An interdisciplinary comparison of these methods, drawing upon common sets of test problems from medicine and geophysical/enviromental sciences, is also addressed. This volume coherently summarizes work carried out in the field of theoretical signal and image processing. It focuses on non-linear and non-parametric models for time series as well as on adaptive methods in image processing.

Kurths, J; Maass, P; Timmer, J

2008-01-01

250

Time-Frequency and Time-Scale-Based Fragile Watermarking Methods for Image Authentication  

Directory of Open Access Journals (Sweden)

Full Text Available Two fragile image watermarking methods are proposed for image authentication. The first method is based on time-frequency analysis and the second one is based on time-scale analysis. For the first method, the watermark is chosen as an arbitrary nonstationary signal with a particular signature in the time-frequency plane. Experimental results show that this technique is very sensitive to many attacks such as cropping, scaling, translation, JPEG compression, and rotation, making it very effective in image authentication. For the second method, based on a wavelet-domain multiresolution analysis, quantization index modulation (QIM embedding scheme and arbitrary frequency-modulated (FM chirp watermarks are used in the implementation. In this blind technique, the original watermark is needed neither for the content integrity verification of the original image nor for the content quality assessment of the distorted image.

Sattar Farook

2010-01-01

251

Coherent temporal imaging with analog time-bandwidth compression  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We introduce the concept of coherent temporal imaging and its combination with the anamorphic stretch transform. The new system can measure both temporal profile of fast waveforms as well as their spectrum in real time and at high-throughput. We show that the combination of coherent detection and warped time-frequency mapping also performs time-bandwidth compression. By reducing the temporal width without sacrificing spectral resolution, it addresses the Big Data problem in ...

Asghari, Mohammad H.; Jalali, Bahram

2013-01-01

252

Real-time particle image velocimetry based on FPGA technology  

International Nuclear Information System (INIS)

Particle image velocimetry (PIV), based on laser sheet, is a method for image processing and calculation of distributed velocity fields.It is well established as a fluid dynamics measurement tool, being applied to liquid, gases and multiphase flows.Images of particles are processed by means of computationally demanding algorithms, what makes its real-time implementation difficult.The most probable displacements are found applying two dimensional cross-correlation function. In this work, we detail how it is possible to achieve real-time visualization of PIV method by designing an adaptive embedded architecture based on FPGA technology.We show first results of a physical field of velocity calculated by this platform system in a real-time approach.

2008-01-01

253

Real-time optical motion correction for diffusion tensor imaging.  

Science.gov (United States)

Head motion is a fundamental problem in brain MRI. The problem is further compounded in diffusion tensor imaging because of long acquisition times, and the sensitivity of the tensor computation to even small misregistration. To combat motion artifacts in diffusion tensor imaging, a novel real-time prospective motion correction method was introduced using an in-bore monovision system. The system consists of a camera mounted on the head coil and a self-encoded checkerboard marker that is attached to the patient's forehead. Our experiments showed that optical prospective motion correction is more effective at removing motion artifacts compared to image-based retrospective motion correction. Statistical analysis revealed a significant improvement in similarity between diffusion data acquired at different time points when prospective correction was used compared to retrospective correction (P<0.001). PMID:21432898

Aksoy, Murat; Forman, Christoph; Straka, Matus; Skare, Stefan; Holdsworth, Samantha; Hornegger, Joachim; Bammer, Roland

2011-08-01

254

Time-resolved PHERMEX image restorations constrained with an additional multiply-exposed image  

International Nuclear Information System (INIS)

There are a number of possible industrial and scientific applications of nanosecond cineradiographs. Although the technology exists to produce closely spaced pulses of x rays for this application, the quality of the time-resolved radiographs is severely limited. The limitations arise from the necessity of using a fluorescent screen to convert the transmitted x rays to light and then using electro-optical imaging systems to gate and to record the images with conventional high-speed cameras. It has been proposed that, in addition to the time-resolved images, a conventional multiply exposed radiograph be obtained. This report uses both PHERMEX and conventional photographic simulations to demonstrate that the additional information supplied by the multiply exposed radiograph can be used to improve the quality of digital image restorations of the time-resolved pictures over what could be achieved with the degraded images alone

1978-01-01

255

Real-Time Ellipsometry-Based Transmission Ultrasound Imaging  

Energy Technology Data Exchange (ETDEWEB)

Ultrasonic imaging is a valuable tool for non-destructive evaluation and medical diagnosis. Reflection mode is exclusively used for medical imaging, and is most frequently used for nondestructive evaluation (NDE) because of the relative speed of acquisition. Reflection mode imaging is qualitative, yielding little information about material properties, and usually only about material interfaces. Transmission imaging can be used in 3D reconstructions to yield quantitative information: sound speed and attenuation. Unfortunately, traditional scanning methods of acquiring transmission data are very slow, requiring on the order of 20 minutes per image. The sensing of acoustic pressure fields as optical images can significantly speed data acquisition. An entire 2D acoustic pressure field can be acquired in under a second. The speed of data acquisition for a 2D view makes it feasible to obtain multiple views of an object. With multiple views, 3D reconstruction becomes possible. A fast, compact (no big magnets or accelerators), inexpensive, 3D imaging technology that uses no ionizing radiation could be a boon to the NDE and medical communities. 2D transmission images could be examined in real time to give the ultrasonic equivalent of a fluoroscope, or accumulated in such a way as to acquire phase and amplitude data over multiple views for 3D reconstruction (for breast cancer imaging, for example). Composite panels produced for the aircraft and automobile industries could be inspected in near real time, and inspection of attenuating materials such as ceramics and high explosives would be possible. There are currently three optical-readout imaging transmission ultrasound technologies available. One is based on frustrated total internal reflection (FTIR) [1,2], one on Fabry-Perot interferometry [3], and another on critical angle modulation [4]. Each of these techniques has its problems. The FTIR based system cannot currently be scaled to large aperture sizes, the Fabry-Perot system has never been fully implemented for area imaging, and the critical angle modulation system is not sensitive enough for medical imaging. We proposed an entirely new way of using acoustic pressure to modulate a light beam. This new technology should be sensitive enough to be useful for medical imaging and have a large enough aperture to speed acquisition by orders of magnitude over point sampling. Unfortunately, we were unable to bring this technology to fruition.

Kallman, J S; Poco, J F; Ashby, A E

2007-02-14

256

Relaxation time T_1, T_2 and proton density images in NMR imaging  

International Nuclear Information System (INIS)

Pure T_1, T_2 and proton density (?) images can be computed from three or more different NMR images. Computed images can be useful for several reasons: a) they are objective, since they are independent of pulse sequence and scan parameters. b) arbitrary composite images can be synthesized from computed images. c) biochemical information can be obtained from relaxation times, so quantitative diagnosis is possible using T_1 and T_2 images. For these reasons, several methods of producing computed images have been tried. However, with these methods, there are several practical problems such as large systematic error and long total scan time. This paper describes how several sets of NMR pulse sequences and scan parameters were investigated, keeping total scan time constant, to find which of them gave computed images with best resolution and minimum systematic error for a given scan time. Pulse sequences and scan parameters were optimized to yield minimum variance of computed images, using the law of error propagation, for a given range of T_1, T_2 and ?. We found that theoretically the combination Inversion Recovery 3 Spin Echo and Saturation Recovery 4 Spin Echo pulse sequence gave the best compromise between scan time and resolution. The effect of slice profile and errors in RF pulses - causes of systematic error - were analyzed in order to find ways to remove or reduce them. Using this method computed T_1, T_2 and ? images were obtained for the human head and for various phantoms. Computed values agreed closely with values measured using analytical methods. We conclude from these results that the combination Inversion Recovery 3 Spin Echo and Saturation Recovery 4 Spin Echo pulse sequence gives the best compromise between scan time, resolution and error. (author)

1987-01-01

257

Bioluminescent assays for high-throughput screening.  

Science.gov (United States)

In the development of high throughput screening (HTS) as a central paradigm of drug discovery, fluorescence has generally been adopted as the favored methodology. Nevertheless, luminescence has maintained a prominent position among certain assay formats, most notably genetic reporters. Recently, there has been growing partiality for luminescent assays across a wider range of applications due to their sensitivity, broad linearity, and robustness to library compounds and complex biological samples. This trend has been fostered by the development of several new assay designs for diverse targets such as kinases, cytochrome p450s, proteases, apoptosis, and cytotoxicity. This review addresses recent progress made in the use of bioluminescent assays for HTS, highlighting new detection capabilities brought about by engineering luciferase genes, enzymes, and substrates. In genetic reporter applications, modifications to the luciferase genes have improved assay sensitivity by substantially increasing expression efficiency and enhanced response dynamics by reducing expression lifetime. The performance of assays based on detection of ATP and luciferin has been enhanced by modifications to the luciferase enzyme that increase its chemical and physical stability. Detection of ATP allows rapid analysis of cell metabolism and enzymatic processes coupled to ATP metabolism. Because luciferins are not naturally associated with mammalian physiology, assays for luciferin detection utilize synthetic derivatives designed to yield luminescence only when coupled with specific target enzymes. Finally, new methods for modulating the specific activity of luciferases are leading to the development of intracellular biosensors for dynamic detection of physiological processes. PMID:17355205

Fan, Frank; Wood, Keith V

2007-02-01

258

Single photon imaging and timing array sensor apparatus and method  

Energy Technology Data Exchange (ETDEWEB)

An apparatus and method are disclosed for generating a three-dimension image of an object or target. The apparatus is comprised of a photon source for emitting a photon at a target. The emitted photons are received by a photon receiver for receiving the photon when reflected from the target. The photon receiver determines a reflection time of the photon and further determines an arrival position of the photon on the photon receiver. An analyzer is communicatively coupled to the photon receiver, wherein the analyzer generates a three-dimensional image of the object based upon the reflection time and the arrival position.

Smith, R. Clayton (Boulder, CO)

2003-06-24

259

Magneto-optical system for high speed real time imaging  

Science.gov (United States)

A new magneto-optical system has been developed to expand the range of high speed real time magneto-optical imaging. A special source for the external magnetic field has also been designed, using a pump solenoid to rapidly excite the field coil. Together with careful modifications of the cryostat, to reduce eddy currents, ramping rates reaching 3000 T/s have been achieved. Using a powerful laser as the light source, a custom designed optical assembly, and a high speed digital camera, real time imaging rates up to 30 000 frames per seconds have been demonstrated.

Baziljevich, M.; Barness, D.; Sinvani, M.; Perel, E.; Shaulov, A.; Yeshurun, Y.

2012-08-01

260

In vitro antagonism of bioluminescent fungi by Trichoderma harzianum.  

Science.gov (United States)

Two species of bioluminescent fungi, Panellus stypticus and Omphalotus olearius were placed in contact with three different strains of interfungal pathogenic Trichoderma harzianum. Subsequent light emission by the luminous fungi and advance of the interfungal pathogens were compared. Relative differences among the pathogens were reflected in their rate of mycelial advance, the total area over which they produced spores upon the host fungi, and decreases in host bioluminescence. After ten days differences in the total surface areas of spore production varied from 1 to 53 per cent. Differences in the reduction of bioluminescence of the same material ranged over 2 orders of magnitude. Final reduction in luminescence ranged over 6 orders of magnitude. A marked reduction in bioluminescence was observed to precede the advance of spore production. The greatest reduction in luminescence was correlated with the presence of T. harzianum hyphae. Two strains of T. harzianum, NRRL 1698 and ATCC 58674, were effective against both bioluminescent fungi within the study period while a third strain, NRRL 13019, was only effective against Omphalotus olearius. PMID:1922266

Bermudes, D; Boraas, M E; Nealson, K H

1991-07-01

 
 
 
 
261

Experimental ultrasound system for real-time synthetic imaging  

DEFF Research Database (Denmark)

Digital signal processing is being employed more and more in modern ultrasound scanners. This has made it possible to do dynamic receive focusing for each sample and implement other advanced imaging methods. The processing, however, has to be very fast and cost-effective at the same time. Dedicated chips are used in order to do real time processing. This often makes it difficult to implement radically different imaging strategies on one platform and makes the scanners less accessible for research purposes. Here flexibility is the prime concern, and the storage of data from all transducer elements over 5 to 10 seconds is needed to perform clinical evaluation of synthetic and 3D imaging. This paper describes a real-time system specifically designed for research purposes. The purpose of the system is to make it possible to acquire multi-channel data in real-time from clinical multi-element ultrasound transducers, and to enable real-time or near realtime processing of the acquired data. The system will be capable of performing the processing for the currently available imaging methods, and will make it possible to perform initial trials in a clinical environment with new imaging modalities for synthetic aperture imaging, 2D and 3D B-mode and velocity imaging. The system can be used with 128 element transducers and can excite 128 channels and receive and sample data from 64 channels simultaneously at 40 MHz with 12 bits precision. Data can be processed in real time using the system's 80 signal processing units or it can be stored directly in RAM. The system has 24 GBytes RAM and can thus store 8 seconds of multi-channel data. It is fully software programmable and its signal processing units can also be reconfigured under software control. The control of the system is done over an Ethernet using C and Matlab. Programs for doing e.g. B-mode imaging can directly be written in Matlab and executed on the system over the net from any workstation running Matlab. The overall system concept is presented and an example ofa 20 lines script for doing phased array B-mode imaging is presented.

Jensen, Jørgen Arendt; Pedersen, Henrik Møller

1999-01-01

262

Robust real-time instrument tracking in ultrasound images  

Science.gov (United States)

Minimally invasive surgery in combination with ultrasound (US) imaging imposes high demands on the surgeon's hand-eye-coordination capabilities. A possible solution to reduce these requirements is minimally invasive robotic surgery in which the instrument is guided by visual servoing towards the goal defined by the surgeon in the US image. This approach requires robust tracking of the instrument in the US image sequences which is known to be difficult due to poor image quality. This paper presents algorithms and results of first tracking experiments. Adaptive thresholding based on Otsu's method allows to cope with large intensity variations of the instrument echo. Median filtering of the binary image and subsequently applied morphological operations suppress noise and echo artefacts. A fast run length code based labelling algorithm allows for real-time labelling of the regions. A heuristic exploiting region size and region velocity helps to overcome ambiguities. The overall computation time is less than 20 ms per frame on a standard PC. The tracking algorithm requires no information about texture and shape which are known to be very unreliable in US image sequences. Experimental results for two different instrument materials (polyvinyl chloride and polyurethane) are given, showing the performance of the proposed approach. Choosing the appropriate material, trajectories are smooth and only few outliers occur.

Ortmaier, Tobias; Vitrani, Marie-Aude; Morel, Guillaume; Pinault, Samuel

2005-04-01

263

Real-time image denoising algorithm in teleradiology systems  

Science.gov (United States)

Denoising of medical images in wavelet domain has potential application in transmission technologies such as teleradiology. This technique becomes all the more attractive when we consider the progressive transmission in a teleradiology system. The transmitted images are corrupted mainly due to noisy channels. In this paper, we present a new real time image denoising scheme based on limited restoration of bit-planes of wavelet coefficients. The proposed scheme exploits the fundamental property of wavelet transform - its ability to analyze the image at different resolution levels and the edge information associated with each sub-band. The desired bit-rate control is achieved by applying the restoration on a limited number of bit-planes subject to the optimal smoothing. The proposed method adapts itself to the preference of the medical expert; a single parameter can be used to balance the preservation of (expert-dependent) relevant details against the degree of noise reduction. The proposed scheme relies on the fact that noise commonly manifests itself as a fine-grained structure in image and wavelet transform allows the restoration strategy to adapt itself according to directional features of edges. The proposed approach shows promising results when compared with unrestored case, in context of error reduction. It also has capability to adapt to situations where noise level in the image varies and with the changing requirements of medical-experts. The applicability of the proposed approach has implications in restoration of medical images in teleradiology systems. The proposed scheme is computationally efficient.

Gupta, Pradeep Kumar; Kanhirodan, Rajan

2006-02-01

264

An instant scan technique for real-time MR imaging  

International Nuclear Information System (INIS)

A recently developed instant scan technique (Advanced NMR Systems, Inc., Woburn, Mass.) yields a complete, motion artifact-free MR image in just 1/25 second. The MR imaging system uses a high-field (2.0-T) magnet and other technical innovations to achieve both high-resolution and high signal-to-noise ratio images of adult humans. The method presented samples of a rapidly modulated spin-echo and thus maintains the conventional abilities to manipulate T1 and T2 contrast. The technique can generate volumetric movie data of the heart in a single suspension of the breath and offers the potential for continuous, real-time MR imaging

1986-12-05

265

IMPLEMENTATION OF IMAGE PROCESSING IN REAL TIME CAR PARKING SYSTEM  

Directory of Open Access Journals (Sweden)

Full Text Available Car parking lots are an important object class in many traffic and civilian applications. With the problems of increasing urban trafficcongestion and the ever increasing shortage of space, these car parking lots are needed to be well equipped with automatic parkingInformation and Guidance systems. Goals of intelligent parking lot management include counting the number of parked cars, and identifyingthe available location. This work proposes a new system for providing parking information and guidance using image processing. The proposed system includes counting the number of parked vehicles, and dentifying the stalls available. The system detects cars through images instead of using electronic sensors embedded on the floor. A camera is installed at the entry point of the parking lot. It capturesimage sequences. The image sequences are then analyzed using digital image processing for vehicle detection and according to the status ofvehicle occupancy inside, real time guidance and information is provided to the incoming driver.

SAYANTI BANERJEE,

2011-02-01

266

Application of Wavelet Transform for Image Denoising of Spatially and Time Variable Astronomical Imaging Systems  

Directory of Open Access Journals (Sweden)

Full Text Available We report on our efforts to formulate algorithms for image signal processing with the spatially and time variant Point-Spread Function (PSF and inhomogeneous noise of real imaging systems. In this paper we focus on application of the wavelet transform for denoising of the astronomical images with complicated conditions. They influence above all accuracy of the measurements and the new source detection ability. Our aim is to test the usefulness ofWavelet transform (as the standard image processing technique for astronomical purposes.

M. Blažek

2011-01-01

267

Real-time artifact-free image upscaling.  

Science.gov (United States)

The problem of creating artifact-free upscaled images appearing sharp and natural to the human observer is probably more interesting and less trivial than it may appear. The solution to the problem, often referred to also as "single-image super-resolution," is related both to the statistical relationship between low-resolution and high-resolution image sampling and to the human perception of image quality. In many practical applications, simple linear or cubic interpolation algorithms are applied for this task, but the results obtained are not really satisfactory, being affected by relevant artifacts like blurring and jaggies. Several methods have been proposed to obtain better results, involving simple heuristics, edge modeling, or statistical learning. The most powerful ones, however, present a high computational complexity and are not suitable for real-time applications, while fast methods, even if edge adaptive, are not able to provide artifacts-free images. In this paper, we describe a new upscaling method (iterative curvature-based interpolation) based on a two-step grid filling and an iterative correction of the interpolated pixels obtained by minimizing an objective function depending on the second-order directional derivatives of the image intensity. We show that the constraints used to derive the function are related with those applied in another well-known interpolation method, providing good results but computationally heavy (i.e., new edge-directed interpolation (NEDI). The high quality of the images enlarged with the new method is demonstrated with objective and subjective tests, while the computation time is reduced of one to two orders of magnitude with respect to NEDI so that we were able, using a graphics processing unit implementation based on the nVidia Compute Unified Device Architecture technology, to obtain real-time performances. PMID:21926002

Giachetti, Andrea; Asuni, Nicola

2011-10-01

268

Real-time co-registered imaging using a novel hand-held optical imager  

Science.gov (United States)

Several hand-held based optical imaging devices have been developed towards breast imaging, which are portable, patient-comfortable, and use non-ionizing radiation. The devices developed to date are limited in that they have flat probe faces and are incapable of real-time coregistration (as needed for 3-D tomographic imaging). A hand-held based optical imager has been developed in our lab, which has unique features of (i) simultaneous over sequential source illumination, which enables rapid data acquisition, (ii) a flexible probe face, which enables it to contour to any tissue curvature, and (iii) self coregistration facilities towards 3-D tomographic imaging. Real-time coregistration is demonstrated using the imager via fluorescence-enhanced studies in the continuous-wave mode, performed on slab phantoms (filled with 1% Liposyn solution) and in vitro samples (chicken breast). Additionally, preliminary studies were conducted using curved phantoms. In all cases, a 0.45-cc target filled with 1 ?M Indocyanine green was used to represent a tumor. Real-time 2-D surface images of the phantom were obtained via multiple scans at different target depths. Preliminary surface imaging studies demonstrated that the summation of multiple scans distinctly differentiated the target from artifacts (up to 3 cm deep), which was not possible from individual scans.

Erickson, Sarah J.; Regalado, Steven; Ge, Jiajia; Godavarty, Anuradha

2009-02-01

269

Real-time Optical Motion Correction for Diffusion Tensor Imaging  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Head motion is a fundamental problem in brain MRI. The problem is further compounded in diffusion tensor imaging (DTI) because of long acquisition times, and the sensitivity of the tensor computation to even small misregistration. To combat motion artifacts in DTI, a novel real-time prospective motion correction method was introduced using an in-bore monovision system. The system consists of a camera mounted on the head coil and a self-encoded checkerboard marker that is attached to the patie...

Aksoy, Murat; Forman, Christoph; Straka, Matus; Skare, Stefan; Holdsworth, Samantha; Hornegger, Joachim; Bammer, Roland

2011-01-01

270

3D imaging using time-correlated single photon counting  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This project investigates a laser radar system. The system is based on the principles of time-correlated single photon counting, and by measuring the times-of-flight of reflected photons it can find range profiles and perform three-dimensional imaging of scenes. Because of the photon counting technique the resolution and precision that the system can achieve is very high compared to analog systems. These properties make the system interesting for many military applications. For example, the s...

Neimert-andersson, Thomas

2010-01-01

271

Time-resolved fast neutron imaging: simulation of detector performance  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have analyzed and compared the performance of two novel fast-neutron imaging methods with time-of-flight spectroscopy capability. Using MCNP and GEANT code simulations of neutron and charged-particle transport in the detectors, key parameters such as detection efficiency, the amount of energy deposited in the converter and the spatial resolution of both detector variants have been evaluated.

Vartsky, D.; Mor, I.; Goldberg, M. B.; Mardor, I.; Feldman, G.; Bar, D.; Shor, A.; Dangendorf, V.; Laczko, G.; Breskin, A.; Chechik, R.

2004-01-01

272

Time-resolved fast neutron imaging: simulation of detector performance  

Energy Technology Data Exchange (ETDEWEB)

We have analyzed and compared the performance of two novel fast-neutron imaging methods with time-of-flight spectroscopy capability. Key parameters such as detection efficiency, the amount of energy deposited in the converter and the spatial resolution of both detector variants have been simulated by means of neutron and charged-particle transport codes.

Vartsky, D. [Soreq NRC, Yavne 81800 (Israel)]. E-mail: david@soreq.gov.il; Mor, I. [Soreq NRC, Yavne 81800 (Israel); Goldberg, M.B. [Soreq NRC, Yavne 81800 (Israel); Mardor, I. [Soreq NRC, Yavne 81800 (Israel); Feldman, G. [Soreq NRC, Yavne 81800 (Israel); Bar, D. [Soreq NRC, Yavne 81800 (Israel); Shor, A. [Soreq NRC, Yavne 81800 (Israel); Dangendorf, V. [Physikalisch-Technische Bundesanstalt, Braunschweig (Germany); Laczko, G. [Physikalisch-Technische Bundesanstalt, Braunschweig (Germany); Breskin, A. [Weizmann Institute of Science, Rehovot (Israel); Chechik, R. [Weizmann Institute of Science, Rehovot (Israel)

2005-04-21

273

Time-resolved fast neutron imaging: simulation of detector performance  

International Nuclear Information System (INIS)

We have analyzed and compared the performance of two novel fast-neutron imaging methods with time-of-flight spectroscopy capability. Key parameters such as detection efficiency, the amount of energy deposited in the converter and the spatial resolution of both detector variants have been simulated by means of neutron and charged-particle transport codes

2005-04-21

274

FPGA implementation of real-time digital image stabilization  

Science.gov (United States)

In order to overcome image shakes in the video of the camera mounted on a movable platform and to implement the image stabilization in real-time, a FPGA platform of real-time digital image stabilization (DIS) based on bit-plane matching was realized. Firstly, the local motion vectors were estimated by using Gray Code bit-plane matching. Then, the global motion vector was generated with a median filter method. Following that, a low pass filter was applied on the previous global motion vectors to get the current motion compensation vector. At last, the stabilized video was obtained by compensation the original one. A multi-bits concurrent matching method was used when estimating the local motion vectors using the bit-plane, which increased the parallelism of the FPGA and speeded up the matching velocity. The FPGA implementation of the function modules of video capture control, image buffering, motion estimation, motion filtering, and motion compensation were detailed out. The experiment results indicate that the proposed method can stabilize the 25 fps video with 720×576 pixels in real-time, and can be applied in real-time applications.

Li, Gang

2014-02-01

275

Toxin-antitoxin-stabilized reporter plasmids for biophotonic imaging of Group A streptococcus.  

Science.gov (United States)

Bioluminescence is a rapid and cost-efficient optical imaging technology that allows the detection of bacteria in real-time during disease development. Here, we report a novel strategy to generate a wide range of bioluminescent group A streptococcus (GAS) strains by using a toxin-antitoxin-stabilized plasmid. The bacterial luciferin-luciferase operon (lux) or the firefly luciferase gene (ffluc) was introduced into GAS via a stabilized plasmid. The FFluc reporter gave significantly stronger bioluminescent signals than the Lux reporter, and was generally more stable. Plasmid-based luciferase reporters could easily be introduced into a variety of GAS strains and the signals correlated linearly with viable cell counts. Co-expression of the streptococcal ?-?-? toxin-antitoxin operon provided segregational stability in the absence of antibiotics for at least 17 passages in vitro and up to 7 days in a mouse infection model. In addition, genome-integrated reporter constructs were also generated by site-specific recombination, but were found to be technically more challenging. The quick and efficient generation of various M-type GAS strains expressing plasmid-based luciferase reporters with comparable and quantifiable bioluminescence signals allows for comparative analysis of different GAS strains in vitro and in vivo. PMID:24061415

Loh, Jacelyn M S; Proft, Thomas

2013-11-01

276

Image domain moving target tracking with advanced image registration and time-differencing techniques  

Science.gov (United States)

Advanced image registration technique with sub-pixel accuracy has been developed and applied for TD (time-differencing) process [1]. The TD process can help to suppress heavy background clutter for improved moving target detection. After processing a CFAR (constant false alarm rate) thresholding detector on the time-differenced image frames, we have developed and applied an image domain moving target tracking (IDMTT) process for robust moving target tracking. The IDMTT process uses a unique location feature by mapping and associating the real moving targets in the previous time-differenced frame with the ghost moving targets in the current time-differenced frame. The accurate location mapping and associating information between time frames is provided by the registration process. Preliminary tests for the IDMTT process are promising. Robust moving target tracking can be achieved even under quite low signal-to-clutter noise-ratio (SCNR = 0.5).

Chen, Hai-Wen; Braunreiter, Denis

2009-05-01

277

A miniature real-time volumetric ultrasound imaging system  

Science.gov (United States)

Progress made in the development of a miniature real-time volumetric ultrasound imaging system is presented. This system is targeted for use in a 5-mm endoscopic channel and will provide real-time, 30-mm deep, volumetric images. It is being developed as a clinically useful device, to demonstrate a means of integrating the front-end electronics with the transducer array, and to demonstrate the advantages of the capacitive micromachined ultrasonic transducer (CMUT) technology for medical imaging. Presented here is the progress made towards the initial implementation of this system, which is based on a two-dimensional, 16x16 CMUT array. Each CMUT element is 250 um by 250 um and has a 5 MHz center frequency. The elements are connected to bond pads on the back side of the array with 400-um long through-wafer interconnects. The transducer array is flip-chip bonded to a custom-designed integrated circuit that comprises the front-end electronics. The result is that each transducer element is connected to a dedicated pulser and low-noise preamplifier. The pulser generates 25-V, 100-ns wide, unipolar pulses. The preamplifier has an approximate transimpedance gain of 500 kOhm and 3-dB bandwidth of 10 MHz. In the first implementation of the system, one element at a time can be selected for transmit and receive and thus synthetic aperture images can be generated. In future implementations, 16 channels will be active at a given time. These channels will connect to an FPGA-based data acquisition system for real-time image reconstruction.

Wygant, Ira O.; Yeh, David T.; Zhuang, Xuefeng; Nikoozadeh, Amin; Oralkan, Omer; Ergun, Arif S.; Karaman, Mustafa; Khuri-Yakub, Butrus T.

2005-04-01

278

Bubble masks for time-encoded imaging of fast neutrons.  

Energy Technology Data Exchange (ETDEWEB)

Time-encoded imaging is an approach to directional radiation detection that is being developed at SNL with a focus on fast neutron directional detection. In this technique, a time modulation of a detected neutron signal is induced-typically, a moving mask that attenuates neutrons with a time structure that depends on the source position. An important challenge in time-encoded imaging is to develop high-resolution two-dimensional imaging capabilities; building a mechanically moving high-resolution mask presents challenges both theoretical and technical. We have investigated an alternative to mechanical masks that replaces the solid mask with a liquid such as mineral oil. Instead of fixed blocks of solid material that move in pre-defined patterns, the oil is contained in tubing structures, and carefully introduced air gaps-bubbles-propagate through the tubing, generating moving patterns of oil mask elements and air apertures. Compared to current moving-mask techniques, the bubble mask is simple, since mechanical motion is replaced by gravity-driven bubble propagation; it is flexible, since arbitrary bubble patterns can be generated by a software-controlled valve actuator; and it is potentially high performance, since the tubing and bubble size can be tuned for high-resolution imaging requirements. We have built and tested various single-tube mask elements, and will present results on bubble introduction and propagation as a function of tubing size and cross-sectional shape; real-time bubble position tracking; neutron source imaging tests; and reconstruction techniques demonstrated on simple test data as well as a simulated full detector system.

Brubaker, Erik; Brennan, James S.; Marleau, Peter; Nowack, Aaron B.; Steele, John; Sweany, Melinda; Throckmorton, Daniel J.

2013-09-01

279

Real-time synthetic aperture imaging: opportunities and challenges  

DEFF Research Database (Denmark)

Synthetic aperture (SA) ultrasound imaging has not been introduced in commercial scanners mainly due to the computational cost associated with the hardware implementation of this imaging modality. SA imaging redefines the term beamformed line. Since the acquired information comes from all points in the region of interest it is possible to beamform the signals along a desired path, thus, improving the estimation of blood flow. The transmission of coded excitations makes it possible to achieve higher contrast and larger penetration depth compared to "conventional" scanners. This paper presents the development and implementation of the signal processing stages employed in SA imaging: compression of received data acquired using codes, and beamforming. The goal was to implement the system using commercially available field programmable gate arrays. The compression filter operates on frequency modulated pulses with duration of up to 50 mus sampled at 70 MHz. The beamformer can process data from 256 channels at a pulse repetition frequency of 5000 Hz and produces 192 lines of 1024 complex samples in real time. The lines are described by their origin, direction, length and distance between two samples in 3D. This parametric description makes it possible to quickly change the image geometry during scanning, thus enabling adaptive imaging and precise flow estimation. The paper addresses problems such as large bandwidth and computational load and gives the solutions that have been adopted for the implementation.

Nikolov, Svetoslav; Tomov, Borislav Gueorguiev

2006-01-01

280

Vegetation classification in eastern China using time series NDVI images  

Science.gov (United States)

The SPOT/VGT NDVI (S10) time series data of eastern China (1998-2005) are smoothed with two methods, the moving average and the Savitzky-Golay filter, after they are downloaded from the official website of VITO. Then the monthly maximal NDVI images (total 93 images) are extracted from 279 NDVI (S10) images and the Principal Component Analysis (PCA) is applied on the 93 images. There are 3 components that each explains more than 1% of the variance, in which the principal components 1, 2 and 3 explain respectively 93.25%, 2.77% and 1.21% of the variance in the original 93 maximum NDVI images. The principal component 1 is interpreted as the "climate" component, and principal components 2 and 3 are interpreted as the "growth season" and "non-growth season" components respectively. Principal components 1, 2 and 3 are composed to a 3-band color image which is classified into 7 classes (including 18 subclasses) by ISODATA. The overall accuracy of classification in five samples is 83.6%, and the kappa index is 0.82. Finally, the unique intra-annual NDVI curve of each vegetation class is displayed.

Han, Guifeng; Xu, Jianhua

2007-11-01

 
 
 
 
281

Diffuse Imaging: Creating Optical Images With Unfocused Time-Resolved Illumination and Sensing  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Conventional imaging uses steady-state illumination and light sensing with focusing optics; variations of the light field with time are not exploited. We develop a signal processing framework for estimating the reflectance of a Lambertian planar surface in a known position using omnidirectional, time-varying illumination and unfocused, time-resolved sensing in place of traditional optical elements such as lenses and mirrors. Our model associates time sampling of the intensity of light inciden...

Kirmani, Ahmed; Jeelani, Haris; Goyal, Vivek K.

2011-01-01

282

Time delay between images of the lensed quasar UM673  

CERN Multimedia

We study brightness variations in the double lensed quasar UM673 (Q0142-100) with the aim of measuring the time delay between its two images. In the paper we combine our previously published observational data of UM673 obtained during the 2003 - 2005 seasons at the Maidanak Observatory with archival and recently observed Maidanak and CTIO UM673 data. We analyze the V, R and I-band light curves of the A and B images of UM673, which cover ten observational seasons from August 2001 to November 2010. We also analyze the time evolution of the difference in magnitudes between images A and B of UM673 over more than ten years. We find that the quasar exhibits both short-term (with amplitude of \\sim 0.1 mag in the R band) and high-amplitude (\\sim 0.3 mag) long-term variability on timescales of about several months and several years, respectively. These brightness variations are used to constrain the time delay between the images of UM673. From cross-correlation analysis of the A and B quasar light curves and error ana...

Koptelova, E; Chiueh, T; Artamonov, B P; Oknyanskij, V L; Nuritdinov, S N; Burkhonov, O; Akhunov, T; Bruevich, V V; Ezhkova, O V; Gusev, A S; Sergeyev, A A; Ehgamberdiev, Sh A; Ibragimov, M A

2012-01-01

283

Time gate, optical layout, and wavelength effects on ballistic imaging.  

Science.gov (United States)

A method to distinguish a hidden object from a perturbing environment is to use an ultrashort femtosecond pulse of light and a time-resolved detection. To separate ballistic light containing information on a hidden object from multiscattered light coming from the surrounding environment that scrambles the signal, an optical Kerr gate can be used. It consists of a carbon disulfide (CS(2)) cell in which birefringence is optically induced. An imaging beam passes through the studied medium while a pump pulse is used to open the gate. The time-delayed scattered light is excluded from measurements by the gate, and the multiple-scattering scrambling effect is reduced. In previous works, the two beams had the same wavelength. We propose a new two-color experimental setup for ballistic imaging in which a second harmonic is generated and used for the image, while the fundamental is used for gate switching. This setup allows one to obtain better resolution by using a spectral filtering to eliminate noise from the pump pulse, instead of a spatial filtering. This new setup is suitable for use in ballistic imaging of dense sprays, multidiffusive, and large enough to show scattered light time delays greater than the gate duration (tau=1.3 ps). PMID:19721685

Idlahcen, Saďd; Méčs, Loďc; Rozé, Claude; Girasole, Thierry; Blaisot, Jean-Bernard

2009-09-01

284

Image reconstruction in time-of-flight positron emission tomography  

International Nuclear Information System (INIS)

Five algorithms for image reconstruction in time-of-flight assisted positron emission tomography have been studied. These algorithms include three approaches previously described in the literature and two new methods recently developed in our institute. Computer simulation studies have been performed to evaluate the relative merits of these various techniques. Performance indices such as computational efficiency, reconstructed image resolution, and signal-to-noise ratio have been investigated. Results from the analysis suggest that the two new methods may offer some potential advantages over other algorithms. (Auth.)

1986-01-01

285

Terahertz time-domain spectroscopy and imaging of artificial RNA  

DEFF Research Database (Denmark)

We use terahertz time-domain spectroscopy (THz-TDS) to measure the far-infrared dielectric function of two artificial RNA single strands, composed of polyadenylic acid (poly-A) and polycytidylic acid (poly-C). We find a significant difference in the absorption between the two types of RNA strands, and we show that we can use this difference to record images of spot arrays of the RNA strands. Under controlled conditions it is possible to use the THz image to distinguish between the two RNA strands. We discuss the requirements to sample preparation imposed by the lack of sharp spectral features in the absorption spectra.

Jepsen, Peter Uhd

2005-01-01

286

Real-time full field laser Doppler imaging  

Science.gov (United States)

We present a full field laser Doppler imaging instrument that enables real-time in vivo assessment of blood flow in dermal tissue and skin. The instrument monitors the blood perfusion in an area of about 50cm2 with 480 × 480 pixels per frame at a rate of 12-14 frames per second. Smaller frames can be monitored at much higher frame rates. We recorded the microcirculation in healthy skin before, during and after arterial occlusion. In initial clinical case studies, we imaged the microcirculation in burned skin and monitored the recovery of blood flow in a skin flap during reconstructive surgery indicating the high potential of LDI for clinical applications.

Leutenegger, Marcel; Harbi, Pascal; Thacher, Tyler; Raffoul, Wassim; Lasser, Theo

2012-06-01

287

Imaging ?-Galactosidase Activity In Vivo Using Sequential Reporter-Enzyme Luminescence  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Bioluminescence using the reporter enzyme firefly luciferase (Fluc) and the substrate luciferin enables noninvasive optical imaging of living animals with extremely high sensitivity. This type of analysis enables studies of gene expression, tumor growth, and cell migration over time in live animals that were previously not possible. However, a major limitation of this system is that Fluc activity is restricted to the intracellular environment, which precludes important applications of in vivo...

2009-01-01

288

Early Time Points Perfusion Imaging: Relative Time of Arrival, Maximum Derivatives and Fractional Derivatives  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Time of arrival (TOA) of a bolus of contrast agent to the tissue voxel is a reference time point critical for the Early Time Points Perfusion Imaging Method (ET) to make relative cerebral blood flow (rCBF) maps. Due to the low contrast to noise (CNR) condition at TOA, other useful reference time points known as relative time of arrival data points (rTOA) are investigated. Candidate rTOA's include the time to reach the maximum derivative, the maximum second derivative, and the maximum fraction...

2011-01-01

289

Real-time digital X-ray subtraction imaging  

International Nuclear Information System (INIS)

A diagnostic anatomical X-ray apparatus comprising a converter and a television camera for converting an X-ray image of a subject into a series of television fields of video signals is described in detail. A digital memory system stores and integrates the video signals over a time interval corresponding to a plurality of successive television fields. The integrated video signals are recovered from storage and fed to a digital or analogue subtractor, the resulting output being displayed on a television monitor. Thus the display represents on-going changes in the anatomical X-ray image. In a modification, successive groups of fields are stored and integrated in three memories, cyclically, and subtractions are performed between successive pieces of integrated signals to provide a display of successive alterations in the X-ray image. For investigations of the heart, the integrating interval should be of the order of one cardiac cycle. (author)

1979-01-01

290

Cellular Neural Network for Real Time Image Processing  

Science.gov (United States)

Since their introduction in 1988, Cellular Nonlinear Networks (CNNs) have found a key role as image processing instruments. Thanks to their structure they are able of processing individual pixels in a parallel way providing fast image processing capabilities that has been applied to a wide range of field among which nuclear fusion. In the last years, indeed, visible and infrared video cameras have become more and more important in tokamak fusion experiments for the twofold aim of understanding the physics and monitoring the safety of the operation. Examining the output of these cameras in real-time can provide significant information for plasma control and safety of the machines. The potentiality of CNNs can be exploited to this aim. To demonstrate the feasibility of the approach, CNN image processing has been applied to several tasks both at the Frascati Tokamak Upgrade (FTU) and the Joint European Torus (JET).

Vagliasindi, G.; Arena, P.; Fortuna, L.; Mazzitelli, G.; Murari, A.

2008-03-01

291

Cellular Neural Network for Real Time Image Processing  

International Nuclear Information System (INIS)

Since their introduction in 1988, Cellular Nonlinear Networks (CNNs) have found a key role as image processing instruments. Thanks to their structure they are able of processing individual pixels in a parallel way providing fast image processing capabilities that has been applied to a wide range of field among which nuclear fusion. In the last years, indeed, visible and infrared video cameras have become more and more important in tokamak fusion experiments for the twofold aim of understanding the physics and monitoring the safety of the operation. Examining the output of these cameras in real-time can provide significant information for plasma control and safety of the machines. The potentiality of CNNs can be exploited to this aim. To demonstrate the feasibility of the approach, CNN image processing has been applied to several tasks both at the Frascati Tokamak Upgrade (FTU) and the Joint European Torus (JET)

2008-03-12

292

Time-resolved tomographic images of a relativistic electron beam  

International Nuclear Information System (INIS)

We obtained a sequential series of time-resolved tomographic two-dimensional images of a 4.5-MeV, 6-kA, 30-ns electron beam. Three linear fiber-optic arrays of 30 or 60 fibers each were positioned around the beam axis at 00, 610, and 1170. The beam interacting with nitrogen at 20 Torr emitted light that was focused onto the fiber arrays and transmitted to a streak camera where the data were recorded on film. The film was digitized, and two-dimensional images were reconstructed using the maximum-entropy tomographic technique. These images were then combined to produce an ultra-high-speed movie of the electron-beam pulse

1984-08-19

293

Time-resolved tomographic images of a relativistic electron beam  

Energy Technology Data Exchange (ETDEWEB)

We obtained a sequential series of time-resolved tomographic two-dimensional images of a 4.5-MeV, 6-kA, 30-ns electron beam. Three linear fiber-optic arrays of 30 or 60 fibers each were positioned around the beam axis at 0/sup 0/, 61/sup 0/, and 117/sup 0/. The beam interacting with nitrogen at 20 Torr emitted light that was focused onto the fiber arrays and transmitted to a streak camera where the data were recorded on film. The film was digitized, and two-dimensional images were reconstructed using the maximum-entropy tomographic technique. These images were then combined to produce an ultra-high-speed movie of the electron-beam pulse.

Koehler, H.A.; Jacoby, B.A.; Nelson, M.

1984-07-01

294

Magnetic resonance imaging (MRI) and relaxation time mapping of concrete  

Science.gov (United States)

The use of Magnetic Resonance Imaging (MRI) of water in concrete is presented. This thesis will approach the problem of MR imaging of concrete by attempting to design new methods, suited to concrete materials, rather than attempting to force the material to suit the method. A number of techniques were developed, which allow the spatial observation of water in concrete in up to three dimensions, and permits the determination of space resolved moisture content, as well as local NMR relaxation times. These methods are all based on the Single-Point Imaging (SPI) method. The development of these new methods will be described, and the techniques validated using phantom studies. The study of one-dimensional moisture transport in drying concrete was performed using SPI. This work examined the effect of initial mixture proportions and hydration time on the drying behaviour of concrete, over a period of three months. Studies of drying concrete were also performed using spatial mapping of the spin-lattice (T1) and effective spin-spin (T2*) relaxation times, thereby permitting the observation of changes in the water occupied pore surface-to-volume ratio (S/V) as a function of drying. Results of this work demonstrated changes in the S/V due to drying, hydration and drying induced microcracking. Three-dimensional MRI of concrete was performed using SPRITE (Single-Point Ramped Imaging with T1 Enhancement) and turboSPI (turbo Single Point Imaging). While SPRITE allows for weighting of MR images using T 1 and T2*, turboSPI allows T2 weighting of the resulting images. Using relaxation weighting it was shown to be possible to discriminate between water contained within a hydrated cement matrix, and water in highly porous aggregates, used to produce low-density concrete. Three dimensional experiments performed using SPRITE and turboSPI examined the role of self-dessication, drying, initial aggregate saturation and initial mixture conditions on the transport of moisture between porous aggregates and the hydrated matrix. The results demonstrate that water is both added and removed from the aggregates, depending upon the physical conditions. The images also appear to show an influx of cement products into cracks in the solid aggregate. (Abstract shortened by UMI.)

Beyea, Steven Donald

2001-07-01

295

Near real-time disturbance detection using satellite image time series  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Near real-time monitoring of ecosystem disturbances is critical for rapidly assessing and addressing impacts on carbon dynamics, biodiversity, and socio-ecological processes. Satellite remote sensing enables cost-effective and accurate monitoring at frequent time steps over large areas. Yet, generic methods to detect disturbances within newly captured satellite images are lacking. We propose a multi-purpose time-series-based disturbance detection approach that identifies and models stable his...

Verbesselt, J. P.; Zeileis, A.; Herold, M.

2012-01-01

296

Real-time single-cell imaging of protein secretion  

Science.gov (United States)

Protein secretion, a key intercellular event for transducing cellular signals, is thought to be strictly regulated. However, secretion dynamics at the single-cell level have not yet been clarified because intercellular heterogeneity results in an averaging response from the bulk cell population. To address this issue, we developed a novel assay platform for real-time imaging of protein secretion at single-cell resolution by a sandwich immunoassay monitored by total internal reflection microscopy in sub-nanolitre-sized microwell arrays. Real-time secretion imaging on the platform at 1-min time intervals allowed successful detection of the heterogeneous onset time of nonclassical IL-1? secretion from monocytes after external stimulation. The platform also helped in elucidating the chronological relationship between loss of membrane integrity and IL-1? secretion. The study results indicate that this unique monitoring platform will serve as a new and powerful tool for analysing protein secretion dynamics with simultaneous monitoring of intracellular events by live-cell imaging.

Shirasaki, Yoshitaka; Yamagishi, Mai; Suzuki, Nobutake; Izawa, Kazushi; Nakahara, Asahi; Mizuno, Jun; Shoji, Shuichi; Heike, Toshio; Harada, Yoshie; Nishikomori, Ryuta; Ohara, Osamu

2014-01-01

297

A Colour Image Quantization Algorithm for Time-Constrained Applications  

Directory of Open Access Journals (Sweden)

Full Text Available Many techniques have been proposed to quantize a digital colour image in order to reduce the representative number of colours to be suitable for presenting on different types of display screens. In addition, the techniques have been used to significantly reduce the amount of image data required to transfer over a communication network. Most of the published techniques are targetted for implementing on a general purpose multitasking computer with low restriction on time and resource utilizations. The drawback of these techniques relies on the fact that they cannot fulfill the requirement of some applications for real-time constraint and limited resources. In addition, most of the techniques are too complex for hardware realization. In this paper, an algorithm which is more suitable for time critical applications with an additional feature of simplicity to implement on FPGA (Field Programmable Gate Array platforms is proposed and the details of its implementation and experimentation are presented. The dominate point of the proposed algorithm relies on the fact that it utilizes the weighted sum of the nearest distance along the axis under consideration, which is nontrivial to calculate, instead of the squared Euclidean distance to find the axis to split during. Also, the proposed algorithm has proved that by reducing the number of subspaces to be considered during the variance representative value calculation from 8 to 2 subspaces, the quality of quantized images are comparable to the previously proposed approaches. This makes it possible to further speed up the computational time of the quantization algorithm.

Wattanapong KURDTHONGMEE

2005-06-01

298

Time-of-flight imaging of invisibility cloaks  

CERN Multimedia

As invisibility cloaking has recently become experimental reality, it is interesting to explore ways to reveal remaining imperfections. In essence, the idea of most invisibility cloaks is to recover the optical path lengths without an object (to be made invisible) by a suitable arrangement around that object. Optical path length is proportional to the time of flight of a light ray or to the optical phase accumulated by a light wave. Thus, time-of-flight images provide a direct and intuitive tool for probing imperfections. Indeed, recent phase-sensitive experiments on the carpet cloak have already made early steps in this direction. In the macroscopic world, time-of-flight images could be measured directly by light detection and ranging (LIDAR). Here, we show calculated time-of-flight images of the conformal Gaussian carpet cloak, the conformal grating cloak, the cylindrical free-space cloak, and of the invisible sphere. All results are obtained by using a ray-velocity equation of motion derived from Fermat's ...

Halimeh, Jad C

2011-01-01

299

Real-time photoacoustic imaging with optical ultrasound detection  

Science.gov (United States)

Optical ultrasound detection has become an attractive alternative to piezoelectric ultrasound detection for photoacoustic imaging. The favorable properties of optical detection are high resolution, complete optical and acoustical transparency. Recently, it has been shown that optical phase contrast full field detection in combination with a CCD-camera can be used to record acoustic fields. This allows to obtain two-dimensional photoacoustic projection images in real-time. The present work shows an extension of the technique towards full three-dimensional photoacoustic tomography. The specifications of the detection system, resolution and sensitivity, are 66?m and 3.4kPa. The reconstruction of the initial three dimensional pressure distribution is a two step process. First of all, projection images of the initial pressure distribution are acquired. This is done by back propagating the observed wave pattern in frequency space. In the second step the inverse Radon transform is applied to the obtained projection dataset to reconstruct the initial three dimensional pressure distribution. Simulations and experiments are performed to show the overall applicability of this technique for real-time photoacoustic imaging.

Nuster, R.; Paltauf, G.

2012-02-01

300

Lightweight distributed computing for intraoperative real-time image guidance  

Science.gov (United States)

In order to provide real-time intraoperative guidance, computer assisted surgery (CAS) systems often rely on computationally expensive algorithms. The real-time constraint is especially challenging if several components such as intraoperative image processing, soft tissue registration or context aware visualization are combined in a single system. In this paper, we present a lightweight approach to distribute the workload over several workstations based on the OpenIGTLink protocol. We use XML-based message passing for remote procedure calls and native types for transferring data such as images, meshes or point coordinates. Two different, but typical scenarios are considered in order to evaluate the performance of the new system. First, we analyze a real-time soft tissue registration algorithm based on a finite element (FE) model. Here, we use the proposed approach to distribute the computational workload between a primary workstation that handles sensor data processing and visualization and a dedicated workstation that runs the real-time FE algorithm. We show that the additional overhead that is introduced by the technique is small compared to the total execution time. Furthermore, the approach is used to speed up a context aware augmented reality based navigation system for dental implant surgery. In this scenario, the additional delay for running the computationally expensive reasoning server on a separate workstation is less than a millisecond. The results show that the presented approach is a promising strategy to speed up real-time CAS systems.

Suwelack, Stefan; Katic, Darko; Wagner, Simon; Spengler, Patrick; Bodenstedt, Sebastian; Röhl, Sebastian; Dillmann, Rüdiger; Speidel, Stefanie

2012-02-01

 
 
 
 
301

Imaging nuclear decays with Optical Time Projection Chamber  

International Nuclear Information System (INIS)

A novel type of gaseous ionization detector - Optical Time Projection Chamber (OTPC) - developed to study rare nuclear decays is presented. The OTPC records tracks of charged particles ionizing a counting gas by optical imaging of the light generated by electrons multiplied in the amplification structures. By combining an electron drift-time profile measured by a photomultiplier and a CCD camera image we reconstruct three-dimensional trajectories of particles, energies and charges. The capabilities of the OTPC detector to study various decay modes are demonstrated by observation of beta-delayed proton emission from 13O, two-alpha break-up of 8Be, triple-alpha decay of 12C excited states and two-proton radioactivity of 45Fe

2007-11-30

302

Real-time in vivo imaging of p16Ink4a gene expression: a new approach to study senescence stress signaling in living animals  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Oncogenic proliferative signals are coupled to a variety of growth inhibitory processes. In cultured primary human fibroblasts, for example, ectopic expression of oncogenic Ras or its downstream mediator initiates cellular senescence, the state of irreversible cell cycle arrest, through up-regulation of cyclin-dependent kinase (CDK inhibitors, such as p16INK4a. To date, much of our current knowledge of how human p16INK4a gene expression is induced by oncogenic stimuli derives from studies undertaken in cultured primary cells. However, since human p16INK4a gene expression is also induced by tissue culture-imposed stress, it remains unclear whether the induction of human p16INK4a gene expression in tissue-cultured cells truly reflects an anti-cancer process or is an artifact of tissue culture-imposed stress. To eliminate any potential problems arising from tissue culture imposed stress, we have recently developed a bioluminescence imaging (BLI system for non-invasive and real-time analysis of human p16INK4a gene expression in the context of a living animal. Here, we discuss the molecular mechanisms that direct p16INK4a gene expression in vivo and its potential for tumor suppression.

Takahashi Akiko

2010-01-01

303

Predicting Clinical Image Delivery Time by Monitoring PACS Queue Behavior  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The expectation of rapid image retrieval from PACS users contributes to increased information technology (IT) infrastructure investments to increase performance as well as continuing demands upon PACS administrators to respond to “slow” system performance. The ability to provide predicted delivery times to a PACS user may curb user expectations for “fastest” response especially during peak hours. This, in turn, could result in a PACS infrastructure tailored to more realistic performan...

King, Nelson E.; Documet, Jorge; Liu, Brent

2006-01-01

304

Time-lapse imaging rendszerek kvalitatív és kvantitatív elemzése  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Vizsgálataim során a Long-Term Scan rendszerrel felvett képek elemzését végeztem, mely rendszer segítségével több sejtciklus alatt nyerünk betekintést a sejtpopulációk életfolyamataiba. A Long-Term Scan rendszer a time-lapse imaging módszert felhasználva el?re meghatározott id?közönként készíti el a felvételeket a vizsgált sejtekr?l. Ez a módszer akkor el?nyös, ha nincs szükségünk a vizsgálat teljes id?tartama alatt az összes képre, hanem csak az egymásh...

2011-01-01

305

Mechanisms in the Quantum Yield of Cypridina Bioluminescence.  

Science.gov (United States)

The influence of temperature, pH, salts, and reactant concentrations on the bioluminescent oxidation of Cypridina luciferin catalyzed by Cypridina luciferase indicated a highest quantum yield phi (einsteins per mole of luciferin oxidized) of 0.31 in H2O, ...

O. Shimomura F. H. Johnson

1970-01-01

306

A bioluminescent signal system: detection of chemical toxicants in water.  

Science.gov (United States)

Prototype technologies of a bioluminescent signal system (BSS) based on the luminous bacterium Photobacterium phosphoreum and three enzymatic bioluminescence systems have been proposed for detecting and signalling the presence of toxicants in water systems. A number of pesticides, mostly known as poisonous substances, similar in their structures and physicochemical properties, have been taken as model compounds of chemical agents. The effect of toxicants (organophosphates, derivatives of dithiocarbamide acid, and pyrethroid preparations) on the bioluminescence of the four systems has been analysed. EC(50) and EC(80) have been determined and compared to the maximum permissible concentration for each of the analysed substances. The triple-enzyme systems with ADH and trypsin have been shown to be more sensitive to organophosphorous compounds (0.13-11 mg/L), while the triple-enzyme system with trypsin is highly sensitive to lipotropic poison, a derivative of dithiocarbamine acid (0.03 mg/L). Sensitivities of the triple-enzyme systems to pyrethroid preparations are similar to those of luminous bacteria (0.9-5 mg/L). The results can be used to construct an alarm-test bioluminescence system for detecting chemical toxicants, based on intact bacteria or enzyme systems. PMID:17603816

Vetrova, E; Esimbekova, E; Remmel, N; Kotova, S; Beloskov, N; Kratasyuk, V; Gitelson, I

2007-01-01

307

Imaging gene expression in real-time using aptamers  

Energy Technology Data Exchange (ETDEWEB)

Signal transduction pathways are usually activated by external stimuli and are transient. The downstream changes such as transcription of the activated genes are also transient. Real-time detection of promoter activity is useful for understanding changes in gene expression, especially during cell differentiation and in development. A simple and reliable method for viewing gene expression in real time is not yet available. Reporter proteins such as fluorescent proteins and luciferase allow for non-invasive detection of the products of gene expression in living cells. However, current reporter systems do not provide for real-time imaging of promoter activity in living cells. This is because of the long time period after transcription required for fluorescent protein synthesis and maturation. We have developed an RNA reporter system for imaging in real-time to detect changes in promoter activity as they occur. The RNA reporter uses strings of RNA aptamers that constitute IMAGEtags (Intracellular MultiAptamer GEnetic tags), which can be expressed from a promoter of choice. The tobramycin, neomycin and PDC RNA aptamers have been utilized for this system and expressed in yeast from the GAL1 promoter. The IMAGEtag RNA kinetics were quantified by RT-qPCR. In yeast precultured in raffinose containing media the GAL1 promoter responded faster than in yeast precultured in glucose containing media. IMAGEtag RNA has relatively short half-life (5.5 min) in yeast. For imaging, the yeast cells are incubated with their ligands that are labeled with fluorescent dyes. To increase signal to noise, ligands have been separately conjugated with the FRET (Förster resonance energy transfer) pairs, Cy3 and Cy5. With these constructs, the transcribed aptamers can be imaged after activation of the promoter by galactose. FRET was confirmed with three different approaches, which were sensitized emission, acceptor photobleaching and donor lifetime by FLIM (fluorescence lifetime imaging microscopy). Real-time transcription was measured by FLIM-FRET, which was detected by the decrease in donor lifetime resulting from ligand binding to IMAGEtags that were newly synthesized from the activated GAL1 promoter. The FRET signal was specific for transcribed IMAGEtags.

Shin, Il Chung

2011-12-13

308

Imaging gene expression in real-time using aptamers  

Energy Technology Data Exchange (ETDEWEB)

Signal transduction pathways are usually activated by external stimuli and are transient. The downstream changes such as transcription of the activated genes are also transient. Real-time detection of promoter activity is useful for understanding changes in gene expression, especially during cell differentiation and in development. A simple and reliable method for viewing gene expression in real time is not yet available. Reporter proteins such as fluorescent proteins and luciferase allow for non-invasive detection of the products of gene expression in living cells. However, current reporter systems do not provide for real-time imaging of promoter activity in living cells. This is because of the long time period after transcription required for fluorescent protein synthesis and maturation. We have developed an RNA reporter system for imaging in real-time to detect changes in promoter activity as they occur. The RNA reporter uses strings of RNA aptamers that constitute IMAGEtags (Intracellular MultiAptamer GEnetic tags), which can be expressed from a promoter of choice. The tobramycin, neomycin and PDC RNA aptamers have been utilized for this system and expressed in yeast from the GAL1 promoter. The IMAGEtag RNA kinetics were quantified by RT-qPCR. In yeast precultured in raffinose containing media the GAL1 promoter responded faster than in yeast precultured in glucose containing media. IMAGEtag RNA has relatively short half-life (5.5 min) in yeast. For imaging, the yeast cells are incubated with their ligands that are labeled with fluorescent dyes. To increase signal to noise, ligands have been separately conjugated with the FRET (Förster resonance energy transfer) pairs, Cy3 and Cy5. With these constructs, the transcribed aptamers can be imaged after activation of the promoter by galactose. FRET was confirmed with three different approaches, which were sensitized emission, acceptor photobleaching and donor lifetime by FLIM (fluorescence lifetime imaging microscopy). Real-time transcription was measured by FLIM-FRET, which was detected by the decrease in donor lifetime resulting from ligand binding to IMAGEtags that were newly synthesized from the activated GAL1 promoter. The FRET signal was specific for transcribed IMAGEtags.

Shin, Il Chung [Ames Laboratory

2012-11-02

309

Charge-coupled imagers for time-resolved macromolecular crystallography  

Energy Technology Data Exchange (ETDEWEB)

There exists considerable promise for the use of charge-coupled device (CCD) imagers in the fast recording of parts of macromolecular crystal Laue diffraction patterns. As part of this development CCD tests have been made with direct detection of Laue patterns from a small molecule test crystal and a protein crystal. Merging {ital R} factors (on intensity), for strong reflections, of 3% have been obtained. A time-slicing scheme for a CCD camera is discussed based on the stacking of slices held in storage in the CCD in the submillisecond time resolution range.

Allinson, N.M. (Department of Electronics, University of York, York, YO1 5DD (United Kingdom)); Colapietro, M. (Department of Chemistry, University of Rome, La Sapienza, Rome (Italy)); Helliwell, J.R. (Department of Chemistry, University of Manchester, Manchester M13 9PL, United Kingdom and SERC, Daresbury Laboratory, Daresbury, Warrington, WA4 4AD (United Kingdom)); Moon, K.J. (Department of Electronics, University of York, York, YO1 5DD (United Kingdom)); Thompson, A.W. (SERC, Daresbury Laboratory, Daresbury, Warrington, WA4 4AD (United Kingdom)); Weisgerber, S. (Department of Chemistry, University of Manchester, Manchester, M13 9PL (United Kingdom))

1992-01-01

310

Real-time image restoration for iris recognition systems.  

Science.gov (United States)

In the field of biometrics, it has been reported that iris recognition techniques have shown high levels of accuracy because unique patterns of the human iris, which has very many degrees of freedom, are used. However, because conventional iris cameras have small depth-of-field (DOF) areas, input iris images can easily be blurred, which can lead to lower recognition performance, since iris patterns are transformed by the blurring caused by optical defocusing. To overcome these problems, an autofocusing camera can be used. However, this inevitably increases the cost, size, and complexity of the system. Therefore, we propose a new real-time iris image-restoration method, which can increase the camera's DOF without requiring any additional hardware. This paper presents five novelties as compared to previous works: 1) by excluding eyelash and eyelid regions, it is possible to obtain more accurate focus scores from input iris images; 2) the parameter of the point spread function (PSF) can be estimated in terms of camera optics and measured focus scores; therefore, parameter estimation is more accurate than it has been in previous research; 3) because the PSF parameter can be obtained by using a predetermined equation, iris image restoration can be done in real-time; 4) by using a constrained least square (CLS) restoration filter that considers noise, performance can be greatly enhanced; and 5) restoration accuracy can also be enhanced by estimating the weight value of the noise-regularization term of the CLS filter according to the amount of image blurring. Experimental results showed that iris recognition errors when using the proposed restoration method were greatly reduced as compared to those results achieved without restoration or those achieved using previous iris-restoration methods. PMID:18179073

Kang, Byung Jun; Park, Kang Ryoung

2007-12-01

311

Image Fusion Real-time System Based on FPGA and Multi-DSP  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In order to solve complex algorithm that is difficult to achieve real-time processing of Multiband image fusion within large amount of data, a real-time image fusion system based on FPGA and multi-DSP is designed. Five-band image acquisition, image registration, image fusion and display output can be done within the system w...

Feng Qu; Bochao Liu; Jian Zhao; Qiang Sun

2013-01-01

312

Real-time image stabilization for arbitrary motion blurred image based on opto-electronic hybrid joint transform correlator.  

Science.gov (United States)

An efficient approach was put forward to keep real-time image stabilization based on opto-electronic hybrid processing, by which image motion vector can be effectively detected and point spread function (PSF) was accurately modeled instantaneously, it will alleviate greatly the complexity of image restoration algorithm. The approach applies to arbitrary motion blurred images. We have also constructed an image stabilization measurement system. The experimental results show that the proposed method has advantages of real time and preferable effect. PMID:21643332

Qian, Yixian; Li, Yong; Shao, Jie; Miao, Hua

2011-05-23

313

High-temperature real-time ultrasonic imaging  

International Nuclear Information System (INIS)

Ultrasonic instrumentation capable of real-time imaging of materials at temperatures up to 288"0C (590"0F) is described. The research and development of this unique instrumentation was sponsored by the Electric Power Research Institute of Palo Alto, California, for use in nuclear power plants. The instrumentation developed will permit continuous surveillance of piping while the power plant is in operation. The instrumentation utilizes a combination of high-temperature materials to fabricate a unique piezoelectric transmitter and a high-temperature electromagnetic acoustic transducer (EMAT). The high-temperature EMAT operates at 2.5 MHz, which is well above preceding models of about 800 kHz. Use of unique high-temperature materials to permit scanning of material volume is combined with an imaging system to allow time-lapse image information. This paper traces the theory and describes material properties of interest and reports on test results for a development system that has been in continuous operation on a field test site for two years. Future applications and development plans are outlined

1984-01-01

314

Real-time neutron monitoring method using an imaging plate  

Energy Technology Data Exchange (ETDEWEB)

A novel system for real-time radiation monitoring in reactor or accelerator facilities has been studied using an imaging plate. The authors made a feasibility study on a new neutron detection system using both photostimulated luminescence (PSL) and prompt luminescence (PL) generated in a neutron imaging plate (NIP) when the NIP is irradiated by neutrons. A readout system consisting of a semiconductor laser and a photomultiplier tube was fabricated for the purpose. It was confirmed that the system can measure both PSL and PL, where Am-Li was used as a neutron source. It may be possible to establish a new wide-range neutron monitoring system using the developed system as a PL mode normally, and as a PSL mode in case of intense neutron dose that cannot be measured in a PL mode because of saturation of the detection system. (author)

Sakasai, Kaoru; Katagiri, Masaki; Kishimoto, Maki; Fujii, Yoshio [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

1999-07-01

315

Real-time neutron monitoring method using an imaging plate  

International Nuclear Information System (INIS)

A novel system for real-time radiation monitoring in reactor or accelerator facilities has been studied using an imaging plate. The authors made a feasibility study on a new neutron detection system using both photostimulated luminescence (PSL) and prompt luminescence (PL) generated in a neutron imaging plate (NIP) when the NIP is irradiated by neutrons. A readout system consisting of a semiconductor laser and a photomultiplier tube was fabricated for the purpose. It was confirmed that the system can measure both PSL and PL, where Am-Li was used as a neutron source. It may be possible to establish a new wide-range neutron monitoring system using the developed system as a PL mode normally, and as a PSL mode in case of intense neutron dose that cannot be measured in a PL mode because of saturation of the detection system. (author)

1999-04-19

316

Ready for prime time? Dual tracer PET and SPECT imaging.  

Science.gov (United States)

Dual isotope single photon emission computed tomography (SPECT) and dual tracer positron emission tomography (PET) imaging have great potential in clinical and molecular applications in the pediatric as well as the adult populations in many areas of brain, cardiac, and oncologic imaging as it allows the exploration of different physiological and molecular functions (e.g., perfusion, neurotransmission, metabolism, apoptosis, angiogenesis) under the same physiological and physical conditions. This is crucial when the physiological functions studied depend on each other (e.g., perfusion and metabolism) hence requiring simultaneous assessment under identical conditions, and can reduce greatly the quantitation errors associated with physical factors that can change between acquisitions (e.g., human subject or animal motion, change in the attenuation map as a function of time) as is detailed in this editorial. The clinical potential of simultaneous dual isotope SPECT, dual tracer PET and dual SPECT/PET imaging are explored and summarized. In this issue of AJNMMI (http://www.ajnmmi.us), Chapman et al. explore the feasibility of simultaneous and sequential SPECT/PET imaging and conclude that down-scatter and crosstalk from 511 keV photons preclude obtaining useful SPECT information in the presence of PET radiotracers. They report on an alternative strategy that consists of performing sequential SPECT and PET studies in hybrid microPET/SPECT/CT scanners, now widely available for molecular imaging. They validate their approach in a phantom consisting of a 96-well plate with variable (99m)Tc and (18)F concentrations and illustrate the utility of such approaches in two sequential SPECT-PET/CT studies that include (99m)Tc-MAA/(18)F-NaF and (99m)Tc-Pentetate/(18)F-NaF. These approaches will need to be proven reproducible, accurate and robust to variations in the experimental conditions before they can be accepted by the molecular imaging community and be implemented in routine molecular microPET and microSPECT explorations. Although currently not accepted as standard procedures in the molecular imaging community, such approaches have the potential to open the way to new SPECT/PET explorations that allow studying molecular mechanisms and pathways in the living animal under similar physiological conditions. Although still premature for the clinical setting, these approaches can be extended to clinical research once proven accurate and precise in vivo in small and large animal models. PMID:23145358

Fakhri, Georges El

2012-01-01

317

Early time points perfusion imaging: relative time of arrival, maximum derivatives and fractional derivatives.  

Science.gov (United States)

Time of arrival (TOA) of a bolus of contrast agent to the tissue voxel is a reference time point critical for the Early Time Points Perfusion Imaging Method (ET) to make relative cerebral blood flow (rCBF) maps. Due to the low contrast to noise (CNR) condition at TOA, other useful reference time points known as relative time of arrival data points (rTOA) are investigated. Candidate rTOA's include the time to reach the maximum derivative, the maximum second derivative, and the maximum fractional derivative. Each rTOA retains the same relative time distance from TOA for all tissue flow levels provided that ET's basic assumption is met, namely, no contrast agent has a chance to leave the tissue before the time of rTOA. The ET's framework insures that rCBF estimates by different orders of the derivative are theoretically equivalent to each other and monkey perfusion imaging results supported the theory. In rCBF estimation, maximum values of higher order fractional derivatives may be used to replace the maximum derivative which runs a higher risk of violating ET's assumption. Using the maximum values of the derivative of orders ranging from 1 to 1.5 to 2, estimated rCBF results were found to demonstrate a gray-white matter ratio of approximately 3, a number consistent with flow ratio reported in the literature. PMID:21600995

Kwong, Kenneth K; Wu, Ona; Chan, Suk-Tak; Nelissen, Koen; Kholodov, Mykhaylo; Chesler, David A

2011-08-01

318

Real time thermal imaging of high temperature semiconductor melts  

Science.gov (United States)

A real time thermal imaging system with temperature resolution better than + or - 1 C and spatial resolution of better than 0.5 mm was developed and applied to the analysis of melt surface thermal field distributions in both Czochralski and liquid encapsulated Czochralski (LEC) growth configurations. The melt is viewed in near normal incidence by a high resolution charge coupled device camera to which is attached a very narrow bandpass filter. The resulting image is digitized and processed using a pipelined pixel processor operating at an effective 40 million operations per second thus permitting real time high frequency spatial and temporal filtering of the high temperature scene. A multi-pixel averaging algorithm was developed which permits localized, low noise sensing of temperature variations at any location in the hot zone as a function of time. This signial is used to implement initial elements of a feedforward growth control scheme which is aimed at reducing disturbances to the melt caused by the batch nature of the growth process. The effect of magnetic melt stabilization on radial melt temperature distributions was measured using this technique. Problems associated with residual internal reflections and non-optimized path geometry are discussed.

Wargo, Michael J.

1988-01-01

319

Whole-body, real-time preclinical imaging of quantum dot fluorescence with time-gated detection  

Science.gov (United States)

We describe a wide-field preclinical imaging system optimized for time-gated detection of quantum dot fluorescence emission. As compared to continuous wave measurements, image contrast was substantially improved by suppression of short-lifetime background autofluorescence. Real-time (8 frames?s) biological imaging of subcutaneous quantum dot injections is demonstrated simultaneously in multiple living mice.

May, Andrzej; Bhaumik, Srabani; Gambhir, Sanjiv S.; Zhan, Chun; Yazdanfar, Siavash

2009-01-01

320

Recursive time-varying filter banks for subband image coding  

Science.gov (United States)

Filter banks and wavelet decompositions that employ recursive filters have been considered previously and are recognized for their efficiency in partitioning the frequency spectrum. This paper presents an analysis of a new infinite impulse response (IIR) filter bank in which these computationally efficient filters may be changed adaptively in response to the input. The filter bank is presented and discussed in the context of finite-support signals with the intended application in subband image coding. In the absence of quantization errors, exact reconstruction can be achieved and by the proper choice of an adaptation scheme, it is shown that IIR time-varying filter banks can yield improvement over conventional ones.

Smith, Mark J. T.; Chung, Wilson C.

1992-01-01

 
 
 
 
321

In-Vivo Real-Time X-ray ?-Imaging  

International Nuclear Information System (INIS)

The technique of X-ray transmission imaging is available for more than 100 years and it is still one of the fastest and easiest ways how to study the internal structure of living biological samples. The advances in semiconductor technology in last years make possible to fabricate new types of X-ray detectors with direct conversion of interacting X-ray photon to an electric signal. Especially semiconductor pixel detectors seem to be very promising. Compared to the film technique they bring single-quantum and real-time digital information about the studied object with high resolution, high sensitivity and broad dynamic range. These pixel detector-based imaging stand promising as a new tool in the field of small animal imaging, for cancer research and for observation of dynamic processes inside organisms. These detectors open up for instance new possibilities for researchers to perform non-invasive studies of tissue for mutations or pathologies and to monitor disease progression or response to therapy

2007-11-26

322

Real-time in-vivo ?-imaging with Medipix2  

International Nuclear Information System (INIS)

An X-ray micro-radiographic system based on the Medipix2 semiconductor pixel detector for dynamic high spatial resolution and for high contrast imaging has been developed. Our system is based on a micro-focus and nano-focus X-ray tube and the hybrid single-photon counting silicon pixel detector Medipix2 (matrix 256x256 sq. pixels of 55 ?m pitch). This compact table-top system stands promising as a new tool in the field of small animal imaging as well as in the in-vivo observation of dynamic processes inside living organisms. The main advantages of these Medipix2 pixel detectors include: high sensitivity to low-energy X-ray photons; position sensitive and noiseless single-photon detection with preselected photon energies; single-quantum counting in each pixel performed by digital counter (therefore there is no dark current); digital integration (providing unlimited dynamic range and absolute linearity in device response to number of photons, high sensitivity and high contrast); real-time digital information, high-speed digital communication and data transfer. We improve the picture quality with the help of statistical data analysis and extended the calibration of individual pixels response. 2D and 3D radiographic images of samples demonstrate the potential and applicability of our system for precise in-vivo X-ray high-resolution dynamic diagnostic and biological studies. Obtained results are shown on small animal and organic samples.

2009-08-01

323

Real-time in-vivo ?-imaging with Medipix2  

Science.gov (United States)

An X-ray micro-radiographic system based on the Medipix2 semiconductor pixel detector for dynamic high spatial resolution and for high contrast imaging has been developed. Our system is based on a micro-focus and nano-focus X-ray tube and the hybrid single-photon counting silicon pixel detector Medipix2 (matrix 256×256 sq. pixels of 55 ?m pitch). This compact table-top system stands promising as a new tool in the field of small animal imaging as well as in the in-vivo observation of dynamic processes inside living organisms. The main advantages of these Medipix2 pixel detectors include: high sensitivity to low-energy X-ray photons; position sensitive and noiseless single-photon detection with preselected photon energies; single-quantum counting in each pixel performed by digital counter (therefore there is no dark current); digital integration (providing unlimited dynamic range and absolute linearity in device response to number of photons, high sensitivity and high contrast); real-time digital information, high-speed digital communication and data transfer. We improve the picture quality with the help of statistical data analysis and extended the calibration of individual pixels response. 2D and 3D radiographic images of samples demonstrate the potential and applicability of our system for precise in-vivo X-ray high-resolution dynamic diagnostic and biological studies. Obtained results are shown on small animal and organic samples.

Dammer, J.; Frallicciardi, P. M.; Jakubek, J.; Jakubek, M.; Pospisil, S.; Prenerova, E.; Vavrik, D.; Volter, L.; Weyda, F.; Zemek, R.

2009-08-01

324

Bacillus anthracis diagnostic detection and rapid antibiotic susceptibility determination using 'bioluminescent' reporter phage.  

Science.gov (United States)

Genetically modified phages have the potential to detect pathogenic bacteria from clinical, environmental, or food-related sources. Herein we assess an engineered 'bioluminescent' reporter phage (Wß::luxAB) as a clinical diagnostic tool for Bacillus anthracis, the etiological agent of anthrax. Wß::luxAB is able to rapidly (within minutes) detect a panel of B. anthracis strains by transducing a bioluminescent phenotype. The reporter phage displays species specificity by its inability, or significantly reduced ability, to detect members of the closely related Bacillus cereus group and other common bacterial pathogens. Using spiked clinical specimens, Wß::luxAB detects B. anthracis within 5 h at clinically relevant concentrations, and provides antibiotic susceptibility information that mirrors the CLSI method, except that data are obtained at least 5-fold faster. Although anthrax is a treatable disease, a positive patient prognosis is dependent on timely diagnosis and appropriate therapy. Wß::luxAB rapidly detects B. anthracis and determines antibiotic efficacy, properties that will help patient outcome. PMID:23994352

Schofield, David A; Sharp, Natasha J; Vandamm, Joshua; Molineux, Ian J; Spreng, Krista A; Rajanna, Chythanya; Westwater, Caroline; Stewart, George C

2013-11-01

325

Distribution of bioluminescent fungi across old-growth and secondary tropical rain forest in Costa Rica  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Most research on bioluminescent fungi is concentrated on their taxonomic relationships, while the basics of their natural history and ecological relationships are poorly understood. In this study, we compared the distribution of bioluminescent fungi between old-growth and secondary forest as related to four different soil types at the tropical rainforest of La Selva Biological Station in Costa Rica. The study was conducted during the wet season of 2009. Bioluminescent fungi were sought follow...

Carolina Seas-Carvajal; Gerardo Avalos

2013-01-01

326

Mechanisms of bioluminescence, chemiluminescence and of their regulation. Progress report, one year period through March 1976  

Energy Technology Data Exchange (ETDEWEB)

Progress is reported on a 10-yr study of the production and role of excited states in biological systems and the mechanisms involved in bioluminescence and chemoluminescence. An hypothesis of the origin of bioluminescence is presented that is based on the mixed function oxygenase reaction. Techniques of absolute measurements of light intensities and spectral composition were applied in studies of bioluminescence of marine dinoflagellates and the chemiluminescence of carcinogenic polycyclic aromatic hydrocarbons as the result of enzymatic hydroxylation. (CH)

Seliger, H.H.

1976-01-01

327

Use of Bioluminescence for Detection of Genetically Engineered Microorganisms Released into the Environment  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The persistence and movement of strain JS414 of Xanthomonas campestris pv. campestris, which was genetically engineered to bioluminesce, were monitored during a limited field introduction. Bioluminescence and traditional dilution plate counts were determined. Strain JS414 was applied to cabbage plants and surrounding soil by mist inoculation, by wound inoculation, by scattering infested debris among plants, and by incorporating bacteria into the soil. Bioluminescent X. campestris pv. campestr...

1992-01-01

328

Evaluation of scintillator afterglow for use in a combined optical and PET imaging tomograph  

International Nuclear Information System (INIS)

The design of a dual modality imaging system for small animal optical and positron emission tomography imaging (OPET) is underway. Its detector must be capable of imaging high energy ?-rays from PET while also resolving optical wavelength photons from bioluminescence. GSO, high purity GSO, BGO, LSO, LYSO, and LaBr scintillators were investigated for their use in the OPET detector. Of specific interest were scintillators with low afterglow, since afterglow photons in the decay of the larger ?-ray events are indistinguishable from the photons generated by bioluminescence. Samples from these crystals were coupled to a photomultiplier tube (PMT) and produced scintillation light from ?-ray events originating from a positron source. The PMT output was directed to a special signal processing circuit that allowed measurement of single photons at different times in the decay of the scintillation. GSO and BGO exhibited optimal performance for use in the OPET system due to their low afterglow. LSO, LYSO, and LaBr were determined unsuitable for use with the current OPET design due to their significant afterglow components. The effect of the afterglow of GSO on the detection of the bioluminescence signal-to-noise ratio (SNR) was evaluated for the OPET system

2006-12-20

329

Evaluation of scintillator afterglow for use in a combined optical and PET imaging tomograph  

Energy Technology Data Exchange (ETDEWEB)

The design of a dual modality imaging system for small animal optical and positron emission tomography imaging (OPET) is underway. Its detector must be capable of imaging high energy {gamma}-rays from PET while also resolving optical wavelength photons from bioluminescence. GSO, high purity GSO, BGO, LSO, LYSO, and LaBr scintillators were investigated for their use in the OPET detector. Of specific interest were scintillators with low afterglow, since afterglow photons in the decay of the larger {gamma}-ray events are indistinguishable from the photons generated by bioluminescence. Samples from these crystals were coupled to a photomultiplier tube (PMT) and produced scintillation light from {gamma}-ray events originating from a positron source. The PMT output was directed to a special signal processing circuit that allowed measurement of single photons at different times in the decay of the scintillation. GSO and BGO exhibited optimal performance for use in the OPET system due to their low afterglow. LSO, LYSO, and LaBr were determined unsuitable for use with the current OPET design due to their significant afterglow components. The effect of the afterglow of GSO on the detection of the bioluminescence signal-to-noise ratio (SNR) was evaluated for the OPET system.

Douraghy, Ali [Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, UCLA School of Medicine, A136, 700 Westwood Plaza, Los Angeles, CA 90095-1770 (United States)]. E-mail: adouraghy@mednet.ucla.edu; Prout, David L. [Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, UCLA School of Medicine, A136, 700 Westwood Plaza, Los Angeles, CA 90095-1770 (United States); Silverman, Robert W. [Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, UCLA School of Medicine, A136, 700 Westwood Plaza, Los Angeles, CA 90095-1770 (United States); Chatziioannou, Arion F. [Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, UCLA School of Medicine, A136, 700 Westwood Plaza, Los Angeles, CA 90095-1770 (United States)

2006-12-20

330

Abdominal MR imaging using a HASTE sequence : image comparison on the different echo times  

Energy Technology Data Exchange (ETDEWEB)

To determine the optimal parameters of abdominal HASTE imaging by means of a comparison of intermediate and long TE (echo time). We evaluated 30 consecutive patients who had undergone liver MR during a three-month period. Twelve patients were diagnosed as normal, four as having liver cirrhosis, and 14 were found to be suffering form hepatic hemangioma. On the basis of measured signal intensity of the liver, spleen, pancreas and gallbladder, and of fat, muscle, hemangioma, and background, we calculated the ratios of signal to noise (S/N), signal difference to noise (SD/N), and signal intensity (SI). Image quality was compared using these three ratios, and using two HASTE sequences with TEs of 90 msec and 134 msec, images were qualitatively evaluated. S/N ratio of the liver was higher when TE was 90 msec(p<.05), though S/N, SD/N and SI rations of the spleen, gallbladder, and pancreas-and of hemangiom-were higher when TE was 134 msec (p<.05). However, in muscle, all these three ratios were higher at a TE of 90 msec. SD/N ratio and SI of fat were higher at a TE of 134 msec. Overall image quality was better at a TE of 134 msec than at one of 90msec. A HASTE sequence with a TE of 134msec showed greater tissue contrast and stronger T2-weighted images than one with a TE of 90msec.

Park, Kwang Bo; Lee, Moon Gyu; Lim, Tae Hwan; Jeong, Yoong Ki; Ha, Hyun Kwon; Kim, Pyo Nyun; Auh, Yong Ho [Ulsan Univ. College of Medicine, Asan Medical Center, Seoul (Korea, Republic of)

1999-11-01

331

Visible light induced ocular delayed bioluminescence as a possible origin of negative afterimage  

CERN Document Server

The delayed luminescence of biological tissues is an ultraweak reemission of absorbed photons after exposure to external monochromatic or white light illumination. Recently, Wang, B\\'okkon, Dai and Antal (Brain Res. 2011) presented the first experimental proof of the existence of spontaneous ultraweak biophoton emission and visible light induced delayed ultraweak photon emission from in vitro freshly isolated rat's whole eye, lens, vitreous humor and retina. Here, we suggest that the photobiophysical source of negative afterimage can also occur within the eye by delayed bioluminescent photons. In other words, when we stare at a colored (or white) image for few seconds, external photons can induce excited electronic states within different parts of the eye that is followed by a delayed reemission of absorbed photons for several seconds. Finally, these reemitted photons can be absorbed by nonbleached photoreceptors that produce a negative afterimage. Although this suggests the photobiophysical source of negativ...

Bokkon, I; Wang, C; Dai, J; Salari, V; Grass, F; Antal, I

2011-01-01

332

Monitoring neural activity with bioluminescence during natural behavior.  

Science.gov (United States)

Existing techniques for monitoring neural activity in awake, freely behaving vertebrates are invasive and difficult to target to genetically identified neurons. We used bioluminescence to non-invasively monitor the activity of genetically specified neurons in freely behaving zebrafish. Transgenic fish with the Ca(2+)-sensitive photoprotein green fluorescent protein (GFP)-Aequorin in most neurons generated large and fast bioluminescent signals that were related to neural activity, neuroluminescence, which could be recorded continuously for many days. To test the limits of this technique, we specifically targeted GFP-Aequorin to the hypocretin-positive neurons of the hypothalamus. We found that neuroluminescence generated by this group of approximately 20 neurons was associated with periods of increased locomotor activity and identified two classes of neural activity corresponding to distinct swim latencies. Our neuroluminescence assay can report, with high temporal resolution and sensitivity, the activity of small subsets of neurons during unrestrained behavior. PMID:20305645

Naumann, Eva A; Kampff, Adam R; Prober, David A; Schier, Alexander F; Engert, Florian

2010-04-01

333

Incorporating MRI structural information into bioluminescence tomography: system, heterogeneous reconstruction and in vivo quantification  

Science.gov (United States)

Combining two or more imaging modalities to provide complementary information has become commonplace in clinical practice and in preclinical and basic biomedical research. By incorporating the structural information provided by computed tomography (CT) or magnetic resonance imaging (MRI), the ill poseness nature of bioluminescence tomography (BLT) can be reduced significantly, thus improve the accuracies of reconstruction and in vivo quantification. In this paper, we present a small animal imaging system combining multi-view and multi-spectral BLT with MRI. The independent MRI-compatible optical device is placed at the end of the clinical MRI scanner. The small animal is transferred between the light tight chamber of the optical device and the animal coil of MRI via a guide rail during the experiment. After the optical imaging and MRI scanning procedures are finished, the optical images are mapped onto the MRI surface by interactive registration between boundary of optical images and silhouette of MRI. Then, incorporating the MRI structural information, a heterogeneous reconstruction algorithm based on finite element method (FEM) with L 1 normalization is used to reconstruct the position, power and region of the light source. In order to validate the feasibility of the system, we conducted experiments of nude mice model implanted with artificial light source and quantitative analysis of tumor inoculation model with MDA-231-GFP-luc. Preliminary results suggest the feasibility and effectiveness of the prototype system.

Zhang, Jun; Chen, Duofang; Liang, Jimin; Xue, Huadan; Lei, Jing; Wang, Qin; Chen, Dongmei; Meng, Ming; Jin, Zhengyu; Tian, Jie

2014-01-01

334

A Double-Threshold Technique for Fast Time-Correspondence Imaging  

CERN Multimedia

We present a robust imaging method based on time-correspondence imaging and normalized ghost imaging (GI) that sets two thresholds to select the reference frame exposures for image reconstruction. This double-threshold time-correspondence imaging protocol always gives better quality and signal-to-noise ratio than previous GI schemes, and is insensitive to surrounding noise. Moreover, only simple add and minus operations are required while less data storage space and computing time are consumed, thus faster imaging speeds are attainable. The protocol offers a general approach applicable to all GI techniques, and marks a further step forward towards real-time practical applications of correlation imaging.

Li, Ming-Fei; Liu, Xue-Feng; Yao, Xu-Ri; Luo, Kai-Hong; Fan, Heng; Wu, Ling-An

2013-01-01

335

Extrinsic and intrinsic nervous control of bioluminescence in ophiuroids (Echinodermata)  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A diversity of organisms is endowed with the ability to emit light; this phenomenon is called bioluminescence and occurs mainly in marine organisms. Among echinoderms, some ophiuroid species (Echinodermata, Ophiuroidea) do possess this amazing capability of light production and are used as a study model in our laboratory. The present work investigates nervous control mechanisms of light emission at different levels such as extrinsic and intrinsic controls in three ophiuroid species: Amphiura ...

Vanderlinden, Christine

2006-01-01

336

A Novel Bioluminescent Protease Assay Using Engineered Firefly Luciferase  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Proteases play important roles in a variety of disease processes. Understanding their biological functions underpins the efforts of drug discovery. We have developed a bioluminescent protease assay using a circularly permuted form of firefly luciferase, wherein the native enzyme termini were joined by a peptide containing a protease site of interest. Protease cleavage of these mutant luciferases greatly activates the enzyme, typically over 100 fold. The mutant luciferase substrates are easily...

Wigdal, Susan S.; Anderson, Jessica L.; Vidugiris, Gediminas J.; Shultz, John; Wood, Keith V.; Fan, Frank

2008-01-01

337

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum  

Digital Repository Infrastructure Vision for European Research (DRIVER)

4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system is valuable for the pre-clinical in vivo evaluation of putative antitumor compounds.

Lim, Ed; Modi, Kshitij D.; Kim, Jaebeom

2009-01-01

338

Time-resolved multiphoton imaging of basal cell carcinoma  

Science.gov (United States)

We investigated human cutaneous basal cell carcinoma ex-vivo samples by combined time resolved two photon intrinsic fluorescence and second harmonic generation microscopy. Morphological and spectroscopic differences were found between malignant skin and corresponding healthy skin tissues. In comparison with normal healthy skin, cancer tissue showed a different morphology and a mean fluorescence lifetime distribution slightly shifted towards higher values. Topical application of delta-aminolevulinic acid to the lesion four hours before excision resulted in an enhancement of the fluorescence signal arising from malignant tissue, due to the accumulation of protoporphyrines inside tumor cells. Contrast enhancement was prevalent at tumor borders by both two photon fluorescence microscopy and fluorescence lifetime imaging. Fluorescence-based images showed a good correlation with conventional histopathological analysis, thereby supporting the diagnostic accuracy of this novel method. Combined morphological and lifetime analysis in the study of ex-vivo skin samples discriminated benign from malignant tissues, thus offering a reliable, non-invasive tool for the in-vivo analysis of inflammatory and neoplastic skin lesions.

Cicchi, R.; Sestini, S.; De Giorgi, V.; Stambouli, D.; Carli, P.; Massi, D.; Pavone, F. S.

2007-03-01

339

Application of time reversal acoustics focusing for nonlinear imaging ms  

Science.gov (United States)

Time reversal acoustic (TRA) focusing of ultrasound appears to be an effective tool for nonlinear imaging in industrial and medical applications because of its ability to efficiently concentrate ultrasonic energy (close to diffraction limit) in heterogeneous media. In this study, we used two TRA systems to focus ultrasonic beams with different frequencies in coinciding focal points, thus causing the generation of ultrasonic waves with combination frequencies. Measurements of the intensity of these combination frequency waves provide information on the nonlinear parameter of medium in the focal region. Synchronized stirring of two TRA focused beams enables obtaining 3-D acoustic nonlinearity images of the object. Each of the TRA systems employed an aluminum resonator with piezotransducers glued to its facet. One of the free facets of each resonator was submerged into a water tank and served as a virtual phased array capable of ultrasound focusing and beam steering. To mimic a medium with spatially varying acoustical nonlinearity a simplest model such as a microbubble column in water was used. Microbubbles were generated by electrolysis of water using a needle electrode. An order of magnitude increase of the sum frequency component was observed when the ultrasound beams were focused in the area with bubbles.

Sarvazyan, Armen; Sutin, Alexander

2001-05-01

340

Adaptive smoothing in real-time image stabilization  

Science.gov (United States)

When using the conventional fixed smoothing factor to display the stabilized video, we have the issue of large undefined black border regions (BBR) when camera is fast panning and zooming. To minimize the size of BBR and also provide smooth visualization to the display, this paper discusses several novel methods that have demonstrated on a real-time platform. These methods include an IIR filter, a single Kalman filter and an interactive multi-model filter. The fundamentals of these methods are to adapt the smoothing factor to the motion change from time to time to ensure small BBR and least jitters. To further remove the residual BBR, the pixels inside the BBR are composited from the previous frames. To do that, we first store the previous images and their corresponding frame-to-frame (F2F) motions in a FIFO queue, and then start filling the black pixels from valid pixels in the nearest neighbor frame based on the F2F motion. If a matching is found, then the search is stopped and continues to the next pixel. If the search is exhausted, the pixel remains black. These algorithms have been implemented and tested in a TI DM6437 processor.

Wu, Shunguang; Zhang, David C.; Zhang, Yuzheng; Basso, James; Melle, Michael

2012-05-01

 
 
 
 
341

Chern numbers hiding in time-of-flight images  

Energy Technology Data Exchange (ETDEWEB)

We present a technique for detecting topological invariants--Chern numbers--from time-of-flight images of ultracold atoms. We show that the Chern numbers of integer quantum Hall states of lattice fermions leave their fingerprints in the atoms' momentum distribution. We analytically demonstrate that the number of local maxima in the momentum distribution is equal to the Chern number in two limiting cases, for large hopping anisotropy and in the continuum limit. In addition, our numerical simulations beyond these two limits show that these local maxima persist for a range of parameters. Thus, an everyday observable in cold atom experiments can serve as a useful tool to characterize and visualize quantum states with nontrivial topology.

Zhao Erhai; Satija, Indubala I. [School of Physics, Astronomy, and Computational Sciences, George Mason University, Fairfax, Virginia 22030 (United States); Joint Quantum Institute, National Institute of Standards and Technology and University of Maryland, Gaithersburg, Maryland 20899 (United States); Bray-Ali, Noah; Williams, Carl J.; Spielman, I. B. [Joint Quantum Institute, National Institute of Standards and Technology and University of Maryland, Gaithersburg, Maryland 20899 (United States)

2011-12-15

342

Digital image processing for real-time neutron radiography and its applications  

International Nuclear Information System (INIS)

The present paper describes several digital image processing approaches for the real-time neutron radiography (neutron television-NTV), such as image integration, adaptive smoothing and image enhancement, which have beneficial effects on image improvements, and also describes how to use these techniques for applications. Details invisible in direct images of NTV are able to be revealed by digital image processing, such as reversed image, gray level correction, gray scale transformation, contoured image, subtraction technique, pseudo color display and so on. For real-time application a contouring operation and an averaging approach can also be utilized effectively. (author)

1989-03-01

343

Radiofrequency transmission line for bioluminescent Vibrio sp. irradiation  

Science.gov (United States)

We present the study and the analyses of a transmission line for radiofrequency (RF) irradiation of bacteria belonging to Vibrio harveyi-related strain PS1, a bioluminescent bacterium living in symbiosis with many marine organisms. The bioluminescence represents a new biologic indicator which is useful for studying the behaviour of living samples in the presence of RF waves due to the modern communication systems. A suitable transmission line, used as an irradiating cell and tested up to the maximum frequency used by the global system for mobile communications and universal mobile telecommunications system transmissions, was characterized. In this experiment, the RF voltage applied to the transmission line was 1 V. Due to short dimensions of the line and the applied high frequencies, standing waves were produced in addition to progressing waves and the electric field strength varies particularly along the longitudinal direction. The magnetic field map was not strongly linked to the electric one due to the presence of standing waves and of the outgoing irradiation. RF fields were measured by two homemade suitable probes able to diagnostic fields of high frequency. The field measurements were performed without any specimens inside the line. Being our sample made of living matter, the real field was modified and its value was estimated by a simulation code. The bioluminescence experiments were performed only at 900 MHz for two different measured electric fields, 53 and 140 V/m. The light emission was measured right from the beginning and after 7 and 25 h. Under RF irradiation, we found that the bioluminescence activity decreased. Compared with the control sample, the diminution was 6.8% and 44% after 7 and 25 h of irradiation, respectively, both with the low or high field. No changes of the survival factor for all the samples were observed. Besides, to understand the emission processes, we operated the deconvolution of the spectra by two Gaussian curves. The Gaussian peaks were approximately centered at 460 nm and 490 nm. The 490 nm peak was higher than the control one. Under RF, the 490 nm peak decreased compared to the 460 nm one. The decreasing was stronger for the sample in the higher field. The ratio of the emission area of the 490 nm to 460 nm was 5 for the control sample. It decreased up to 1.6 for the samples under RF. The bioluminescence improves the DNA repair by photoreactivation, and there is evidence that photolyase is preferentially activated by blue/violet light. Our finding suggests that RF exposure may stimulate DNA repair by shifting the emission spectra from blue/green (490 nm) to blue/violet (460 nm).

Nassisi, V.; Alifano, P.; Talŕ, A.; Velardi, L.

2012-07-01

344

BLProt: prediction of bioluminescent proteins based on support vector machine and relieff feature selection  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Bioluminescence is a process in which light is emitted by a living organism. Most creatures that emit light are sea creatures, but some insects, plants, fungi etc, also emit light. The biotechnological application of bioluminescence has become routine and is considered essential for many medical and general technological advances. Identification of bioluminescent proteins is more challenging due to their poor similarity in sequence. So far, no specific method has been reported to identify bioluminescent proteins from primary sequence. Results In this paper, we propose a novel predictive method that uses a Support Vector Machine (SVM and physicochemical properties to predict bioluminescent proteins. BLProt was trained using a dataset consisting of 300 bioluminescent proteins and 300 non-bioluminescent proteins, and evaluated by an independent set of 141 bioluminescent proteins and 18202 non-bioluminescent proteins. To identify the most prominent features, we carried out feature selection with three different filter approaches, ReliefF, infogain, and mRMR. We selected five different feature subsets by decreasing the number of features, and the performance of each feature subset was evaluated. Conclusion BLProt achieves 80% accuracy from training (5 fold cross-validations and 80.06% accuracy from testing. The performance of BLProt was compared with BLAST and HMM. High prediction accuracy and successful prediction of hypothetical proteins suggests that BLProt can be a useful approach to identify bioluminescent proteins from sequence information, irrespective of their sequence similarity. The BLProt software is available at http://www.inb.uni-luebeck.de/tools-demos/bioluminescent%20protein/BLProt

Hazrati Mehrnaz

2011-08-01

345

Evaluation of full time and half time acquired cardiac perfusion images and its correlation with coronary angiography  

International Nuclear Information System (INIS)

Full text: The myocardial perfusion study takes a longer time to complete. A reduction in acquisition time would mean reduced patient motion related artifacts, improvement in camera efficiency and reduction in cost. Iterative reconstruction algorithms produce more accurate images with fewer artifacts. Materials and Methods: Seventy three patients undergoing myocardial perfusion imaging were selected for additional half time acquisition. Patients with suspected or known coronary artery disease who have undergone coronary angiography recently were preferably included. Images were analysed in 4 groups - full time FBP, half time FBP, half time OSEM and half time OSEM. Three independent observers blinded to the clinical data and the acquisition protocol analysed images for change in image quality between these groups. Semiquantitative parameters of summed stress score, summed rest score, summed difference score and left ventricular ejection fraction were also compared using appropriate statistical methods. Results: No difference was noted in SSS, SRS, SDS and LVEF calculated for full time and half time. However, significant difference was found between SSS, SRS and SDS calculated for FBP and OSEM processed half time studies and no significant difference for LVEF calculated for these two groups. Significant change in image quality was noted by 2 observers only in 1.4% and 2.7% of cases. A true positivity rate of 88% was seen in comparison with coronary angiography. Conclusion: Gated myocardial perfusion SPECT images acquired in half the routine scan time provides equal diagnostic information compared to a conventional full time study, regardless of the processing protocol

2010-01-01

346

Forming rotated SAR images by real-time motion compensation.  

Energy Technology Data Exchange (ETDEWEB)

Proper waveform parameter selection allows collecting Synthetic Aperture Radar (SAR) phase history data on a rotated grid in the Fourier Space of the scene being imaged. Subsequent image formation preserves the rotated geometry to allow SAR images to be formed at arbitrary rotation angles without the use of computationally expensive interpolation or resampling operations. This should be useful where control of image orientation is desired such as generating squinted stripmaps and VideoSAR applications, among others.

Doerry, Armin Walter

2012-12-01

347

A Remote Laboratory for Real-Time Digital Image Processing on Embedded Systems  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The purpose of this paper is to present a Remote Laboratory on embedded systems focused in real-time digital image processing. This laboratory consists of a Main Web Server and several Workstations which are designed for digital image retrieval from a CMOS Image Sensor and real-time image processing on a Digital Signal Processor development platform. The Main Web Server redirects the authorised remote users to available Workstations in order to execute and verify their image processing algori...

Athanasios Kalantzopoulos; Dimitrios Markonis; Evangelos Zigouris

2009-01-01

348

Real-time image processing of TOF range images using a reconfigurable processor system  

Science.gov (United States)

During the last years, Time-of-Flight sensors achieved a significant impact onto research fields in machine vision. In comparison to stereo vision system and laser range scanners they combine the advantages of active sensors providing accurate distance measurements and camera-based systems recording a 2D matrix at a high frame rate. Moreover low cost 3D imaging has the potential to open a wide field of additional applications and solutions in markets like consumer electronics, multimedia, digital photography, robotics and medical technologies. This paper focuses on the currently implemented 4-phase-shift algorithm in this type of sensors. The most time critical operation of the phase-shift algorithm is the arctangent function. In this paper a novel hardware implementation of the arctangent function using a reconfigurable processor system is presented and benchmarked against the state-of-the-art CORDIC arctangent algorithm. Experimental results show that the proposed algorithm is well suited for real-time processing of the range images of TOF cameras.

Hussmann, S.; Knoll, F.; Edeler, T.

2011-06-01

349

Microwave Imaging for Breast Cancer Detection : Comparison of Tomographic Imaging Algorithms using Single-Frequency and Time-Domain Data  

DEFF Research Database (Denmark)

Still more research groups are promoting microwave imaging as a viable supplement or substitution to more conventional imaging modalities. A widespread approach for microwave imaging of the breast is tomographic imaging in which one seeks to reconstruct the distributions of permittivity and conductivity in the breast. In this paper two nonlinear tomographic algorithms are compared â?? one is a single-frequency algorithm and the other is a time-domain algorithm.

Rubæk, Tonny; Fhager, Andreas

2011-01-01

350

Real-time image-processing algorithm for markerless tumour tracking using X-ray fluoroscopic imaging.  

Science.gov (United States)

Objective: To ensure accuracy in respiratory-gating treatment, X-ray fluoroscopic imaging is used to detect tumour position in real time. Detection accuracy is strongly dependent on image quality, particularly positional differences between the patient and treatment couch. We developed a new algorithm to improve the quality of images obtained in X-ray fluoroscopic imaging and report the preliminary results. Methods: Two oblique X-ray fluoroscopic images were acquired using a dynamic flat panel detector (DFPD) for two patients with lung cancer. The weighting factor was applied to the DFPD image in respective columns, because most anatomical structures, as well as the treatment couch and port cover edge, were aligned in the superior-inferior direction when the patient lay on the treatment couch. The weighting factors for the respective columns were varied until the standard deviation of the pixel values within the image region was minimized. Once the weighting factors were calculated, the quality of the DFPD image was improved by applying the factors to multiframe images. Results: Applying the image-processing algorithm produced substantial improvement in the quality of images, and the image contrast was increased. The treatment couch and irradiation port edge, which were not related to a patient's position, were removed. The average image-processing time was 1.1?ms, showing that this fast image processing can be applied to real-time tumour-tracking systems. Conclusion: These findings indicate that this image-processing algorithm improves the image quality in patients with lung cancer and successfully removes objects not related to the patient. Advances in knowledge: Our image-processing algorithm might be useful in improving gated-treatment accuracy. PMID:24661056

Mori, S

2014-05-01

351

A new, rapid and sensitive bioluminescence assay for drug screening on Leishmania.  

Science.gov (United States)

We validated a new method, based on luciferine/luciferase bioluminescence, for drug screening on promastigotes of different Leishmania species. Results obtained with this new, rapid, reproducible, and reliable method are in good accordance with results obtained by the conventional MTT assay. This bioluminescence assay has a lower detection limit. PMID:24055386

Paloque, Lucie; Vidal, Nicolas; Casanova, Magali; Dumčtre, Aurélien; Verhaeghe, Pierre; Parzy, Daniel; Azas, Nadine

2013-12-01

352

Bioluminescent nanosensors for protease detection based upon gold nanoparticle-luciferase conjugatesw†  

Science.gov (United States)

This communication reports the use of click chemistry to site-specifically conjugate bioluminescent Renilla luciferase proteins to gold nanoparticles (Au NPs) for sensing protease activity. The bioluminescent emission from luciferase was efficiently quenched by Au NPs, but significantly recovered after the proteolytic cleavage.

Kim, Young-Pil; Daniel, Weston L.; Xia, Zuyong; Xie, Hexin; Mirkin, Chad A.; Rao, Jianghong

2014-01-01

353

Monitoring Therapeutic Treatments against Burkholderia Infections Using Imaging Techniques  

Directory of Open Access Journals (Sweden)

Full Text Available Burkholderia mallei, the etiologic agent of glanders, are Category B select agents with biothreat potential, and yet effective therapeutic treatments are lacking. In this study, we showed that CpG administration increased survival, demonstrating protection in the murine glanders model. Bacterial recovery from infected lungs, liver and spleen was significantly reduced in CpG-treated animals as compared with non-treated mice. Reciprocally, lungs of CpG-treated infected animals were infiltrated with higher levels of neutrophils and inflammatory monocytes, as compared to control animals. Employing the B. mallei bioluminescent strain CSM001 and the Neutrophil-Specific Fluorescent Imaging Agent, bacterial dissemination and neutrophil trafficking were monitored in real-time using multimodal in vivo whole body imaging techniques. CpG-treatment increased recruitment of neutrophils to the lungs and reduced bioluminescent bacteria, correlating with decreased bacterial burden and increased protection against acute murine glanders. Our results indicate that protection of CpG-treated animals was associated with recruitment of neutrophils prior to infection and demonstrated, for the first time, simultaneous real time in vivo imaging of neutrophils and bacteria. This study provides experimental evidence supporting the importance of incorporating optimized in vivo imaging methods to monitor disease progression and to evaluate the efficacy of therapeutic treatment during bacterial infections.

Tiffany M. Mott

2013-05-01

354

Real-time image acquisition and deblurring for underwater gravel extraction by smartphone  

Directory of Open Access Journals (Sweden)

Full Text Available Gravel size distribution is an important aspect of stream investigation. Using water photography to determine such distribution is a simple and cost-effective approach for gathering instream gravel information. However, good-quality images of underwater gravels in shallow areas are difficult to acquire because of the flow- and wind-induced perturbation at water surface. Thus, two Lucy–Richardson iterations are applied on an averaged image to obtain a deblurred image for gravel extraction. A Matlab code for multi-frame image averaging and image deblurring is implemented on a laptop computer. Underwater gravel images are acquired using a video camera and processed offline. Thus, the usability of the images acquired during field investigation cannot be determined immediately. However, returning to the investigated streams for additional data gathering would be costly, and the cameras may accidentally be dropped into the water. This paper presents multi-frame image averaging and image deblurring smartphone-based approaches for underwater gravel extraction. A waterproof smartphone is used to acquire the images, on which image deblurring is immediately conducted to test whether the images can be used for gravel extraction. The averaged image of using mean-based filter is derived during real-time image acquisition. The deblurred image is derived block-by-block because of limited memory capacity of smartphones. The time consumed for acquiring 1500 frame images with size of 1280 × 720 pixels is approximately 6 min by Sony Xperia smartphones. Image averaging can be performed in real time during image acquisition. Image deblurring is accomplished accurately and is consistent with results of the Matlab code. The processing time for image deblurring is approximately 12 min. A compact system for underwater gravel investigation using smartphones is successfully developed in this study. Image acquisition and deblurring are completed in real time at the investigated fields. Thus, we can immediately test whether the acquired images are usable for gravel extraction, thereby improving investigation efficiency significantly. 

Ming-Fu Chen

2014-02-01

355

Interpreting response time effects in functional imaging studies.  

Science.gov (United States)

It has been suggested that differential neural activity in imaging studies is most informative if it is independent of response time (RT) differences. However, others view RT as a behavioural index of key cognitive processes, which is likely linked to underlying neural activity. Here, we reconcile these views using the effort and engagement framework developed by Taylor, Rastle, and Davis (2013) and data from the domain of reading aloud. We propose that differences in neural engagement should be independent of RT, whereas, differences in neural effort should co-vary with RT. We illustrate these different mechanisms using data from an fMRI study of neural activity during reading aloud of regular words, irregular words, and pseudowords. In line with our proposals, activation revealed by contrasts designed to tap differences in neural engagement (e.g., words are meaningful and therefore engage semantic representations more than pseudowords) survived correction for RT, whereas activation for contrasts designed to tap differences in neural effort (e.g., it is more difficult to generate the pronunciation of pseudowords than words) correlated with RT. However, even for contrasts designed to tap neural effort, activity remained after factoring out the RT-BOLD response correlation. This may reveal unpredicted differences in neural engagement (e.g., learning phonological forms for pseudowords>words) that could further the development of cognitive models of reading aloud. Our framework provides a theoretically well-grounded and easily implemented method for analysing and interpreting RT effects in neuroimaging studies of cognitive processes. PMID:24904992

Taylor, J S H; Rastle, Kathleen; Davis, Matthew H

2014-10-01

356

Real time neutron reflectometry using neutron optical imaging  

International Nuclear Information System (INIS)

We will describe recent improvements to the SPEAR reflectometer at the Manuel Lujan Jr. Neutron Scattering Center at Los Alamos. One of the changes consists of wider convergent, incident-beam, collimation to take advantage of optical imaging for specular scattering. In addition, the instrument now views a partially coupled liquid hydrogen moderator as opposed to the decoupled moderator that was previous in-place. While the wavelength distribution is poorer, it matches the time (wavelength) resolution of the reflectometer more closely with the angular resolution. Since the integrated intensity of the partially coupled moderator is higher than the decoupled moderator, we show a similar gain in incident beam flux on the sample without loss of the ability to separate fringes. The increases in intensity from the moderator gain and the improved collimation combine to allow us to measure reflectivities with good statistics down to 10"-"4 in a matter of minutes and reflectivities of 10"-"6 in an hour. Examples of measurements showing the gain in data accumulation rates are presented. (author)

2001-03-01

357

Enhanced firefly bioluminescent assay of adenosine 5'-triphosphate using liposomes containing cationic cholesterols.  

Science.gov (United States)

Cationic liposomes composed of phosphatidylcholine and cationic cholesterols were prepared by extrusion technique. Dimethylaminoethyl-carbamoyl cholesterol (DMAE-chol) and diethylaminoethyl-carbamoyl cholesterol (DEAE-chol) were synthesized as a cationic cholesterol. Cationic liposomes containing DMAE-chol and DEAE-chol enhanced the intensity of maximum light emission from the firefly bioluminescent (BL) reaction. The sensitivity for ATP in the presence of cationic liposomes containing DMAE-chol and DEAE-chol was improved by a factor of 10 times compared to that in water alone. The detection limit for ATP upon using cationic liposomes was 1.0 pmol/L in an aqueous standard solution. The BL enhancement in the presence of cationic liposomes could be explained in terms of BL emitters and electrostatic interaction between the liposomes surface and BL reactants. PMID:11512141

Nakata, N; Kamidate, T

2001-01-01

358

Real-time 3D image registration for functional MRI.  

Science.gov (United States)

Subject head movements are one of the main practical difficulties with brain functional MRI. A fast, accurate method for rotating and shifting a three-dimensional (3D) image using a shear factorization of the rotation matrix is described. Combined with gradient descent (repeated linearization) on a least squares objective function, 3D image realignment for small movements can be computed as rapidly as whole brain images can be acquired on current scanners. Magn Reson Med 42:1014-1018, 1999. PMID:10571921

Cox, R W; Jesmanowicz, A

1999-12-01

359

Time-resolved image analysis for turbulent flows:  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Classical Particle Image Velocimetry (PIV) uses two representations of the particle image distribution to determine the displacement of the particle image pattern by spatial cross-correlation. The accuracy and the robustness are however limited by the fact that only two representations at t and t +?t are present. Thus, only a first order approximation of the velocity can be estimated. To enhance the precision in estimating the flow velocity, multi-pulse or multi-frame techniques were already...

Ka?hler, C. J.; Cierpka, C.; Scharnowski, S.; Manhart, M.; Sciacchitano, A.; Lynch, K.; Scarano, F.; Wieneke, B.; Willert, C.; Jeon, Y. J.; Chatellier, L.; Augereau, L.; Tremblais, B.; David, L.

2013-01-01

360

Time-resolved image analysis for turbulent flows Conference paper  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Classical Particle Image Velocimetry (PIV) uses two representations of the particle image distribution to determine the displacement of the particle image pattern by spatial cross-correlation. The accuracy and the robustness are however limited by the fact that only two representations at t and t +?t are present. Thus, only a first order approximation of the velocity can be estimated. To enhance the precision in estimating the flow velocity, multi-pulse or multi-frame techniques were already...

Ka?hler, Christian; Cierpka, Christian; Scharnowski, Sven; Manhart, Michael; Sciacchitano, Andrea; Lynch, Kyle; Scarano, Fulvio; Wieneke, Bernhard; Willert, Christian; Jeon, Youn; Chatellier, Ludovic; Augereau, Bertrand; Tremblais, Benoit; David, Laurent

2013-01-01

 
 
 
 
361

When should we recommend use of dual time-point and delayed time-point imaging techniques in FDG PET?  

Energy Technology Data Exchange (ETDEWEB)

FDG PET and PET/CT are now widely used in oncological imaging for tumor characterization, staging, restaging, and response evaluation. However, numerous benign etiologies may cause increased FDG uptake indistinguishable from that of malignancy. Multiple studies have shown that dual time-point imaging (DTPI) of FDG PET may be helpful in differentiating malignancy from benign processes. However, exceptions exist, and some studies have demonstrated significant overlap of FDG uptake patterns between benign and malignant lesions on delayed time-point images. In this review, we summarize our experience and opinions on the value of DTPI and delayed time-point imaging in oncology, with a review of the relevant literature. We believe that the major value of DTPI and delayed time-point imaging is the increased sensitivity due to continued clearance of background activity and continued FDG accumulation in malignant lesions, if the same diagnostic criteria (as in the initial standard single time-point imaging) are used. The specificity of DTPI and delayed time-point imaging depends on multiple factors, including the prevalence of malignancies, the patient population, and the cut-off values (either SUV or retention index) used to define a malignancy. Thus, DTPI and delayed time-point imaging would be more useful if performed for evaluation of lesions in regions with significant background activity clearance over time (such as the liver, the spleen, the mediastinum), and if used in the evaluation of the extent of tumor involvement rather than in the characterization of the nature of any specific lesion. Acute infectious and non-infectious inflammatory lesions remain as the major culprit for diminished diagnostic performance of these approaches (especially in tuberculosis-endemic regions). Tumor heterogeneity may also contribute to inconsistent performance of DTPI. The authors believe that selective use of DTPI and delayed time-point imaging will improve diagnostic accuracy and interpretation confidence in FDG PET imaging. (orig.)

Cheng, Gang [Philadelphia VA Medical Center, Department of Radiology, Philadelphia, PA (United States); Hospital of the University of Pennsylvania, Department of Radiology, Philadelphia, PA (United States); Torigian, Drew A.; Alavi, Abass [Hospital of the University of Pennsylvania, Department of Radiology, Philadelphia, PA (United States); Zhuang, Hongming [Children' s Hospital of Philadelphia, Department of Radiology, Philadelphia, PA (United States)

2013-05-15

362

A VLSI Processor Design of Real-Time Data Compression for High-Resolution Imaging Radar  

Science.gov (United States)

For the high-resolution imaging radar systems, real-time data compression of raw imaging data is required to accomplish the science requirements and satisfy the given communication and storage constraints. The Block Adaptive Quantizer (BAQ) algorithm and its associated VLSI processor design have been developed to provide a real-time data compressor for high-resolution imaging radar systems.

Fang, W.

1994-01-01

363

Real Time Deconvolution of In-Vivo Ultrasound Images  

DEFF Research Database (Denmark)

The axial resolution in medical ultrasound is directly linked to the emitted ultrasound frequency, which, due to tissue attenuation, is selected based on the depth of scanning. The resolution is etermined by the transducers impulse response, which limits the attainable resolution to be between one and two wavelengths. This can be improved by deconvolution, which increase the bandwidth and equalizes the phase to increase resolution under the constraint of the electronic noise in the received signal. A fixed interval Kalman filter based deconvolution routine written in C is employed. It uses a state based model for the ultrasound pulse and can include a depth varying pulse and spatially varying signal-to-noise ration. An autoregressive moving average (ARMA) model of orders 8 and 9 is used for the pulse, and the ARMA parameters are determined as a function of depth using a minimum variance algorithm using averaging over several RF lines. In vivo data from a 3 MHz mechanically rotating probe is used and the received signal is sampled at 20 MHz and 12 bits. In-vivo data acquired from a 16th week old fetus is used along with a scan from the liver and right kidney of a 27 years old male. The axial resolution has been determined from the in-vivo liver image using the auto-covariance function. From the envelope of the estimated pulse the axial resolution at Full-Width-Half-Max is 0.581 mm corresponding to 1.13 l at 3 MHz. The algorithm increases the resolution to 0.116 mm or 0.227 l corresponding to a factor of 5.1. The basic pulse can be estimated in roughly 0.176 seconds on a single CPU core on an Intel i5 CPU running at 1.8 GHz. An in-vivo image consisting of 100 lines of 1600 samples can be processed in roughly 0.1 seconds making it possible to perform real-time deconvolution on ultrasound data by using dual or quad core CPUs for frame-rates of 20-40 Hz.

Jensen, Jørgen Arendt

2013-01-01

364

Density resolutionary optimization of real time radiotherapy portal imagings  

International Nuclear Information System (INIS)

Objective: Electronic portal imaging devices (EPIDs) are widely used as a replacement of portal films for patient position verification, but the image quality is not always optimal. Because of very low density resolution, the portal imaging is difficult to be used clinically. In this study, several transforming models and the optimization exposure or acquisition conditions were studied for optimization portal imaging, which based on DicomRT platform built by ourselves. Methods: 6 MV X-ray from Varian 21EX linac was used to generate portal images by Portal Vision aSi500 amorphous silicon detector image acquisition system. The density resolution study was based on the number of the lines which could be seen in the image of a special Las Vegas image quality test board. The optimization calculating models were focused on equalization after stretch transforming discrete wavelet transform (DWT) and Butter worth high pass filters. The calculation was performed in Matlab language. Results: The optimal numbers of MU, average frames and reset number were 4 - 5, 3 - 4 and 2 - 3, respectively. The density resolution of optimized imaging via equalization after stretch transforming, DWT and Butter worth high pass filter transforming was markedly improved. The bone structure could be definitely distinguished. The number of lines distinguished in Las Vegas image via equalization after stretch transforming, DWT and Better worth high pass filter transforming was 3, 4 and 5, respectively. Conclusions: The proposed transforming systems, including DWT edge detection and Butter worth high pass filter transform, are suitable for improving density resolving power of MV X-ray portal image. (authors)

2010-03-01

365

Inhibitory effect of lipoic acid on firefly luciferase bioluminescence  

International Nuclear Information System (INIS)

Lipoic acid was found to inhibit the firefly luciferin-luciferase reaction. The inhibition is competitive and is the strongest known (Ki 0.026 ± 0.013 ?M) compared with other reported inhibitors. Considering the structure-activity correlations, the mechanism of inhibition may originate from the sulfur atom and carboxyl moiety of lipoic acid giving it structural specificity. Subsequent addition of lipoic acid and nitric oxide accelerated the inhibition in vitro, suggesting that lipoic acid may have a functional role in regulating firefly bioluminescence

2004-10-15

366

Toxicity assessment of Hanford Site wastes by bacterial bioluminescence  

International Nuclear Information System (INIS)

This paper examines the toxicity of the nonradioactive component of low-level wastes stored in tanks on the Hanford reservation. The use of a faster, cheaper bioassay to replace the 96 hour fish acute toxicity test is examined. The new bioassay is based on loss of bioluminescence of Photobacter phosphoreum (commonly called Microtox) following exposure to toxic materials. This bioassay is calibrated and compares well to the standard fish acute toxicity test for characterization of Hanford Wastes. 4 refs., 11 figs., 11 tabs

1991-10-06

367

Luciferase-dependent oxygen consumption by bioluminescent vibrios  

Energy Technology Data Exchange (ETDEWEB)

Oxygen uptake due to luciferase in two luminous Vibrio species was estimated in vivo by utilizing inhibitors having specificities for luciferase (decanol) and cytochromes (cyanide). Cyanide titration of respiration revealed a component of oxygen uptake less sensitive to cyanide which was completely inhibitable by low concentrations of decanol. From this it was estimated that in vivo luciferase is responsible for less than 12% (Vibrio harveyi) or 20% (Vibrio fischeri) of the total respiration. From these data in vivo bioluminescent quantum yields are estimated to be not lower than 1.7 and 2.6%, respectively.

Makemson, J.C.

1986-02-01

368

Bioluminescęncia de fungos: distribuiçăo, funçăo e mecanismo de emissăo de luz Fungi bioluminescence: distribution, function and mechanism of light emission  

Directory of Open Access Journals (Sweden)

Full Text Available The emission of light by living organisms, bioluminescence, has been studied since the nineteenth century. However, some bioluminescent systems, such as fungi, remain poorly understood. The emitter, the two enzymes involved, and the reaction mechanism have not yet been unraveled. Moreover, the ecological role and evolutionary significance for fungal luminescence is also unknown. It is hoped that comprehensive research on fungal bioluminescent systems will generate knowledge and tools for academic and applied sciences. This review discusses the distribution of bioluminescent fungi on Earth, attempts to elucidate the mechanism involved in light emission, and presents preliminary results on the evolution and ecological role of fungal