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Sample records for time bioluminescence imaging

  1. Optimisation of acquisition time in bioluminescence imaging

    Taylor, Shelley L.; Mason, Suzannah K. G.; Glinton, Sophie; Cobbold, Mark; Styles, Iain B.; Dehghani, Hamid

    2015-03-01

    Decreasing the acquisition time in bioluminescence imaging (BLI) and bioluminescence tomography (BLT) will enable animals to be imaged within the window of stable emission of the bioluminescent source, a higher imaging throughput and minimisation of the time which an animal is anaesthetised. This work investigates, through simulation using a heterogeneous mouse model, two methods of decreasing acquisition time: 1. Imaging at fewer wavelengths (a reduction from five to three); and 2. Increasing the bandwidth of filters used for imaging. The results indicate that both methods are viable ways of decreasing the acquisition time without a loss in quantitative accuracy. Importantly, when choosing imaging wavelengths, the spectral attenuation of tissue and emission spectrum of the source must be considered, in order to choose wavelengths at which a high signal can be achieved. Additionally, when increasing the bandwidth of the filters used for imaging, the bandwidth must be accounted for in the reconstruction algorithm.

  2. Bioluminescence Imaging

    Sadikot, Ruxana T.; Blackwell, Timothy S.

    2005-01-01

    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...

  3. Real-Time Bioluminescence Imaging of Nitroreductase in Mouse Model.

    Feng, Ping; Zhang, Huateng; Deng, Quankun; Liu, Wei; Yang, Linghui; Li, Guobo; Chen, Guo; Du, Lupei; Ke, Bowen; Li, Minyong

    2016-06-01

    Nitroreductase (NTR) is an endogenous reductase overexpressed in hypoxic tumors; however, its precise detection in living cells and animals remains a considerable challenge. Herein, we developed three reaction-based probes and a related bioluminescence assay for the real-time NTR detection. The high sensitivity and selectivity of probe 3, combined with its remarkable potential of bioluminescence imaging, affords a valuable approach for in vivo imaging of NTR in a tumor model mouse. PMID:27197544

  4. Bioanalytical Applications of Real-Time ATP Imaging Via Bioluminescence

    Jason Alan Gruenhagen

    2003-12-12

    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 Cd

  5. Bioluminescence microscopy using a short focal-length imaging lens

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

    2014-01-01

    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....

  6. Infection routes of Aeromonas salmonicida in rainbow trout monitored in vivo by real-time bioluminescence imaging

    Bartkova, Simona; Kokotovic, Branko; Dalsgaard, Inger

    2016-01-01

    Recent development of imaging tools has facilitated studies of pathogen infections in vivo in real time. This trend can be exemplified by advances in bioluminescence imaging (BLI), an approach that helps to visualize dissemination of pathogens within the same animal over several time points. Here...

  7. BIOLUMINESCENCE IMAGING: PROGRESS AND APPLICATIONS

    Badr, Christian E.; Tannous, Bakhos A

    2011-01-01

    Application of bioluminescence imaging has grown tremendously in the past decade and has significantly contributed to the core conceptual advances in biomedical research. This technology provides valuable means for monitoring of different biological processes for immunology, oncology, virology and neuroscience. In this review, we will discuss current trends in bioluminescence and its application in different fields with emphasis on cancer research.

  8. Real-time monitoring of cariogenic bacteria via bioluminescent imaging: A biodontic hypothesis

    Jafar Kolahi

    2016-01-01

    Full Text Available Introduction: Dental caries (tooth decay remains one of the most common chronic infectious disease in the world. Disclosure of camouflaged cariogenic bacteria will be a great motivation for better oral hygiene. The Hypothesis: At present, lux transposon cassette, Tn4001 luxABCDE Kmr, is available that could be used for stable bioluminescent transformation of a wide range of gram-positive bacteria, e.g. Streptococcus mutans and Lactobacillus. After this step, sensitive charge-coupled device (CCD camera could be used to detect the low levels of light emitted from bioluminescent cariogenic bacteria. Living imaging software would be used for analysis and three-dimensional (3D reconstruction of images. Evaluation of the Hypothesis: Entrance of transgenic organisms into the oral cavity should be done with great caution. Ethical consideration is necessary and primary animal studies are required. The main limitation of this technique will be oxygen. As mentioned previously, bioluminescent reactions need oxygen. Hence, bioluminescent imaging cannot be used for anaerobic bacteria, e.g., Streptococcus sobrinus.

  9. Bioluminescence imaging in live cells and animals.

    Tung, Jack K; Berglund, Ken; Gutekunst, Claire-Anne; Hochgeschwender, Ute; Gross, Robert E

    2016-04-01

    The use of bioluminescent reporters in neuroscience research continues to grow at a rapid pace as their applications and unique advantages over conventional fluorescent reporters become more appreciated. Here, we describe practical methods and principles for detecting and imaging bioluminescence from live cells and animals. We systematically tested various components of our conventional fluorescence microscope to optimize it for long-term bioluminescence imaging. High-resolution bioluminescence images from live neurons were obtained with our microscope setup, which could be continuously captured for several hours with no signs of phototoxicity. Bioluminescence from the mouse brain was also imaged noninvasively through the intact skull with a conventional luminescence imager. These methods demonstrate how bioluminescence can be routinely detected and measured from live cells and animals in a cost-effective way with common reagents and equipment. PMID:27226972

  10. Bioluminescence imaging characteristics and application

    Bioluminescence imaging (BLI) by luciferase gene marked cells or DNA, in the presence of ATP and oxygen, catalytic oxidation reaction of fluorescein luminescence. So that it can directly monitor in vivo cell activity and gene behavior. In this paper, by comparing the BLI and MRI, PET, radiography of the similarities and differences, as well as about their cancer, stem cells and immune cells transportation, apoptosis and other aspects of the application, in order to better provide the basis for promoting the application of BLI. (authors)

  11. Noninvasive Bioluminescence Imaging in Small Animals

    Zinn, Kurt R.; Chaudhuri, Tandra R.; Szafran, April Adams; O’Quinn, Darrell; Weaver, Casey; Dugger, Kari; Lamar, Dale; Kesterson, Robert A.; Wang, Xiangdong; Frank, Stuart J.

    2008-01-01

    There has been a rapid growth of bioluminescence imaging applications in small animal models in recent years, propelled by the availability of instruments, analysis software, reagents, and creative approaches to apply the technology in molecular imaging. Advantages include the sensitivity of the technique as well as its efficiency, relatively low cost, and versatility. Bioluminescence imaging is accomplished by sensitive detection of light emitted following chemical reaction of the luciferase...

  12. In vivo cell tracking with bioluminescence imaging

    Molecular imaging is a fast growing biomedical research that allows the visual representation, characterization and quantification of biological processes at the cellular and subcellular levels within intact living organisms. In vivo tracking of cells is an indispensable technology for development and optimization of cell therapy for replacement or renewal of damaged or diseased tissue using transplanted cells, often autologous cells. With outstanding advantages of bioluminescence imaging, the imaging approach is most commonly applied for in vivo monitoring of transplanted stem cells or immune cells in order to assess viability of administered cells with therapeutic efficacy in preclinical small animal models. In this review, a general overview of bioluminescence is provided and recent updates of in vivo cell tracking using the bioluminescence signal are discussed

  13. In vivo cell tracking with bioluminescence imaging

    Kim, Jung Eun; Kalimuthu, Senthilkumar; Ahn, Byeong Cheol [Dept. of Nuclear Medicine, Kyungpook National University School of Medicine and Hospital, Daegu (Korea, Republic of)

    2015-03-15

    Molecular imaging is a fast growing biomedical research that allows the visual representation, characterization and quantification of biological processes at the cellular and subcellular levels within intact living organisms. In vivo tracking of cells is an indispensable technology for development and optimization of cell therapy for replacement or renewal of damaged or diseased tissue using transplanted cells, often autologous cells. With outstanding advantages of bioluminescence imaging, the imaging approach is most commonly applied for in vivo monitoring of transplanted stem cells or immune cells in order to assess viability of administered cells with therapeutic efficacy in preclinical small animal models. In this review, a general overview of bioluminescence is provided and recent updates of in vivo cell tracking using the bioluminescence signal are discussed.

  14. Bioluminescence imaging of Chlamydia muridarum ascending infection in mice.

    Jessica Campbell

    Full Text Available Chlamydial pathogenicity in the upper genital tract relies on chlamydial ascending from the lower genital tract. To monitor chlamydial ascension, we engineered a luciferase-expressing C. muridarum. In cells infected with the luciferase-expressing C. muridarum, luciferase gene expression and enzymatic activity (measured as bioluminescence intensity correlated well along the infection course, suggesting that bioluminescence can be used for monitoring chlamydial replication. Following an intravaginal inoculation with the luciferase-expressing C. muridarum, 8 of 10 mice displayed bioluminescence signal in the lower with 4 also in the upper genital tracts on day 3 after infection. By day 7, all 10 mice developed bioluminescence signal in the upper genital tracts. The bioluminescence signal was maintained in the upper genital tract in 6 and 2 mice by days 14 and 21, respectively. The bioluminescence signal was no longer detectable in any of the mice by day 28. The whole body imaging approach also revealed an unexpected airway infection following the intravaginal inoculation. Although the concomitant airway infection was transient and did not significantly alter the genital tract infection time courses, caution should be taken during data interpretation. The above observations have demonstrated that C. muridarum can not only achieve rapid ascending infection in the genital tract but also cause airway infection following a genital tract inoculation. These findings have laid a foundation for further optimizing the C. muridarum intravaginal infection murine model for understanding chlamydial pathogenic mechanisms.

  15. Bioluminescence in vivo imaging of autoimmune encephalomyelitis predicts disease

    Steinman Lawrence

    2008-02-01

    Full Text Available Abstract Background Experimental autoimmune encephalomyelitis is a widely used animal model to understand not only multiple sclerosis but also basic principles of immunity. The disease is scored typically by observing signs of paralysis, which do not always correspond with pathological changes. Methods Experimental autoimmune encephalomyelitis was induced in transgenic mice expressing an injury responsive luciferase reporter in astrocytes (GFAP-luc. Bioluminescence in the brain and spinal cord was measured non-invasively in living mice. Mice were sacrificed at different time points to evaluate clinical and pathological changes. The correlation between bioluminescence and clinical and pathological EAE was statistically analyzed by Pearson correlation analysis. Results Bioluminescence from the brain and spinal cord correlates strongly with severity of clinical disease and a number of pathological changes in the brain in EAE. Bioluminescence at early time points also predicts severity of disease. Conclusion These results highlight the potential use of bioluminescence imaging to monitor neuroinflammation for rapid drug screening and immunological studies in EAE and suggest that similar approaches could be applied to other animal models of autoimmune and inflammatory disorders.

  16. Bioluminescent system for dynamic imaging of cell and animal behavior

    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.

  17. Bioluminescent system for dynamic imaging of cell and animal behavior

    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

    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.

  18. 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

    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.)

  19. In Vivo Bioluminescence Imaging of the Murine Pathogen Citrobacter rodentium

    Wiles, Siouxsie; Pickard, Karen M.; Peng, Katian; MacDonald, Thomas T.; Frankel, Gad

    2006-01-01

    Citrobacter rodentium is a natural mouse pathogen related to enteropathogenic and enterohemorrhagic Escherichia coli. We have previously utilized bioluminescence imaging (BLI) to determine the in vivo colonization dynamics of C. rodentium. However, due to the oxygen requirement of the bioluminescence system and the colonic localization of C. rodentium, in vivo localization studies were performed using harvested organs. Here, we report the detection of bioluminescent C. rodentium and commensal...

  20. Bioluminescence imaging of estrogen receptor activity during breast cancer progression.

    Vantaggiato, Cristina; Dell'Omo, Giulia; Ramachandran, Balaji; Manni, Isabella; Radaelli, Enrico; Scanziani, Eugenio; Piaggio, Giulia; Maggi, Adriana; Ciana, Paolo

    2016-01-01

    Estrogen receptors (ER) are known to play an important regulatory role in mammary gland development as well as in its neoplastic transformation. Although several studies highlighted the contribution of ER signaling in the breast transformation, little is known about the dynamics of ER state of activity during carcinogenesis due to the lack of appropriate models for measuring the extent of receptor signaling in time, in the same animal. To this aim, we have developed a reporter mouse model for the non-invasive in vivo imaging of ER activity: the ERE-Luc reporter mouse. ERE-Luc is a transgenic mouse generated with a firefly luciferase (Luc) reporter gene driven by a minimal promoter containing an estrogen responsive element (ERE). This model allows to measure receptor signaling in longitudinal studies by bioluminescence imaging (BLI). Here, we have induced sporadic mammary cancers by treating systemically ERE-Luc reporter mice with DMBA (9,10-dimethyl 1,2-benzanthracene) and measured receptor signaling by in vivo imaging in individual animals from early stage until a clinically palpable tumor appeared in the mouse breast. We showed that DMBA administration induces an increase of bioluminescence in the whole abdominal area 6 h after treatment, the signal rapidly disappears. Several weeks later, strong bioluminescence is observed in the area corresponding to the mammary glands. In vivo and ex vivo imaging analysis demonstrated that this bioluminescent signal is localized in the breast area undergoing neoplastic transformation. We conclude that this non-invasive assay is a novel relevant tool to identify the activation of the ER signaling prior the morphological detection of the neoplastic transformation. PMID:27069764

  1. Bioluminescence.

    Jones, M. Gail

    1993-01-01

    Describes bioluminescence and the chemistry of how it occurs. Presents information for conducting the following classroom activities: (1) firefly mimic; (2) modeling deep-sea fish; (3) sea fireflies; and (4) the chemistry of light. (PR)

  2. Bioluminescence: a versatile technique for imaging cellular and molecular features

    Paley, Miranda A.

    2016-01-01

    Bioluminescence is a ubiquitous imaging modality for visualizing biological processes in vivo. This technique employs visible light and interfaces readily with most cell and tissue types, making it a versatile technology for preclinical studies. Here we review basic bioluminescence imaging principles, along with applications of the technology that are relevant to the medicinal chemistry community. These include noninvasive cell tracking experiments, analyses of protein function, and methods to visualize small molecule metabolites. In each section, we also discuss how bioluminescent tools have revealed insights into experimental therapies and aided drug discovery. Last, we highlight the development of new bioluminescent tools that will enable more sensitive and multi-component imaging experiments and, thus, expand our broader understanding of living systems.

  3. Bioluminescence Imaging to Detect Late Stage Infection of African Trypanosomiasis.

    Burrell-Saward, Hollie; Ward, Theresa H

    2016-01-01

    Human African trypanosomiasis (HAT) is a multi-stage disease that manifests in two stages; an early blood stage and a late stage when the parasite invades the central nervous system (CNS). In vivo study of the late stage has been limited as traditional methodologies require the removal of the brain to determine the presence of the parasites. Bioluminescence imaging is a non-invasive, highly sensitive form of optical imaging that enables the visualization of a luciferase-transfected pathogen in real-time. By using a transfected trypanosome strain that has the ability to produce late stage disease in mice we are able to study the kinetics of a CNS infection in a single animal throughout the course of infection, as well as observe the movement and dissemination of a systemic infection. Here we describe a robust protocol to study CNS infections using a bioluminescence model of African trypanosomiasis, providing real time non-invasive observations which can be further analyzed with optional downstream approaches. PMID:27284970

  4. Interactive graphic editing tools in bioluminescent imaging simulation

    Li, Hui; Tian, Jie; Luo, Jie; Wang, Ge; Cong, Wenxiang

    2005-04-01

    It is a challenging task to accurately describe complicated biological tissues and bioluminescent sources in bioluminescent imaging simulation. Several graphic editing tools have been developed to efficiently model each part of the bioluminescent simulation environment and to interactively correct or improve the initial models of anatomical structures or bioluminescent sources. There are two major types of graphic editing tools: non-interactive tools and interactive tools. Geometric building blocks (i.e. regular geometric graphics and superquadrics) are applied as non-interactive tools. To a certain extent, complicated anatomical structures and bioluminescent sources can be approximately modeled by combining a sufficient large number of geometric building blocks with Boolean operators. However, those models are too simple to describe the local features and fine changes in 2D/3D irregular contours. Therefore, interactive graphic editing tools have been developed to facilitate the local modifications of any initial surface model. With initial models composed of geometric building blocks, interactive spline mode is applied to conveniently perform dragging and compressing operations on 2D/3D local surface of biological tissues and bioluminescent sources inside the region/volume of interest. Several applications of the interactive graphic editing tools will be presented in this article.

  5. Continuous, real-time bioimaging of chemical bioavailability and toxicology using autonomously bioluminescent human cell lines

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

    2013-05-01

    Bioluminescent imaging is an emerging biomedical surveillance strategy that uses external cameras to detect in vivo light generated in small animal models of human physiology or in vitro light generated in tissue culture or tissue scaffold mimics of human anatomy. The most widely utilized of reporters is the firefly luciferase (luc) gene; however, it generates light only upon addition of a chemical substrate, thus only generating intermittent single time point data snapshots. To overcome this disadvantage, we have demonstrated substrate-independent bioluminescent imaging using an optimized bacterial bioluminescence (lux) system. The lux reporter produces bioluminescence autonomously using components found naturally within the cell, thereby allowing imaging to occur continuously and in real-time over the lifetime of the host. We have validated this technology in human cells with demonstrated chemical toxicological profiling against exotoxin exposures at signal strengths comparable to existing luc systems (~1.33 × 107 photons/second). As a proof-in-principle demonstration, we have engineered breast carcinoma cells to express bioluminescence for real-time screening of endocrine disrupting chemicals and validated detection of 17β-estradiol (EC50 = ~ 10 pM). These and other applications of this new reporter technology will be discussed as potential new pathways towards improved models of target chemical bioavailability, toxicology, efficacy, and human safety.

  6. Filtering and deconvolution for bioluminescence imaging of small animals

    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)

  7. The Expanding Toolbox of In Vivo Bioluminescent Imaging

    Xu, Tingting; Close, Dan; Handagama, Winode; Marr, Enolia; Sayler, Gary; Ripp, Steven

    2016-01-01

    In vivo bioluminescent imaging (BLI) permits the visualization of engineered bioluminescence from living cells and tissues to provide a unique perspective toward the understanding of biological processes as they occur within the framework of an authentic in vivo environment. The toolbox of in vivo BLI includes an inventory of luciferase compounds capable of generating bioluminescent light signals along with sophisticated and powerful instrumentation designed to detect and quantify these light signals non-invasively as they emit from the living subject. The information acquired reveals the dynamics of a wide range of biological functions that play key roles in the physiological and pathological control of disease and its therapeutic management. This mini review provides an overview of the tools and applications central to the evolution of in vivo BLI as a core technology in the preclinical imaging disciplines. PMID:27446798

  8. The Expanding Toolbox of In Vivo Bioluminescent Imaging.

    Xu, Tingting; Close, Dan; Handagama, Winode; Marr, Enolia; Sayler, Gary; Ripp, Steven

    2016-01-01

    In vivo bioluminescent imaging (BLI) permits the visualization of engineered bioluminescence from living cells and tissues to provide a unique perspective toward the understanding of biological processes as they occur within the framework of an authentic in vivo environment. The toolbox of in vivo BLI includes an inventory of luciferase compounds capable of generating bioluminescent light signals along with sophisticated and powerful instrumentation designed to detect and quantify these light signals non-invasively as they emit from the living subject. The information acquired reveals the dynamics of a wide range of biological functions that play key roles in the physiological and pathological control of disease and its therapeutic management. This mini review provides an overview of the tools and applications central to the evolution of in vivo BLI as a core technology in the preclinical imaging disciplines. PMID:27446798

  9. Space application research of EMCCDs for bioluminescence imaging

    Zhang, Tao

    The detection of bioluminescense is widely used on the ground, while the detection of bioluminescence in space is still at the stage of detecting bright bioluminescense. With the rapid development of research in Space Life Sciences, it will be necessary to develop a detection technology to detect weak bioluminescense. Compared to other low-light detection techniques for ground, there are more advantages of EMCCDs for space application. Build a space bioluminescence imaging detection system, analysis the feasibility and capability of its will be significant. Co-Author:Xie Zongbao,Zheng Weibo

  10. Computer-aided photometric analysis of dynamic digital bioluminescent images

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

    2003-04-01

    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.

  11. Development of Quantification Method for Bioluminescence Imaging

    Optical molecular luminescence imaging is widely used for detection and imaging of bio-photons emitted by luminescent luciferase activation. The measured photons in this method provide the degree of molecular alteration or cell numbers with the advantage of high signal-to-noise ratio. To extract useful information from the measured results, the analysis based on a proper quantification method is necessary. In this research, we propose a quantification method presenting linear response of measured light signal to measurement time. We detected the luminescence signal by using lab-made optical imaging equipment of animal light imaging system (ALIS) and different two kinds of light sources. One is three bacterial light-emitting sources containing different number of bacteria. The other is three different non-bacterial light sources emitting very weak light. By using the concept of the candela and the flux, we could derive simplified linear quantification formula. After experimentally measuring light intensity, the data was processed with the proposed quantification function. We could obtain linear response of photon counts to measurement time by applying the pre-determined quantification function. The ratio of the re-calculated photon counts and measurement time present a constant value although different light source was applied. The quantification function for linear response could be applicable to the standard quantification process. The proposed method could be used for the exact quantitative analysis in various light imaging equipment with presenting linear response behavior of constant light emitting sources to measurement time

  12. Infection with adenovirus-mediated luciferase reporter gene in mesenchymal stem cells and bioluminescence imaging

    Objective: To construct adenovirus vector containing firefly luciferase reporter gene (Ad-Luc) and infect bone marrow mesenchymal stem cells (BMSC), then to take bioluminescence imaging in vitro and in vivo for identification. Methods: The luciferase gene was amplified with PCR from psiCHECK-2 plasmid and cloned into the adenoviral shuttle vector (pShuttle-CMV). It was confirmed by Nhe Ⅰ/Xba Ⅰ digestion and sequencing. PShuttle-CMV-Luc and backbone vector (pAdeno) were homologous recombined. Then the recombinant plasmid was packaged in HEK293 cells and the virus titer was detected. The BMSC were infected by the recombinant adenovirus. The bioluminescence imaging in vitro was performed to determine the best multiplicity of infection (MOI), and the relationship between bioluminescence intensity and MOI was analyzed by curve fitting regression analysis. Viability was evaluated via Trypan blue staining. The transfected BMSC (1 × 106) were implanted into the muscles of forelimb of SD rats,and then tracked by bioluminescence imaging in vivo. Cell viability was compared using two-way repeated measures analysis of variance between groups. Results: Enzyme digestion and sequence analysis indicated that Ad-Luc was successfully constructed. The virus titer was 1 × 1010 plaque forming unit (PFU)/ml. The bioluminescence detection in vitro showed that Ad-Luc could infect BMSC high efficiently to express luciferase and the best MOI was 50. The bioluminescence intensity enhanced with increase of MOI (R2 =0.98). No statistically significant difference was found in cell viability between transfected and untransfected BMSC at 1, 3, 5, 7 d. The cell survival rates were (92.5±2.3)% vs (94.1±1.8)%, (91.4±0.9)% vs (92.7±2.0)%, (92.1±1.6)% vs (93.3± 2.4)%, (91.9 ± 1.5)% vs (93.0 ± 3.1)%, respectively (F=4.38, P>0.05). The bioluminescence imaging in vivo showed that BMSC survived 1, 3, 7 d after implantation. However, bioluminescence signal decreased gradually over time

  13. Hyperspectral and multispectral bioluminescence optical tomography for small animal imaging

    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

  14. In Vivo Mouse Bioluminescence Tomography with Radionuclide-Based Imaging Validation

    Lu, Yujie; Machado, Hidevaldo B.; Bao, Qinan; Stout, David; Herschman, Harvey; Chatziioannou, Arion F.

    2010-01-01

    Introduction Bioluminescence imaging, especially planar bioluminescence imaging, has been extensively applied in in vivo preclinical biological research. Bioluminescence tomography (BLT) has the potential to provide more accurate imaging information due to its 3D reconstruction compared with its planar counterpart. Methods In this work, we introduce a positron emission tomography (PET) radionuclide imaging-based strategy to validate the BLT results. X-ray computed tomography, PET, spectrally ...

  15. In vivo fluorescence imaging of the reticuloendothelial system using quantum dots in combination with bioluminescent tumour monitoring

    We characterised in vivo fluorescence imaging (FLI) of the reticuloendothelial system using quantum dots (QD) and investigated its use in combination with in vivo bioluminescence imaging (BLI). In vivo FLI was performed in five mice repeatedly after the intravenous administration of QD without conjugation to targeting ligands. Ex vivo FLI of the excised organs was performed 24 h after QD injection in three mice. Seven days after intravenous inoculation of luciferase-expressing model cells of a haematological malignancy, mice were injected with the QD or saline (n = 5 each), and combined BLI/FLI was performed repeatedly. Additional five mice inoculated with the tumour cells were examined by in vivo BLI/FLI, and the structures harbouring bioluminescent foci were determined by ex vivo BLI. The utility of combining FLI with bioluminescent tumour monitoring was evaluated. In vivo FLI after QD injection allowed long-term, repeated observation of the reticuloendothelial system in individual mice, although fluorescence intensity and image contrast gradually decreased over time. Ex vivo FLI verified selective accumulation in reticuloendothelial structures. The administration of QD did not affect whole-body bioluminescent signal intensities during longitudinal tumour monitoring. In vivo BLI/FLI, accompanied by fusion of both images, improved the accuracy and confidence level of the localisation of the bioluminescent foci. In vivo FLI using QD provides an overview of the reticuloendothelial system in living mice. In combination with bioluminescent tumour monitoring, fluorescent reticuloendothelial imaging is expected to provide valuable information for lesion localisation. (orig.)

  16. Impact of Anesthesia Protocols on In Vivo Bioluminescent Bacteria Imaging Results

    Chuzel, Thomas; Sanchez, Violette; Vandamme, Marc; Martin, Stéphane; Flety, Odile; Pager, Aurélie; Chabanel, Christophe; Magnier, Luc; Foskolos, Marie; Petit, Océane; Rokbi, Bachra; Chereul, Emmanuel

    2015-01-01

    Infectious murine models greatly benefit from optical imaging using bioluminescent bacteria to non-invasively and repeatedly follow in vivo bacterial infection. In this context, one of the most critical parameters is the bioluminescence sensitivity to reliably detect the smallest number of bacteria. Another critical point is the anesthetic approaches that have been demonstrated to impact the bioluminescence flux emission in studies with luciferase-transfected tumor cells. However, this impact...

  17. Assessing laser-tissue damage with bioluminescent imaging

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

    2006-07-01

    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 (λ=10.6 µm, 0.679 to 2.262 W/cm2, cw, unfocused Gaussian beam, ωL=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

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

    CHATZIIOANNOU, ARION

    2011-12-21

    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.

  19. Confocal Bioluminescence Imaging for Living Tissues with a Caged Substrate of Luciferin.

    Hattori, Mitsuru; Kawamura, Genki; Kojima, Ryosuke; Kamiya, Mako; Urano, Yasuteru; Ozawa, Takeaki

    2016-06-21

    Fluorescence imaging can elucidate morphological organization and coordinal networks, but its background luminescence degrades the image contrast. Our confocal bioluminescence imaging system uses a luciferase caged substrate, with light passing through multipinhole arrays, causing bioluminescence at a focal plane. After a charge-coupled device camera captures luminescence, the imaging system acquires confocal images of multilayered cells with depth information, supporting quantitative analysis of spatial cellular localization in living tissues. PMID:27216493

  20. In vivo bioluminescence and reflectance imaging of multiple organs in bioluminescence reporter mice by bundled-fiber-coupled microscopy.

    Ando, Yoriko; Sakurai, Takashi; Koida, Kowa; Tei, Hajime; Hida, Akiko; Nakao, Kazuki; Natsume, Mistuo; Numano, Rika

    2016-03-01

    Bioluminescence imaging (BLI) is used in biomedical research to monitor biological processes within living organisms. Recently, fiber bundles with high transmittance and density have been developed to detect low light with high resolution. Therefore, we have developed a bundled-fiber-coupled microscope with a highly sensitive cooled-CCD camera that enables the BLI of organs within the mouse body. This is the first report of in vivo BLI of the brain and multiple organs in luciferase-reporter mice using bundled-fiber optics. With reflectance imaging, the structures of blood vessels and organs can be seen clearly with light illumination, and it allowed identification of the structural details of bioluminescence images. This technique can also be applied to clinical diagnostics in a low invasive manner. PMID:27231601

  1. In vivo bioluminescence and reflectance imaging of multiple organs in bioluminescence reporter mice by bundled-fiber-coupled microscopy

    Ando, Yoriko; Sakurai, Takashi; Koida, Kowa; Tei, Hajime; Hida, Akiko; Nakao, Kazuki; Natsume, Mistuo; Numano, Rika

    2016-01-01

    Bioluminescence imaging (BLI) is used in biomedical research to monitor biological processes within living organisms. Recently, fiber bundles with high transmittance and density have been developed to detect low light with high resolution. Therefore, we have developed a bundled-fiber-coupled microscope with a highly sensitive cooled-CCD camera that enables the BLI of organs within the mouse body. This is the first report of in vivo BLI of the brain and multiple organs in luciferase-reporter mice using bundled-fiber optics. With reflectance imaging, the structures of blood vessels and organs can be seen clearly with light illumination, and it allowed identification of the structural details of bioluminescence images. This technique can also be applied to clinical diagnostics in a low invasive manner. PMID:27231601

  2. ATP binding cassette transporters modulate both coelenterazine- and D-luciferin- based bioluminescence imaging

    Huang, Ruimin; Vider, Jelena; Serganova, Inna; Blasberg, Ronald G.

    2011-01-01

    Bioluminescence imaging (BLI) of luciferase reporters provides a cost-effective and sensitive means to image biological processes. However, transport of luciferase substrates across the cell membrane does affect BLI-readout-intensity from intact living cells.

  3. Characterization of sevoflurane effects on Per2 expression using ex vivo bioluminescence imaging of the suprachiasmatic nucleus in transgenic rats.

    Matsuo, Izumi; Iijima, Norio; Takumi, Ken; Higo, Shimpei; Aikawa, Satoko; Anzai, Megumi; Ishii, Hirotaka; Sakamoto, Atsuhiro; Ozawa, Hitoshi

    2016-06-01

    The inhalation anesthetic sevoflurane suppresses Per2 expression in the suprachiasmatic nucleus (SCN) in rodents. Here, we investigated the intra-SCN regional specificity, time-dependency, and pharmacological basis of sevoflurane-effects. Bioluminescence image was taken from the SCN explants of mPer2 promoter-destabilized luciferase transgenic rats, and each small regions of interest (ROI) of the image was analyzed. Sevoflurane suppressed bioluminescence in all ROIs, suggesting that all regions in the SCN are sensitive to sevoflurane. Clear time-dependency in sevoflurane effects were also observed; application during the trough phase of the bioluminescence cycle suppressed the subsequent increase in bioluminescence and resulted in a phase delay of the cycle; sevoflurane applied during the middle of the ascending phase induced a phase advance; sevoflurane on the descending phase showed no effect. These results indicate that the sevoflurane effect may depend on the intrinsic state of circadian machinery. Finally, we examined the involvement of GABAergic signal transduction in the sevoflurane effect. Co-application of both GABAA and GABAB receptor antagonists completely blocked the effect of sevoflurane on the bioluminescence rhythm, suggesting that sevoflurane inhibits Per2 expression via GABAergic signal transduction. Current study elucidated the anesthetic effects on the molecular mechanisms of circadian rhythm. PMID:26696094

  4. U-SPECT-BioFluo: an integrated radionuclide, bioluminescence, and fluorescence imaging platform

    2014-01-01

    Background In vivo bioluminescence, fluorescence, and single-photon emission computed tomography (SPECT) imaging provide complementary information about biological processes. However, to date these signatures are evaluated separately on individual preclinical systems. In this paper, we introduce a fully integrated bioluminescence-fluorescence-SPECT platform. Next to an optimization in logistics and image fusion, this integration can help improve understanding of the optical imaging (OI) resul...

  5. Tomographic bioluminescence imaging by an iteratively re-weighted minimization

    Wu, Ping; Liu, Kai; Xue, Zhenwen; Guo, Wei; Qin, Chenghu; Tian, Jie

    2012-03-01

    Tomographic bioluminescence imaging (TBI), with visible light emission in living organisms, is an effective way of molecular imaging, which allows for the study of ongoing tumor biological processes in vivo and non-invasively. This newly developed technology enables three-dimensional accuracy localization and quantitative analysis of the target tumor cells in small animal via reconstructing the images acquired by the high-resolution imaging system. Due to the difficulty of reconstruction, which is often referred to an ill-posed inverse problem, continuous efforts are still made to find more practical and efficient approaches. In this paper, an iteratively re-weighted minimization (IRM) has been applied to reconstruct the entire source distribution, which is known as sparse signals, inside the target tissue with the limited outgoing photon density on its boundary. By introducing a weight function into the objective function, we convert the lp norm problem into a more simple form of l2 norm to reduce the computational complexity. The weight function is updated in each iterative step to compute the final optimal solution more efficiently. This method is proved to be robust to different parameters, and mouse experiments are conducted to validate the feasibility of IRM approach, which is also reliable at whole-body imaging.

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

    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

  7. Monitoring and quantitative assessment of tumor burden using in vivo bioluminescence imaging

    In vivo bioluminescence imaging (BLI) is a sensitive imaging modality that is rapid and accessible, and may comprise an ideal tool for evaluating tumor growth. In this study, the kinetic of tumor growth has been assessed in C26 colon carcinoma bearing BALB/c mouse model. The ability of BLI to noninvasively quantitate the growth of subcutaneous tumors transplanted with C26 cells genetically engineered to stably express firefly luciferase and herpes simplex virus type-1 thymidine kinase (C26/tk-luc). A good correlation (R 2=0.998) of photon emission to the cell number was found in vitro. Tumor burden and tumor volume were monitored in vivo over time by quantitation of photon emission using Xenogen IVIS 50 and standard external caliper measurement, respectively. At various time intervals, tumor-bearing mice were imaged to determine the correlation of in vivo BLI to tumor volume. However, a correlation of BLI to tumor volume was observed when tumor volume was smaller than 1000 mm3 (R 2=0.907). γ Scintigraphy combined with [131I]FIAU was another imaging modality used for verifying the previous results. In conclusion, this study showed that bioluminescence imaging is a powerful and quantitative tool for the direct assay to monitor tumor growth in vivo. The dual reporter genes transfected tumor-bearing animal model can be applied in the evaluation of the efficacy of new developed anti-cancer drugs

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

    2013-01-01

    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 и in vivo was carried out on IVIS-Spectrum system (Caliper Life Sciences, USA). Pri...

  9. High resolution in vitro bioluminescence imaging using a multimodal optical system

    Bioluminescence in vitro studies are usually performed with dedicated microscopes. In this work, we developed a novel image recovery algorithm and a multimodal system prototype to perform bioluminescence microscopy. We performed a feasibility study using GEANT4 Monte Carlo (MC) simulation of bioluminescent cells acquired at low SNR frames and processed using a Super Resolution Regularization Algorithm (SRRA). The method was also tested using in vitro cell acquisition. The results obtained with MC simulations showed an improvement in the spatial resolution from 90 μm to 10 μm and from 110 μm to 13 μm for in vitro imaging of mesothelioma cells

  10. High resolution in vitro bioluminescence imaging using a multimodal optical system

    Altabella, L.; Gigliotti, C. R.; Perani, L.; Crippa, M. P.; Boschi, F.; Spinelli, A. E.

    2016-01-01

    Bioluminescence in vitro studies are usually performed with dedicated microscopes. In this work, we developed a novel image recovery algorithm and a multimodal system prototype to perform bioluminescence microscopy. We performed a feasibility study using GEANT4 Monte Carlo (MC) simulation of bioluminescent cells acquired at low SNR frames and processed using a Super Resolution Regularization Algorithm (SRRA). The method was also tested using in vitro cell acquisition. The results obtained with MC simulations showed an improvement in the spatial resolution from 90 μ m to 10 μ m and from 110 μ m to 13 μ m for in vitro imaging of mesothelioma cells.

  11. U-SPECT-BioFluo: an integrated radionuclide, bioluminescence, and fluorescence imaging platform

    Van Oosterom, M.N.; Kreuger, R.; Buckle, T.; Mahn, W.A.; Bunschoten, A.; Josephson, L.; Van Leeuwen, F.W.B.; Beekman, F.J.

    2014-01-01

    Background: In vivo bioluminescence, fluorescence, and single-photon emission computed tomography (SPECT) imaging provide complementary information about biological processes. However, to date these signatures are evaluated separately on individual preclinical systems. In this paper, we introduce a

  12. Bioluminescence Imaging of Chlamydia muridarum Ascending Infection in Mice

    Jessica Campbell; Yumeng Huang; Yuanjun Liu; Robert Schenken; Bernard Arulanandam; Guangming Zhong

    2014-01-01

    Chlamydial pathogenicity in the upper genital tract relies on chlamydial ascending from the lower genital tract. To monitor chlamydial ascension, we engineered a luciferase-expressing C. muridarum. In cells infected with the luciferase-expressing C. muridarum, luciferase gene expression and enzymatic activity (measured as bioluminescence intensity) correlated well along the infection course, suggesting that bioluminescence can be used for monitoring chlamydial replication. Following an intrav...

  13. A luciferin analogue generating near-infrared bioluminescence achieves highly sensitive deep-tissue imaging.

    Kuchimaru, Takahiro; Iwano, Satoshi; Kiyama, Masahiro; Mitsumata, Shun; Kadonosono, Tetsuya; Niwa, Haruki; Maki, Shojiro; Kizaka-Kondoh, Shinae

    2016-01-01

    In preclinical cancer research, bioluminescence imaging with firefly luciferase and D-luciferin has become a standard to monitor biological processes both in vitro and in vivo. However, the emission maximum (λmax) of bioluminescence produced by D-luciferin is 562 nm where light is not highly penetrable in biological tissues. This emphasizes a need for developing a red-shifted bioluminescence imaging system to improve detection sensitivity of targets in deep tissue. Here we characterize the bioluminescent properties of the newly synthesized luciferin analogue, AkaLumine-HCl. The bioluminescence produced by AkaLumine-HCl in reactions with native firefly luciferase is in the near-infrared wavelength ranges (λmax=677 nm), and yields significantly increased target-detection sensitivity from deep tissues with maximal signals attained at very low concentrations, as compared with D-luciferin and emerging synthetic luciferin CycLuc1. These characteristics offer a more sensitive and accurate method for non-invasive bioluminescence imaging with native firefly luciferase in various animal models. PMID:27297211

  14. A luciferin analogue generating near-infrared bioluminescence achieves highly sensitive deep-tissue imaging

    Kuchimaru, Takahiro; Iwano, Satoshi; Kiyama, Masahiro; Mitsumata, Shun; Kadonosono, Tetsuya; Niwa, Haruki; Maki, Shojiro; Kizaka-Kondoh, Shinae

    2016-01-01

    In preclinical cancer research, bioluminescence imaging with firefly luciferase and D-luciferin has become a standard to monitor biological processes both in vitro and in vivo. However, the emission maximum (λmax) of bioluminescence produced by D-luciferin is 562 nm where light is not highly penetrable in biological tissues. This emphasizes a need for developing a red-shifted bioluminescence imaging system to improve detection sensitivity of targets in deep tissue. Here we characterize the bioluminescent properties of the newly synthesized luciferin analogue, AkaLumine-HCl. The bioluminescence produced by AkaLumine-HCl in reactions with native firefly luciferase is in the near-infrared wavelength ranges (λmax=677 nm), and yields significantly increased target-detection sensitivity from deep tissues with maximal signals attained at very low concentrations, as compared with D-luciferin and emerging synthetic luciferin CycLuc1. These characteristics offer a more sensitive and accurate method for non-invasive bioluminescence imaging with native firefly luciferase in various animal models. PMID:27297211

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

    1985-01-01

    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...

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

    Flor-Henry Michel

    2004-11-01

    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.

  17. In vivo bioluminescence imaging of Burkholderia mallei respiratory infection and treatment in the mouse model

    Shane eMassey

    2011-08-01

    Full Text Available Bioluminescent imaging (BLI technology is a powerful tool for monitoring infectious disease progression and treatment approaches. BLI is particularly useful for tracking fastidious intracellular pathogens that might be difficult to recover from certain organs. Burkholderia mallei, the causative agent of glanders, is a facultative intracellular pathogen and has been classified by the CDC as a Category B select agent due to its highly infectious nature and potential use as a biological weapon. Very little is known regarding pathogenesis or treatment of glanders. We investigated the use of bioluminescent reporter constructs to monitor the dynamics of infection as well as the efficacy of therapeutics for B. mallei in real time. A stable luminescent reporter B. mallei strain was created using the pUTmini-Tn5::luxKm2 plasmid and used to monitor glanders in the BALB/c murine model. Mice were infected via the intranasal route with 5x103 bacteria and monitored by BLI at 24, 48 and 72 h. We verified that our reporter construct maintained similar virulence and growth kinetics compared to wild-type B. mallei and confirmed that it maintains luminescent stability in the presence or absence of antibiotic selection. The luminescent signal was initially seen in the lungs, and progressed to the liver and spleen over the course of infection. We demonstrated that antibiotic treatment 24 h post-infection resulted in reduction of bioluminescence that can be attributed to decreased bacterial burden in target organs. These findings suggest that BLI can be used to monitor disease progression and efficacy of therapeutics during glanders infections. Finally, we report an alternative method to mini-Tn5::luxKm2 transposon using mini-Tn7-lux elements that insert site-specifically at known genomic attachment sites and that can also be used to tag bacteria.

  18. Bioluminescence imaging of fungal biofilm development in live animals.

    Vande Velde, Greetje; Kucharíková, Soňa; Van Dijck, Patrick; Himmelreich, Uwe

    2014-01-01

    Fungal biofilms formed on various types of medical implants represent a major problem for hospitalized patients. These biofilms and related infections are usually difficult to treat because of their resistance to the classical antifungal drugs. Animal models are indispensable for investigating host-pathogen interactions and for identifying new antifungal targets related to biofilm development. A limited number of animal models is available that can be used for testing novel antifungal drugs in vivo against C. albicans, one of the most common pathogens causing fungal biofilms. Fungal load in biofilms in these models is traditionally analyzed postmortem, requiring host sacrifice and enumeration of microorganisms from individual biofilms in order to evaluate the amount of colony forming units and the efficacy of antifungal treatment. Bioluminescence imaging (BLI) made compatible with small animal models for in vivo biofilm formation is a valuable noninvasive tool to follow-up biofilm development and its treatment longitudinally, reducing the number of animals needed for such studies. Due to the nondestructive and noninvasive nature of BLI, the imaging procedure can be repeated in the same animal, allowing follow-up of the biofilm growth in vivo without removing the implanted device or detaching the biofilm from its substrate. The method described here introduces BLI of C. albicans biofilm formation in vivo on subcutaneously implanted catheters in mice. One of the main challenges to overcome for BLI of fungi is the hampered intracellular substrate delivery through the fungal cell wall, which is managed by using extracellularly located Gaussia luciferase. Although detecting a quantifiable in vivo BLI signal from biofilms formed on the inside of implanted catheters is challenging, BLI proved to be a practical tool in the study of fungal biofilms. This method describing the use of BLI for in vivo follow-up of device-related fungal biofilm formation has the potential for

  19. Bioluminescence Imaging Reveals Dynamics of Beta Cell Loss in the Non-Obese Diabetic (NOD) Mouse Model

    John Virostko; Armandla Radhika; Greg Poffenberger; Dula, Adrienne N.; Moore, Daniel J.; Alvin C Powers

    2013-01-01

    We generated a mouse model (MIP-Luc-VU-NOD) that enables non-invasive bioluminescence imaging (BLI) of beta cell loss during the progression of autoimmune diabetes and determined the relationship between BLI and disease progression. MIP-Luc-VU-NOD mice displayed insulitis and a decline in bioluminescence with age which correlated with beta cell mass, plasma insulin, and pancreatic insulin content. Bioluminescence declined gradually in female MIP-Luc-VU-NOD mice, reaching less than 50% of the ...

  20. Integrated visualization of multi-angle bioluminescence imaging and micro CT

    Kok, P.; Dijkstra, J.; Botha, C.P.; Post, F.H.; Kaijzel, E.; Que, I.; Löwik, C.W.G.M.; Reiber, J.H.C.; Lelieveldt, B.P.F.

    2007-01-01

    This paper explores new methods to visualize and fuse multi-2D bioluminescence imaging (BLI) data with structural imaging modalities such as micro CT and MR. A geometric, back-projection-based 3D reconstruction for superficial lesions from multi-2D BLI data is presented, enabling a coarse estimate o

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

    Liu Zhihui

    2008-08-01

    Full Text Available Abstract Background Interleukin 1 beta (IL-1β plays an important role in a number of chronic and acute inflammatory diseases. To understand the role of IL-1β 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-1β 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-1β mRNA and pro-IL-1β 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-1β gene expression in the inflammatory process and evaluation the effect of anti-inflammatory drug in vivo.

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

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

    2007-02-01

    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.

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

    Daniell Henry

    2011-06-01

    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 μm 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 μm. 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

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

    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

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

    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

    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.

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

    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

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

    Lee, Byeong Il; Kim, Hyeon Sik; Jeong, Hye Jin; Lee, Hyung Jae; Moon, Seung Min; Kwon, Seung Young; Jeong, Shin Young; Bom, Hee Seung; Min, Jung Joon [Chonnam National University Hospital, Gwangju (Korea, Republic of); Choi, Eun Seo [Chosun University, Gwangju (Korea, Republic of)

    2009-08-15

    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

  8. Bioluminescence in vivo imaging of autoimmune encephalomyelitis predicts disease

    Steinman Lawrence; Ho Peggy; Luo Jian; Wyss-Coray Tony

    2008-01-01

    Abstract Background Experimental autoimmune encephalomyelitis is a widely used animal model to understand not only multiple sclerosis but also basic principles of immunity. The disease is scored typically by observing signs of paralysis, which do not always correspond with pathological changes. Methods Experimental autoimmune encephalomyelitis was induced in transgenic mice expressing an injury responsive luciferase reporter in astrocytes (GFAP-luc). Bioluminescence in the brain and spinal co...

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

    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

  10. Application of Bioluminescence Imaging for In Vivo Monitoring of Fungal Infections

    Matthias Brock

    2012-01-01

    Full Text Available Fungi can cause severe invasive infections especially in the immunocompromised host. Patient populations at risk are increasing due to ongoing developments in cancer treatment and transplantation medicine. Only limited diagnostic tools and few antifungals are available, rendering a significant number of invasive fungal infections life threatening. To reduce mortality rates, a better understanding of the infection processes is urgently required. Bioluminescence imaging (BLI is a powerful tool for such purposes, since it allows visualisation of temporal and spatial progression of infections in real time. BLI has been successfully used to monitor infections caused by various microorganisms, in particular bacteria. However, first studies have also been performed on the fungi Candida albicans and Aspergillus fumigatus. Although BLI was, in principle, suitable to study the infection process, some limitations remained. Here, different luciferase systems are introduced, and current approaches are summarised. Finally, suggestions for further improvements of BLI to monitor fungal infections are provided.

  11. 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

    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

  12. Rapid and Quantitative Assessment of Cancer Treatment Response Using In Vivo Bioluminescence Imaging

    Alnawaz Rehemtulla

    2000-01-01

    Full Text Available Current assessment of orthotopic tumor models in animals utilizes survival as the primary therapeutic end point. In vivo bioluminescence imaging (BLI is a sensitive imaging modality that is rapid and accessible, and may comprise an ideal tool for evaluating antineoplastic therapies [1 ]. Using human tumor cell lines constitutively expressing luciferase, the kinetics of tumor growth and response to therapy have been assessed in intraperitoneal [2], subcutaneous, and intravascular [3] cancer models. However, use of this approach for evaluating orthotopic tumor models has not been demonstrated. In this report, the ability of BLI to noninvasively quantitate the growth and therapeuticinduced cell kill of orthotopic rat brain tumors derived from 9L gliosarcoma cells genetically engineered to stably express firefly luciferase (9LLuc was investigated. Intracerebral tumor burden was monitored over time by quantitation of photon emission and tumor volume using a cryogenically cooled CCD camera and magnetic resonance imaging (MRI, respectively. There was excellent correlation (r=0.91 between detected photons and tumor volume. A quantitative comparison of tumor cell kill determined from serial MRI volume measurements and BLI photon counts following 1,3-bis(2-chloroethyl-1-nitrosourea (BCNU treatment revealed that both imaging modalities yielded statistically similar cell kill values (P=.951. These results provide direct validation of BLI imaging as a powerful and quantitative tool for the assessment of antineoplastic therapies in living animals.

  13. Noninvasive bioluminescence imaging of the dynamics of sanguinarine induced apoptosis via activation of reactive oxygen species

    Dai, Yunpeng; Shi, Yaru; Zeng, Qi; Wang, Fu

    2016-01-01

    Most chemotherapeutic drugs exert their anti-tumor effects primarily by triggering a final pathway leading to apoptosis. Noninvasive imaging of apoptotic events in preclinical models would greatly facilitate the development of apoptosis-inducing compounds and evaluation of their therapeutic efficacy. Here we employed a cyclic firefly luciferase (cFluc) reporter to screen potential pro-apoptotic compounds from a number of natural agents. We demonstrated that sanguinarine (SANG) could induce apoptosis in a dose- and time-dependent manner in UM-SCC-22B head and neck cancer cells. Moreover, SANG-induced apoptosis was associated with the generation of reactive oxygen species (ROS) and activation of c-Jun-N-terminal kinase (JNK) and nuclear factor-kappaB (NF-κB) signal pathways. After intravenous administration with SANG in 22B-cFluc xenograft models, a dramatic increase of luminescence signal can be detected as early as 48 h post-treatment, as revealed by longitudinal bioluminescence imaging in vivo. Remarkable apoptotic cells reflected from ex vivo TUNEL staining confirmed the imaging results. Importantly, SANG treatment caused distinct tumor growth retardation in mice compared with the vehicle-treated group. Taken together, our results showed that SANG is a candidate anti-tumor drug and noninvasive imaging of apoptosis using cFluc reporter could provide a valuable tool for drug development and therapeutic efficacy evaluation. PMID:26968950

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

    O'Neill, Karen

    2010-02-01

    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.

  15. Efficacy of Lantibiotic Treatment of Staphylococcus aureus-Induced Skin Infections, Monitored by In Vivo Bioluminescent Imaging.

    van Staden, Anton Du Preez; Heunis, Tiaan; Smith, Carine; Deane, Shelly; Dicks, Leon M T

    2016-07-01

    Staphylococcus aureus is a bacterial pathogen responsible for the majority of skin and soft tissue infections. Antibiotics are losing their efficacy as treatment for skin and soft tissue infections as a result of increased resistance in a variety of pathogens, including S. aureus It is thus imperative to explore alternative antimicrobial treatments to ensure future treatment options for skin and soft tissue infections. A select few lantibiotics, a group of natural defense peptides produced by bacteria, inhibit the growth of numerous clinical S. aureus isolates, including methicillin-resistant strains. In this study, the antimicrobial activities of nisin, clausin, and amyloliquecidin, separately administered, were compared to that of a mupirocin-based ointment, which is commonly used as treatment for S. aureus-induced skin infections. Full-thickness excisional wounds, generated on the dorsal surfaces of mice, were infected with a bioluminescent strain of S. aureus (strain Xen 36). The infections were monitored in real time using in vivo bioluminescent imaging. Lantibiotic treatments significantly reduced the bioluminescence of S. aureus Xen 36 to a level similar to that recorded with mupirocin treatment. Wound closure, however, was more pronounced during lantibiotic treatment. Lantibiotics thus have the potential to be used as an alternative treatment option for S. aureus-induced skin infections. PMID:27067340

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

    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.)

  17. Recovering 3D tumor locations from 2D bioluminescence images and registration with CT images

    Huang, Xiaolei; Metaxas, Dimitris N.; Menon, Lata G.; Mayer-Kuckuk, Philipp; Bertino, Joseph R.; Banerjee, Debabrata

    2006-02-01

    In this paper, we introduce a novel and efficient algorithm for reconstructing the 3D locations of tumor sites from a set of 2D bioluminescence images which are taken by a same camera but after continually rotating the object by a small angle. Our approach requires a much simpler set up than those using multiple cameras, and the algorithmic steps in our framework are efficient and robust enough to facilitate its use in analyzing the repeated imaging of a same animal transplanted with gene marked cells. In order to visualize in 3D the structure of the tumor, we also co-register the BLI-reconstructed crude structure with detailed anatomical structure extracted from high-resolution microCT on a single platform. We present our method using both phantom studies and real studies on small animals.

  18. Live Cell Bioluminescence Imaging in Temporal Reaction of G Protein-Coupled Receptor for High-Throughput Screening and Analysis.

    Hattori, Mitsuru; Ozawa, Takeaki

    2016-01-01

    G protein-coupled receptors (GPCRs) are notable targets of basic therapeutics. Many screening methods have been established to identify novel agents for GPCR signaling in a high-throughput manner. However, information related to the temporal reaction of GPCR with specific ligands remains poor. We recently developed a bioluminescence method for the quantitative detection of the interaction between GPCR and β-arrestin using split luciferase complementation. To monitor time-course variation of the interactions, a new imaging system contributes to the accurate evaluation of drugs for GPCRs in a high-throughput manner. PMID:27424906

  19. Design and development of high bioluminescent resonance energy transfer efficiency hybrid-imaging constructs.

    Kumar, Manoj; Kovalski, Letícia; Broyles, David; Hunt, Eric A; Daftarian, Pirouz; Dikici, Emre; Daunert, Sylvia; Deo, Sapna K

    2016-04-01

    Here we describe the design and construction of an imaging construct with high bioluminescent resonance energy transfer (BRET) efficiency that is composed of multiple quantum dots (QDs; λem = 655 nm) self-assembled onto a bioluminescent protein, Renilla luciferase (Rluc). This is facilitated by the streptavidin-biotin interaction, allowing the facile formation of a hybrid-imaging construct (HIC) comprising up to six QDs (acceptor) grafted onto a light-emitting Rluc (donor) core. The resulting assembly of multiple acceptors surrounding a donor permits this construct to exhibit high resonance energy transfer efficiency (∼64.8%). The HIC was characterized using fluorescence excitation anisotropy measurements and high-resolution transmission electron microscopy. To demonstrate the application of our construct, a generation-5 (G5) polyamidoamine dendrimer (PAMAM) nanocarrier was loaded with our HIC for in vitro and in vivo imaging. We envision that this design of multiple acceptors and bioluminescent donor will lead to the development of new BRET-based systems useful in sensing, imaging, and other bioanalytical applications. PMID:26772160

  20. Quantitative bioluminescence imaging of transgene expression in intact porcine antral follicles in vitro

    Jung, Song-Yi; Willard, Scott T

    2014-01-01

    Background The porcine oocyte maturation in vivo occurs within the ovarian follicle and is regulated by the interactions between oocytes and surrounding follicular components, including theca, granulosa, and cumulus cells, and follicular fluid. Therefore, the antral follicle is an essential microenvironment for efficient oocyte maturation and its developmental competence. Quantitative bioluminescence imaging of firefly luciferase reporter genes in an intact antral follicle would allow investi...

  1. Design and Synthesis of an Alkynyl Luciferin Analogue for Bioluminescence Imaging.

    Steinhardt, Rachel C; O'Neill, Jessica M; Rathbun, Colin M; McCutcheon, David C; Paley, Miranda A; Prescher, Jennifer A

    2016-03-01

    Herein, the synthesis and characterization of an alkyne-modified luciferin is reported. This bioluminescent probe was accessed using C-H activation methodology and was found to be stable in solution and capable of light production with firefly luciferase. The luciferin analogue was also cell permeant and emitted more redshifted light than d-luciferin, the native luciferase substrate. Based on these features, the alkynyl luciferin will be useful for a variety of imaging applications. PMID:26784889

  2. Bioluminescence imaging of β cells and intrahepatic insulin gene activity under normal and pathological conditions.

    Tokio Katsumata

    Full Text Available In diabetes research, bioluminescence imaging (BLI has been applied in studies of β-cell impairment, development, and islet transplantation. To develop a mouse model that enables noninvasive imaging of β cells, we generated a bacterial artificial chromosome (BAC transgenic mouse in which a mouse 200-kbp genomic fragment comprising the insulin I gene drives luciferase expression (Ins1-luc BAC transgenic mouse. BLI of mice was performed using the IVIS Spectrum system after intraperitoneal injection of luciferin, and the bioluminescence signal from the pancreatic region analyzed. When compared with MIP-Luc-VU mice [FVB/N-Tg(Ins1-lucVUPwrs/J] expressing luciferase under the control of the 9.2-kbp mouse insulin I promoter (MIP, the bioluminescence emission from Ins1-luc BAC transgenic mice was enhanced approximately 4-fold. Streptozotocin-treated Ins1-luc BAC transgenic mice developed severe diabetes concomitant with a sharp decline in the BLI signal intensity in the pancreas. Conversely, mice fed a high-fat diet for 8 weeks showed an increase in the signal, reflecting a decrease or increase in the β-cell mass. Although the bioluminescence intensity of the islets correlated well with the number of isolated islets in vitro, the intensity obtained from a living mouse in vivo did not necessarily reflect an absolute quantification of the β-cell mass under pathological conditions. On the other hand, adenovirus-mediated gene transduction of β-cell-related transcription factors in Ins1-luc BAC transgenic mice generated luminescence from the hepatic region for more than 1 week. These results demonstrate that BLI in Ins1-luc BAC transgenic mice provides a noninvasive method of imaging islet β cells and extrapancreatic activity of the insulin gene in the liver under normal and pathological conditions.

  3. Construction of a bioluminescence reporter plasmid for Francisella tularensis

    Bina, Xiaowen R.; Miller, Mark A.; James E Bina

    2010-01-01

    A Francisella tularensis shuttle vector that constitutively expresses the Photorhabdus luminescens lux operon in type A and type B strains of F. tularensis was constructed. The bioluminescence reporter plasmid was introduced into the live vaccine strain of F. tularensis and used to follow F. tularensis growth in a murine intranasal challenge model in real time by bioluminescence imaging. The results show that the new bioluminescence reporter plasmid represents a useful tool for tularemia rese...

  4. Firefly Luciferase Mutants Allow Substrate-Selective Bioluminescence Imaging in the Mouse Brain.

    Adams, Spencer T; Mofford, David M; Reddy, G S Kiran Kumar; Miller, Stephen C

    2016-04-11

    Bioluminescence imaging is a powerful approach for visualizing specific events occurring inside live mice. Animals can be made to glow in response to the expression of a gene, the activity of an enzyme, or the growth of a tumor. But bioluminescence requires the interaction of a luciferase enzyme with a small-molecule luciferin, and its scope has been limited by the mere handful of natural combinations. Herein, we show that mutants of firefly luciferase can discriminate between natural and synthetic substrates in the brains of live mice. When using adeno-associated viral (AAV) vectors to express luciferases in the brain, we found that mutant luciferases that are inactive or weakly active with d-luciferin can light up brightly when treated with the aminoluciferins CycLuc1 and CycLuc2 or their respective FAAH-sensitive luciferin amides. Further development of selective luciferases promises to expand the power of bioluminescence and allow multiple events to be imaged in the same live animal. PMID:26991209

  5. Investigating real-time activation of adenosine receptors by bioluminescence resonance energy transfer technique

    Huang, Yimei; Yang, Hongqin; Zheng, Liqin; Chen, Jiangxu; Wang, Yuhua; Li, Hui; Xie, Shusen

    2013-02-01

    Adenosine receptors play important roles in many physiological and pathological processes, for example regulating myocardial oxygen consumption and the release of neurotransmitters. The activations of adenosine receptors have been studied by some kinds of techniques, such as western blot, immunohistochemistry, etc. However, these techniques cannot reveal the dynamical response of adenosine receptors under stimulation. In this paper, bioluminescence resonance energy transfer technique was introduced to study the real-time activation of adenosine receptors by monitoring the dynamics of cyclic adenosine monophosphate (cAMP) level. The results showed that there were significant differences between adenosine receptors on real-time responses under stimulation. Moreover, the dynamics of cAMP level demonstrated that competition between adenosine receptors existed. Taken together, our study indicates that monitoring the dynamics of cAMP level using bioluminescence resonance energy transfer technique could be one potential approach to investigate the mechanism of competitions between adenosine receptors.

  6. A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo.

    Chu, Jun; Oh, Younghee; Sens, Alex; Ataie, Niloufar; Dana, Hod; Macklin, John J; Laviv, Tal; Welf, Erik S; Dean, Kevin M; Zhang, Feijie; Kim, Benjamin B; Tang, Clement Tran; Hu, Michelle; Baird, Michelle A; Davidson, Michael W; Kay, Mark A; Fiolka, Reto; Yasuda, Ryohei; Kim, Douglas S; Ng, Ho-Leung; Lin, Michael Z

    2016-07-01

    Orange-red fluorescent proteins (FPs) are widely used in biomedical research for multiplexed epifluorescence microscopy with GFP-based probes, but their different excitation requirements make multiplexing with new advanced microscopy methods difficult. Separately, orange-red FPs are useful for deep-tissue imaging in mammals owing to the relative tissue transmissibility of orange-red light, but their dependence on illumination limits their sensitivity as reporters in deep tissues. Here we describe CyOFP1, a bright, engineered, orange-red FP that is excitable by cyan light. We show that CyOFP1 enables single-excitation multiplexed imaging with GFP-based probes in single-photon and two-photon microscopy, including time-lapse imaging in light-sheet systems. CyOFP1 also serves as an efficient acceptor for resonance energy transfer from the highly catalytic blue-emitting luciferase NanoLuc. An optimized fusion of CyOFP1 and NanoLuc, called Antares, functions as a highly sensitive bioluminescent reporter in vivo, producing substantially brighter signals from deep tissues than firefly luciferase and other bioluminescent proteins. PMID:27240196

  7. Autonomously Bioluminescent Mammalian Cells For Continuous And Real-Time Monitoring Of Cytotoxicity

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

    2013-01-01

    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 fac...

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

    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

  9. Bioluminescence imaging of leukemia cell lines in vitro and in mouse xenografts: effects of monoclonal and polyclonal cell populations on intensity and kinetics of photon emission

    Christoph Sandra

    2013-01-01

    Full Text Available Abstract Background We investigated the utility of bioluminescence imaging (BLI using firefly luciferase in monoclonal and polyclonal populations of leukemia cells in vitro and in vivo. Methods Monoclonal and polyclonal human lymphoid and myeloid leukemia cell lines transduced with firefly luciferase were used for BLI. Results Kinetics and dynamics of bioluminescence signal were cell line dependent. Luciferase expression decreased significantly over time in polyclonal leukemia cells in vitro. Transplantation of polyclonal luciferase-tagged cells in mice resulted in inconsistent signal intensity. After selection of monoclonal cell populations, luciferase activity was stable, equal kinetic and dynamic of bioluminescence intensity and strong correlation between cell number and light emission in vitro were observed. We obtained an equal development of leukemia burden detected by luciferase activity in NOD-scid-gamma mice after transplantation of monoclonal populations. Conclusion The use of monoclonal leukemia cells selected for stable and equal luciferase activity is recommended for experiments in vitro and xenograft mouse models. The findings are highly significant for bioluminescence imaging focused on pre-clinical drug development.

  10. Detection and quantitation of circulating tumor cell dynamics by bioluminescence imaging in an orthotopic mammary carcinoma model.

    Laura Sarah Sasportas

    Full Text Available Circulating tumor cells (CTCs have been detected in the bloodstream of both early-stage and advanced cancer patients. However, very little is know about the dynamics of CTCs during cancer progression and the clinical relevance of longitudinal CTC enumeration. To address this, we developed a simple bioluminescence imaging assay to detect CTCs in mouse models of metastasis. In a 4T1 orthotopic metastatic mammary carcinoma mouse model, we demonstrated that this quantitative method offers sensitivity down to 2 CTCs in 0.1-1mL blood samples and high specificity for CTCs originating from the primary tumor, independently of their epithelial status. In this model, we simultaneously monitored blood CTC dynamics, primary tumor growth, and lung metastasis progression over the course of 24 days. Early in tumor development, we observed low numbers of CTCs in blood samples (10-15 cells/100 µL and demonstrated that CTC dynamics correlate with viable primary tumor growth. To our knowledge, these data represent the first reported use of bioluminescence imaging to detect CTCs and quantify their dynamics in any cancer mouse model. This new assay is opening the door to the study of CTC dynamics in a variety of animal models. These studies may inform clinical decision on the appropriate timing of blood sampling and value of longitudinal CTC enumeration in cancer patients.

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

    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

  12. Quantification of bioluminescence images of point source objects using diffusion theory models

    A simple approach for estimating the location and power of a bioluminescent point source inside tissue is reported. The strategy consists of using a diffuse reflectance image at the emission wavelength to determine the optical properties of the tissue. Following this, bioluminescence images are modelled using a single point source and the optical properties from the reflectance image, and the depth and power are iteratively adjusted to find the best agreement with the experimental image. The forward models for light propagation are based on the diffusion approximation, with appropriate boundary conditions. The method was tested using Monte Carlo simulations, Intralipid tissue-simulating phantoms and ex vivo chicken muscle. Monte Carlo data showed that depth could be recovered within 6% for depth 4-12 mm, and the corresponding relative source power within 12%. In Intralipid, the depth could be estimated within 8% for depth 4-12 mm, and the relative source power, within 20%. For ex vivo tissue samples, source depths of 4.5 and 10 mm and their relative powers were correctly identified

  13. Accounting for systematic errors in bioluminescence imaging to improve quantitative accuracy

    Taylor, Shelley L.; Perry, Tracey A.; Styles, Iain B.; Cobbold, Mark; Dehghani, Hamid

    2015-07-01

    Bioluminescence imaging (BLI) is a widely used pre-clinical imaging technique, but there are a number of limitations to its quantitative accuracy. This work uses an animal model to demonstrate some significant limitations of BLI and presents processing methods and algorithms which overcome these limitations, increasing the quantitative accuracy of the technique. The position of the imaging subject and source depth are both shown to affect the measured luminescence intensity. Free Space Modelling is used to eliminate the systematic error due to the camera/subject geometry, removing the dependence of luminescence intensity on animal position. Bioluminescence tomography (BLT) is then used to provide additional information about the depth and intensity of the source. A substantial limitation in the number of sources identified using BLI is also presented. It is shown that when a given source is at a significant depth, it can appear as multiple sources when imaged using BLI, while the use of BLT recovers the true number of sources present.

  14. Remote detection of human toxicants in real time using a human-optimized, bioluminescent bacterial luciferase gene cassette bioreporter

    Close, Dan; Webb, James; Ripp, Steven; Patterson, Stacey; Sayler, Gary

    2012-06-01

    Traditionally, human toxicant bioavailability screening has been forced to proceed in either a high throughput fashion using prokaryotic or lower eukaryotic targets with minimal applicability to humans, or in a more expensive, lower throughput manner that uses fluorescent or bioluminescent human cells to directly provide human bioavailability data. While these efforts are often sufficient for basic scientific research, they prevent the rapid and remote identification of potentially toxic chemicals required for modern biosecurity applications. To merge the advantages of high throughput, low cost screening regimens with the direct bioavailability assessment of human cell line use, we re-engineered the bioluminescent bacterial luciferase gene cassette to function autonomously (without exogenous stimulation) within human cells. Optimized cassette expression provides for fully endogenous bioluminescent production, allowing continuous, real time monitoring of the bioavailability and toxicology of various compounds in an automated fashion. To access the functionality of this system, two sets of bioluminescent human cells were developed. The first was programed to suspend bioluminescent production upon toxicological challenge to mimic the non-specific detection of a toxicant. The second induced bioluminescence upon detection of a specific compound to demonstrate autonomous remote target identification. These cells were capable of responding to μM concentrations of the toxicant n-decanal, and allowed for continuous monitoring of cellular health throughout the treatment process. Induced bioluminescence was generated through treatment with doxycycline and was detectable upon dosage at a 100 ng/ml concentration. These results demonstrate that leveraging autonomous bioluminescence allows for low-cost, high throughput direct assessment of toxicant bioavailability.

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

    Steven Ripp

    2010-12-01

    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.

  16. 18F-fluoride PET imaging in a nude rat model of bone metastasis from breast cancer: Comparison with 18F-FDG and bioluminescence imaging

    Introduction: Clinically-relevant animal models and appropriate imaging diagnostic tools are essential to study cancer and develop novel therapeutics. We evaluated a model of bone metastasis in nude rats by micro-PET and bioluminescence imaging. Methods: A bone metastasis model was produced by intracardiac injection of osteotropic MDA-MB-231Bo-Luc human breast cancer cells into nude rats. Bioluminescence imaging and micro-PET scans using 18F-FDG and 18F-fluoride were acquired serially for 5 weeks. We correlated bioluminescence imaging, 18F-FDG and 18F-fluoride PET images, and histological slides. Results: Multiple bone metastases were successfully evaluated by bioluminescence imaging and 18F-FDG and 18F-fluoride PET scans. Bioluminescence photon flux increased exponentially on weekly follow-up. 18F-FDG PET revealed increased FDG uptake at the spine and bilaterally in the hind legs in week 2 images, and showed a progressive pattern up to 4 weeks that correlated with bioluminescence imaging. 18F-fluoride PET showed minimal abnormal findings in week 2 images, but it showed an irregular pattern at the spine from week 3 or 4 images. On quantitative analysis with standardized uptake values, a pattern of gradual increase was observed from week 2 to week 4 in both 18F-FDG PET and fluoride PET. Histopathological examination confirmed the formation of osteolytic metastasis and necrosis of the distal femur, which appeared as a photon defect on PET scans. Conclusion: Developing bone metastasis from breast cancer in a nude rat model was successfully evaluated with an animal PET imaging system and bioluminescence imaging. This nude rat model of bone metastasis, which can be evaluated by PET imaging, may be a valuable tool for evaluating early responses to novel therapeutics

  17. Novel bioluminescent quantitative detection of nucleic acid amplification in real-time.

    Olga A Gandelman

    Full Text Available BACKGROUND: The real-time monitoring of polynucleotide amplification is at the core of most molecular assays. This conventionally relies on fluorescent detection of the amplicon produced, requiring complex and costly hardware, often restricting it to specialised laboratories. PRINCIPAL FINDINGS: Here we report the first real-time, closed-tube luminescent reporter system for nucleic acid amplification technologies (NAATs enabling the progress of amplification to be continuously monitored using simple light measuring equipment. The Bioluminescent Assay in Real-Time (BART continuously reports through bioluminescent output the exponential increase of inorganic pyrophosphate (PPi produced during the isothermal amplification of a specific nucleic acid target. BART relies on the coupled conversion of inorganic pyrophosphate (PPi produced stoichiometrically during nucleic acid synthesis to ATP by the enzyme ATP sulfurylase, and can therefore be coupled to a wide range of isothermal NAATs. During nucleic acid amplification, enzymatic conversion of PPi released during DNA synthesis into ATP is continuously monitored through the bioluminescence generated by thermostable firefly luciferase. The assay shows a unique kinetic signature for nucleic acid amplifications with a readily identifiable light output peak, whose timing is proportional to the concentration of original target nucleic acid. This allows qualitative and quantitative analysis of specific targets, and readily differentiates between negative and positive samples. Since quantitation in BART is based on determination of time-to-peak rather than absolute intensity of light emission, complex or highly sensitive light detectors are not required. CONCLUSIONS: The combined chemistries of the BART reporter and amplification require only a constant temperature maintained by a heating block and are shown to be robust in the analysis of clinical samples. Since monitoring the BART reaction requires only a

  18. Bioluminescence imaging of cord blood derived mesenchymal stem cell transplanatation into myocardium

    The conventional method of analyzing myocardial cell transplanation relies on postmortem histology. We sought to demonstrate the feasibility of longitudinal monitoring transplanted cell survival in living animals using optical imaging techniques. Umblical cord blood was collected upon delivery with informed consent. Umblical mononuclear cells were obtained by negative immuno-depletion of CD3, CD14, CD19, CD38, CD66b, and glycophorin- A positive cells, followed by Ficoll- Paque density gradient centrifugation, and plated in non-coated tissue culture flasks in expansion medium. Cells were allowed to adhere overnight, thereafter non-adherent cells were washed out with medium changes. After getting the MSCs, they were transfected [multiplicity of infection (MOl) = 40) with Ad-CMV-Fluc overnight. Rats (n=4) underwent intramyocardial injection of 5 x 105 MSCs expressing firefly luciferase (Fluc) reporter gene. Optical bioluminescence imaging was performed using the charged-coupled device camera (Xenogen) from the 1st day of transplantion. Cardiac bioluminescence signals were present from 2nd day of transplantation. Cardiac signals were clearly present at day 2 (9.2x103p/s/cm2/sr). The signal reduced from day 3. The locations, magnitude, and survival duration of cord blood derived MSCs were monitored noninvasively. With further development, molecular imaging studies should add critical insights into cardiac cell transplantation

  19. Bioluminescence imaging of cord blood derived mesenchymal stem cell transplanatation into myocardium

    Min, Jung Joon; Ahn, Young Keun; Moon, Sung Min; Lim, Sang Yup; Yun, Kyung Ho; Heo, Young Jun; Song, Ho Chun; Jeong, Myung Ho; Bom, Hee Seung [School of Medicine, Chonnam National University, Gwangju (Korea, Republic of)

    2004-07-01

    The conventional method of analyzing myocardial cell transplanation relies on postmortem histology. We sought to demonstrate the feasibility of longitudinal monitoring transplanted cell survival in living animals using optical imaging techniques. Umblical cord blood was collected upon delivery with informed consent. Umblical mononuclear cells were obtained by negative immuno-depletion of CD3, CD14, CD19, CD38, CD66b, and glycophorin- A positive cells, followed by Ficoll- Paque density gradient centrifugation, and plated in non-coated tissue culture flasks in expansion medium. Cells were allowed to adhere overnight, thereafter non-adherent cells were washed out with medium changes. After getting the MSCs, they were transfected [multiplicity of infection (MOl) = 40) with Ad-CMV-Fluc overnight. Rats (n=4) underwent intramyocardial injection of 5 x 10{sup 5} MSCs expressing firefly luciferase (Fluc) reporter gene. Optical bioluminescence imaging was performed using the charged-coupled device camera (Xenogen) from the 1st day of transplantion. Cardiac bioluminescence signals were present from 2nd day of transplantation. Cardiac signals were clearly present at day 2 (9.2x10{sup 3}p/s/cm{sup 2}/sr). The signal reduced from day 3. The locations, magnitude, and survival duration of cord blood derived MSCs were monitored noninvasively. With further development, molecular imaging studies should add critical insights into cardiac cell transplantation.

  20. Relation between deep bioluminescence and oceanographic variables: A statistical analysis using time-frequency decompositions

    Martini, S.; Nerini, D.; Tamburini, C.

    2014-09-01

    We consider the statistical analysis of a 1.7-year high-frequency sampled time series, between 2009 and 2010, recorded at the ANTARES observatory in the deep NW Mediterranean Sea (2475 m depth). The objective was to estimate relationships between bioluminescence and environmental time series (temperature, salinity and current speed). As this entire dataset is characterized by non-linearity and non-stationarity, two time-frequency decomposition methods (wavelet and Hilbert-Huang) were used. These mathematical methods are dedicated to the analysis of a signal at various time and frequencies scales. This work propose some statistical tools dedicated to the study of relationships between two time series. Our study highlights three events of high bioluminescence activity in March 2009, December 2009 and March 2010. We demonstrate that the two events occurring in March 2009 and 2010 are correlated to the arrival of newly formed deep water masses at frequencies of approximately 4.8×10-7 (period of 24.1 days). In contrast, the event in December 2009 is only correlated with current speed at frequencies of approximately 1.9×10-6 (period of 6.0 days). The use of both wavelet and Hilbert-Huang transformations has proven to be successful for the analysis of multivariate time series. These methods are well-suited in a context of the increasing number of long time series recorded in oceanography.

  1. Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models

    Han, Wei; Li, Hui; Segal, Brahm H.; Blackwell, Timothy S.

    2012-01-01

    NADPH oxidase is a critical enzyme that mediates antibacterial and antifungal host defense. In addition to its role in antimicrobial host defense, NADPH oxidase has critical signaling functions that modulate the inflammatory response 1. Thus, the development of a method to measure in "real-time" the kinetics of NADPH oxidase-derived ROS generation is expected to be a valuable research tool to understand mechanisms relevant to host defense, inflammation, and injury.

  2. Bioluminescence : the potential of a non-invasive bio-optical imaging technique and improvement of animal research

    Hesselink, J. W.; van Dam, G. M.

    2007-01-01

    Bioluminescence is an optical imaging technique that exploits the emission of photons at specific wavelengths based on energy-dependent reactions catalysed by luciferases. The technique makes it possible to monitor measure, and track biological processes in living animals. A short review is presente

  3. Noninvasive monitoring of placenta-specific transgene expression by bioluminescence imaging.

    Xiujun Fan

    Full Text Available BACKGROUND: Placental dysfunction underlies numerous complications of pregnancy. A major obstacle to understanding the roles of potential mediators of placental pathology has been the absence of suitable methods for tissue-specific gene manipulation and sensitive assays for studying gene functions in the placentas of intact animals. We describe a sensitive and noninvasive method of repetitively tracking placenta-specific gene expression throughout pregnancy using lentivirus-mediated transduction of optical reporter genes in mouse blastocysts. METHODOLOGY/PRINCIPAL FINDINGS: Zona-free blastocysts were incubated with lentivirus expressing firefly luciferase (Fluc and Tomato fluorescent fusion protein for trophectoderm-specific infection and transplanted into day 3 pseudopregnant recipients (GD3. Animals were examined for Fluc expression by live bioluminescence imaging (BLI at different points during pregnancy, and the placentas were examined for tomato expression in different cell types on GD18. In another set of experiments, blastocysts with maximum photon fluxes in the range of 2.0E+4 to 6.0E+4 p/s/cm(2/sr were transferred. Fluc expression was detectable in all surrogate dams by day 5 of pregnancy by live imaging, and the signal increased dramatically thereafter each day until GD12, reaching a peak at GD16 and maintaining that level through GD18. All of the placentas, but none of the fetuses, analyzed on GD18 by BLI showed different degrees of Fluc expression. However, only placentas of dams transferred with selected blastocysts showed uniform photon distribution with no significant variability of photon intensity among placentas of the same litter. Tomato expression in the placentas was limited to only trophoblast cell lineages. CONCLUSIONS/SIGNIFICANCE: These results, for the first time, demonstrate the feasibility of selecting lentivirally-transduced blastocysts for uniform gene expression in all placentas of the same litter and early

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

    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

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

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

    2005-01-01

    The feasibility and limits in performing tomographic bioluminescence imaging with a combined optical-PET (OPET) system was 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 °C. The TOAST finite-element code was employed to simulate the diffuse transport of photons emitted from biolumi...

  6. In Vivo Bioluminescent Imaging of ATP-Binding Cassette Transporter-Mediated Efflux at the Blood-Brain Barrier.

    Bakhsheshian, Joshua; Wei, Bih-Rong; Hall, Matthew D; Simpson, R Mark; Gottesman, Michael M

    2016-01-01

    We provide a detailed protocol for imaging ATP-binding cassette subfamily G member 2 (ABCG2) function at the blood-brain barrier (BBB) of transgenic mice. D-Luciferin is specifically transported by ABCG2 found on the apical side of endothelial cells at the BBB. The luciferase-luciferin enzymatic reaction produces bioluminescence, which allows a direct measurement of ABCG2 function at the BBB. Therefore bioluminescence imaging (BLI) correlates with ABCG2 function at the BBB and this can be measured by administering luciferin in a mouse model that expresses luciferase in the brain parenchyma. BLI allows for a relatively low-cost alternative for studying transporter function in vivo compared to other strategies such as positron emission tomography. This method for imaging ABCG2 function at the BBB can be used to investigate pharmacokinetic inhibition of the transporter. PMID:27424909

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

    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

  8. Metabolic imaging in microregions of tumors and normal tissues with bioluminescence and photon counting

    A method has been developed for metabolic imaging on a microscopic level in tumors, tumor spheroids, and normal tissues. The technique makes it possible to determine the spatial distribution of glucose, lactate, and ATP in absolute terms at similar locations within tissues or cell aggregates. The substrate distributions are registered in serial cryostat sections from tissue cryobiopsies or from frozen spheroids with the use of bioluminescence reactions. The light emission is measured directly by a special imaging photon counting system enabling on-line image analysis. The technique has been applied to human breast cancer xenografts, to spheroids originating from a human colon adenocarcinoma, and to skeletal rat muscle. Preliminary data obtained indicate that heterogeneities in the substrate distributions measured are much more pronounced in tumors than in normal tissue. There was no obvious correlation among the three quantities measured at similar locations within the tissues. The distribution of ATP corresponded well with the histological structure of larger spheroids; values were low in the necrotic center and high in the viable rim of these cell aggregates

  9. In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging

    Cao, Juan; Hou, Shike; Ding, Hui; Liu, Ziquan; Song, Meijuan; Qin, Xiaojing; Wang, Xue; Yu, Mengyang; Sun, Zhiguang; Liu, Jinyang; Sun, Shuli; Xiao, Peixin

    2016-01-01

    Recently, mesenchymal stem cells (MSCs) are increasingly used as a panacea for multiple types of disease short of effective treatment. Dozens of clinical trials published demonstrated strikingly positive therapeutic effects of MSCs. However, as a specific agent, little research has focused on the dynamic distribution of MSCs after in vivo administration. In this study, we track systemically transplanted allogeneic bone marrow mesenchymal stem cells (BMSCs) in normal rats through bioluminescence imaging (BLI) in real time. Ex vivo organ imaging, immunohistochemistry (IHC), and RT-PCR were conducted to verify the histological distribution of BMSCs. Our results showed that BMSCs home to the dorsal skin apart from the lungs and kidneys after tail vein injection and could not be detected 14 days later. Allogeneic BMSCs mainly appeared not at the parenchymatous organs but at the subepidermal connective tissue and adipose tissue in healthy rats. There were no significant MSCs-related adverse effects except for transient decrease in neutrophils. These findings will provide experimental evidences for a better understanding of the biocharacteristics of BMSCs.

  10. Physicochemical characterization and in vivo bioluminescence imaging of nanostructured lipid carriers for targeting the brain: apomorphine as a model drug

    Hsu, Shu-Hui; Wen, Chih-Jen; Al-Suwayeh, S. A.; Chang, Hui-Wen; Yen, Tzu-Chen; Fang, Jia-You

    2010-10-01

    Nanostructured lipid carriers (NLCs) were prepared to investigate whether the duration of brain targeting and accumulation of drugs in the brain can be improved by intravenous delivery. NLCs were developed using cetyl palmitate as the lipid matrix, squalene as the cationic surfactant, and Pluronic F68, polysorbate 80 and polyethylene glycol as the interfacial additives. Solid lipid nanoparticles (SLNs) and lipid emulsions (LEs) were also prepared for comparison. An anti-Parkinson's drug, apomorphine, was used as the model drug. Nuclear magnetic resonance and differential scanning calorimetry showed possible interactions between the solid and liquid lipids in the inner core. The lipid nanoparticles with different compositions were characterized by mean size, zeta potential, apomorphine encapsulation and in vitro drug release. NLCs were 370-430 nm in size, which was between the sizes of the SLNs and LEs. A cationic surfactant was used to produce a positive surface charge of 42-50 mV. The base form of apomorphine was successfully entrapped by NLCs with an entrapment percentage of > 60%. The loading of apomorphine in nanoparticles resulted in a slower release behavior compared to the aqueous solution, with LEs showing the lowest release. In vivo real-time bioluminescence imaging of the rat brain revealed that NLCs could be targeted, through certain vessels, to selected brain regions. This effect was further confirmed by imaging the entire brain and brain slices. The results indicated that NLCs with moderate additives are a promising controlled-release and drug-targeting system.

  11. Physicochemical characterization and in vivo bioluminescence imaging of nanostructured lipid carriers for targeting the brain: apomorphine as a model drug

    Nanostructured lipid carriers (NLCs) were prepared to investigate whether the duration of brain targeting and accumulation of drugs in the brain can be improved by intravenous delivery. NLCs were developed using cetyl palmitate as the lipid matrix, squalene as the cationic surfactant, and Pluronic F68, polysorbate 80 and polyethylene glycol as the interfacial additives. Solid lipid nanoparticles (SLNs) and lipid emulsions (LEs) were also prepared for comparison. An anti-Parkinson's drug, apomorphine, was used as the model drug. Nuclear magnetic resonance and differential scanning calorimetry showed possible interactions between the solid and liquid lipids in the inner core. The lipid nanoparticles with different compositions were characterized by mean size, zeta potential, apomorphine encapsulation and in vitro drug release. NLCs were 370-430 nm in size, which was between the sizes of the SLNs and LEs. A cationic surfactant was used to produce a positive surface charge of 42-50 mV. The base form of apomorphine was successfully entrapped by NLCs with an entrapment percentage of > 60%. The loading of apomorphine in nanoparticles resulted in a slower release behavior compared to the aqueous solution, with LEs showing the lowest release. In vivo real-time bioluminescence imaging of the rat brain revealed that NLCs could be targeted, through certain vessels, to selected brain regions. This effect was further confirmed by imaging the entire brain and brain slices. The results indicated that NLCs with moderate additives are a promising controlled-release and drug-targeting system.

  12. In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging.

    Cao, Juan; Hou, Shike; Ding, Hui; Liu, Ziquan; Song, Meijuan; Qin, Xiaojing; Wang, Xue; Yu, Mengyang; Sun, Zhiguang; Liu, Jinyang; Sun, Shuli; Xiao, Peixin; Lv, Qi; Fan, Haojun

    2016-01-01

    Recently, mesenchymal stem cells (MSCs) are increasingly used as a panacea for multiple types of disease short of effective treatment. Dozens of clinical trials published demonstrated strikingly positive therapeutic effects of MSCs. However, as a specific agent, little research has focused on the dynamic distribution of MSCs after in vivo administration. In this study, we track systemically transplanted allogeneic bone marrow mesenchymal stem cells (BMSCs) in normal rats through bioluminescence imaging (BLI) in real time. Ex vivo organ imaging, immunohistochemistry (IHC), and RT-PCR were conducted to verify the histological distribution of BMSCs. Our results showed that BMSCs home to the dorsal skin apart from the lungs and kidneys after tail vein injection and could not be detected 14 days later. Allogeneic BMSCs mainly appeared not at the parenchymatous organs but at the subepidermal connective tissue and adipose tissue in healthy rats. There were no significant MSCs-related adverse effects except for transient decrease in neutrophils. These findings will provide experimental evidences for a better understanding of the biocharacteristics of BMSCs. PMID:27610137

  13. Physicochemical characterization and in vivo bioluminescence imaging of nanostructured lipid carriers for targeting the brain: apomorphine as a model drug

    Hsu, Shu-Hui [Department of Pharmacy, Chia Nan University of Pharmacy and Science, Tainan 717, Taiwan (China); Wen, Chih-Jen; Yen, Tzu-Chen [Animal Molecular Imaging Center, Chang Gung Memorial Hospital, Kweishan, Taoyuan 333, Taiwan (China); Al-Suwayeh, S A; Fang, Jia-You [Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh (Saudi Arabia); Chang, Hui-Wen, E-mail: fajy@mail.cgu.edu.tw [Pharmaceutics Laboratory, Graduate Institute of Natural Products, Chang Gung University, Kweishan, Taoyuan 333, Taiwan (China)

    2010-10-08

    Nanostructured lipid carriers (NLCs) were prepared to investigate whether the duration of brain targeting and accumulation of drugs in the brain can be improved by intravenous delivery. NLCs were developed using cetyl palmitate as the lipid matrix, squalene as the cationic surfactant, and Pluronic F68, polysorbate 80 and polyethylene glycol as the interfacial additives. Solid lipid nanoparticles (SLNs) and lipid emulsions (LEs) were also prepared for comparison. An anti-Parkinson's drug, apomorphine, was used as the model drug. Nuclear magnetic resonance and differential scanning calorimetry showed possible interactions between the solid and liquid lipids in the inner core. The lipid nanoparticles with different compositions were characterized by mean size, zeta potential, apomorphine encapsulation and in vitro drug release. NLCs were 370-430 nm in size, which was between the sizes of the SLNs and LEs. A cationic surfactant was used to produce a positive surface charge of 42-50 mV. The base form of apomorphine was successfully entrapped by NLCs with an entrapment percentage of > 60%. The loading of apomorphine in nanoparticles resulted in a slower release behavior compared to the aqueous solution, with LEs showing the lowest release. In vivo real-time bioluminescence imaging of the rat brain revealed that NLCs could be targeted, through certain vessels, to selected brain regions. This effect was further confirmed by imaging the entire brain and brain slices. The results indicated that NLCs with moderate additives are a promising controlled-release and drug-targeting system.

  14. Assessment of chitosan-affected metabolic response by peroxisome proliferator-activated receptor bioluminescent imaging-guided transcriptomic analysis.

    Chia-Hung Kao

    Full Text Available Chitosan has been widely used in food industry as a weight-loss aid and a cholesterol-lowering agent. Previous studies have shown that chitosan affects metabolic responses and contributes to anti-diabetic, hypocholesteremic, and blood glucose-lowering effects; however, the in vivo targeting sites and mechanisms of chitosan remain to be clarified. In this study, we constructed transgenic mice, which carried the luciferase genes driven by peroxisome proliferator-activated receptor (PPAR, a key regulator of fatty acid and glucose metabolism. Bioluminescent imaging of PPAR transgenic mice was applied to report the organs that chitosan acted on, and gene expression profiles of chitosan-targeted organs were further analyzed to elucidate the mechanisms of chitosan. Bioluminescent imaging showed that constitutive PPAR activities were detected in brain and gastrointestinal tract. Administration of chitosan significantly activated the PPAR activities in brain and stomach. Microarray analysis of brain and stomach showed that several pathways involved in lipid and glucose metabolism were regulated by chitosan. Moreover, the expression levels of metabolism-associated genes like apolipoprotein B (apoB and ghrelin genes were down-regulated by chitosan. In conclusion, these findings suggested the feasibility of PPAR bioluminescent imaging-guided transcriptomic analysis on the evaluation of chitosan-affected metabolic responses in vivo. Moreover, we newly identified that downregulated expression of apoB and ghrelin genes were novel mechanisms for chitosan-affected metabolic responses in vivo.

  15. Development of bioluminescent Salmonella strains for use in food safety

    Bailey R Hartford

    2008-01-01

    Full Text Available Abstract Background Salmonella can reside in healthy animals without the manifestation of any adverse effects on the carrier. If raw products of animal origin are not handled properly during processing or cooked to a proper temperature during preparation, salmonellosis can occur. In this research, we developed bioluminescent Salmonella strains that can be used for real-time monitoring of the pathogen's growth on food products. To accomplish this, twelve Salmonella strains from the broiler production continuum were transformed with the broad host range plasmid pAKlux1, and a chicken skin attachment model was developed. Results Salmonella strains carrying pAKlux1 constitutively expressed the luxCDABE operon and were therefore detectable using bioluminescence. Strains were characterized in terms of bioluminescence properties and plasmid stability. To assess the usefulness of bioluminescent Salmonella strains in food safety studies, we developed an attachment model using chicken skin. The effect of washing on attachment of Salmonella strains to chicken skin was tested using bioluminescent strains, which revealed the attachment properties of each strain. Conclusion This study demonstrated that bioluminescence is a sensitive and effective tool to detect Salmonella on food products in real-time. Bioluminescence imaging is a promising technology that can be utilized to evaluate new food safety measures for reducing Salmonella contamination on food products.

  16. Bioluminescence imaging of transplanted human endothelial colony-forming cells in an ischemic mouse model.

    Ding, Jie; Zhao, Zhen; Wang, Chao; Wang, Cong-Xiao; Li, Pei-Cheng; Qian, Cheng; Teng, Gao-Jun

    2016-07-01

    Ischemic strokes are devastating events responsible for high mortality and morbidity worldwide each year. Endothelial colony-forming cell (ECFC) therapy holds promise for stroke treatment; however, grafted ECFCs need to be monitored better understand their biological behavior in vivo, so as to evaluate their safety and successful delivery. The objectives of this study are to visualize the fate of infused human cord blood derived ECFCs via bioluminescence imaging (BLI) in an ischemic stroke mouse model and to determine the therapeutic effects of ECFC transplantation. ECFCs derived from human umbilical cord blood were infected with lentivirus carrying enhanced green fluorescent protein (eGFP) and firefly luciferase (Luc2) double fusion reporter gene. Labeled ECFCs were grafted into a photothrombotic ischemic stroke mouse model via intra-arterial injection though the left cardiac ventricle. The homing of infused cells and functional recovery of stroke mice were evaluated using BLI, neurological scoring, and immunohistochemistry. Significantly, BLI signals were highest in the brain on day 1 and decreased steadily until day 14. GFP-positive cells were also found surrounding infarct border zones in brain sections using immunohistochemical staining, suggesting that ECFCs properly homed to the ischemic brain tissue. Using a modified neurological severity score assay and histological analysis of brain slices with CD31 immunostaining in brain tissue, double cortin analysis, and the TdT-mediated dUTP nick end labeling (TUNEL) assay, we demonstrated functional restoration, improved angiogenesis, neurogenesis, and decreased apoptosis in ischemic mice after ECFC infusion. Collectively, our data support that ECFCs may be a promising therapeutic agent for stroke. PMID:27038754

  17. A new multicolor bioluminescence imaging platform to investigate NF-κB activity and apoptosis in human breast cancer cells.

    Laura Mezzanotte

    Full Text Available BACKGROUND: Evaluation of novel drugs for clinical development depends on screening technologies and informative preclinical models. Here we developed a multicolor bioluminescent imaging platform to simultaneously investigate transcription factor NF-κB signaling and apoptosis. METHODS: The human breast cancer cell line (MDA-MB-231 was genetically modified to express green, red and blue light emitting luciferases to monitor cell number and viability, NF-κB promoter activity and to perform specific cell sorting and detection, respectively. The pro-luciferin substrate Z-DEVD-animoluciferin was employed to determine apoptotic caspase 3/7 activity. We used the cell line for the in vitro evaluation of natural compounds and in vivo optical imaging of tumor necrosis factor TNFα-induced NF-κB activation. RESULTS: Celastrol, resveratrol, sulphoraphane and curcumin inhibited the NF-κB promoter activity significantly and in a dose dependent manner. All compounds except resveratrol induced caspase 3/7 dependent apoptosis. Multicolor bioluminescence in vivo imaging allowed the investigation of tumor growth and NF-κB induction in a mouse model of breast cancer. CONCLUSION: Our new method provides an imaging platform for the identification, validation, screening and optimization of compounds acting on NF-κB signaling and apoptosis both in vitro and in vivo.

  18. A New Multicolor Bioluminescence Imaging Platform to Investigate NF-κB Activity and Apoptosis in Human Breast Cancer Cells

    Mezzanotte, Laura; An, Na; Mol, Isabel M.; Löwik, Clemens W. G. M.; Kaijzel, Eric L.

    2014-01-01

    Background Evaluation of novel drugs for clinical development depends on screening technologies and informative preclinical models. Here we developed a multicolor bioluminescent imaging platform to simultaneously investigate transcription factor NF-κB signaling and apoptosis. Methods The human breast cancer cell line (MDA-MB-231) was genetically modified to express green, red and blue light emitting luciferases to monitor cell number and viability, NF-κB promoter activity and to perform specific cell sorting and detection, respectively. The pro-luciferin substrate Z-DEVD-animoluciferin was employed to determine apoptotic caspase 3/7 activity. We used the cell line for the in vitro evaluation of natural compounds and in vivo optical imaging of tumor necrosis factor TNFα-induced NF-κB activation. Results Celastrol, resveratrol, sulphoraphane and curcumin inhibited the NF-κB promoter activity significantly and in a dose dependent manner. All compounds except resveratrol induced caspase 3/7 dependent apoptosis. Multicolor bioluminescence in vivo imaging allowed the investigation of tumor growth and NF-κB induction in a mouse model of breast cancer. Conclusion Our new method provides an imaging platform for the identification, validation, screening and optimization of compounds acting on NF-κB signaling and apoptosis both in vitro and in vivo. PMID:24465597

  19. Three-dimensional localization of in vivo bioluminescent source based on multispectral imaging

    Feng, Jinchao; Jia, Kebin; Tian, Jie; Yan, Guorui; Zhu, Shouping

    2009-02-01

    Bioluminescence tomography (BLT) is a novel in vivo technique in small animal studies, which can reveal the molecular and cellular information at the whole-body small animal level. At present, there is an increasing interest in multispectral bioluminescence tomography, since multispectral data acquisition could improve the BLT performance significantly. In view to the ill-posedness of BLT problem, we develop an optimal permissible source region strategy to constrain the possible solution of the source by utilizing spectrum character of bioluminescent source. Then a linear system to link the measured data with the unknown light source variables is established by utilizing the optimal permissible region strategy based on adaptive finite element analysis. Furthermore, singular value decomposition analysis is used for data dimensionality reduction and improving computational efficiency in multispectral case. The reconstructed speed and stability benefit from adaptive finite element, the permissible region strategy and singular value decomposition. In the numerical simulation, the heterogeneous phantom experiment has been used to evaluate the performance of the proposed algorithm with the Monte Carlo based synthetic data. The reconstruction results demonstrate the merits and potential of our methodology for localizing bioluminescent source.

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

    Hai-juan WANG

    2012-05-01

    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 200μl 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, β=0.972; R2=0.991, β=0.996; R2=0.351, β=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.

  1. Comprehensive assessment of host responses to ionizing radiation by nuclear factor-κB bioluminescence imaging-guided transcriptomic analysis.

    Chung-Ta Chang

    Full Text Available The aim of this study was to analyze the host responses to ionizing radiation by nuclear factor-κB (NF-κB bioluminescence imaging-guided transcriptomic tool. Transgenic mice carrying the NF-κB-driven luciferase gene were exposed to a single dose of 8.5 Gy total-body irradiation. In vivo imaging showed that a maximal NF-κB-dependent bioluminescent intensity was observed at 3 h after irradiation and ex vivo imaging showed that liver, intestine, and brain displayed strong NF-κB activations. Microarray analysis of these organs showed that irradiation altered gene expression signatures in an organ-specific manner and several pathways associated with metabolism and immune system were significantly altered. Additionally, the upregulation of fatty acid binding protein 4, serum amyloid A2, and serum amyloid A3 genes, which participate in both inflammation and lipid metabolism, suggested that irradiation might affect the cross pathways of metabolism and inflammation. Moreover, the alteration of chemokine (CC-motif ligand 5, chemokine (CC-motif ligand 20, and Jagged 1 genes, which are involved in the inflammation and enterocyte proliferation, suggested that these genes might be involved in the radiation enteropathy. In conclusion, this report describes the comprehensive evaluation of host responses to ionizing radiation. Our findings provide the fundamental information about the in vivo NF-κB activity and transcriptomic pattern after irradiation. Moreover, novel targets involved in radiation injury are also suggested.

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

    Conley, Nicholas R.

    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

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

    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

  4. Development of a Multicolor Bioluminescence Imaging Platform to Simultaneously Investigate Transcription Factor NF-κB Signaling and Apoptosis.

    Knol-Blankevoort, Vicky T; Mezzanotte, Laura; Rabelink, Martijn J W E; Löwik, Clemens W G M; Kaijzel, Eric L

    2016-01-01

    Here we describe a novel multicolor bioluminescent imaging platform that enables us to simultaneously investigate transcription factor nuclear factor-κB (NF-κB) signalling and apoptosis. We genetically modified the human breast cancer cell line MDA-MB-231 to express green, red, and blue light-emitting luciferases to monitor cell number and viability, NF-κB promoter activity, and to enable specific cell sorting and detection, respectively. Z-DEVD-animoluciferin, the pro-luciferin substrate, was used to determine apoptotic caspase 3/7 activity. We used this multicolored cell line for the in vitro evaluation of natural compounds and in vivo optical imaging of tumor necrosis factor (TNFα)-induced NF-κB activation (Mezzanotte et al., PLoS One 9:e85550, 2014). PMID:27424911

  5. Destabilized bioluminescent proteins

    Allen, Michael S.; Rakesh, Gupta; Gary, Sayler S.

    2007-07-31

    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.

  6. A Bone Metastasis Nude Mouse Model Created by Ultrasound Guided Intracardiac Injection of Breast Cancer Cells: the Micro-CT, MRI and Bioluminescence Imaging Analysis

    The purpose of this study was to develop a nude mouse model of bone metastasis by performing intracardiac injection of breast cancer cells under ultrasonography guidance and we wanted to evaluate the development and the distribution of metastasis in vivo using micro-CT, MRI and bioluminescence imaging. Animal experiments were performed in 6-week-old female nude mice. The animals underwent left ventricular injection of 2x105 MDA-MB-231Bo-Luc cells. After injection of the tumor cells, serial bioluminescence imaging was performed for 7 weeks. The findings of micro-CT, MRI and the histology were correlated with the 'hot' lesions seen on the bioluminescence imaging. Metastasis was found in 62.3% of the animals. Two weeks after intracardiac injection, metastasis to the brain, spine and femur was detected with bioluminescence imaging with an increasing intensity by week 7. Micro-CT scan confirmed multiple osteolytic lesions at the femur, spine and skull. MRI and the histology were able to show metastasis in the brain and extraskeletal metastasis around the femur. The intracardiac injection of cancer cells under ultrasonography guidance is a safe and highly reproducible method to produce bone metastasis in nude mice. This bone metastasis nude mouse model will be useful to study the mechanism of bone metastasis and to validate new therapeutics

  7. A Bone Metastasis Nude Mouse Model Created by Ultrasound Guided Intracardiac Injection of Breast Cancer Cells: the Micro-CT, MRI and Bioluminescence Imaging Analysis

    Park, Young Jin; Song, Eun Hye; Kim, Seol Hwa; Song, Ho Taek; Suh, Jin Suck [Yonsei University College of Medicine, Seoul (Korea, Republic of); Choi, Sang Hyun [Korean Minjok Leadership Academy, Heongsung (Korea, Republic of)

    2011-01-15

    The purpose of this study was to develop a nude mouse model of bone metastasis by performing intracardiac injection of breast cancer cells under ultrasonography guidance and we wanted to evaluate the development and the distribution of metastasis in vivo using micro-CT, MRI and bioluminescence imaging. Animal experiments were performed in 6-week-old female nude mice. The animals underwent left ventricular injection of 2x105 MDA-MB-231Bo-Luc cells. After injection of the tumor cells, serial bioluminescence imaging was performed for 7 weeks. The findings of micro-CT, MRI and the histology were correlated with the 'hot' lesions seen on the bioluminescence imaging. Metastasis was found in 62.3% of the animals. Two weeks after intracardiac injection, metastasis to the brain, spine and femur was detected with bioluminescence imaging with an increasing intensity by week 7. Micro-CT scan confirmed multiple osteolytic lesions at the femur, spine and skull. MRI and the histology were able to show metastasis in the brain and extraskeletal metastasis around the femur. The intracardiac injection of cancer cells under ultrasonography guidance is a safe and highly reproducible method to produce bone metastasis in nude mice. This bone metastasis nude mouse model will be useful to study the mechanism of bone metastasis and to validate new therapeutics

  8. In vivo bioluminescence imaging of hyperglycemia exacerbating stem cells on choroidal neovascularization in mice

    Gao, Xiang; Wang, Yu; Hou, Hui-Yuan; Lyu, Yang; Wang, Hai-Yan; Yao, Li-Bo; Zhang, Jian; Cao, Feng; Wang, Yu-Sheng

    2016-01-01

    AIM To investigate the influence of hyperglycemia on the severity of choroidal neovascularization (CNV), especially the involvement of bone marrow-derived cells (BMCs) and underlying mechanisms. METHODS BMCs from firefly luciferase (Fluc)/green fluorescent protein (GFP) double transgenic mice were transplanted into C57BL/6J wide-type mice. The recipient mice were injected intraperitoneally with streptozotocin (STZ) daily for 5 consecutive days to induce diabetes mellitus (DM), followed by CNV laser photocoagulation. The BMCs recruitment in CNV exposed to hyperglycemia was firstly examined in Fluc/GFP chimeric mice by in vivo optical bioluminescence imaging (BLI) and in vitro Fluc assays. The CNV severity was evaluated by H&E staining and choroidal flatmount. The expression of vascular endothelial growth factor (VEGF) and stromal cell derived factor-1 (SDF-1) was detected by Western Blot. RESULTS BLI showed that the BMCs exerted dynamic effects in CNV model in Fluc/GFP chimeric mice exposed to hyperglycemia. The signal intensity of transplanted Fluc+GFP+ BMCs in the DM chimeric mice was significantly higher than that in the control chimeric mice with CNV induction at days 5, 7, 14 and 21 (121861.67±9948.81 vs 144998.33±13787.13 photons/second/cm2/sr for control and DM mice, P5d<0.05; 178791.67±30350.8 vs 240166.67±22605.3, P7d<0.05; 124176.67±16253.52 vs 196376.67±18556.79, P14d<0.05; 97951.60±10343.09 vs 119510.00±14383.76, P21d<0.05), which was consistent with in vitro Fluc assay at day 7 [relative light units of Fluc (RLU1)], 215.00±52.05 vs 707.33±88.65, P<0.05; RLU1/ relative light units of renilla luciferase (RLU2), 0.90±0.17 vs 1.83±0.17, P<0.05]. The CNVs in the DM mice were wider than those in the control group at days 5, 7, 14 and 21 (147.83±17.36 vs 220.33±20.17 µm, P5d<0.05; 212.17±24.63 vs 326.83±19.49, P7d<0.05; 163.17±18.24 vs 265.17±20.55, P14d<0.05; 132.00±10.88 vs 205.33±12.98, P21d<0.05). The average area of CNV in the DM

  9. A Bioluminescence Assay System for Imaging Metal Cationic Activities in Urban Aerosols.

    Kim, Sung-Bae; Naganawa, Ryuichi; Murata, Shingo; Nakayama, Takayoshi; Miller, Simon; Senda, Toshiya

    2016-01-01

    A bioluminescence-based assay system was fabricated for an efficient determination of the activities of air pollutants. The following four components were integrated into this assay system: (1) an 8-channel assay platform uniquely designed for simultaneously sensing multiple optical samples, (2) single-chain probes illuminating toxic chemicals or heavy metal cations from air pollutants, (3) a microfluidic system for circulating medium mimicking the human body, and (4) the software manimulating the above system. In the protocol, we briefly introduce how to integrate the components into the system and the application to the illumination of the metal cationic activities in air pollutants. PMID:27424913

  10. Improved Reconstruction Quality of Bioluminescent Images by Combining SP3 Equations and Bregman Iteration Method

    Qiang Wu

    2013-01-01

    Full Text Available Bioluminescence tomography (BLT has a great potential to provide a powerful tool for tumor detection, monitoring tumor therapy progress, and drug development; developing new reconstruction algorithms will advance the technique to practical applications. In the paper, we propose a BLT reconstruction algorithm by combining SP3 equations and Bregman iteration method to improve the quality of reconstructed sources. The numerical results for homogeneous and heterogeneous phantoms are very encouraging and give significant improvement over the algorithms without the use of SP3 equations and Bregman iteration method.

  11. Inflammatory modulating effects of low level laser therapy on iNOS expression by means of bioluminescence imaging

    Moriyama, Yumi; Moriyama, Eduardo H.; Blackmore, Kristina; Akens, Margarete K.; Lilge, Lothar

    2005-09-01

    This study investigates the efficacy of low level laser therapy (LLLT) in modulating inducible nitric oxide synthase (iNOS) expression as molecular marker of the inflammation signaling pathway. LLLT was mediated by different therapeutic wavelengths using transgenic animals with the luciferase gene under control of the iNOS gene expression. Inflammation in 30 transgenic mice (iNOS-luc mice, from FVB strain) was induced by intra-articular injection of Zymosan-A in both knee joints. Four experimental groups were treated with one of four different wavelengths (λ=635, 785, 808 and 905nm) and one not laser-irradiated control group. Laser treatment (25 mW cm-2, 5 J cm-2) was applied to the knees 15 minutes after inflammation induction. Measurements of iNOS expression were performed at multiple times (0, 3, 5, 7, 9 and 24h) post-LLLT by measuring the bioluminescence signal using a highly sensitive charge-coupled device (CCD) camera. The responsivity of BLI was sufficient to demonstrate a significant increase in bioluminescence signals after laser irradiation of 635nm when compared to non-irradiated animals and the other LLLT treated groups, showing the wavelength-dependence of LLLT on iNOS expression during the acute inflammatory process.

  12. Chemiluminescence and bioluminescence microbe detection

    Taylor, R. E.; Chappelle, E.; Picciolo, G. L.; Jeffers, E. L.; Thomas, R. R.

    1978-01-01

    Automated biosensors for online use with NASA Water Monitoring System employs bioluminescence and chemiluminescence techniques to rapidly measure microbe contamination of water samples. System eliminates standard laboratory procedures requiring time duration of 24 hours or longer.

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

    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 (Δ45HSV1-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 Δ45HSV1-tk/GFP/luciferase (Δ45-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-Δ45-TGL cells had cytoplasmic green fluorescence. The in vitro BLI was 7- and 13-fold higher in U87-NES-TGL and U87-Δ45-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-Δ45-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-Δ45-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

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

    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

    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

  15. Quantitative imaging of D-2-hydroxyglutarate (D2HG in selected histological tissue areas by a novel bioluminescence technique

    Nadine Fabienne Voelxen

    2016-03-01

    Full Text Available AbstractPatients with malignant gliomas have a poor prognosis with average survival of less than one year. Whereas in other tumor entities the characteristics of tumor metabolism are successfully used for therapeutic approaches, such developments are very rare in brain tumors, notably in gliomas. One metabolic feature characteristic of gliomas, in particular diffuse astrocytomas and oligodendroglial tumors, is the variable content of D-2-hydroxyglutarate (D2HG, a metabolite, which was discovered first in this tumor entity. D2HG is generated in large amounts due to various gain-of–function mutations in the isocitrate dehydrogenases IDH-1 and IDH-2. Meanwhile, D2HG has been detected in several other tumor entities including intrahepatic bile-duct cancer, chondrosarcoma, acute myeloid leukemia, and angioimmunoblastic T-cell lymphoma. D2HG is barely detectable in healthy tissue (< 0.1 mM, but its concentration increases up to 35 mM in malignant tumor tissues. Consequently, the oncometabolite D2HG has gained increasing interest in the field of tumor metabolism. To facilitate its quantitative measurement without loss of spatial resolution at a microscopical level, we have developed a novel bioluminescence assay for determining D2HG in sections of snap-frozen tissue. The assay was verified independently by photometric tests and liquid chromatography / mass spectrometry (LC/MS. The novel technique allows the microscopically resolved determination of D2HG in a concentration range of 0 – 10 µmol/g tissue (wet weight. In combination with the already established bioluminescence imaging techniques for ATP, glucose, pyruvate, and lactate, the novel D2HG assay enables a comparative characterization of the metabolic profile of individual tumors in a further dimension.

  16. 一种生物在体荧光成像的自适应分割算法%Adaptive Segmentation Algorithm of Bioluminescent Image

    常志军; 杨鑫

    2011-01-01

    Bioluminescence imaging is regarded as an imaging modality with a high performance, low cost and good prospect in molecular imaging technique. This paper proposes a new adaptive segmentation algorithm, which is based on the characteristics and the application requirements of the bioluminescent images. The adaptive segmentation is realized by performing the normalized processing, connectivity operation and the interested regions distinguishing on bioluminescent images. Experimental results show that this algorithm can get better segmentation results in the ease of weak signal, low signal-to-noise ratio and multiple light sources, so it is a kind of effective segmentation algorithm of bioluminescent images.%生物在体荧光成像是新兴分子影像技术中性能高、费用低、前景好的一种成像模态.针对生物在体荧光图像的特点和应用需求,提出一种全新的自适应图像分割算法.通过对荧光图像的归一化处理、连通性操作、感兴趣区域区分实现自适应分割.实验结果表明,该算法能够在弱信号、低信噪比、多光源的情况下得到较理想的分割结果,是一种有效的荧光图像分割算法.

  17. Repeated and Widespread Evolution of Bioluminescence in Marine Fishes.

    Davis, Matthew P; Sparks, John S; Smith, W Leo

    2016-01-01

    Bioluminescence is primarily a marine phenomenon with 80% of metazoan bioluminescent genera occurring in the world's oceans. Here we show that bioluminescence has evolved repeatedly and is phylogenetically widespread across ray-finned fishes. We recover 27 independent evolutionary events of bioluminescence, all among marine fish lineages. This finding indicates that bioluminescence has evolved many more times than previously hypothesized across fishes and the tree of life. Our exploration of the macroevolutionary patterns of bioluminescent lineages indicates that the present day diversity of some inshore and deep-sea bioluminescent fish lineages that use bioluminescence for communication, feeding, and reproduction exhibit exceptional species richness given clade age. We show that exceptional species richness occurs particularly in deep-sea fishes with intrinsic bioluminescent systems and both shallow water and deep-sea lineages with luminescent systems used for communication. PMID:27276229

  18. Repeated and Widespread Evolution of Bioluminescence in Marine Fishes

    Davis, Matthew P.; Sparks, John S.; Smith, W. Leo

    2016-01-01

    Bioluminescence is primarily a marine phenomenon with 80% of metazoan bioluminescent genera occurring in the world’s oceans. Here we show that bioluminescence has evolved repeatedly and is phylogenetically widespread across ray-finned fishes. We recover 27 independent evolutionary events of bioluminescence, all among marine fish lineages. This finding indicates that bioluminescence has evolved many more times than previously hypothesized across fishes and the tree of life. Our exploration of the macroevolutionary patterns of bioluminescent lineages indicates that the present day diversity of some inshore and deep-sea bioluminescent fish lineages that use bioluminescence for communication, feeding, and reproduction exhibit exceptional species richness given clade age. We show that exceptional species richness occurs particularly in deep-sea fishes with intrinsic bioluminescent systems and both shallow water and deep-sea lineages with luminescent systems used for communication. PMID:27276229

  19. Observations of in situ deep-sea marine bioluminescence with a high-speed, high-resolution sCMOS camera

    Phillips, Brennan T.; Gruber, David F.; Vasan, Ganesh; Roman, Christopher N.; Pieribone, Vincent A.; Sparks, John S.

    2016-05-01

    Observing and measuring marine bioluminescence in situ presents unique challenges, characterized by the difficult task of approaching and imaging weakly illuminated bodies in a three-dimensional environment. To address this problem, a scientific complementary-metal-oxide-semiconductor (sCMOS) microscopy camera was outfitted for deep-sea imaging of marine bioluminescence. This system was deployed on multiple platforms (manned submersible, remotely operated vehicle, and towed body) in three oceanic regions (Western Tropical Pacific, Eastern Equatorial Pacific, and Northwestern Atlantic) to depths up to 2500 m. Using light stimulation, bioluminescent responses were recorded at high frame rates and in high resolution, offering unprecedented low-light imagery of deep-sea bioluminescence in situ. The kinematics of light production in several zooplankton groups was observed, and luminescent responses at different depths were quantified as intensity vs. time. These initial results signify a clear advancement in the bioluminescent imaging methods available for observation and experimentation in the deep-sea.

  20. Susceptibility of the wild-derived inbred CAST/Ei mouse to infection by orthopoxviruses analyzed by live bioluminescence imaging

    Classical inbred mice are extensively used for virus research. However, we recently found that some wild-derived inbred mouse strains are more susceptible than classical strains to monkeypox virus. Experiments described here indicated that the 50% lethal dose of vaccinia virus (VACV) and cowpox virus (CPXV) were two logs lower in wild-derived inbred CAST/Ei mice than classical inbred BALB/c mice, whereas there was little difference in the susceptibility of the mouse strains to herpes simplex virus. Live bioluminescence imaging was used to follow spread of pathogenic and attenuated VACV strains and CPXV virus from nasal passages to organs in the chest and abdomen of CAST/Ei mice. Luminescence increased first in the head and then simultaneously in the chest and abdomen in a dose-dependent manner. The spreading kinetics was more rapid with VACV than CPXV although the peak photon flux was similar. These data suggest advantages of CAST/Ei mice for orthopoxvirus studies. - Highlights: • Wild-derived inbred CAST/Ei mice are susceptible to vaccinia virus and cowpox virus. • Morbidity and mortality from orthopoxviruses are greater in CAST/Ei than BALB/c mice. • Morbidity and mortality from herpes simplex virus type 1 are similar in both mice. • Imaging shows virus spread from nose to lungs, abdominal organs and brain. • Vaccinia virus spreads more rapidly than cowpox virus

  1. Susceptibility of the wild-derived inbred CAST/Ei mouse to infection by orthopoxviruses analyzed by live bioluminescence imaging

    Americo, Jeffrey L.; Sood, Cindy L.; Cotter, Catherine A.; Vogel, Jodi L.; Kristie, Thomas M.; Moss, Bernard, E-mail: bmoss@nih.gov; Earl, Patricia L., E-mail: pearl@nih.gov

    2014-01-20

    Classical inbred mice are extensively used for virus research. However, we recently found that some wild-derived inbred mouse strains are more susceptible than classical strains to monkeypox virus. Experiments described here indicated that the 50% lethal dose of vaccinia virus (VACV) and cowpox virus (CPXV) were two logs lower in wild-derived inbred CAST/Ei mice than classical inbred BALB/c mice, whereas there was little difference in the susceptibility of the mouse strains to herpes simplex virus. Live bioluminescence imaging was used to follow spread of pathogenic and attenuated VACV strains and CPXV virus from nasal passages to organs in the chest and abdomen of CAST/Ei mice. Luminescence increased first in the head and then simultaneously in the chest and abdomen in a dose-dependent manner. The spreading kinetics was more rapid with VACV than CPXV although the peak photon flux was similar. These data suggest advantages of CAST/Ei mice for orthopoxvirus studies. - Highlights: • Wild-derived inbred CAST/Ei mice are susceptible to vaccinia virus and cowpox virus. • Morbidity and mortality from orthopoxviruses are greater in CAST/Ei than BALB/c mice. • Morbidity and mortality from herpes simplex virus type 1 are similar in both mice. • Imaging shows virus spread from nose to lungs, abdominal organs and brain. • Vaccinia virus spreads more rapidly than cowpox virus.

  2. A Dual-Color Far-Red to Near-Infrared Firefly Luciferin Analogue Designed for Multiparametric Bioluminescence Imaging**

    Jathoul, A. P.; Grounds, H.; Anderson, J.; Pule, M. A.

    2014-01-01

    Red-shifted bioluminescent emitters allow improved in vivo tissue penetration and signal quantification, and have led to the development of beetle luciferin analogues that elicit red-shifted bioluminescence with firefly luciferase. However, unlike natural luciferin, none have been shown to emit different colors with different luciferases. We have synthesized and tested the first dual color, far-red to near infrared (nIR) emitting analogue of beetle luciferin, which akin to natural luciferin e...

  3. Intracellular Self-Assembly of Cyclic d-Luciferin Nanoparticles for Persistent Bioluminescence Imaging of Fatty Acid Amide Hydrolase.

    Yuan, Yue; Wang, Fuqiang; Tang, Wei; Ding, Zhanling; Wang, Lin; Liang, Lili; Zheng, Zhen; Zhang, Huafeng; Liang, Gaolin

    2016-07-26

    Fatty acid amide hydrolase (FAAH) overexpression induces several disorder symptoms in nerve systems, and therefore long-term tracing of FAAH activity in vivo is of high importance but remains challenging. Current bioluminescence (BL) methods are limited in detecting FAAH activity within 5 h. Herein, by rational design of a latent BL probe (d-Cys-Lys-CBT)2 (1), we developed a "smart" method of intracellular reduction-controlled self-assembly and FAAH-directed disassembly of its cyclic d-luciferin-based nanoparticles (i.e., 1-NPs) for persistent BL imaging of FAAH activity in vitro, in cells, and in vivo. Using aminoluciferin methyl amide (AMA), Lys-amino-d-luciferin (Lys-Luc), and amino-d-luciferin (NH2-Luc) as control BL probes, we validated that the persistent BL of 1 from luciferase-expressing cells or tumors was controlled by the activity of intracellular FAAH. With the property of long-term tracing of FAAH activity in vivo of 1, we envision that our BL precursor 1 could probably be applied for in vivo screening of FAAH inhibitors and the diagnosis of their related diseases (or disorders) in the future. PMID:27348334

  4. 生物发光成像的特点及应用%Bioluminescence imaging characteristics and application

    杨丽平; 赵敬湘; 裴雪涛

    2009-01-01

    生物发光成像(BLI)是通过荧光素酶基因标记细胞或DNA,在ATP及氧气存在条件下,催化荧光素的氧化反应而发光,从而能够直接监控活体内的细胞活动和基因行为.该文通过比较BLI与MRI,PET、放射成像的异同,以及BLI在肿瘤、干细胞和免疫细胞运输、细胞凋亡等方面的应用,为更好地推广BLI的应用提供依据.%Bioluminescence imaging (BLI) by luciferase gene marked cells or DNA, in the presence of ATP and oxygen, catalytic oxidation reaction of fluorescein luminescence. So that it can directly monitor in vivo cell activity and gene behavior. In this paper, by comparing the BLI and MRI, PET, radiography of the similarities and differences, as well as about their cancer, stem cells and immune cells transportation, apoptosis and other aspects of the application, in order to better provide the basis for promoting the application of BLI.

  5. Real-Time Monitoring of Bacterial Infection In Vivo: Development of Bioluminescent Staphylococcal Foreign-Body and Deep-Thigh-Wound Mouse Infection Models

    Kuklin, Nelly A.; Pancari, Gregory D.; Tobery, Timothy W.; Cope, Leslie; Jackson, Jesse; Gill, Charles; Overbye, Karen; Francis, Kevin P.; Yu, Jun; Montgomery, Donna; Anderson, Annaliesa S.; McClements, William; Jansen, Kathrin U.

    2003-01-01

    Staphylococcal infections associated with catheter and prosthetic implants are difficult to eradicate and often lead to chronic infections. Development of novel antibacterial therapies requires simple, reliable, and relevant models for infection. Using bioluminescent Staphylococcus aureus, we have adapted the existing foreign-body and deep-wound mouse models of staphylococcal infection to allow real-time monitoring of the bacterial colonization of catheters or tissues. This approach also enab...

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

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

    2013-07-01

    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

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

    Kiddle Guy

    2012-04-01

    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

  8. Rational development of an attenuated recombinant cyprinid herpesvirus 3 vaccine using prokaryotic mutagenesis and in vivo bioluminescent imaging.

    Boutier, Maxime; Ronsmans, Maygane; Ouyang, Ping; Fournier, Guillaume; Reschner, Anca; Rakus, Krzysztof; Wilkie, Gavin S; Farnir, Frédéric; Bayrou, Calixte; Lieffrig, François; Li, Hong; Desmecht, Daniel; Davison, Andrew J; Vanderplasschen, Alain

    2015-02-01

    Cyprinid herpesvirus 3 (CyHV 3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candidate, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vaccine candidate. The safety and efficacy of this strain were studied using an in vivo bioluminescent imaging system (IVIS), qPCR, and histopathological examination, which demonstrated that it enters fish via skin infection similar to the wild type strain. However, compared to the parental wild type strain, the vaccine candidate replicated at lower levels and spread less efficiently to secondary sites of infection. Transmission experiments allowing water contamination with or without additional physical contact between fish demonstrated that the vaccine candidate has a reduced ability to spread from vaccinated fish to naïve sentinel cohabitants. Finally, IVIS analyses demonstrated that the vaccine candidate induces a protective mucosal immune response at the portal of entry. Thus, the present study is the first to report the rational development of a recombinant attenuated vaccine against CyHV 3 for mass vaccination of carp. We also demonstrated the relevance of the CyHV 3 carp model for studying alloherpesvirus transmission and mucosal immunity in teleost skin. PMID:25700279

  9. Development of a Chinchilla Model To Allow Direct, Continuous, Biophotonic Imaging of Bioluminescent Nontypeable Haemophilus influenzae during Experimental Otitis Media

    Novotny, Laura A.; Mason, Kevin M.; Bakaletz, Lauren O.

    2005-01-01

    We transformed a nontypeable Haemophilus influenzae clinical isolate with a plasmid containing the luxCDABE operon driven by the H. influenzae outer membrane protein P2 promoter. Herein, we demonstrate the ability to detect bioluminescence and to monitor infection within the nasopharynges, eustachian tubes, and middle ears of chinchillas after intranasal and transbullar challenges.

  10. Molecular Imaging Using Fluorescence and Bioluminescence to Reveal Tissue Response to Laser-Mediated Thermal Injury

    Mackanos, Mark A.; Jansen, E. Duco; Contag, Christopher H.

    For decades biological investigation has focused on a reductionist approach, which has greatly advanced our understanding of the biological process, but has also served to move the analysis further and further away from the living body. This was necessary as we sought to identify the cells, genes, mutations and/or etiological agents that were associated with a given process. The information generated through these approaches can now be used to advance more integrative strategies in which specific cellular and molecular events can be studied in context of the functional circulation and intact organ systems of living animals, and humans. Essential tools for integrative analyses of biology include imaging modalities that enable visualization of structure and function in the living body. The relatively recent development of molecular probes as exogenous contrast agents and reporter genes that encode proteins with unique properties that can be distinguished from tissues and cells has ushered in a new set of approaches that are being called molecular imaging.

  11. Bioluminescent imaging of ABCG2 efflux activity at the blood-placenta barrier

    Kumar, Jeyan S.; Bih-Rong Wei; Madigan, James P.; R Mark Simpson; Hall, Matthew D; Gottesman, Michael M.

    2016-01-01

    Physiologic barriers such as the blood placenta barrier (BPB) and the blood brain barrier protect the underlying parenchyma from pathogens and toxins. ATP-binding cassette (ABC) transporters are transmembrane proteins found at these barriers, and function to efflux xenobiotics and maintain chemical homeostasis. Despite the plethora of ex vivo and in vitro data showing the function and expression of ABC transporters, no imaging modality exists to study ABC transporter activity in vivo at the B...

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

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

    2013-01-01

    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 rege...

  13. Monitoring the Growth of an Orthotopic Tumour Xenograft Model: Multi-Modal Imaging Assessment with Benchtop MRI (1T), High-Field MRI (9.4T), Ultrasound and Bioluminescence

    Stuckey, Daniel J.; David, Anna L.; Pedley, R. Barbara; Lythgoe, Mark F.; Siow, Bernard; Walker-Samuel, Simon

    2016-01-01

    Background Research using orthotopic and transgenic models of cancer requires imaging methods to non-invasively quantify tumour burden. As the choice of appropriate imaging modality is wide-ranging, this study aimed to compare low-field (1T) magnetic resonance imaging (MRI), a novel and relatively low-cost system, against established preclinical techniques: bioluminescence imaging (BLI), ultrasound imaging (US), and high-field (9.4T) MRI. Methods A model of colorectal metastasis to the liver was established in eight mice, which were imaged with each modality over four weeks post-implantation. Tumour burden was assessed from manually segmented regions. Results All four imaging systems provided sufficient contrast to detect tumours in all of the mice after two weeks. No significant difference was detected between tumour doubling times estimated by low-field MRI, ultrasound imaging or high-field MRI. A strong correlation was measured between high-field MRI estimates of tumour burden and all the other modalities (p < 0.001, Pearson). Conclusion These results suggest that both low-field MRI and ultrasound imaging are accurate modalities for characterising the growth of preclinical tumour models. PMID:27223614

  14. MEASUREMENTS OF BIOLUMINESCENCE IN DEEP SEA

    Chikawa, M.; Kitamura, T; Nakagawa, Nakagawa; Yamamoto, I.; Wada, T.; Okei, K; Yamashita, Y.

    1996-01-01

    [Abstract] We have designed and built a photon counting system which measures low intensities of bioluminescence in deep sea. The system comprises a CCD-TV camera, two-dimensional image intensifier and video cassette recorder. Using this system we measured the vertical profile of bioluminescence in situ at the Suruga Trough and Nankai Trough to a depth of 3600 m and analyzed cultivated them.

  15. 腺病毒介导荧光素酶报告基因感染间充质干细胞的研究%Infection with adenovirus-mediated luciferase reporter gene in mesenchymal stem cells and bioluminescence imaging

    王一帆; 夏睿; 郭玉林; 郜发宝

    2013-01-01

    .PShuttle-CMV-Luc and backbone vector (pAdeno) were homologous recombined.Then the recombinant plasmid was packaged in HEK293 cells and the virus titer was detected.The BMSC were infected by the recombinant adenovirus.The bioluminescence imaging in vitro was performed to determine the best multiplicity of infection (MOI),and the relationship between bioluminescence intensity and MOI was analyzed by curve fitting regression analysis.Viability was evaluated via Trypan blue staining.The transfected BMSC (l× 106) were implanted into the muscles of forelimb of SD rats,and then tracked by bioluminescence imaging in vivo.Cell viability was compared using two-way repeated measures analysis of variance between groups.Results Enzyme digestion and sequence analysis indicated that Ad-Luc was successfully constructed.The virus titer was 1 × 1010 plaque forming unit (PFU)/ml.The bioluminescence detection in vitro showed that Ad-Luc could infect BMSC high efficiently to express luciferase and the best MOI was 50.The bioluminescence intensity enhanced with increase of MOI (R2 =0.98).No statistically significant difference was found in cell viability between transfected and untransfected BMSC at 1,3,5,7 d.The cell survival rates were (92.5±2.3)% vs (94.1±1.8)%,(91.4±0.9)% vs (92.7±2.0)%,(92.1±1.6)% vs (93.3± 2.4) %,(91.9 ± 1.5) % vs (93.0 ± 3.1) %,respectively (F =4.38,P > 0.05).The bioluminescence imaging in vivo showed that BMSC survived 1,3,7 d after implantation.However,bioluminescence signal decreased gradually over time.Conclusion It is feasible to apply the optical reporter gene imaging for tracing transplanted stem cells in vitro and in vivo due to the effective transformation of luciferase reporter gene into BMSC by adenovirus vector.

  16. Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian

    Background: The fertility performance of animals is still a mystery and the full comprehension of mammalian gametes maturation and early embryonic development remains to be elucidated. The recent development in nanotechnology offers a new opportunity for real-time study of reproductive cells in thei...

  17. Influence of antibiotic pressure on bacterial bioluminescence, with emphasis on Staphylococcus aureus

    Daghighi, Seyedmojtaba; Sjollema, Jelmer; Harapanahalli, Akshay; Dijkstra, Rene J. B.; van der Mei, Henny C.; Busscher, Henk J.

    2015-01-01

    Bioluminescence imaging is used for longitudinal evaluation of bacteria in live animals. Clear relations exist between bacterial numbers and their bioluminescence. However, bioluminescence images of Staphylococcus aureus Xen29, S. aureus Xen36 and Escherichia coli Xen14 grown on tryptone soy agar in

  18. Spectrally resolved bioluminescence tomography using the reciprocity approach

    Dehghani, Hamid; Davis, Scott C.; Pogue, Brian W.

    2008-01-01

    Spectrally resolved bioluminescence optical tomography is an approach to recover images of, for example, Luciferase activity within a volume using multiwavelength emission data from internal bioluminescence sources. The underlying problem of uniqueness associated with nonspectrally resolved intensity-based bioluminescence tomography is demonstrated and it is shown that using a non-negative constraint inverse algorithm, an accurate solution for the source distribution can be calculated from th...

  19. Bioluminescent Probe for Detecting Mercury(II) in Living Mice.

    Jiang, Tianyu; Ke, Bowen; Chen, Hui; Wang, Weishan; Du, Lupei; Yang, Keqian; Li, Minyong

    2016-08-01

    A novel bioluminescence probe for mercury(II) was obtained on the basis of the distinct deprotection reaction of dithioacetal to decanal, so as to display suitable sensitivity and selectivity toward mercury(II) over other ions with bacterial bioluminescence signal. These experimental results indicated such a probe was a novel promising method for mercury(II) bioluminescence imaging in environmental and life sciences ex vivo and in vivo. PMID:27412583

  20. Autonomous bioluminescent expression of the bacterial luciferase gene cassette (lux in a mammalian cell line.

    Dan M Close

    Full Text Available The bacterial luciferase (lux gene cassette consists of five genes (luxCDABE whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multi-bicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo.Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH(2 was identified as the limiting bioluminescence substrate in the mammalian cell environment even after the addition of a constitutively expressed flavin reductase gene (frp from Vibrio harveyi. FMNH(2 supplementation led to a 151-fold increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes. When injected subcutaneously into nude mice, in vivo optical imaging permitted near instantaneous light detection that persisted independently for the 60 min length of the assay with negligible background.The speed, longevity, and self-sufficiency of lux expression in the mammalian cellular environment provides a viable and powerful alternative for real-time target visualization not currently offered by existing bioluminescent and fluorescent imaging technologies.

  1. Quantum dot-NanoLuc bioluminescence resonance energy transfer enables tumor imaging and lymph node mapping in vivo.

    Kamkaew, Anyanee; Sun, Haiyan; England, Christopher G; Cheng, Liang; Liu, Zhuang; Cai, Weibo

    2016-05-19

    A small luciferase protein (Nluc) was conjugated to QDs as a bioluminescence resonance energy transfer (BRET) pair. The conjugate showed 76% BRET efficiency and lymph node mapping was successfully performed. The cRGD peptide was conjugated to QD-Nluc for tumor targeting. The self-illuminating QD-Nluc showed excellent energy transfer in a living system and offered an optimal tumor-to-background ratio (>85). PMID:27157466

  2. Dynamic Observation on In vivo Bioluminescence Imaging of Experimental Metastatic Animal Models in Nude Mice%实验性肿瘤细胞转移动物模型的活体成像观察

    闫明霞; 朱淼鑫; 刘蕾; 李静; 林河春; 赵方瑜; 姚明

    2012-01-01

    Objective To observe the tumor metastasis in deep organisms of the nude mice by in vivo bioluminescence imaging system. Methods The SMMC-7721-GFP/Luc cells with different concentrations were intravenously inoculated into the tail vein and spleen of the BALB/c-nu/nu mice, the distribution and expression of luciferase in nude mice were monitored by in vivo bioluminescence imaging system. Results The experimental metastatic animal models had been successfully established. The distribution and expression of luciferase ascended with cell concentration increased and decreased with the passage of time. Conclusion The in vivo bioluminescence imaging system may monitor the in vivo growth and metastasis of tumors and provide for studying the mechanisms of tumor metastasis and development of anticancer drug.%目的 利用小动物活体成像系统观察肿瘤细胞在动物体内的转移情况.方法 分别将不同浓度的绿色荧光蛋白(GPF)和荧光素酶(luciferase,Luc)双标的SMMC-7721细胞接种入裸小鼠尾静脉和脾,建立实验性转移动物模型,采用活体成像技术监测不同浓度的细胞在小鼠体内的转移情况,动态观察同一细胞于不同时间点在小鼠体内的转移情况.结果 成功建立了尾静脉接种肺转移及脾内接种肝转移的实验性转移动物模型,经小动物活体成像系统检测发现,随着接种细胞浓度的增加,荧光素的表达面积和强度逐渐增加,二者呈正比关系;随着接种时间的延长,荧光素的表达面积和强度逐渐减弱,二者呈成反比关系.结论 活体荧光成像系统可较好地观测肿瘤在动物体内深部脏器的转移情况,它将为肿瘤转移机制、抗转移治疗等研究提供有益的帮助.

  3. Real-time monitoring of bacterial infection in vivo: development of bioluminescent staphylococcal foreign-body and deep-thigh-wound mouse infection models.

    Kuklin, Nelly A; Pancari, Gregory D; Tobery, Timothy W; Cope, Leslie; Jackson, Jesse; Gill, Charles; Overbye, Karen; Francis, Kevin P; Yu, Jun; Montgomery, Donna; Anderson, Annaliesa S; McClements, William; Jansen, Kathrin U

    2003-09-01

    Staphylococcal infections associated with catheter and prosthetic implants are difficult to eradicate and often lead to chronic infections. Development of novel antibacterial therapies requires simple, reliable, and relevant models for infection. Using bioluminescent Staphylococcus aureus, we have adapted the existing foreign-body and deep-wound mouse models of staphylococcal infection to allow real-time monitoring of the bacterial colonization of catheters or tissues. This approach also enables kinetic measurements of bacterial growth and clearance in each infected animal. Persistence of infection was observed throughout the course of the study until termination of the experiment at day 16 in a deep-wound model and day 21 in the foreign-body model, providing sufficient time to test the effects of antibacterial compounds. The usefulness of both animal models was assessed by using linezolid as a test compound and comparing bioluminescent measurements to bacterial counts. In the foreign-body model, a three-dose antibiotic regimen (2, 5, and 24 h after infection) resulted in a decrease in both luminescence and bacterial counts recovered from the implant compared to those of the mock-treated infected mice. In addition, linezolid treatment prevented the formation of subcutaneous abscesses, although it did not completely resolve the infection. In the thigh model, the same treatment regimen resulted in complete resolution of the luminescent signal, which correlated with clearance of the bacteria from the thighs. PMID:12936968

  4. Effect of electromagnetic fields on the bacteria bioluminescent activity

    The effect of electromagnetic field with frequency from 36.2 to 55.9 GHz on bioluminescence activity of bacterium were investigated. Electromagnetic field results in decrease of bioluminescence, which depends from frequency. The electromagnetic field adaptation time is higher of intrinsic time parameters of bioluminescence system. The effect has nonthermal nature. It is suggested that electromagnetic field influence connects with structure rearrangements near cell emitter. 8 refs.; 3 figs

  5. aequorine bioluminescence response to calcium in vitro and in cerebral cortex

    Tricoire, Ludovic

    2006-01-01

    During my PhD, I investigated in vitro the calcium-dependent bioluminescence of thephotoprotein aequorin and then used its bioluminescence to image neuronal activities in theneocortical network. This genetically encoded calcium sensor can be expressed in specific cell types and its bioluminescence is not toxic and exhibit a high signal/noise ratio.I first search for mutations modifying aequorin bioluminescence, using a randommutagenesis and in vitro evolution approach. I isolated mutants show...

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

    Rasko John EJ

    2010-11-01

    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.

  7. Bioluminescence in the high Arctic during the polar night

    Berge, Jørgen; Båtnes, Anna Solvang; Johnsen, Geir; Blackwell, Susan; Mark A. Moline

    2012-01-01

    This study examines the composition and activity of the planktonic community during the polar night in the high Arctic Kongsfjord, Svalbard. Our results are the first published evidence of bioluminescence among zooplankton during the Arctic polar night. The observations were collected by a bathyphotometer detecting bioluminescence, integrated into an autonomous underwater vehicle, to determine the concentration and intensity of bioluminescent flashes as a function of time of day and depth. To...

  8. Bioluminescence imaging of point sources implanted in small animals post mortem: evaluation of a method for estimating source strength and depth

    The performance of a simple approach for the in vivo reconstruction of bioluminescent point sources in small animals was evaluated. The method uses the diffusion approximation as a forward model of light propagation from a point source in a homogeneous tissue to find the source depth and power. The optical properties of the tissue are estimated from reflectance images obtained at the same location on the animal. It was possible to localize point sources implanted in mice, 2-8 mm deep, to within 1 mm. The same performance was achieved for sources implanted in rat abdomens when the effects of tissue surface curvature were eliminated. The source power was reconstructed within a factor of 2 of the true power for the given range of depths, even though the apparent brightness of the source varied by several orders of magnitude. The study also showed that reconstructions using optical properties measured in situ were superior to those based on data in the literature

  9. High-throughput and quantitative approaches for measuring circadian rhythms in cyanobacteria using bioluminescence.

    Shultzaberger, Ryan K; Paddock, Mark L; Katsuki, Takeo; Greenspan, Ralph J; Golden, Susan S

    2015-01-01

    The temporal measurement of a bioluminescent reporter has proven to be one of the most powerful tools for characterizing circadian rhythms in the cyanobacterium Synechococcus elongatus. Primarily, two approaches have been used to automate this process: (1) detection of cell culture bioluminescence in 96-well plates by a photomultiplier tube-based plate-cycling luminometer (TopCount Microplate Scintillation and Luminescence Counter, Perkin Elmer) and (2) detection of individual colony bioluminescence by iteratively rotating a Petri dish under a cooled CCD camera using a computer-controlled turntable. Each approach has distinct advantages. The TopCount provides a more quantitative measurement of bioluminescence, enabling the direct comparison of clock output levels among strains. The computer-controlled turntable approach has a shorter set-up time and greater throughput, making it a more powerful phenotypic screening tool. While the latter approach is extremely useful, only a few labs have been able to build such an apparatus because of technical hurdles involved in coordinating and controlling both the camera and the turntable, and in processing the resulting images. This protocol provides instructions on how to construct, use, and process data from a computer-controlled turntable to measure the temporal changes in bioluminescence of individual cyanobacterial colonies. Furthermore, we describe how to prepare samples for use with the TopCount to minimize experimental noise and generate meaningful quantitative measurements of clock output levels for advanced analysis. PMID:25662451

  10. In vivo functional calcium imaging of induced or spontaneous activity in the fly brain using a GFP-apoaequorin-based bioluminescent approach.

    Minocci, Daiana; Carbognin, Elena; Murmu, Meena Sriti; Martin, Jean-René

    2013-07-01

    Different optical imaging techniques have been developed to study neuronal activity with the goal of deciphering the neural code underlying neurophysiological functions. Because of several constraints inherent in these techniques as well as difficulties interpreting the results, the majority of these studies have been dedicated more to sensory modalities than to the spontaneous activity of the central brain. Recently, a novel bioluminescence approach based on GFP-aequorin (GA) (GFP: Green fluorescent Protein), has been developed, allowing us to functionally record in-vivo neuronal activity. Taking advantage of the particular characteristics of GA, which does not require light excitation, we report that we can record induced and/or the spontaneous Ca(2+)-activity continuously over long periods. Targeting GA to the mushrooms-bodies (MBs), a structure implicated in learning/memory and sleep, we have shown that GA is sensitive enough to detect odor-induced Ca(2+)-activity in Kenyon cells (KCs). It has been possible to reveal two particular peaks of spontaneous activity during overnight recording in the MBs. Other peaks of spontaneous activity have been recorded in flies expressing GA pan-neurally. Similarly, expression in the glial cells has revealed that these cells exhibit a cell-autonomous Ca(2+)-activity. These results demonstrate that bioluminescence imaging is a useful tool for studying Ca(2+)-activity in neuronal and/or glial cells and for functional mapping of the neurophysiological processes in the fly brain. These findings provide a framework for investigating the biological meaning of spontaneous neuronal activity. This article is part of a Special Issue entitled: 12th European Symposium on Calcium. PMID:23287020

  11. In vivo bioluminescence tomography with a blocking-off finite-difference SP3 method and MRI∕CT coregistration

    Klose, Alexander D.; Beattie, Bradley J.; Dehghani, Hamid; Vider, Lena; Le, Carl; Ponomarev, Vladimir; Blasberg, Ronald

    2009-01-01

    Purpose: Bioluminescence imaging is a research tool for studying gene expression levels in small animal models of human disease. Bioluminescence light, however, is strongly scattered in biological tissue and no direct image of the light-emitting reporter probe’s location can be obtained. Therefore, the authors have developed a linear image reconstruction method for bioluminescence tomography (BLT) that recovers the three-dimensional spatial bioluminescent source distribution in small animals.

  12. Stimulated bioluminescence by fluid shear stress associated with pipe flow

    Dinoflagellate can be stimulated bioluminescence by hydrodynamic agitation. Two typical dinoflagellate (Lingulodinium polyedrum and Pyrocystis noctiluca) was choosed to research stimulated bioluminescence. The bioluminescence intensity and shear stress intensity were measured using fully developed pipe flow. There is shear stress threshold to agitate organism bioluminescence. From these experiment, the response thresholds of the stimulated bioluminscence always occurred in laminar flows at a shear stress level of 0.6-3 dyn/cm2. At the same time, the spectral characteristc of dinoflagellate was recorded, the wavelength of them is about 470nm, and the full width at half maximum is approximate 30nm.

  13. Stimulated bioluminescence by fluid shear stress associated with pipe flow

    Cao Jing; Wang Jiangan; Wu Ronghua, E-mail: caojing981@126.com [Col. of Electronic Eng., Naval University of Engineering, Wuhan 430033 (China)

    2011-01-01

    Dinoflagellate can be stimulated bioluminescence by hydrodynamic agitation. Two typical dinoflagellate (Lingulodinium polyedrum and Pyrocystis noctiluca) was choosed to research stimulated bioluminescence. The bioluminescence intensity and shear stress intensity were measured using fully developed pipe flow. There is shear stress threshold to agitate organism bioluminescence. From these experiment, the response thresholds of the stimulated bioluminscence always occurred in laminar flows at a shear stress level of 0.6-3 dyn/cm{sup 2}. At the same time, the spectral characteristc of dinoflagellate was recorded, the wavelength of them is about 470nm, and the full width at half maximum is approximate 30nm.

  14. An advanced preclinical mouse model for acute myeloid leukemia using patients' cells of various genetic subgroups and in vivo bioluminescence imaging.

    Vick, Binje; Rothenberg, Maja; Sandhöfer, Nadine; Carlet, Michela; Finkenzeller, Cornelia; Krupka, Christina; Grunert, Michaela; Trumpp, Andreas; Corbacioglu, Selim; Ebinger, Martin; André, Maya C; Hiddemann, Wolfgang; Schneider, Stephanie; Subklewe, Marion; Metzeler, Klaus H; Spiekermann, Karsten; Jeremias, Irmela

    2015-01-01

    Acute myeloid leukemia (AML) is a clinically and molecularly heterogeneous disease with poor outcome. Adequate model systems are required for preclinical studies to improve understanding of AML biology and to develop novel, rational treatment approaches. Xenografts in immunodeficient mice allow performing functional studies on patient-derived AML cells. We have established an improved model system that integrates serial retransplantation of patient-derived xenograft (PDX) cells in mice, genetic manipulation by lentiviral transduction, and essential quality controls by immunophenotyping and targeted resequencing of driver genes. 17/29 samples showed primary engraftment, 10/17 samples could be retransplanted and some of them allowed virtually indefinite serial transplantation. 5/6 samples were successfully transduced using lentiviruses. Neither serial transplantation nor genetic engineering markedly altered sample characteristics analyzed. Transgene expression was stable in PDX AML cells. Example given, recombinant luciferase enabled bioluminescence in vivo imaging and highly sensitive and reliable disease monitoring; imaging visualized minimal disease at 1 PDX cell in 10000 mouse bone marrow cells and facilitated quantifying leukemia initiating cells. We conclude that serial expansion, genetic engineering and imaging represent valuable tools to improve the individualized xenograft mouse model of AML. Prospectively, these advancements enable repetitive, clinically relevant studies on AML biology and preclinical treatment trials on genetically defined and heterogeneous subgroups. PMID:25793878

  15. Lighting up bioluminescence with coelenterazine: strategies and applications.

    Jiang, Tianyu; Du, Lupei; Li, Minyong

    2016-04-13

    Bioluminescence-based techniques, such as bioluminescence imaging, BRET and dual-luciferase reporter assay systems, have been widely used to examine a myriad of biological processes. Coelenterazine (CTZ), a luciferin or light-producing compound found in bioluminescent organisms, has sparked great curiosity and interest in searching for analogues with improved photochemical properties. This review summarizes the current development of coelenterazine analogues, their bioluminescence properties, and the rational design of caged coelenterazine towards biotargets, as well as their applications in bioassays. It should be emphasized that the design of caged luciferins can provide valuable insight into detailed molecular processes in organisms and will be a trend in the development of bioluminescent molecules. PMID:27009907

  16. Multispectral Bioluminescence Tomography: Methodology and Simulation

    Ge Wang

    2006-02-01

    Full Text Available Bioluminescent imaging has proven to be a valuable tool for monitoring physiological and pathological activities at cellular and molecular levels in living small animals. Using biological techniques, target cells can be tagged with reporters encoding several kinds of luciferase enzymes, which generate characteristic photons in a wide spectrum covering the infrared range. Part of the diffused light can reach the body surface of the small animal, be separated into several spectral bands using appropriate filters, and collected by a sensitive CCD camera. Here we present a bioluminescence tomography (BLT method for a bioluminescent source reconstruction from multispectral data measured on the external surface, and demonstrate the advantages of multispectral BLT in a numerical study using a heterogeneous mouse chest phantom. The results show that the multispectral approach significantly improves the accuracy and stability of the BLT reconstruction even if the data are highly noisy.

  17. 活体生物发光成像技术及其在病毒感染研究中的应用%Bioluminescence in-vivo imaging technology and its application in the study of viral infection-A review

    柴凡; 周耘裔; 肖庚富

    2011-01-01

    生物发光是动物活体光学成像技术之一,因其反应灵敏、操作简单、数据精确,而被广泛地应用于生命科学研究多个领域,观测活体动物体内病毒复制、肿瘤生长等生命过程.生物发光技术采用荧光素酶基因标记细胞或病毒,与外源注射的底物荧光素发生反应,在冷CCD成像系统下显像并进行数据记录、分析.本文简要介绍活体生物发光成像这一新技术的原理,综述了其在发现病毒新的复制位点、研究干扰素及药物对病毒的抑制作用、展示病毒潜伏感染与再激活的历程等方面的应用,并结合自己工作对该技术及其发展前景进行评述.%Bioluminescence imaging is one of the bio-optical imaging techniques which report definite biological event in living animals with genetic modification.With high sensitivity, simple operation and high precision, it is particularly applied in observing vital processes like viral infection and tumor growth in vivo.We summarize the principle of bioluminescence imaging, introduce its application in finding virus replication site, study of interferon( IFN) inhibiting-virus effect and real-time visualization of viral latent infection and reactivation, and preview the trend of bioluminescence imaging technological development.

  18. Bioluminescent imaging of vaccinia virus infection in immunocompetent and immunodeficient rats as a model for human smallpox.

    Liu, Qiang; Fan, Changfa; Zhou, Shuya; Guo, Yanan; Zuo, Qin; Ma, Jian; Liu, Susu; Wu, Xi; Peng, Zexu; Fan, Tao; Guo, Chaoshe; Shen, Yuelei; Huang, Weijin; Li, Baowen; He, Zhengming; Wang, Youchun

    2015-01-01

    Due to the increasing concern of using smallpox virus as biological weapons for terrorist attack, there is renewed interest in studying the pathogenesis of human smallpox and development of new therapies. Animal models are highly demanded for efficacy and safety examination of new vaccines and therapeutic drugs. Here, we demonstrated that both wild type and immunodeficient rats infected with an engineered vaccinia virus carrying Firefly luciferase reporter gene (rTV-Fluc) could recapitulate infectious and clinical features of human smallpox. Vaccinia viral infection in wild type Sprague-Dawley (SD) rats displayed a diffusible pattern in various organs, including liver, head and limbs. The intensity of bioluminescence generated from rTV-Fluc correlated well with viral loads in tissues. Moreover, neutralizing antibodies had a protective effect against virus reinfection. The recombination activating gene 2 (Rag2) knockout rats generated by transcription activator-like effector nucleases (TALENs) technology were further used to examine the infectivity of the rTV-Fluc in immunodeficient populations. Here we demonstrated that Rag2-/- rats were more susceptible to rTV-Fluc than SD rats with a slower virus clearance rate. Therefore, the rTV-Fluc/SD rats and rTV-Fluc/Rag2-/- rats are suitable visualization models, which recapitulate wild type or immunodeficient populations respectively, for testing human smallpox vaccine and antiviral drugs. PMID:26235050

  19. Space-Time Quantum Imaging

    Ronald E. Meyers

    2015-03-01

    Full Text Available We report on an experimental and theoretical investigation of quantum imaging where the images are stored in both space and time. Ghost images of remote objects are produced with either one or two beams of chaotic laser light generated by a rotating ground glass and two sensors measuring the reference field and bucket field at different space-time points. We further observe that the ghost images translate depending on the time delay between the sensor measurements. The ghost imaging experiments are performed both with and without turbulence. A discussion of the physics of the space-time imaging is presented in terms of quantum nonlocal two-photon analysis to support the experimental results. The theoretical model includes certain phase factors of the rotating ground glass. These experiments demonstrated a means to investigate the time and space aspects of ghost imaging and showed that ghost imaging contains more information per measured photon than was previously recognized where multiple ghost images are stored within the same ghost imaging data sets. This suggests new pathways to explore quantum information stored not only in multi-photon coincidence information but also in time delayed multi-photon interference. The research is applicable to making enhanced space-time quantum images and videos of moving objects where the images are stored in both space and time.

  20. 6-hydroxydopamine-induced Parkinson's disease-like degeneration generates acute microgliosis and astrogliosis in the nigrostriatal system but no bioluminescence imaging-detectable alteration in adult neurogenesis.

    Fricke, Inga B; Viel, Thomas; Worlitzer, Maik M; Collmann, Franziska M; Vrachimis, Alexis; Faust, Andreas; Wachsmuth, Lydia; Faber, Cornelius; Dollé, Frédéric; Kuhlmann, Michael T; Schäfers, Klaus; Hermann, Sven; Schwamborn, Jens C; Jacobs, Andreas H

    2016-05-01

    Parkinson's disease is a slowly progressing neurodegenerative disorder caused by loss of dopaminergic neurons in the substantia nigra (SN), leading to severe impairment in motor and non-motor functions. Endogenous subventricular zone (SVZ) neural stem cells constantly give birth to new cells that might serve as a possible source for regeneration in the adult brain. However, neurodegeneration is accompanied by neuroinflammation and dopamine depletion, potentially compromising regeneration. We therefore employed in vivo imaging methods to study striatal deafferentation (N-ω-fluoropropyl-2β-carbomethoxy-3β-(4-[(123) I]iodophenyl)nortropane single photon emission computed tomography, DaTscan(™) ) and neuroinflammation in the SN and striatum (N,N-diethyl-2-(2-(4-(2-[(18) F]fluoroethoxy)phenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidin-3-yl)acetamide positron emission tomography, [(18) F]DPA-714 PET) in the intranigral 6-hydroxydopamine Parkinson's disease mouse model. Additionally, we transduced cells in the SVZ with a lentivirus encoding firefly luciferase and followed migration of progenitor cells in the SVZ-olfactory bulb axis via bioluminescence imaging under disease and control conditions. We found that activation of microglia in the SN is an acute process accompanying the degeneration of dopaminergic cell bodies in the SN. Dopaminergic deafferentation of the striatum does not influence the generation of doublecortin-positive neuroblasts in the SVZ, but generates chronic astrogliosis in the nigrostriatal system. PMID:26950181

  1. Nanostructured bioluminescent sensor for rapidly detecting thrombin.

    Chen, Longyan; Bao, Yige; Denstedt, John; Zhang, Jin

    2016-03-15

    Thrombin plays a key role in thrombosis and hemostasis. The abnormal level of thrombin in body fluids may lead to different diseases, such as rheumatoid arthritis, glomerulonephritis, etc. Detection of thrombin level in blood and/or urine is one of important methods for medical diagnosis. Here, a bioluminescent sensor is developed for non-invasively and rapidly detecting thrombin in urine. The sensor is assembled through conjugating gold nanoparticles (Au NPs) and a recombinant protein containing Renilla luciferase (pRluc) by a peptide, which is thrombin specific substrate. The luciferase-catalyzed bioluminescence can be quenched by peptide-conjugating Au NPs. In the presence of thrombin, the short peptide conjugating luciferase and Au NPs is digested and cut off, which results in the recovery of bioluminescence due to the release of luciferase from Au NPs. The bioluminescence intensity at 470 nm is observed, and increases with increasing concentration of thrombin. The bioluminescence intensity of this designed sensor is significantly recovered when the thrombin digestion time lasts for 10 min. In addition, a similar linear relationship between luminescence intensity and the concentration of thrombin is found in the range of 8 nM to 8 μM in both buffer and human urine spiked samples. The limit of detection is as low as 80 pM. It is anticipated that our nanosensor could be a promising tool for clinical diagnosis of thrombin in human urine. PMID:26397418

  2. 艾灸抑制小鼠皮下移植瘤生长的活体成像观察%Bioluminescence imaging evaluation of the inhibitory effect of moxibustion in mice bear-ing subcutaneous cancer cell lines

    周俊梅; 韦芳; 刘素君; 张必萌

    2014-01-01

    目的:观察艾灸对小鼠皮下移植瘤的抑制作用。方法:通过慢病毒转染,构建能稳定表达荧光素酶基因的乳腺癌细胞株,建立小鼠皮下移植瘤模型后通过随机区组方法设为对照组和治疗组,治疗组又分为关元组、大椎组。治疗组艾灸隔天一次,每次10min,共7次,对照组不做处理。通过活体成像系统以及体外测量,监测肿瘤生长情况。治疗终点,处死小鼠,HE染色观察肿瘤的病理形态学变化。结果:建立能稳定表达荧光素酶基因的乳腺癌细胞株及小鼠皮下移植瘤模型,细胞数与荧光强度呈线性关系( r=0.899)。活体成像显示:治疗组小鼠荧光强度较对照组明显减弱。体外测量,治疗组肿瘤体积及重量明显小于对照组(P%Objective:To investigate the inhibitory effect of moxibustion in mice bearing subcutaneous cancer cell lines. Methods:Breast cancer cell line 4T1 with stable expression of luciferase gene(4T1/luc)were established by transfection via lentiviral vector. 4T1/luc cells were inoculated into the right leg of mice to prepare subcutaneous tumor model. A total of 15 tumor-bearing mouse were randomly divided into the following groups:the model control group and treatment group,the later includes guanyuan group and dazhui group. Mouse in dazhui and guanyuan group were treated with moxibustion,once another day for 10min,for a total of 7 times. The control group did nothing. The dynamic growth of subcutaneous tumor was observed using bioluminescence imaging system in vivo. At the end of the treatment,morphology of transplanted tumor tissue was observed by HE staining. Results:The stable 4T1/luc cells and subcutaneous tumor model were obtained. Bioluminescence imaging showed that fluorescent intensity of treated group was apparently lower than that of the control group. Compare with control group,the tomor volume and weight of treated group were significantly decreased(P 0

  3. Bioluminescence imaging evaluation of the inhibitory effect of moxibustion in mice bear-ing subcutaneous cancer cell lines%艾灸抑制小鼠皮下移植瘤生长的活体成像观察

    周俊梅; 韦芳; 刘素君; 张必萌

    2014-01-01

    目的:观察艾灸对小鼠皮下移植瘤的抑制作用。方法:通过慢病毒转染,构建能稳定表达荧光素酶基因的乳腺癌细胞株,建立小鼠皮下移植瘤模型后通过随机区组方法设为对照组和治疗组,治疗组又分为关元组、大椎组。治疗组艾灸隔天一次,每次10min,共7次,对照组不做处理。通过活体成像系统以及体外测量,监测肿瘤生长情况。治疗终点,处死小鼠,HE染色观察肿瘤的病理形态学变化。结果:建立能稳定表达荧光素酶基因的乳腺癌细胞株及小鼠皮下移植瘤模型,细胞数与荧光强度呈线性关系( r=0.899)。活体成像显示:治疗组小鼠荧光强度较对照组明显减弱。体外测量,治疗组肿瘤体积及重量明显小于对照组(P%Objective:To investigate the inhibitory effect of moxibustion in mice bearing subcutaneous cancer cell lines. Methods:Breast cancer cell line 4T1 with stable expression of luciferase gene(4T1/luc)were established by transfection via lentiviral vector. 4T1/luc cells were inoculated into the right leg of mice to prepare subcutaneous tumor model. A total of 15 tumor-bearing mouse were randomly divided into the following groups:the model control group and treatment group,the later includes guanyuan group and dazhui group. Mouse in dazhui and guanyuan group were treated with moxibustion,once another day for 10min,for a total of 7 times. The control group did nothing. The dynamic growth of subcutaneous tumor was observed using bioluminescence imaging system in vivo. At the end of the treatment,morphology of transplanted tumor tissue was observed by HE staining. Results:The stable 4T1/luc cells and subcutaneous tumor model were obtained. Bioluminescence imaging showed that fluorescent intensity of treated group was apparently lower than that of the control group. Compare with control group,the tomor volume and weight of treated group were significantly decreased(P 0

  4. la bioluminescence de l'aequorine en réponse au calcium In vitro et dans le Cortex cerebral

    Tricoire, Ludovic

    2006-01-01

    During my PhD, I investigated in vitro the calcium-dependent bioluminescence of thephotoprotein aequorin and then used its bioluminescence to image neuronal activities in theneocortical network. This genetically encoded calcium sensor can be expressed in specific cell types and its bioluminescence is not toxic and exhibit a high signal/noise ratio.I first search for mutations modifying aequorin bioluminescence, using a randommutagenesis and in vitro evolution approach. I isolated mutants show...

  5. A Multichannel Bioluminescence Determination Platform for Bioassays.

    Kim, Sung-Bae; Naganawa, Ryuichi

    2016-01-01

    The present protocol introduces a multichannel bioluminescence determination platform allowing a high sample throughput determination of weak bioluminescence with reduced standard deviations. The platform is designed to carry a multichannel conveyer, an optical filter, and a mirror cap. The platform enables us to near-simultaneously determine ligands in multiple samples without the replacement of the sample tubes. Furthermore, the optical filters beneath the multichannel conveyer are designed to easily discriminate colors during assays. This optical system provides excellent time- and labor-efficiency to users during bioassays. PMID:27424912

  6. Evaluation of the inflammatory potential of implant materials in a mouse model by bioluminescent imaging of intravenously injected bone marrow cells.

    Rais, Bushra; Köster, Mario; Rahim, Muhammad Imran; Pils, Marina; Seitz, Jan-Marten; Hauser, Hansjörg; Wirth, Dagmar; Mueller, Peter P

    2016-09-01

    To evaluate the inflammatory potential of implants a bioluminescent imaging assay was developed using luciferase-expressing bone marrow cells that were injected into the blood circulation of wild-type mice. After subcutaneous implantation of titanium discs as an example for a clinically established biocompatible material, the luminosity was modest. Similarly, low luminosity signals were generated by pure magnesium implants that were used to represent metallic alloys that are presently under investigation as novel degradable implant materials. Increased luminosity was observed in response to degradable polymeric PLGA implants. Surgical wounds induced a basic luminescent response even in the absence of an implant. However, the material-independent response to injury could be minimized using injectable microparticle suspensions. In parallel with the resorption of biodegradable microparticles, the signal induced by PLGA declined faster when compared to non-degradable polystyrene suspensions. By using an interferon type I inducible Mx2 promoter construct to drive luciferase gene expression, the highest luminosity was observed in response to bacteria, indicating that the system could also be employed to monitor implant infections. Overall, labeled bone marrow cells yielded specific, well-defined localized signals that correlated with the inflammatory responses to implants. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2149-2158, 2016. PMID:27102724

  7. Optimization and technology extension of bioluminescence imaging in vivo in small animals%小动物生物发光活体成像的条件优化与技术扩展研究

    王豫; 关华; 宋曼; 王晓迪; 高毅; 周平坤

    2013-01-01

    目的 为发掘生物发光活体成像技术的灵敏性和应用潜力,进行在体检测条件的优化和技术扩展.方法 将稳定高表达荧光素酶的乳腺癌细胞(4T-1-Luc+)和肝癌细胞(Huh-7-Luc+)接种于BALB/c 小鼠体内,通过活体成像仪进行生物发光检测灵敏度和毛发的影响分析,利用生物发光结合X线扩展活体成像技术.结果 毛发对小鼠皮下接种的肿瘤细胞的生物发光成像有干扰作用,在局部剃除毛发小鼠内体可以检测到50个细胞的光信号值.通过建立X结合发光的检测模型,实现了既能检测发光信号值强度大小,又能比较清晰地观测到部分脏器和发光体(病变组织)的部位.结论 实现了生物发光活体成像技术的检测条件优化,建立了X线结合发光新的检测模式,为扩展小动物活体成像的应用范围奠定了基础.%Objective To explore the sensitivity and potential applicability of bioluminescence in vivo imaging technique and to achieve optimization of detection conditions and technique extension . Methods Breast cancer cells 4T-L-Luc or liver cancer cells Huh -7 -Luc+ with stable expression of luciferase were implanted subcutaneously into BALB /c mice. The sensitivity and effect on the hair were investigated using the small animal in vivo imager of bioluminescent signals. Technique extension was performed by combining bioluminescence with X -ray. Results The hair as showed are obvious interference with the detection of bioluminescent signals emitted from the cancer cells implanted in BALB /c mice. The optical signal could be detected in the cellular mass of 50 cells when local hair was removed. The imaging model established by combining X-ray with bioluminescence could not only detect the strength of optical signals emitted , but also clearly locate some organs and the origin of optical signals or pathological tissue . Conclusion Optimization of bioluminescence imaging condition has been achieved . A imaging

  8. Bioluminescence lights the way to food safety

    Brovko, Lubov Y.; Griffiths, Mansel W.

    2003-07-01

    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.

  9. Time-resolved molecular imaging

    Xu, Junliang; Blaga, Cosmin I.; Agostini, Pierre; DiMauro, Louis F.

    2016-06-01

    Time-resolved molecular imaging is a frontier of ultrafast optical science and physical chemistry. In this article, we review present and future key spectroscopic and microscopic techniques for ultrafast imaging of molecular dynamics and show their differences and connections. The advent of femtosecond lasers and free electron x-ray lasers bring us closer to this goal, which eventually will extend our knowledge about molecular dynamics to the attosecond time domain.

  10. Circadian regulation of bioluminescence in Gonyaulax involves translational control.

    Morse, D.; Milos, P M; Roux, E.; Hastings, J. W.

    1989-01-01

    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 ...

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

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

    2013-01-01

    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 64Cu to yield 64Cu-NOTA-TRC105-800CW. Flow cytometry analysis revealed no difference in CD105 binding affinity/specificity between TRC105 and NOTA-TRC105-800CW. Serial bio...

  12. Expression of a humanized viral 2A-mediated lux operon efficiently generates autonomous bioluminescence in human cells.

    Tingting Xu

    Full Text Available 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.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.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

  13. Action of γ-radiation on bioluminescence of Noctiluca miliaris

    Results of the study in the action of various doses of irradiation on the bioluminescence of Noctiluca miliaris are presented. The doses are found that stimulate the bioluminescence and the dose - effect curves are obtained. It has been shown that stimulation of Noctiluca luminescence by γ-radiation is not of a constant character and extinguishes after a period of time determined by a dose rate

  14. Bioluminescent bacteria: lux genes as environmental biosensors

    Nunes-Halldorson Vânia da Silva; Duran Norma Letícia

    2003-01-01

    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...

  15. Targeted gene therapy and in vivo bioluminescent imaging for monitoring postsurgical recurrence and metastasis in mouse models of liver cancer.

    He, Q; Yao, C L; Li, L; Xin, Z; Jing, Z K; Li, L X

    2016-01-01

    We investigated the effects of combined targeted gene therapy on recurrence and metastasis after liver cancer resection in nude mice. Twenty BALB/C mice were randomly divided into control and treatment groups with 10 mice in each group and a male/female ratio of 1:1. Luciferase gene-labeled human primary hepatic carcinoma cell line MHCC97-H was then used to prepare a carcinoma model. An optical in vivo imaging technique (OIIT) was used 10 days later to detect the distribution of tumor cells, followed by partial liver resection and gene therapy. In the treatment group, 100 mL phosphate-buffered saline (PBS) containing 1 x 1012 rAAV/AFP/IL-24 gene viral vectors was injected into liver sections and peritumoral posterior peritoneal tissues; in the control group, the same amount of PBS containing 1 x 1012 empty viral vectors was injected at the same sites. OIIT was then used to detect the in vivo tumor metastasis 21 days later. Luciferase gene-labeled human primary hepatic carcinoma cell line MHCC97-H successfully infected 20 nude mice, and OIIT showed that the two groups exhibited metastasis after local tumor resection, but there were more tumor cells in the control group (P AFP/IL-24 gene therapy can inhibit recurrence after liver cancer resection. PMID:27525931

  16. Measurement of Bacterial Bioluminescence Intensity and Spectrum: Current Physical Techniques and Principles.

    Jia, Kun; Ionescu, Rodica Elena

    2016-01-01

    : Bioluminescence is light production by living organisms, which can be observed in numerous marine creatures and some terrestrial invertebrates. More specifically, bacterial bioluminescence is the "cold light" produced and emitted by bacterial cells, including both wild-type luminescent and genetically engineered bacteria. Because of the lively interplay of synthetic biology, microbiology, toxicology, and biophysics, different configurations of whole-cell biosensors based on bacterial bioluminescence have been designed and are widely used in different fields, such as ecotoxicology, food toxicity, and environmental pollution. This chapter first discusses the background of the bioluminescence phenomenon in terms of optical spectrum. Platforms for bacterial bioluminescence detection using various techniques are then introduced, such as a photomultiplier tube, charge-coupled device (CCD) camera, micro-electro-mechanical systems (MEMS), and complementary metal-oxide-semiconductor (CMOS) based integrated circuit. Furthermore, some typical biochemical methods to optimize the analytical performances of bacterial bioluminescent biosensors/assays are reviewed, followed by a presentation of author's recent work concerning the improved sensitivity of a bioluminescent assay for pesticides. Finally, bacterial bioluminescence as implemented in eukaryotic cells, bioluminescent imaging, and cancer cell therapies is discussed. PMID:25981856

  17. Real-Time Imaging of Gene Delivery and Expression with DNA Nanoparticle Technologies

    Sun, Wenchao; Ziady, Assem G.

    The construction of safe, efficient, and modifiable synthetic DNA nanoparticles is an emerging technology that has achieved important milestones of success in the past 5 years. Advances in chemical conjugation, purification, and controlled synthesis have allowed researchers to produce uniform and stable particles, whose physical characteristics can be well characterized and monitored. As a result of these improvements, DNA nanoparticles have now been cleared for clinical testing, and show good potential for human gene therapy. A very important recent development in the study of DNA nanoparticles is the use of small-animal imaging. Real-time imaging has become a valuable technique for tracking particle biodistribution and gene transfer efficacy. In this chapter, we discuss how bioluminescent, positron emission tomography, and magnetic resonance imaging can be used separately or in concert to study particle delivery, localization, and magnitude of gene expression in vivo.

  18. Bioluminescence tomography with Gaussian prior

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

    2010-01-01

    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....

  19. Microtiter plate tests for segregation of bioluminescent bacteria.

    Šimkus, Remigijus; Meškienė, Rita; Ledas, Žilvinas; Baronas, Romas; Meškys, Rolandas

    2016-02-01

    It has been recently shown that bioluminescence imaging can be usefully applied to provide new insights into bacterial self-organization. In this work we employ bioluminescence imaging to record images of nutrient rich liquid cultures of the lux-gene reporter Escherichia coli in microtiter plate wells. The images show that patterns of inhomogenous bioluminescence form along the three-phase contact lines. The paper analyzes the dependencies of the average number of luminous aggregates (clouds) on various environmental factors. In particular, our results show that optimal (neutral) pH and high aeration rates determine the highest mean number of clouds, and that spatiotemporal patterns do not form in the pH buffered suspensions. In addition, a sigmoidal (switch-like) dependence of the number of aggregates on the rate of aeration was observed. The obtained bioluminescence imaging data was interpreted by employing the Keller-Segel-Fisher (KSF) model of chemotaxis and logistic growth, adapted to systems of metabolically flexible (two-state) bacteria. The modified KSF model successfully simulated the observed switch-like responses. The results of the microtiter plate tests and their simulations indicate that the segregation of bacteria with different activities proceeds in the three-phase contact line region. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26039821

  20. 活体荧光成像对裸鼠肝癌细胞系MHCC97-H原位移植模型的动态量化分析%Dynamic and Quantitative Analysis of Orthotopically Transplanted Nude Mouse Model with MHCC97-H Cells using Bioluminescent Imaging Technology

    曹阳; 韩炜; 刘洋; 张勇; 郭欣; 陈勇

    2013-01-01

    from liver to abdominal cavity, and the total photon flux emitting from tumor cells increased expotetially over time. Then the tumor was confirmed by pathological examination. Conclusion: Dynamic and quantitative analysis used bioluminescent imaging technology can accurately reflect the volume, growth and metastasis of tumors in orthotopically transplanted nude mouse model for HCC, and provide sensitive access to developing the research of mechanism of oncogenesis, growth and metastasis of tumor, and anti-cancer study.

  1. The rapid bioluminescence assay method for content of bacteria in dehydrated vegetable and condiment before radiation

    The microbial colony-forming unit (cfu) in dehydrated vegetable and condiment was determined by using ATP bioluminescence method. The result showed that bioluminescence of ATP was correlative to the microbial cfu obviously. The detecting time was within 1-2 h. This method could be applied to determine micro load of products before irradiation sterilization. (authors)

  2. In vitro validation of bioluminescent monitoring of disease progression and therapeutic response in leukaemia model animals

    The application of in vivo bioluminescence imaging to non-invasive, quantitative monitoring of tumour models relies on a positive correlation between the intensity of bioluminescence and the tumour burden. We conducted cell culture studies to investigate the relationship between bioluminescent signal intensity and viable cell numbers in murine leukaemia model cells. Interleukin-3 (IL-3)-dependent murine pro-B cell line Ba/F3 was transduced with firefly luciferase to generate cells expressing luciferase stably under the control of a retroviral long terminal repeat. The luciferase-expressing cells were transduced with p190 BCR-ABL to give factor-independent proliferation. The cells were cultured under various conditions, and bioluminescent signal intensity was compared with viable cell numbers and the cell cycle stage. The Ba/F3 cells showed autonomous growth as well as stable luciferase expression following transduction with both luciferase and p190 BCR-ABL, and in vivo bioluminescence imaging permitted external detection of these cells implanted into mice. The bioluminescence intensities tended to reflect cell proliferation and responses to imatinib in cell culture studies. However, the luminescence per viable cell was influenced by the IL-3 concentration in factor-dependent cells and by the stage of proliferation and imatinib concentration in factor-independent cells, thereby impairing the proportionality between viable cell number and bioluminescent signal intensity. Luminescence per cell tended to vary in association with the fraction of proliferating cells. Although in vivo bioluminescence imaging would allow non-invasive monitoring of leukaemia model animals, environmental factors and therapeutic interventions may cause some discrepancies between tumour burden and bioluminescence intensity. (orig.)

  3. In vitro validation of bioluminescent monitoring of disease progression and therapeutic response in leukaemia model animals

    Inoue, Yusuke; Okubo, Toshiyuki [University of Tokyo, Department of Radiology, Institute of Medical Science, Tokyo (Japan); Tojo, Arinobu; Sekine, Rieko; Soda, Yasushi; Kobayashi, Seiichiro; Nomura, Akiko; Izawa, Kiyoko [University of Tokyo, Division of Molecular Therapy, Advanced Clinical Research Centre, Tokyo (Japan); Kitamura, Toshio [University of Tokyo, Division of Cellular Therapy, Advanced Clinical Research Centre, Tokyo (Japan); Ohtomo, Kuni [University of Tokyo, Department of Radiology, Graduate School of Medicine, Tokyo (Japan)

    2006-05-15

    The application of in vivo bioluminescence imaging to non-invasive, quantitative monitoring of tumour models relies on a positive correlation between the intensity of bioluminescence and the tumour burden. We conducted cell culture studies to investigate the relationship between bioluminescent signal intensity and viable cell numbers in murine leukaemia model cells. Interleukin-3 (IL-3)-dependent murine pro-B cell line Ba/F3 was transduced with firefly luciferase to generate cells expressing luciferase stably under the control of a retroviral long terminal repeat. The luciferase-expressing cells were transduced with p190 BCR-ABL to give factor-independent proliferation. The cells were cultured under various conditions, and bioluminescent signal intensity was compared with viable cell numbers and the cell cycle stage. The Ba/F3 cells showed autonomous growth as well as stable luciferase expression following transduction with both luciferase and p190 BCR-ABL, and in vivo bioluminescence imaging permitted external detection of these cells implanted into mice. The bioluminescence intensities tended to reflect cell proliferation and responses to imatinib in cell culture studies. However, the luminescence per viable cell was influenced by the IL-3 concentration in factor-dependent cells and by the stage of proliferation and imatinib concentration in factor-independent cells, thereby impairing the proportionality between viable cell number and bioluminescent signal intensity. Luminescence per cell tended to vary in association with the fraction of proliferating cells. Although in vivo bioluminescence imaging would allow non-invasive monitoring of leukaemia model animals, environmental factors and therapeutic interventions may cause some discrepancies between tumour burden and bioluminescence intensity. (orig.)

  4. Effect of irradiation on bioluminescence spectrum of microbial ATP

    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)

  5. A novel reconstruction algorithm for bioluminescent tomography based on Bayesian compressive sensing

    Wang, Yaqi; Feng, Jinchao; Jia, Kebin; Sun, Zhonghua; Wei, Huijun

    2016-03-01

    Bioluminescence tomography (BLT) is becoming a promising tool because it can resolve the biodistribution of bioluminescent reporters associated with cellular and subcellular function through several millimeters with to centimeters of tissues in vivo. However, BLT reconstruction is an ill-posed problem. By incorporating sparse a priori information about bioluminescent source, enhanced image quality is obtained for sparsity based reconstruction algorithm. Therefore, sparsity based BLT reconstruction algorithm has a great potential. Here, we proposed a novel reconstruction method based on Bayesian compressive sensing and investigated its feasibility and effectiveness with a heterogeneous phantom. The results demonstrate the potential and merits of the proposed algorithm.

  6. Accounting for filter bandwidth improves the quantitative accuracy of bioluminescence tomography

    Taylor, Shelley L.; Mason, Suzannah K. G.; Glinton, Sophie L.; Cobbold, Mark; Dehghani, Hamid

    2015-09-01

    Bioluminescence imaging is a noninvasive technique whereby surface weighted images of luminescent probes within animals are used to characterize cell count and function. Traditionally, data are collected over the entire emission spectrum of the source using no filters and are used to evaluate cell count/function over the entire spectrum. Alternatively, multispectral data over several wavelengths can be incorporated to perform tomographic reconstruction of source location and intensity. However, bandpass filters used for multispectral data acquisition have a specific bandwidth, which is ignored in the reconstruction. In this work, ignoring the bandwidth is shown to introduce a dependence of the recovered source intensity on the bandwidth of the filters. A method of accounting for the bandwidth of filters used during multispectral data acquisition is presented and its efficacy in increasing the quantitative accuracy of bioluminescence tomography is demonstrated through simulation and experiment. It is demonstrated that while using filters with a large bandwidth can dramatically decrease the data acquisition time, if not accounted for, errors of up to 200% in quantitative accuracy are introduced in two-dimensional planar imaging, even after normalization. For tomographic imaging, the use of this method to account for filter bandwidth dramatically improves the quantitative accuracy.

  7. Circular polarization observed in bioluminescence

    Wijnberg, Hans; Meijer, E.W.; Hummelen, J.C.; Dekkers, H.P.J.M.; Schippers, P.H.; Carlson, A.D.

    1980-01-01

    While investigating circular polarization in luminescence, and having found it in chemiluminescence, we have studied bioluminescence because it is such a widespread and dramatic natural phenomenon. We report here that left and right lanterns of live larvae of the fireflies, Photuris lucicrescens and

  8. Novel Bioluminescent Activatable Reporter for Src Tyrosine Kinase Activity in Living Mice.

    Leng, Weibing; Li, Dezhi; Chen, Liang; Xia, Hongwei; Tang, Qiulin; Chen, Baoqin; Gong, Qiyong; Gao, Fabao; Bi, Feng

    2016-01-01

    Aberrant activation of the Src kinase is implicated in the development of a variety of human malignancies. However, it is almost impossible to monitor Src activity in an in vivo setting with current biochemical techniques. To facilitate the noninvasive investigation of the activity of Src kinase both in vitro and in vivo, we developed a genetically engineered, activatable bioluminescent reporter using split-luciferase complementation. The bioluminescence of this reporter can be used as a surrogate for Src activity in real time. This hybrid luciferase reporter was constructed by sandwiching a Src-dependent conformationally responsive unit (SH2 domain-Srcpep) between the split luciferase fragments. The complementation bioluminescence of this reporter was dependent on the Src activity status. In our study, Src kinase activity in cultured cells and tumor xenografts was monitored quantitatively and dynamically in response to clinical small-molecular kinase inhibitors, dasatinib and saracatinib. This system was also applied for high-throughput screening of Src inhibitors against a kinase inhibitor library in living cells. These results provide unique insights into drug development and pharmacokinetics/phoarmocodynamics of therapeutic drugs targeting Src signaling pathway enabling the optimization of drug administration schedules for maximum benefit. Using both Firefly and Renilla luciferase imaging, we have successfully monitored Src tyrosine kinase activity and Akt serine/threonine kinase activity concurrently in one tumor xenograft. This dual luciferase reporter imaging system will be helpful in exploring the complex signaling networks in vivo. The strategies reported here can also be extended to study and image other important kinases and the cross-talks among them. PMID:26941850

  9. 活体生物发光成像追踪大鼠跟腱内移植干细胞**☆○%Monitoring transplanted stem cells in rat Achilles tendon by in vivo bioluminescent imaging

    黄德清; Gary Balian

    2013-01-01

      BACKGROUND: The mechanisms for the homing, migration, proliferation and differentiation of transplanted adipose tissue derived stem cel s remain unclear. The in vivo bioluminescent imaging system is a newly developed technique for directly detecting the biological behaviors of transplanted cel s in vivo. OBJECTIVE: To demonstrate the feasibility of using in vivo bioluminescent imaging system to monitor the genetical y modified adipose tissue derived stem cel s transplanted in Achil es tendon of rats. METHODS: Adipose tissue derived stem cel s isolated from the abdominal cavity of Sprague-Dawley rat were transduced with an adenovirus containing the luciferase reporter gene (3×1010/L), to observe the influence of transfection on the adipose tissue derived stem cel s. Subsequently, the transfected cel s were implanted into Achil es tendon defects in rats. The in vivo bioluminescent imaging system was used at days 1, 4, 7 and 14 fol owing transplantation to assess the luciferase expression. The cryosections of repaired Achil es tendon of rats were observed under fluorescence microscope at day 28 postoperatively. RESULTS AND CONCLUSION: No influence on the morphology and proliferation of adipose tissue derived stem cel s was observed after transducing in vitro (P > 0.05). On the repaired Achil es tendon, the luciferase gene expression detected with in vivo bioluminescent imaging system at days 1, 4, 7 and 14 was respectively (1.22±0.43)×106, (1.81±0.76)×106, (1.88±0.69)×106 and (0.89±0.26)×105 counts/s (n=6). Abundant adipose tissue derived stem cel s with luciferase expression were also seen in tendon cryosections of this side under fluorescence microscope at day 28. The luciferase gene expression was not detected in the control side. Experimental findings demonstrate that the in vivo bioluminescent imaging system can successful y monitor the fluorogene modified adipose tissue derived stem cel s that are implanted into the rat Achil es tendon, and

  10. Monitoring transplanted stem cells in rat Achilles tendon by in vivo bioluminescent imaging%活体生物发光成像追踪大鼠跟腱内移植干细胞**☆○

    黄德清; Gary Balian

    2013-01-01

      背景:移植脂肪源干细胞在活体内的归巢、迁移、增殖和分化的机制仍未得到充分阐明。活体生物发光活体成像技术是近来发展起来的一种可以直接检测活细胞在动物体内生物学行为的新的技术方法。目的:探讨用活体生物发光成像技术检测大鼠跟腱内移植的经荧光基因修饰的脂肪源干细胞可行性。方法:分离培养大鼠腹腔来源的脂肪源干细胞,用浓度为3×1010 L-1携带虫荧光素酶的腺病毒载体对其进行转染,观察转染对脂肪源干细胞的影响;将转染的脂肪源干细胞移植到大鼠跟腱缺损处,移植后1,4,7,14 d用活体生物发光成像技术检测移植脂肪源干细胞荧光素酶的表达,移植后28 d跟腱标本冰冻切片在荧光显微镜下观察。结果与结论:在体外,腺病毒转染对脂肪源干细胞的生长和增殖无明显影响(P >0.05)。细胞移植后1,4,7,14 d,活体生物发光成像技术在实验侧修复段跟腱检测到的荧光表达强度分别为(1.22±0.43)×106、(1.81±0.76)×106、(1.88±0.69)×106和(0.89±0.26)×105光子/s(n=6)。而对照侧跟腱修复段未检测到荧光表达;移植后28 d,实验侧跟腱冰冻切片在荧光显微镜下见到大量表达荧光的细胞。表明活体生物发光成像技术可成功追踪大鼠跟腱内移植的经荧光基因修饰的脂肪源干细胞。脂肪源干细胞有望成为肌腱组织工程的种子细胞。%  BACKGROUND: The mechanisms for the homing, migration, proliferation and differentiation of transplanted adipose tissue derived stem cel s remain unclear. The in vivo bioluminescent imaging system is a newly developed technique for directly detecting the biological behaviors of transplanted cel s in vivo. OBJECTIVE: To demonstrate the feasibility of using in vivo bioluminescent imaging system to monitor the genetical y modified adipose tissue derived stem cel s transplanted in

  11. Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level.

    Kenta Saito

    Full Text Available BACKGROUND: Efficient bioluminescence resonance energy transfer (BRET from a bioluminescent protein to a fluorescent protein with high fluorescent quantum yield has been utilized to enhance luminescence intensity, allowing single-cell imaging in near real time without external light illumination. METHODOLOGY/PRINCIPAL FINDINGS: We applied BRET to develop an autoluminescent Ca(2+ indicator, BRAC, which is composed of Ca(2+-binding protein, calmodulin, and its target peptide, M13, sandwiched between a yellow fluorescent protein variant, Venus, and an enhanced Renilla luciferase, RLuc8. Adjusting the relative dipole orientation of the luminescent protein's chromophores improved the dynamic range of BRET signal change in BRAC up to 60%, which is the largest dynamic range among BRET-based indicators reported so far. Using BRAC, we demonstrated successful visualization of Ca(2+ dynamics at the single-cell level with temporal resolution at 1 Hz. Moreover, BRAC signals were acquired by ratiometric imaging capable of canceling out Ca(2+-independent signal drifts due to change in cell shape, focus shift, etc. CONCLUSIONS/SIGNIFICANCE: The brightness and large dynamic range of BRAC should facilitate high-sensitive Ca(2+ imaging not only in single live cells but also in small living subjects.

  12. Persistence of a bioluminescent Staphylococcus aureus strain on and around degradable and non-degradable surgical meshes in a murine model.

    Daghighi, Seyedmojtaba; Sjollema, Jelmer; Jaspers, Valery; de Boer, Leonie; Zaat, Sebastian A J; Dijkstra, Rene J B; van Dam, Gooitzen M; van der Mei, Henny C; Busscher, Henk J

    2012-11-01

    Biomaterials are increasingly used for the restoration of human function, but can become infected as a result of peri- or early post-operative bacterial contamination, although biomaterial-associated infections (BAIs) can also initiate at any time from hematogenous spreading of bacteria from an infection elsewhere in the body. Infecting bacteria in BAIs not only seek shelter in their own protective biofilm matrix, but also hide in surrounding tissue. This study compares staphylococcal persistence on and around a degradable and non-degradable surgical mesh through the use of longitudinal bioluminescence imaging in a murine model, including histological evaluation of surrounding tissue after sacrifice. Surgical meshes were first contaminated with bioluminescent Staphylococcus aureus Xen29 and subsequently subcutaneously implanted in mice. Bioluminescent staphylococci persisted on and around non-degradable meshes during the 28-day course of the study, whereas bioluminescence returned to control levels and bacteria disappeared from surrounding tissues once a degradable mesh had fully dissolved. Thus the application of degradable biomaterials yields major advantages with respect to the prevention of BAIs, as dissolution of the implant not only is associated with elimination of the protective biofilm mode of growth of the infecting organisms, but also allows the immune system to clear the surrounding tissue from infecting organisms. PMID:22824527

  13. Bioluminescence Imaging Cells Labeled with Membrane-Anchored Form of Gaussia Luciferase%膜锚定Gaussia萤光素酶的细胞标记及生物荧光成像

    樊炜; 贾帅争; 王怡; 阎少多; 高博; 彭剑淳; 詹林盛

    2011-01-01

    目的:制备表达膜锚定Gaussia萤光素酶(extGluc)报告基因的慢病毒,用于标记细胞.方法:将报告基因extGluc克隆至慢病毒载体pCCsin.PPT.SFFV.IRES.eGFP.Wpre (VeGFP)中,以聚乙烯亚胺(PEI)介导,将慢病毒包装所需4种质粒(pVeGFP-extGLuc、pMDL、pRey、pVSVG),转染293FT细胞,72 h后收集病毒上清进行浓缩,感染293FT细胞,并用流式细胞仪检测病毒滴度,生物荧光成像和化学发光分析extGluc的表达;之后,用收集的慢病毒感染人单核细胞白血病细胞株U937.结果:对经PCR筛选出的阳性克隆所含质粒进行酶切鉴定,表明extGlu报告基因插入载体中;重组慢病毒包装成功且病毒滴度为5×106 TU/mL;用包装的病毒颗粒感染293FT、细胞,生物荧光成像和化学发光证实extGluc的膜定位,且酶活性与细胞数目呈线性相关;病毒颗粒能够感染悬浮细胞U937.结论:包装了extGluc标记的重组慢病毒,可用于标记细胞,为体内监测细胞迁移、聚集和变化提供了一种方法.%Objective: To produce lentivirus expressing reporter gene membrane-anchored form of Gaussia lu-ciferase (extGluc) used for bioluminescence imaging cells. Methods: Reporter gene extGluc was cloned into pCCsin. PPT.SFFV.IRES.eGFP.Wpre(VeGFP). pVeGFP-extGLuc, pMDL, pRev and pVSVG, which were required for packaging, were cotransfected into 293FT cells mediated by polyethylenimine branched. Lentivirus was collected at 72 h post-transfection, concentrated and then was used to infect 293FT cells. Viral titer was determined by flow cytome-try. Luciferase activity was detected by bioluminescence imaging and chemiluminescence. At last, human monocytic leukemia cell line U937 were infected by viral supernatant. Results: PCR and enzyme digestion results indicated that reporter gene extGLuc was successfully cloned into VeGFP. The lentivirus was packaged successfully. The lentivirus titer was 5xlO6 TU/mL. After infecting 293FT with virus particles, ext

  14. Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging.

    Takai, Akira; Nakano, Masahiro; Saito, Kenta; Haruno, Remi; Watanabe, Tomonobu M; Ohyanagi, Tatsuya; Jin, Takashi; Okada, Yasushi; Nagai, Takeharu

    2015-04-01

    Fluorescence live imaging has become an essential methodology in modern cell biology. However, fluorescence requires excitation light, which can sometimes cause potential problems, such as autofluorescence, phototoxicity, and photobleaching. Furthermore, combined with recent optogenetic tools, the light illumination can trigger their unintended activation. Because luminescence imaging does not require excitation light, it is a good candidate as an alternative imaging modality to circumvent these problems. The application of luminescence imaging, however, has been limited by the two drawbacks of existing luminescent protein probes, such as luciferases: namely, low brightness and poor color variants. Here, we report the development of bright cyan and orange luminescent proteins by extending our previous development of the bright yellowish-green luminescent protein Nano-lantern. The color change and the enhancement of brightness were both achieved by bioluminescence resonance energy transfer (BRET) from enhanced Renilla luciferase to a fluorescent protein. The brightness of these cyan and orange Nano-lanterns was ∼20 times brighter than wild-type Renilla luciferase, which allowed us to perform multicolor live imaging of intracellular submicron structures. The rapid dynamics of endosomes and peroxisomes were visualized at around 1-s temporal resolution, and the slow dynamics of focal adhesions were continuously imaged for longer than a few hours without photobleaching or photodamage. In addition, we extended the application of these multicolor Nano-lanterns to simultaneous monitoring of multiple gene expression or Ca(2+) dynamics in different cellular compartments in a single cell. PMID:25831507

  15. A multithread based new sparse matrix method in bioluminescence tomography

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

    2010-03-01

    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.

  16. Multi-atlas registration and adaptive hexahedral voxel discretization for fast bioluminescence tomography

    Ren, Shenghan; Hu, Haihong; Li, Gen; Cao, Xu; Zhu, Shouping; Chen, Xueli; Liang, Jimin

    2016-01-01

    Bioluminescence tomography (BLT) has been a valuable optical molecular imaging technique to non-invasively depict the cellular and molecular processes in living animals with high sensitivity and specificity. Due to the inherent ill-posedness of BLT, a priori information of anatomical structure is usually incorporated into the reconstruction. The structural information is usually provided by computed tomography (CT) or magnetic resonance imaging (MRI). In order to obtain better quantitative results, BLT reconstruction with heterogeneous tissues needs to segment the internal organs and discretize them into meshes with the finite element method (FEM). It is time-consuming and difficult to handle the segmentation and discretization problems. In this paper, we present a fast reconstruction method for BLT based on multi-atlas registration and adaptive voxel discretization to relieve the complicated data processing procedure involved in the hybrid BLT/CT system. A multi-atlas registration method is first adopted to estimate the internal organ distribution of the imaged animal. Then, the animal volume is adaptively discretized into hexahedral voxels, which are fed into FEM for the following BLT reconstruction. The proposed method is validated in both numerical simulation and an in vivo study. The results demonstrate that the proposed method can reconstruct the bioluminescence source efficiently with satisfactory accuracy.

  17. Multi-atlas registration and adaptive hexahedral voxel discretization for fast bioluminescence tomography.

    Ren, Shenghan; Hu, Haihong; Li, Gen; Cao, Xu; Zhu, Shouping; Chen, Xueli; Liang, Jimin

    2016-04-01

    Bioluminescence tomography (BLT) has been a valuable optical molecular imaging technique to non-invasively depict the cellular and molecular processes in living animals with high sensitivity and specificity. Due to the inherent ill-posedness of BLT, a priori information of anatomical structure is usually incorporated into the reconstruction. The structural information is usually provided by computed tomography (CT) or magnetic resonance imaging (MRI). In order to obtain better quantitative results, BLT reconstruction with heterogeneous tissues needs to segment the internal organs and discretize them into meshes with the finite element method (FEM). It is time-consuming and difficult to handle the segmentation and discretization problems. In this paper, we present a fast reconstruction method for BLT based on multi-atlas registration and adaptive voxel discretization to relieve the complicated data processing procedure involved in the hybrid BLT/CT system. A multi-atlas registration method is first adopted to estimate the internal organ distribution of the imaged animal. Then, the animal volume is adaptively discretized into hexahedral voxels, which are fed into FEM for the following BLT reconstruction. The proposed method is validated in both numerical simulation and an in vivo study. The results demonstrate that the proposed method can reconstruct the bioluminescence source efficiently with satisfactory accuracy. PMID:27446674

  18. Images of time mind, science, reality

    Jaroszkiewicz, George

    2016-01-01

    Have you ever wondered about Time: what it is or how to discuss it? If you have, then you may have been bewildered by the many different views and opinions in many diverse fields to be found, such as physics, mathematics, philosophy, religion, history, and science fiction novels and films. This book will help you unravel fact from fiction. It provides a broad survey of many of these views, these images of time, covering historical, cultural, philosophical, biological, mathematical and physical images of time, including classical and quantum mechanics, special and general relativity and cosmology. This book gives you more than just a review of such images. It provides the reader a basis for judging the scientific soundness of these various images. It develops the reader's critical ability to distinguish Images of Time in terms of its contextual completeness. Differentiating between metaphysical images (which cannot be scientifically validated) and those that could, in principle, be put to empirical test. Showi...

  19. Protein-protein complexation in bioluminescence

    Titushin, Maxim S.; Feng, Yingang; Lee, John; Vysotski, Eugene S.; Liu, Zhi-jie

    2011-01-01

    In this review we summarize the progress made towards understanding the role of protein-protein interactions in the function of various bioluminescence systems of marine organisms, including bacteria, jellyfish and soft corals, with particular focus on methodology used to detect and characterize these interactions. In some bioluminescence systems, protein-protein interactions involve an “accessory protein” whereby a stored substrate is efficiently delivered to the bioluminescent enzyme lucife...

  20. A Nisin Bioassay Based on Bioluminescence

    Wahlström, G.; Saris, P. E. J.

    1999-01-01

    A Lactococcus lactis subsp. lactis strain that can sense the bacteriocin nisin and transduce the signal into bioluminescence was constructed. By using this strain, a bioassay based on bioluminescence was developed for quantification of nisin, for detection of nisin in milk, and for identification of nisin-producing strains. As little as 0.0125 ng of nisin per ml was detected within 3 h by this bioluminescence assay. This detection limit was lower than in previously described methods.

  1. Non-invasive imaging of GFP-luciferase labeled orthotopic prostate cancer model in nude mice using bioluminescence system%可发光可连续检测原位前列腺癌模型的建立

    宋超; 廖文彪; 杨嗣星; 王玲珑

    2012-01-01

    .43 ± 4.56 ) g accordingly.There was a positive linear correlation between in vivo bioluminescent values and ex vivo tumor weight with coefficient being 0.973,P <0.05.Conclusion Our findings demonstrate the ability of the luciferase labeled tumor cells combinated within vivo bioluminesence imaging system is an excellent experimental animal model in tracking the location,magnitude and persistence of luciferase expression cells in human prostate cancer mouse models,which may be usful in non-invasively monitoring on progression of prostate cancer in vivo,even after the metastasis of the tumor.%目的 建立一种可连续定量监测的原位前列腺癌动物模型.方法 利用分子克隆技术构建表达荧光素酶的慢病毒载体,用慢病毒将GFP-Luciferase融合蛋白载体转染到人前列腺癌PC3细胞株;利用稻瘟素Blasticidin筛选获得稳定转染的GFP-Luc-PC3细胞株.将6只雄性裸鼠随机分2组,应用5×106 GFP-Luc-PC3或普通PC3细胞雄性裸鼠皮下注射获得皮下瘤.12周时无菌条件下取荧光素酶标记和未标记的皮下肿瘤组织,将其切成2 mm3大小的组织块,并将组织块随机种植于24只雄性裸鼠前列腺包膜下,实验组、对照组各12只.用IVIS 200光子发射定量分析动态监测肿瘤生长.并于种植后2、4、6、8周分别处死3只裸鼠,取前列腺组织称重.分析肿瘤在体光子值与其重量的相关性.结果 成功建立GFP-Luc-PC3细胞株,该系生长曲线与未标记细胞一致,在荧光素作用下,发光能力与细胞数量呈正相关(r=0.997),其裸鼠皮下成瘤率为100%.原位前列腺癌模型中,所有24只动物均形成前列腺肿瘤,其中GFP-Luc-PC3组可通过IVIS 200系统观察到肿瘤组织的发光情况,PC3组信号不明显.原位肿瘤模型建立后2、4、6、8周,肿瘤在体光子值(光子值/秒,photons/second)依次为(69.13298±2.07900)E+05、(82.66208±1.23100)E+05、(91.942 57±2.32100)E +05、(130.643 40±3.247 00)E+05.

  2. Positive self-image over time

    Santos-Pinto, Luís

    2005-01-01

    This paper incorporates egocentric comparisons into a human capital accumulation model and studies the evolution of positive self image over time. The paper shows that the process of human capital accumulation together with egocentric comparisons imply that positive self image of a cohort is first increasing and then decreasing over time. Additionally, the paper finds that positive self image: (1) peaks earlier in activities where skill depreciation is higher, (2) is smaller in activities whe...

  3. Foraging in the darkness of the Southern Ocean: influence of bioluminescence on a deep diving predator.

    Vacquié-Garcia, Jade; Royer, François; Dragon, Anne-Cécile; Viviant, Morgane; Bailleul, Frédéric; Guinet, Christophe

    2012-01-01

    How non-echolocating deep diving marine predators locate their prey while foraging remains mostly unknown. Female southern elephant seals (SES) (Mirounga leonina) have vision adapted to low intensity light with a peak sensitivity at 485 nm. This matches the wavelength of bioluminescence produced by a large range of marine organisms including myctophid fish, SES's main prey. In this study, we investigated whether bioluminescence provides an accurate estimate of prey occurrence for SES. To do so, four SES were satellite-tracked during their post-breeding foraging trip and were equipped with Time-Depth-Recorders that also recorded light levels every two seconds. A total of 3386 dives were processed through a light-treatment model that detected light events higher than ambient level, i.e. bioluminescence events. The number of bioluminescence events was related to an index of foraging intensity for SES dives deep enough to avoid the influence of natural ambient light. The occurrence of bioluminescence was found to be negatively related to depth both at night and day. Foraging intensity was also positively related to bioluminescence both during day and night. This result suggests that bioluminescence likely provides SES with valuable indications of prey occurrence and might be a key element in predator-prey interactions in deep-dark marine environments. PMID:22952706

  4. Foraging in the darkness of the Southern Ocean: influence of bioluminescence on a deep diving predator.

    Jade Vacquié-Garcia

    Full Text Available How non-echolocating deep diving marine predators locate their prey while foraging remains mostly unknown. Female southern elephant seals (SES (Mirounga leonina have vision adapted to low intensity light with a peak sensitivity at 485 nm. This matches the wavelength of bioluminescence produced by a large range of marine organisms including myctophid fish, SES's main prey. In this study, we investigated whether bioluminescence provides an accurate estimate of prey occurrence for SES. To do so, four SES were satellite-tracked during their post-breeding foraging trip and were equipped with Time-Depth-Recorders that also recorded light levels every two seconds. A total of 3386 dives were processed through a light-treatment model that detected light events higher than ambient level, i.e. bioluminescence events. The number of bioluminescence events was related to an index of foraging intensity for SES dives deep enough to avoid the influence of natural ambient light. The occurrence of bioluminescence was found to be negatively related to depth both at night and day. Foraging intensity was also positively related to bioluminescence both during day and night. This result suggests that bioluminescence likely provides SES with valuable indications of prey occurrence and might be a key element in predator-prey interactions in deep-dark marine environments.

  5. BIOLUMINESCENCE TOMOGRAPHY: BIOMEDICAL BACKGROUND, MATHEMATICAL THEORY, AND NUMERICAL APPROXIMATION

    Weimin Han; Ce Wang

    2008-01-01

    Over the last couple of years molecular imaging has been rapidly developed to study physiological and pathological processes in vivo at the cellular and molecular levels. Among molecular imaging modalities, optical imaging stands out for its unique advantages, especially performance and cost-effectiveness. Bioluminescence tomography (BLT) is an emerging optical imaging mode with promising biomedical advantages. In this survey paper, we explain the biomedical significance of BLT, summarize theoretical results on the analysis and numerical solution of a diffusion based BLT model, and comment on a few extensions for the study of BLT.

  6. Mathematical Study and Numerical Simulation of Multispectral Bioluminescence Tomography

    Weimin Han; Wenxiang Cong; Ge Wang

    2006-01-01

    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 illust...

  7. Experimental Study on Bioluminescence Tomography with Multimodality Fusion

    Lv, Yujie; Tian, Jie; Cong, Wenxiang; Wang, Ge

    2007-01-01

    To verify the influence of a priori information on the nonuniqueness problem of bioluminescence tomography (BLT), the multimodality imaging fusion based BLT experiment is performed by multiview noncontact detection mode, which incorporates the anatomical information obtained by the microCT scanner and the background optical properties based on diffuse reflectance measurements. In the reconstruction procedure, the utilization of adaptive finite element methods (FEMs) and a priori permissible s...

  8. Multi-projection bioluminescence tomography guided system for small animal radiation research platform (SARRP)

    Zhang, Bin; Iordachita, Iulian; Wong, John W.; Wang, Ken Kang-Hsin

    2016-03-01

    Cone beam computed tomography (CBCT) is limited in guiding irradiation for soft tissue targets. As a complementary imaging modality, bioluminescence tomography (BLT) provides strong soft tissue contrast. We developed a dual-use BLT system which consists of an optical assembly, a mobile cart and an independent mouse bed. The system is motorized which can easily dock onto an independent mouse bed operating as a standalone system for longitudinal bioluminescence imaging (BLI)/BLT studies and also dock onto the SARRP for on-line radiation guidance. Our initial tests for the system demonstrate that (i) the imaging depth is 28 mm, (ii) the optical background is sufficiently low and uniform, (iii) the non-uniform response of the optical imaging can be corrected by the flat field correction, and (iv) the imaging acquisition speed was improved by an average of 3.7 times faster than our previous systems. We also presented a geometry calibration procedure to map the planar BLIs acquired at multi-projections onto the surface of the CBCT image. The CBCT is required to generate the mesh for BLT reconstruction and used for treatment planning and radiation delivery. Feasibility study of the geometry calibration was performed on a manual-docking prototype. The mean and maximum mapping accuracy is 0.3 and 0.6 mm. The performance of the proposed motorized dual-use system is expected to be superior to that of the manual-docking prototype because of the mechanism stability. We anticipate the dual-use system as a highly efficient and cost-effective platform to facilitate optical imaging for preclinical radiation research.

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

    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 β-catenin, Tcf4 and β-catenin/Tcf4 transcriptional activity, concomitantly with decreased expression of β-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-α, JNK, NF-κB, c-Jun and c-Fos while increased that of IκBα. 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 β-catenin, cyclin D1, survivin, and endothelial marker CD31 while increased caspase-3 expression. Collectively, our data supports DB as an inhibitor of Wnt/β-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 β-catenin, Tcf4. ► DB attenuated cyclin D1, c-myc, survivin and tumorigenesis in HT29 xenograft mice.

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

    Yeh, Chi-Tai [Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan (China); Center of Excellence for Cancer Research, Taipei Medical University, Taipei, Taiwan (China); Department of Surgery, Taipei Medical University-Shuang Ho Hospital, Taipei, Taiwan (China); Rao, Yerra Koteswara [Institute of Biochemical Sciences and Technology, Chaoyang University of Technology, Taichung, Taiwan (China); Ye, Min [Department of Natural Medicine, School of Pharmaceutical Sciences, Peking University, Beijing (China); Wu, Wen-Shi [Department of Horticulture and Biotechnology, Chinese Culture University, Taipei, Taiwan (China); Chang, Tung-Chen [Department of Surgery, Taipei Medical University-Shuang Ho Hospital, Taipei, Taiwan (China); Wang, Liang-Shun [Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan (China); Division of Thoracic Surgery, Department of Surgery, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan (China); Wu, Chih-Hsiung [Center of Excellence for Cancer Research, Taipei Medical University, Taipei, Taiwan (China); Department of Surgery, Taipei Medical University-Shuang Ho Hospital, Taipei, Taiwan (China); Wu, Alexander T.H., E-mail: chaw1211@tmu.edu.tw [Ph.D. Program for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan (China); Department of Radiation Oncology, Taipei Medical University Hospital, Taipei, Taiwan (China); Tzeng, Yew-Min, E-mail: ymtzeng@cyut.edu.tw [Institute of Biochemical Sciences and Technology, Chaoyang University of Technology, Taichung, Taiwan (China)

    2012-05-15

    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 β-catenin, Tcf4 and β-catenin/Tcf4 transcriptional activity, concomitantly with decreased expression of β-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-α, JNK, NF-κB, c-Jun and c-Fos while increased that of IκBα. 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 β-catenin, cyclin D1, survivin, and endothelial marker CD31 while increased caspase-3 expression. Collectively, our data supports DB as an inhibitor of Wnt/β-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 β-catenin, Tcf4. ► DB attenuated cyclin D1, c-myc, survivin and tumorigenesis in HT29 xenograft mice.

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

    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.

  12. REVIEW OF ENVIRONMENTAL APPLICATIONS OF BIOLUMINESCENCE MEASUREMENTS

    This review of the recent literature on environmental applications of bioluminescence systems will focus on in vivo and in vitro bioluminescence methods that have been utilized to elucidate properties of chemicals, toxic and mutagenic effects, and to estimate biomass. he unifying...

  13. Bacterial bioluminescence and Gumbel statistics: From quorum sensing to correlation

    Delle Side, Domenico; Velardi, Luciano; Nassisi, Vincenzo; Pennetta, Cecilia; Alifano, Pietro; Talà, Adelfia; Salvatore Tredici, Maurizio

    2013-12-01

    We show that, in particular experimental conditions, the time course of the radiant fluxes, measured from a bioluminescent emission of a Vibrio harveyi related strain, collapse after suitable rescaling onto the Gumbel distribution of extreme value theory. We argue that the activation times of the strain luminous emission follow the universal behavior described by this statistical law, in spite of the fact that no extremal process is known to occur.

  14. Understanding bioluminescence in dinoflagellates — how far have we come?

    Martha Valiadi; Debora Iglesias-Rodriguez

    2013-01-01

    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 bioluminescence ...

  15. Bioluminescence tomography based on the phase approximation model

    Cong, W.; Wang, G

    2010-01-01

    A reconstruction method of bioluminescence sources is proposed based on a phase approximation model. Compared with the diffuse approximation, this phase approximation model more correctly predicts bioluminescence photon propagation in biological tissues, so that bioluminescence tomography can accurately locate and quantify the distribution of bioluminescence sources. The compressive sensing (CS) technique is applied to regularize the inverse source reconstruction to enhance numerical stabilit...

  16. Bioluminescent human breast cancer cell lines that permit rapid and sensitive in vivo detection of mammary tumors and multiple metastases in immune deficient mice

    Our goal was to generate xenograft mouse models of human breast cancer based on luciferase-expressing MDA-MB-231 tumor cells that would provide rapid mammary tumor growth; produce metastasis to clinically relevant tissues such as lymph nodes, lung, and bone; and permit sensitive in vivo detection of both primary and secondary tumor sites by bioluminescent imaging. Two clonal cell sublines of human MDA-MB-231 cells that stably expressed firefly luciferase were isolated following transfection of the parental cells with luciferase cDNA. Each subline was passaged once or twice in vivo to enhance primary tumor growth and to increase metastasis. The resulting luciferase-expressing D3H1 and D3H2LN cells were analyzed for long-term bioluminescent stability, primary tumor growth, and distal metastasis to lymph nodes, lungs, bone and soft tissues by bioluminescent imaging. Cells were injected into the mammary fat pad of nude and nude-beige mice or were delivered systemically via intracardiac injection. Metastasis was also evaluated by ex vivo imaging and histologic analysis postmortem. The D3H1 and D3H2LN cell lines exhibited long-term stable luciferase expression for up to 4–6 months of accumulative tumor growth time in vivo. Bioluminescent imaging quantified primary mammary fat pad tumor development and detected early spontaneous lymph node metastasis in vivo. Increased frequency of spontaneous lymph node metastasis was observed with D3H2LN tumors as compared with D3H1 tumors. With postmortem ex vivo imaging, we detected additional lung micrometastasis in mice with D3H2LN mammary tumors. Subsequent histologic evaluation of tissue sections from lymph nodes and lung lobes confirmed spontaneous tumor metastasis at these sites. Following intracardiac injection of the MDA-MB-231-luc tumor cells, early metastasis to skeletal tissues, lymph nodes, brain and various visceral organs was detected. Weekly in vivo imaging data permitted longitudinal analysis of metastasis at

  17. Toward Real Time Uavs' Image Mosaicking

    Mehrdad, S.; Satari, M.; Safdary, M.; Moallem, P.

    2016-06-01

    Anyone knows that sudden catastrophes can instantly do great damage. Fast and accurate acquisition of catastrophe information is an essential task for minimize life and property damage. Compared with other ways of catastrophe data acquisition, UAV based platforms can optimize time, cost and accuracy of the data acquisition, as a result UAVs' data has become the first choice in such condition. In this paper, a novel and fast strategy is proposed for registering and mosaicking of UAVs' image data. Firstly, imprecise image positions are used to find adjoining frames. Then matching process is done by a novel matching method. With keeping Sift in mind, this fast matching method is introduced, which uses images exposure time geometry, SIFT point detector and rBRIEF descriptor vector in order to match points efficiency, and by efficiency we mean not only time efficiency but also elimination of mismatch points. This method uses each image sequence imprecise attitude in order to use Epipolar geometry to both restricting search space of matching and eliminating mismatch points. In consideration of reaching to images imprecise attitude and positions we calibrated the UAV's sensors. After matching process, RANSAC is used to eliminate mismatched tie points. In order to obtain final mosaic, image histograms are equalized and a weighted average method is used to image composition in overlapping areas. The total RMSE over all matching points is 1.72 m.

  18. Increased bioassay sensitivity of bioactive molecule discovery using metal-enhanced bioluminescence

    We report the use of bioluminescence signal enhancement via proximity to deposited silver nanoparticles for bioactive compound discovery. This approach employs a whole-cell bioreporter harboring a plasmid-borne fusion of a specific promoter incorporated with a bioluminescence reporter gene. The silver deposition process was first optimized to provide optimal nanoparticle size in the reaction time dependence with fluorescein. The use of silver deposition of 350 nm particles enabled the doubling of the bioluminescent signal amplitude by the bacterial bioreporter when compared to an untouched non-silver-deposited microtiter plate surface. This recording is carried out in the less optimal but necessary far-field distance. SEM micrographs provided a visualization of the proximity of the bioreporter to the silver nanoparticles. The electromagnetic field distributions around the nanoparticles were simulated using Finite Difference Time Domain, further suggesting a re-excitation of non-chemically excited bioluminescence in addition to metal-enhanced bioluminescence. The possibility of an antiseptic silver effect caused by such a close proximity was eliminated disregarded by the dynamic growth curves of the bioreporter strains as seen using viability staining. As a highly attractive biotechnology tool, this silver deposition technique, coupled with whole-cell sensing, enables increased bioluminescence sensitivity, making it especially useful for cases in which reporter luminescence signals are very weak

  19. Increased bioassay sensitivity of bioactive molecule discovery using metal-enhanced bioluminescence

    Golberg, Karina, E-mail: karingo@bgu.ac.il; Elbaz, Amit [Ben-Gurion University of the Negev, Avram and Stella Goldstein-Goren Department of Biotechnology Engineering (Israel); McNeil, Ronald [The Institute of Fluorescence, University of Maryland Baltimore County (United States); Kushmaro, Ariel [Ben-Gurion University of the Negev, Avram and Stella Goldstein-Goren Department of Biotechnology Engineering (Israel); Geddes, Chris D. [The Institute of Fluorescence, University of Maryland Baltimore County (United States); Marks, Robert S., E-mail: rsmarks@bgu.ac.il [Ben-Gurion University of the Negev, Avram and Stella Goldstein-Goren Department of Biotechnology Engineering (Israel)

    2014-12-15

    We report the use of bioluminescence signal enhancement via proximity to deposited silver nanoparticles for bioactive compound discovery. This approach employs a whole-cell bioreporter harboring a plasmid-borne fusion of a specific promoter incorporated with a bioluminescence reporter gene. The silver deposition process was first optimized to provide optimal nanoparticle size in the reaction time dependence with fluorescein. The use of silver deposition of 350 nm particles enabled the doubling of the bioluminescent signal amplitude by the bacterial bioreporter when compared to an untouched non-silver-deposited microtiter plate surface. This recording is carried out in the less optimal but necessary far-field distance. SEM micrographs provided a visualization of the proximity of the bioreporter to the silver nanoparticles. The electromagnetic field distributions around the nanoparticles were simulated using Finite Difference Time Domain, further suggesting a re-excitation of non-chemically excited bioluminescence in addition to metal-enhanced bioluminescence. The possibility of an antiseptic silver effect caused by such a close proximity was eliminated disregarded by the dynamic growth curves of the bioreporter strains as seen using viability staining. As a highly attractive biotechnology tool, this silver deposition technique, coupled with whole-cell sensing, enables increased bioluminescence sensitivity, making it especially useful for cases in which reporter luminescence signals are very weak.

  20. Interactive Real-time Magnetic Resonance Imaging

    Brix, Lau

    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...... 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...

  1. NanoLuc: A Small Luciferase Is Brightening Up the Field of Bioluminescence.

    England, Christopher G; Ehlerding, Emily B; Cai, Weibo

    2016-05-18

    The biomedical field has greatly benefited from the discovery of bioluminescent proteins. Currently, scientists employ bioluminescent systems for numerous biomedical applications, ranging from highly sensitive cellular assays to bioluminescence-based molecular imaging. Traditionally, these systems are based on Firefly and Renilla luciferases; however, the applicability of these enzymes is limited by their size, stability, and luminescence efficiency. NanoLuc (NLuc), a novel bioluminescence platform, offers several advantages over established systems, including enhanced stability, smaller size, and >150-fold increase in luminescence. In addition, the substrate for NLuc displays enhanced stability and lower background activity, opening up new possibilities in the field of bioluminescence imaging. The NLuc system is incredibly versatile and may be utilized for a wide array of applications. The increased sensitivity, high stability, and small size of the NLuc system have the potential to drastically change the field of reporter assays in the future. However, as with all such technology, NLuc has limitations (including a nonideal emission for in vivo applications and its unique substrate) which may cause it to find restricted use in certain areas of molecular biology. As this unique technology continues to broaden, NLuc may have a significant impact in both preclinical and clinical fields, with potential roles in disease detection, molecular imaging, and therapeutic monitoring. This review will present the NLuc technology to the scientific community in a nonbiased manner, allowing the audience to adopt their own views of this novel system. PMID:27045664

  2. Experimental Study on Bioluminescence Tomography with Multimodality Fusion

    Yujie Lv

    2007-01-01

    Full Text Available To verify the influence of a priori information on the nonuniqueness problem of bioluminescence tomography (BLT, the multimodality imaging fusion based BLT experiment is performed by multiview noncontact detection mode, which incorporates the anatomical information obtained by the microCT scanner and the background optical properties based on diffuse reflectance measurements. In the reconstruction procedure, the utilization of adaptive finite element methods (FEMs and a priori permissible source region refines the reconstructed results and improves numerical robustness and efficiency. The comparison between the absence and employment of a priori information shows that multimodality imaging fusion is essential to quantitative BLT reconstruction.

  3. Actual imaging time in fetal MRI

    Objective: Safety issues in magnetic resonance imaging (MRI) are important, especially in fetal MRI. However, since basic data with respect of the effective exposure time in fetal MRI are not available, this study aimed to determine the actual imaging time during a fetal MRI study. Methods: 100 fetal MRI studies of singleton pregnancies performed on a 1.5 T system were analysed with respect to study duration (from starting the survey scan until the end of study), the number of sequences acquired, and the actual imaging time, which was calculated by adding up scan time of each sequence. Furthermore, each sequence type was analysed regarding the number of acquisitions, specific absorption rates (SAR), and duration. Results: Mean study duration was 34.6 min (range: 14–58 min; standard deviation (SD): 9.7 min), the average number of sequences acquired was 26.6 (range: 11–44, SD: 6.6). Actual scan time averaged 11.4 min (range: 4–19 min, SD: 4.0 min). Ultrafast T2-weighted and steady-state free-precession sequences accounted for 62.3% of actual scan time, and were distributed over the whole duration of the study. Conclusion: Actual imaging time only accounts for 33% of total study time and is not continuous. The remaining time is consumed by the preparation phases of the scanner, and is spent with planning sequences and the eventual repositioning of the coil and/or pregnant woman. These data may help to more accurately estimate the exposure to radiofrequency deposition and noise during fetal MRI studies.

  4. Gemcitabine upregulates ABCG2/BCRP and modulates the intracellular pharmacokinetic profiles of bioluminescence in pancreatic cancer cells.

    Sun, Yue; Gu, Mancang; Zhu, Lixin; Liu, Junying; Xiong, Yang; Wei, Yinghui; Li, Fanzhu

    2016-03-01

    A lack of methods capable of exploring real-time intracellular drug deposition has since limited the investigation of gemcitabine-induced multidrug resistance in vitro and in vivo. Specifically, resistance induced by D-luciferin, a substrate of the breast cancer resistance protein (ABCG2/BCRP), has recently attracted clinical attention, but further investigation has been limited. Herein, the intracellular pharmacokinetic behavior of D-luciferin was investigated in pancreatic cancer cell lines in real time by using bioluminescence imaging. To achieve this feat, BxPC3 and Panc1 pancreatic cancer cells overexpressing firefly luciferase were treated with gemcitabine in a dose and time gradient manner in vitro. The intracellular pharmacokinetic profiles of each group were then determined through the acquisition of bioluminescent signal intensity of D-luciferin in cells. Simultaneously, key pharmacokinetic parameters including area under the curve, elimination rate constant (K), and mean resident time were calculated according to the noncompartment model. ABCG2 protein levels following gemcitabine treatment were detected through western blot, and gemcitabine showed no significant effect on luciferase activity over dimethyl sulfoxide (DMSO) as a control (P>0.05). However, gemcitabine significantly increased K values while suppressing area under the curve and mean resident time compared with DMSO (Pbioluminescent model and its capability to observe the onset of chemoresistance in real time. PMID:26556627

  5. Analytical Applications of Bioluminescence and Chemiluminescence

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

    1975-01-01

    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.

  6. Circadian Control Sheds Light on Fungal Bioluminescence

    Oliveira, Anderson G.; Cassius V. Stevani; Waldenmaier, Hans E.; Viviani, Vadim; Emerson, Jillian M.; Loros, Jennifer J.; Jay C Dunlap

    2015-01-01

    Bioluminescence, the creation and emission of light by organisms, affords insight into the lives of organisms doing it. Luminous living things are widespread and access diverse mechanisms to generate and control luminescence [1-5]. Among the least studied bioluminescent organisms are phylogenetically rare fungi – only 71 species, all within the ~9000 fungi of the temperate and tropical Agaricales Order - are reported from among ~100,000 described fungal species [6,7]. All require oxygen [8] a...

  7. Circadian control sheds light on fungal bioluminescence.

    Oliveira, Anderson G; Stevani, Cassius V; Waldenmaier, Hans E; Viviani, Vadim; Emerson, Jillian M; Loros, Jennifer J; Dunlap, Jay C

    2015-03-30

    Bioluminescence, the creation and emission of light by organisms, affords insight into the lives of organisms doing it. Luminous living things are widespread and access diverse mechanisms to generate and control luminescence [1-5]. Among the least studied bioluminescent organisms are phylogenetically rare fungi-only 71 species, all within the ∼ 9,000 fungi of the temperate and tropical Agaricales order-are reported from among ∼ 100,000 described fungal species [6, 7]. All require oxygen [8] and energy (NADH or NADPH) for bioluminescence and are reported to emit green light (λmax 530 nm) continuously, implying a metabolic function for bioluminescence, perhaps as a byproduct of oxidative metabolism in lignin degradation. Here, however, we report that bioluminescence from the mycelium of Neonothopanus gardneri is controlled by a temperature-compensated circadian clock, the result of cycles in content/activity of the luciferase, reductase, and luciferin that comprise the luminescent system. Because regulation implies an adaptive function for bioluminescence, a controversial question for more than two millennia [8-15], we examined interactions between luminescent fungi and insects [16]. Prosthetic acrylic resin "mushrooms," internally illuminated by a green LED emitting light similar to the bioluminescence, attract staphilinid rove beetles (coleopterans), as well as hemipterans (true bugs), dipterans (flies), and hymenopterans (wasps and ants), at numbers far greater than dark control traps. Thus, circadian control may optimize energy use for when bioluminescence is most visible, attracting insects that can in turn help in spore dispersal, thereby benefitting fungi growing under the forest canopy, where wind flow is greatly reduced. PMID:25802150

  8. Real-time image and video processing

    Kehtarnavaz, Nasser

    2006-01-01

    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

  9. An adaptive regularization parameter choice strategy for multispectral bioluminescence tomography

    Purpose: Bioluminescence tomography (BLT) provides an effective tool for monitoring physiological and pathological activities in vivo. However, the measured data in bioluminescence imaging are corrupted by noise. Therefore, regularization methods are commonly used to find a regularized solution. Nevertheless, for the quality of the reconstructed bioluminescent source obtained by regularization methods, the choice of the regularization parameters is crucial. To date, the selection of regularization parameters remains challenging. With regards to the above problems, the authors proposed a BLT reconstruction algorithm with an adaptive parameter choice rule. Methods: The proposed reconstruction algorithm uses a diffusion equation for modeling the bioluminescent photon transport. The diffusion equation is solved with a finite element method. Computed tomography (CT) images provide anatomical information regarding the geometry of the small animal and its internal organs. To reduce the ill-posedness of BLT, spectral information and the optimal permissible source region are employed. Then, the relationship between the unknown source distribution and multiview and multispectral boundary measurements is established based on the finite element method and the optimal permissible source region. Since the measured data are noisy, the BLT reconstruction is formulated as l2 data fidelity and a general regularization term. When choosing the regularization parameters for BLT, an efficient model function approach is proposed, which does not require knowledge of the noise level. This approach only requests the computation of the residual and regularized solution norm. With this knowledge, we construct the model function to approximate the objective function, and the regularization parameter is updated iteratively. Results: First, the micro-CT based mouse phantom was used for simulation verification. Simulation experiments were used to illustrate why multispectral data were used rather

  10. An adaptive regularization parameter choice strategy for multispectral bioluminescence tomography

    Feng Jinchao; Qin Chenghu; Jia Kebin; Han Dong; Liu Kai; Zhu Shouping; Yang Xin; Tian Jie [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, P. O. Box 2728, Beijing 100190 (China); College of Electronic Information and Control Engineering, Beijing University of Technology, Beijing 100124 (China); Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, P. O. Box 2728, Beijing 100190 (China); Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, P. O. Box 2728, Beijing 100190 (China) and School of Life Sciences and Technology, Xidian University, Xi' an 710071 (China)

    2011-11-15

    Purpose: Bioluminescence tomography (BLT) provides an effective tool for monitoring physiological and pathological activities in vivo. However, the measured data in bioluminescence imaging are corrupted by noise. Therefore, regularization methods are commonly used to find a regularized solution. Nevertheless, for the quality of the reconstructed bioluminescent source obtained by regularization methods, the choice of the regularization parameters is crucial. To date, the selection of regularization parameters remains challenging. With regards to the above problems, the authors proposed a BLT reconstruction algorithm with an adaptive parameter choice rule. Methods: The proposed reconstruction algorithm uses a diffusion equation for modeling the bioluminescent photon transport. The diffusion equation is solved with a finite element method. Computed tomography (CT) images provide anatomical information regarding the geometry of the small animal and its internal organs. To reduce the ill-posedness of BLT, spectral information and the optimal permissible source region are employed. Then, the relationship between the unknown source distribution and multiview and multispectral boundary measurements is established based on the finite element method and the optimal permissible source region. Since the measured data are noisy, the BLT reconstruction is formulated as l{sub 2} data fidelity and a general regularization term. When choosing the regularization parameters for BLT, an efficient model function approach is proposed, which does not require knowledge of the noise level. This approach only requests the computation of the residual and regularized solution norm. With this knowledge, we construct the model function to approximate the objective function, and the regularization parameter is updated iteratively. Results: First, the micro-CT based mouse phantom was used for simulation verification. Simulation experiments were used to illustrate why multispectral data were used

  11. Arterially transplanted mesenchymal stem cells in a mouse reversible unilateral ureteral obstruction model: in vivo bioluminescence imaging and effects on renal fibrosis

    BAI Zhi-ming; DENG Xiang-dong; LI Jin-dong; LI Dong-hui; CAO Hui; LIU Zhen-xiang; ZHANG Jie

    2013-01-01

    Background Chronic kidney disease (CDK) is a worldwide health problem,but there is currently no effective treatment that can completely cure this disease.Recently,studies with mesenchymal stem cells (MSCs) on treating various renal diseases have shown breakthroughs.This study is to observe the homing features of MSCs transplanted via kidney artery and effects on renal fibrosis in a reversible unilateral ureteral obstruction (R-UUO) model.Methods Thirty-six Balb/c mice were divided into UUO group,UUO-MSC group,and sham group randomly,with 12 mice in each group.The MSCs had been infected by a lentiviral vector to express stably the luciferase reporter gene and green fluorescence protein genes (Luc-GFP-MSC).Homing of MSCs was tracked using in vivo imaging system (IVIS) 1,3,14,and 28 days after transplantation.Imaging results were verified by detecting GFP expression in frozen section under a fluorescence microscope.E-cadherin,α-SMA,TGF-β1,and TNF-α mRNA expression in all groups at 1 and 4 weeks after transplantation were analyzed by quantitative PCR.Results Transplanted Luc-GFP-MSCs showed increased Luciferase expression 3 days after transplantation.The expression decreased from 7 days,weakened thereafter and could not be detected 14 days after transplantation.Quantitative PCR results showed that all gene expressions in UUO group and UUO-MSC group at 1 week had no statistical difference,while at 4 weeks,except TGF-β expression (P>0.05),the expression of E-cadherin,α-SMA,and TNF-α in the above two groups have statistical difference (P<0.01).Conclusion IVIS enables fast,noninvasive,and intuitive tracking of MSC homing in vivo.MSCs can be taken home to kidney tissues of the diseased side in R-UUO model,and renal interstitial fibrosis can be improved as well.

  12. Compact snapshot real-time imaging spectrometer

    Kudenov, Michael W.; Dereniak, Eustace L.

    2011-11-01

    The described spectral imaging system, referred to as a Snapshot Hyperspectral Imaging Fourier Transform (SHIFT) spectrometer, is capable of acquiring spectral image data of a scene in a single integration of a camera, is ultra-compact, inexpensive (commercial off-the-shelf), has no moving parts, and can produce datacubes (x, y, λ) in real time. Based on the multiple-image FTS originally developed by A. Hirai [1], the presented device offers significant advantages over his original implementation. Namely, its birefringent nature results in a common-path interferometer which makes the spectrometer insensitive to vibration. Furthermore, it enables the potential of making the instrument ultra-compact, thereby improving the portability of the sensor. By combining a birefringent interferometer with a lenslet array, the entire spectrometer consumes approximately 15×15×20 mm3, excluding the imaging camera. The theory of the birefringent FTS is provided, followed by details of its specific embodiment and a laboratory proof of concept of the sensor. Post-processing is currently accomplished in Matlab, but progress is underway in developing real-time reconstruction capabilities with software programmed on a graphics processing unit (GPU). It is anticipated that processing of >30 datacubes per second can be achieved with modest GPU hardware, with spatial/spectral data of or exceeding 256×256 spatial resolution elements and 60 spectral bands over the visible (400-800 nm) spectrum. Data were collected outdoors, demonstrating the sensor's ability to resolve spectral signatures in standard outdoor lighting and environmental conditions as well as retinal imaging.

  13. Non-invasive bioluminescence imaging to monitor the immunological control of a plasmablastic lymphoma-like B cell neoplasia after hematopoietic cell transplantation.

    Martin Chopra

    Full Text Available To promote cancer research and to develop innovative therapies, refined pre-clinical mouse tumor models that mimic the actual disease in humans are of dire need. A number of neoplasms along the B cell lineage are commonly initiated by a translocation recombining c-myc with the immunoglobulin heavy-chain gene locus. The translocation is modeled in the C.129S1-Igha(tm1(MycJanz/J mouse which has been previously engineered to express c-myc under the control of the endogenous IgH promoter. This transgenic mouse exhibits B cell hyperplasia and develops diverse B cell tumors. We have isolated tumor cells from the spleen of a C.129S1-Igha(tm1(MycJanz/J mouse that spontaneously developed a plasmablastic lymphoma-like disease. These cells were cultured, transduced to express eGFP and firefly luciferase, and gave rise to a highly aggressive, transplantable B cell lymphoma cell line, termed IM380. This model bears several advantages over other models as it is genetically induced and mimics the translocation that is detectable in a number of human B cell lymphomas. The growth of the tumor cells, their dissemination, and response to treatment within immunocompetent hosts can be imaged non-invasively in vivo due to their expression of firefly luciferase. IM380 cells are radioresistant in vivo and mice with established tumors can be allogeneically transplanted to analyze graft-versus-tumor effects of transplanted T cells. Allogeneic hematopoietic stem cell transplantation of tumor-bearing mice results in prolonged survival. These traits make the IM380 model very valuable for the study of B cell lymphoma pathophysiology and for the development of innovative cancer therapies.

  14. Design and implementation of an optical simulation environment for bioluminescent tomography studies

    LI Hui; TIAN Jie; LUO Jie; L(U) Yujie; CONG Wenxiang; WANG Ge

    2007-01-01

    As a challenging task for bioluminescent tomography simulation, a virtual optical environment is needed to solve the forward problem accurately, that is, to achieve a high precision for bioluminescent signal synthesis on the external body surface of a small animal. The molecular optical simulation environment named MOSE is implemented using the C + + programming language and the OpenGL techniques, including a user-friendly interface with interactive tools facilitating users' operations. The accuracy of the virtual optical environment is verified by error analysis of mesh simplification and comparison between MOSE results and experimental data. This virtual optical environment is accurate, flexible and efficient to simulate the photon propagation in complicated tissues, which has a great potential to become a software platform for bioluminescent tomography studies and other molecular imaging applications.

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

    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. (topical review)

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

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

    2014-01-01

    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.

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

    Zhun Xu; Xiaolei Song; Xiaomeng Zhang; Jing Bai

    2007-01-01

    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 ...

  18. Fluorescent and Bioluminescent Reporter Myxoviruses.

    Rostad, Christina A; Currier, Michael C; Moore, Martin L

    2016-01-01

    The advent of virus reverse genetics has enabled the incorporation of genetically encoded reporter proteins into replication-competent viruses. These reporters include fluorescent proteins which have intrinsic chromophores that absorb light and re-emit it at lower wavelengths, and bioluminescent proteins which are luciferase enzymes that react with substrates to produce visible light. The incorporation of these reporters into replication-competent viruses has revolutionized our understanding of molecular virology and aspects of viral tropism and transmission. Reporter viruses have also enabled the development of high-throughput assays to screen antiviral compounds and antibodies and to perform neutralization assays. However, there remain technical challenges with the design of replication-competent reporter viruses, and each reporter has unique advantages and disadvantages for specific applications. This review describes currently available reporters, design strategies for incorporating reporters into replication-competent paramyxoviruses and orthomyxoviruses, and the variety of applications for which these tools can be utilized both in vitro and in vivo. PMID:27527209

  19. Fluorescent and Bioluminescent Reporter Myxoviruses

    Rostad, Christina A.; Currier, Michael C.; Moore, Martin L.

    2016-01-01

    The advent of virus reverse genetics has enabled the incorporation of genetically encoded reporter proteins into replication-competent viruses. These reporters include fluorescent proteins which have intrinsic chromophores that absorb light and re-emit it at lower wavelengths, and bioluminescent proteins which are luciferase enzymes that react with substrates to produce visible light. The incorporation of these reporters into replication-competent viruses has revolutionized our understanding of molecular virology and aspects of viral tropism and transmission. Reporter viruses have also enabled the development of high-throughput assays to screen antiviral compounds and antibodies and to perform neutralization assays. However, there remain technical challenges with the design of replication-competent reporter viruses, and each reporter has unique advantages and disadvantages for specific applications. This review describes currently available reporters, design strategies for incorporating reporters into replication-competent paramyxoviruses and orthomyxoviruses, and the variety of applications for which these tools can be utilized both in vitro and in vivo. PMID:27527209

  20. Luciferase bioluminescence imaging monitoring gene therapeutic effect of apoptosis-inducing ligand for lung cancer A549 cells nude mice transplantation tumor in vivo

    Objective: To detect the expression and effect of human tumor necrosis factor related apoptosis-inducing ligand (hTRAIL) in vivo,by using a novel double expressing adenoviral vector encoding hTRAIL and firefly luciferase (luc) gene (ad-luc-hTRAIL), in which luc was used as reporter gene. Methods: Lung cancer A549 cell xenografts in 16 nude mice models were established in subcutaneous inoculation way, the adenovirus vectors (ad-luc-hTRAIL, ad-hTRAIL, ad-luc) and phosphate buffer saline (PBS) (n=4) as control were injected into tumor respectively. The size of the tumor was measured at different time points (4, 7, 10, 14, 21, 28 d) after injection. The activity of luciferase in surface of the tumor was detected in vivo by using high-sensitivity cooled-charged coupled device (CCD) camera. The expression of hTRAIL was demonstrated by immunohistochemistry staining after sacrificing the animals at different time points, and immunohistochemical scores (IHS) were measured. The apoptosis rate of tumor cells was detected by using TUNEL and calculated. Analysis of variance, the paired t test and linear correlation analysis was used for the statistics. Results: The growing speed of tumour xenografts was more slowly in ad-luc-hTRAIL and ad-hTRAIL groups than PBS group (t=2.71, 2.72, P<0.05). The tumor volumes of ad-luc-hTRAIL, ad-hTRAIL, ad-luc and PBS groups 28 days after injection were (208.4 ± 42.3), (181.5 ±23.9), (403.1 ± 54.0) and (427.0 ± 59.3) mm3, respectively. There was no significant difference between ad-luc group and PBS group (t=2.07, P>0.05). The expression of luciferase in ad-luc-hTRAIL group reached its peak at 7th day (1.37 ± 1.04), and then decreased quickly. The IHS and apoptosis rate in ad-luc-hTRAIL and ad-hTRAIL groups reached their peaks at 7th day, the peak values of IHS were 6.25 ±2.06 and 6.5 ± 2.89, the peak values of apoptosis rate were (60.75 ± 8.06)% and (61.50 ± 8.47)%,respectively. The amount of luciferase expression (absolute number of

  1. Bacteria bioluminescent activity as an indicator of geomagnetic disturbances

    The effect of geomagnetic disturbances and storms on bioluminescence activity of bacterium were investigated. The bioluminescence intensity change depended on amplitude and continuous of geomagnetic storms. It is assumed, that the synchronization of luminous radiation take place in cellos when frequency of geomagnetic disturbances approached to an intrinsic one of a bioluminescence system. High sensitivity of bioluminescence of geomagnetic storms was detected. 5 refs., 4 figs

  2. Generation of a new bioluminescent model for visualisation of mammary tumour development in transgenic mice

    Zagozdzon, Agnieszka M

    2012-05-30

    AbstractBackgroundNumerous transgenic models have been generated to study breast cancer. However, despite many advantages, traditional transgenic models for breast cancer are also burdened with difficulties in early detection and longitudinal observation of transgene-induced tumours, which in most cases are randomly located and occur at various time points. Methods such as palpation followed by mechanical measurement of the tumours are of limited value in transgenic models. There is a crucial need for making these previously generated models suitable for modern methods of tumour visualisation and monitoring, e.g. by bioluminescence-based techniques. This approach was successfully used in the current study.ResultsA new mouse strain (MMTV-Luc2 mice) expressing Luc2 luciferase primarily in mammary tissue in females, with low-level background expression in internal organs, was generated and bred to homozygosity. After these mice were intercrossed with MMTV-PyVT mice, all double transgenic females developed mammary tumours by the age of 10 weeks, the localisation and progression of which could be effectively monitored using the luminescence-based in vivo imaging. Luminescence-based readout allowed for early visualisation of the locally overgrown mammary tissue and for longitudinal evaluation of local progression of the tumours. When sampled ex vivo at the age of 10 weeks, all tumours derived from MMTV-Luc2PyVT females displayed robust bioluminescent signal.ConclusionsWe have created a novel transgenic strain for visualisation and longitudinal monitoring of mammary tumour development in transgenic mice as an addition and\\/or a new and more advanced alternative to manual methods. Generation of this mouse strain is vital for making many of the existing mammary tumour transgenic models applicable for in vivo imaging techniques.

  3. Generation of a new bioluminescent model for visualisation of mammary tumour development in transgenic mice

    Numerous transgenic models have been generated to study breast cancer. However, despite many advantages, traditional transgenic models for breast cancer are also burdened with difficulties in early detection and longitudinal observation of transgene-induced tumours, which in most cases are randomly located and occur at various time points. Methods such as palpation followed by mechanical measurement of the tumours are of limited value in transgenic models. There is a crucial need for making these previously generated models suitable for modern methods of tumour visualisation and monitoring, e.g. by bioluminescence-based techniques. This approach was successfully used in the current study. A new mouse strain (MMTV-Luc2 mice) expressing Luc2 luciferase primarily in mammary tissue in females, with low-level background expression in internal organs, was generated and bred to homozygosity. After these mice were intercrossed with MMTV-PyVT mice, all double transgenic females developed mammary tumours by the age of 10 weeks, the localisation and progression of which could be effectively monitored using the luminescence-based in vivo imaging. Luminescence-based readout allowed for early visualisation of the locally overgrown mammary tissue and for longitudinal evaluation of local progression of the tumours. When sampled ex vivo at the age of 10 weeks, all tumours derived from MMTV-Luc2PyVT females displayed robust bioluminescent signal. We have created a novel transgenic strain for visualisation and longitudinal monitoring of mammary tumour development in transgenic mice as an addition and/or a new and more advanced alternative to manual methods. Generation of this mouse strain is vital for making many of the existing mammary tumour transgenic models applicable for in vivo imaging techniques

  4. Generation of a new bioluminescent model for visualisation of mammary tumour development in transgenic mice

    Zagozdzon Agnieszka M

    2012-05-01

    Full Text Available Abstract Background Numerous transgenic models have been generated to study breast cancer. However, despite many advantages, traditional transgenic models for breast cancer are also burdened with difficulties in early detection and longitudinal observation of transgene-induced tumours, which in most cases are randomly located and occur at various time points. Methods such as palpation followed by mechanical measurement of the tumours are of limited value in transgenic models. There is a crucial need for making these previously generated models suitable for modern methods of tumour visualisation and monitoring, e.g. by bioluminescence-based techniques. This approach was successfully used in the current study. Results A new mouse strain (MMTV-Luc2 mice expressing Luc2 luciferase primarily in mammary tissue in females, with low-level background expression in internal organs, was generated and bred to homozygosity. After these mice were intercrossed with MMTV-PyVT mice, all double transgenic females developed mammary tumours by the age of 10 weeks, the localisation and progression of which could be effectively monitored using the luminescence-based in vivo imaging. Luminescence-based readout allowed for early visualisation of the locally overgrown mammary tissue and for longitudinal evaluation of local progression of the tumours. When sampled ex vivo at the age of 10 weeks, all tumours derived from MMTV-Luc2PyVT females displayed robust bioluminescent signal. Conclusions We have created a novel transgenic strain for visualisation and longitudinal monitoring of mammary tumour development in transgenic mice as an addition and/or a new and more advanced alternative to manual methods. Generation of this mouse strain is vital for making many of the existing mammary tumour transgenic models applicable for in vivo imaging techniques.

  5. Fluorescence and bioluminescence of bacterial luciferase intermediates

    An intermediate in the luciferase-catalyzed bioluminescent oxidation of FMNH2, isolated and purified by chromatography at --200, was postulated to be an oxygenated reduced flavine-luciferase. Maintained and studied at --20 to --300, this material exhibits a relatively weak fluorescence emission peaking at about 505 nm when excited at 370 nm. It may comprise more than one species. Upon continued exposure to light at 370 nm, the intensity of this fluorescence increases, often by a factor of 5 or more, and its emission spectrum is blue shifted to a maximum at about 485 nm. Upon warming this fluorescence is lost and the fluorescence of flavine mononucleotide appears. If warming is carried out in the presence of a long chain aldehyde, bioluminescence occurs, with the appearance of a similar amount of flavine fluorescence. The bioluminescence yield is about the same with irradiated and nonirradiated samples. The bioluminescence emission spectrum corresponds exactly to the fluorescence emission spectrum of the intermediate formed by irradiation, implicating the latter as being structurally close to the emitting species in bioluminescence. (auth)

  6. Papyrus imaging with terahertz time domain spectroscopy

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

    2010-09-01

    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.

  7. Real-Time Imaging of Quantum Entanglement

    Full text: Photonic entanglement of spatial modes is routinely studied in many experiments and offers interesting features for quantum optical and quantum information experiments. To investigate the properties of these complex modes, it is crucial to gain information about the transversal structure with high precision and in an efficient way. We show that modern technology, namely triggered intensified charge coupled device (ICCD) cameras are fast and sensitive enough to image in real-time the effect of the measurement of one photon on the spatial mode of its entangled partner photon. We determine from imaged intensity pattern the number of photons within a certain region, evaluate its error margin and thereby quantitatively verify the non-classicality of the measurements. In addition, the use of the ICCD camera allows us to demonstrate visually the enhanced remote angular sensing and the high flexibility of our setup in creating any desired spatial-mode entanglement. (author)

  8. Discovery of New Substrates for LuxAB Bacterial Bioluminescence.

    Jiang, Tianyu; Wang, Weishan; Wu, Xingkang; Wu, Wenxiao; Bai, Haixiu; Ma, Zhao; Shen, Yuemao; Yang, Keqian; Li, Minyong

    2016-08-01

    In this article, four novel substrates with long halftime have been designed and synthesized successfully for luxAB bacterial bioluminescence. After in vitro and in vivo biological evaluation, these molecules can emit obvious bioluminescence emission with known bacterial luciferase, thus indicating a new promising approach to developing the bacterial bioluminescent system. PMID:26896339

  9. Chemistry and biology of insect bioluminescence

    Colepicolo Neto, P.; Bechara, E.J.H. (Sao Paulo Univ. (Brazil). Inst. de Quimica)

    1984-12-01

    Basic aspects on the Chemistry and Biology of bioluminescence are reviewed, with emphasis on insects. Data from the investigation of Lampyridae (fireflies) are collected from literature. With regard to Elateridae (click beetles) and Phengodidae (rail road worms), the least explored families of luminescent insects, new data are presented on the following aspects: (i) 'in vivo' emission spectra, (ii) chemical nature of the luciferin, (iii) conection between bioluminescence and 'oxygen toxicity' as a result of molecular oxygen storage and (iv) the role of light emission by larvae and pupae.

  10. Chemistry and biology of insect bioluminescence

    Basic aspects on the Chemistry and Biology of bioluminescence are reviewed, with emphasis on insects. Data from the investigation of Lampyridae (fireflies) are collected from literature. With regard to Elateridae (click beetles) and Phengodidae (rail road worms), the least explored families of luminescent insects, new data are presented on the following aspects: (i) 'in vivo' emission spectra, (ii) chemical nature of the luciferin, (iii) conection between bioluminescence and 'oxygen toxicity' as a result of molecular oxygen storage and (iv) the role of light emission by larvae and pupae. (Author)

  11. A two-hour antibiotic susceptibility test by ATP-bioluminescence.

    March Rosselló, Gabriel Alberto; García-Loygorri Jordán de Urries, María Cristina; Gutiérrez Rodríguez, María Purificación; Simarro Grande, María; Orduña Domingo, Antonio; Bratos Pérez, Miguel Ángel

    2016-01-01

    The antibiotic susceptibility test (AST) in Clinical Microbiology laboratories is still time-consuming, and most procedures take 24h to yield results. In this study, a rapid antimicrobial susceptibility test using ATP-bioluminescence has been developed. The design of method was performed using five ATCC collection strains of known susceptibility. This procedure was then validated against standard commercial methods on 10 strains of enterococci, 10 staphylococci, 10 non-fermenting gram negative bacilli, and 13 Enterobacteriaceae from patients. The agreement obtained in the sensitivity between the ATP-bioluminescence method and commercial methods (E-test, MicroScan and VITEK2) was 100%. In summary, the preliminary results obtained in this work show that the ATP-bioluminescence method could provide a fast and reliable AST in two hours. PMID:25979598

  12. Quantification of adipose transfer viability using a novel, bioluminescent murine model.

    Gassman, Andrew A; Kao, Kenneth K; Bradley, James P; Lee, Justine C

    2016-07-01

    Fat grafting has highly variable long-term results. Research efforts to improve the reliability of fat grafting are limited by inefficient methods for evaluation of fat engraftment. In this work, we describe a novel animal model for the quantitative evaluation of fat grafting using in vivo bioluminescence of adipocytes from luciferase-expressing mice. Subcutaneous adipose tissue from GFP and luciferase-expressing FVB mice were obtained. The samples were homogenized, decanted, and injected into the dorsal skin folds of wild-type FVB mice. Viability of the transferred tissue was examined over a 28-day time period with quantitative bioluminescence after luciferin injection. All animals demonstrated viable adipose transfer with bioluminescence detectable on days 0, 1, 7, 14, and 28. This animal model may be used for noninvasive, longitudinal studies for quantification of the fat engraftment process. PMID:27017232

  13. The First Bioluminescence Tomography System for Simultaneous Acquisition of Multiview and Multispectral Data

    Ge Wang

    2006-01-01

    Full Text Available We describe the system design of the first bioluminescence tomography (BLT system for parallel acquisition of multiple bioluminescent views around a mouse in a number of spectral channels simultaneously. The primary component of this BLT system is a novel mirror module and a unique mouse holder. The mirror module consists of a mounting plate and four mirrors with stages. These mirror stages are right triangular blocks symmetrically arranged and attached to the mounting plate such that the hypotenuse surfaces of the triangular blocks all make 45∘ to the plate surface. The cylindrical/polygonal mouse holder has semitransparent rainbow bands on its side surface for the acquisition of spectrally resolved data. Numerical studies and experiments are performed to demonstrate the feasibility of this system. It is shown that bioluminescent signals collected using our system can produce a similar BLT reconstruction quality while reducing the data acquisition time, as compared to the sequential data acquisition mode.

  14. Time-Lapse Imaging of Cell Death.

    Wallberg, Fredrik; Tenev, Tencho; Meier, Pascal

    2016-03-01

    The best approach to distinguish between necrosis and apoptosis is time-lapse video microscopy. This technique enables a biological process to be photographed at regular intervals over a period, which may last from a few hours to several days, and can be applied to cells in culture or in vivo. We have established two time-lapse microscopy methods based on different ways of calculating cell death: semiautomated and automated. In the semiautomated approach, cell death can be visualized by staining with combinations of Alexa Fluor 647-conjugated Annexin V and Sytox Green (SG), or Annexin V(FITC) and Propidium iodide (PI). The automated method is similar except that all cells are labeled with dyes. This allows faster quantification of data. To this end Cell Tracker Green is used to label all cells at time zero in combination with PI and Alexa Fluor 647-conjugated Annexin V. Necrotic cell death is accompanied by either simultaneous labeling with Annexin V and PI or SG (double-positive), or direct PI or SG staining. Additionally, necrotic cells display characteristic morphology, such as cytoplasmic swelling. In contrast to necrosis where membrane permeabilization is an early event, cells that die by apoptosis lose their membrane permeability relatively late. Therefore, the time between Annexin V staining and PI or SG uptake (double-positive) can be used to distinguish necrosis from apoptosis. This protocol describes the analysis of cell death by time-lapse imaging of HT1080 and L929 cells stained with these dyes, but it can be readily adapted to other cell types of interest. PMID:26933245

  15. Bioluminescent Bioreporters Encapsulated in Silica Gel

    Kuncová, Gabriela; Trögl, J.; Demnerová, K.; Ripp, S.; Sayler, G. S.

    -: -, 2008, O08-2 - 1-O08-2 - 4. [XVI International Conference on Bioencapsulation. Dublin (IE), 04.09.2008-06.09.2008] Institutional research plan: CEZ:AV0Z40720504 Keywords : bioluminescent bioreporter * silica gel * biosensor Subject RIV: CE - Biochemistry

  16. Bioluminescence: diplôme 2015

    Giachino, Vincent; Geiser, Martial

    2016-01-01

    Dans le cadre du projet européen BRAAVOO, qui vise à mesurer les concentrations de différents polluants présents dans la mer, nous développons un lecteur de bioluminescence embarqué dans une bouée.

  17. Bioluminescence for determining energy state of plants

    Ching, T. M.

    1975-01-01

    Bioluminescence produced by the luciferin-luciferase system is a very sensitive assay for ATP content in extracts of plant materials. The ATP test for seed and pollen viability and vigor is presented, along with prediction of high growth potential and productivity in new crosses and selections of breeding materials. ATP as an indicator for environmental quality, stresses, and metabolic regulation is also considered.

  18. In vivo bioluminescence tomography based on multi-view projection and 3D surface reconstruction

    Zhang, Shuang; Wang, Kun; Leng, Chengcai; Deng, Kexin; Hu, Yifang; Tian, Jie

    2015-03-01

    Bioluminescence tomography (BLT) is a powerful optical molecular imaging modality, which enables non-invasive realtime in vivo imaging as well as 3D quantitative analysis in preclinical studies. In order to solve the inverse problem and reconstruct inner light sources accurately, the prior structural information is commonly necessary and obtained from computed tomography or magnetic resonance imaging. This strategy requires expensive hybrid imaging system, complicated operation protocol and possible involvement of ionizing radiation. The overall robustness highly depends on the fusion accuracy between the optical and structural information. In this study we present a pure optical bioluminescence tomographic system (POBTS) and a novel BLT method based on multi-view projection acquisition and 3D surface reconstruction. The POBTS acquired a sparse set of white light surface images and bioluminescent images of a mouse. Then the white light images were applied to an approximate surface model to generate a high quality textured 3D surface reconstruction of the mouse. After that we integrated multi-view luminescent images based on the previous reconstruction, and applied an algorithm to calibrate and quantify the surface luminescent flux in 3D.Finally, the internal bioluminescence source reconstruction was achieved with this prior information. A BALB/C mouse with breast tumor of 4T1-fLuc cells mouse model were used to evaluate the performance of the new system and technique. Compared with the conventional hybrid optical-CT approach using the same inverse reconstruction method, the reconstruction accuracy of this technique was improved. The distance error between the actual and reconstructed internal source was decreased by 0.184 mm.

  19. Enhanced Landweber algorithm via Bregman iterations for bioluminescence tomography

    Xia, Yi; Zhang, Meng

    2014-09-01

    Bioluminescence tomography (BLT) is an important optical molecular imaging modality aimed at visualizing physiological and pathological processes at cellular and molecular levels. While the forward process of light propagation is described by the diffusion approximation to radiative transfer equation, BLT is the inverse problem to reconstruct the 3D localization and quantification of internal bioluminescent sources distribution. Due to the inherent ill-posedness of the BLT problem, regularization is generally indispensable to obtain more favorable reconstruction. In particular, total variation (TV) regularization is known to be effective for piecewise-constant source distribution which can permit sharp discontinuities and preserve edges. However, total variation regularization generally suffers from the unsatisfactory staircasing effect. In this work, we introduce the Bregman iterative regularization to alleviate this degeneration and enhance the numerical reconstruction of BLT. Based on the existing Landweber method (LM), we put forward the Bregman-LM-TV algorithm for BLT. Numerical experiments are carried out and preliminary simulation results are reported to evaluate the proposed algorithms. It is found that Bregman-LM-TV can significantly outperform the individual Landweber method for BLT when the source distribution is piecewise-constant.

  20. Modeling and measurement of a whole-cell bioluminescent biosensor based on a single photon avalanche diode.

    Daniel, Ramiz; Almog, Ronen; Ron, Amit; Belkin, Shimshon; Diamand, Yosi Shacahm

    2008-12-01

    Whole-cell biosensors are potential candidates for on-line and in situ environmental monitoring. In this work we present a new design of a whole-cell bioluminescence biosensor for water toxicity detection, based on genetically engineered Escherichia coli bacteria, carrying a recA::luxCDABE promoter-reporter fusion. Sensitive optical detection is achieved using a single photon avalanche photodiode (SPAD) working in the Geiger mode. The present work describes a simple mathematical model for the kinetic process of the bioluminescence based SOS toxin response of E. coli bacteria. We find that initially the bioluminescence signal depends on the time square and we show that the spectral intensity of the bioluminescence signal is inverse proportional to the frequency. We get excellent agreement between the theoretical model and the measured light signal. Furthermore, we present experimental results of the bioluminescent signal measurement using a SPAD and a photomultiplier, and demonstrate improvement of the measurement by applying a matched digital filter. Low intensity bioluminescence signals were measured after the whole-cell sensors were exposed to various toxicant concentrations (5, 15 and 20ppm). PMID:18774705

  1. Real-time adaptive video image enhancement

    Garside, John R.; Harrison, Chris G.

    1999-07-01

    As part of a continuing collaboration between the University of Manchester and British Aerospace, a signal processing array has been constructed to demonstrate that it is feasible to compensate a video signal for the degradation caused by atmospheric haze in real-time. Previously reported work has shown good agreement between a simple physical model of light scattering by atmospheric haze and the observed loss of contrast. This model predicts a characteristic relationship between contrast loss in the image and the range from the camera to the scene. For an airborne camera, the slant-range to a point on the ground may be estimated from the airplane's pose, as reported by the inertial navigation system, and the contrast may be obtained from the camera's output. Fusing data from these two streams provides a means of estimating model parameters such as the visibility and the overall illumination of the scene. This knowledge allows the same model to be applied in reverse, thus restoring the contrast lost to atmospheric haze. An efficient approximation of range is vital for a real-time implementation of the method. Preliminary results show that an adaptive approach to fitting the model's parameters, exploiting the temporal correlation between video frames, leads to a robust implementation with a significantly accelerated throughput.

  2. Real-time Embedded Panoramic Imaging for Spherical Camera System

    Uddin-Al-Hasan, Main

    2013-01-01

    Panoramas or stitched images are used in topographical mapping, panoramic 3D reconstruction, deep space exploration image processing, medical image processing, multimedia broadcasting, system automation, photography and other numerous fields. Generating real-time panoramic images in small embedded computer is of particular importance being lighter, smaller and mobile imaging system. Moreover, this type of lightweight panoramic imaging system is used for different types of industrial or home i...

  3. Using Luciferase to Image Bacterial Infections in Mice

    Chang, Mi Hee; Cirillo, Suat L. G.; Cirillo, Jeffrey D.

    2011-01-01

    Imaging is a valuable technique that can be used to monitor biological processes. In particular, the presence of cancer cells, stem cells, specific immune cell types, viral pathogens, parasites and bacteria can be followed in real-time within living animals 1-2. Application of bioluminescence imaging to the study of pathogens has advantages as compared to conventional strategies for analysis of infections in animal models3-4. Infections can be visualized within individual animals over time,...

  4. Reliability of a bioluminescence ATP assay for detection of bacteria.

    Selan, L.; Berlutti, F; Passariello, C.; Thaller, M C; Renzini, G

    1992-01-01

    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...

  5. Construction of a Bioluminescent Reporter Strain To Detect Polychlorinated Biphenyls

    Layton, A C; Muccini, M.; Ghosh, M. M.; Sayler, G. S.

    1998-01-01

    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/lit...

  6. Cloning and characterization of new bioluminescent proteins

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

    1999-07-01

    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.

  7. Transformation Experiment Using Bioluminescence Genes of "Vibrio fischeri."

    Slock, James

    1995-01-01

    Bioluminescence transformation experiments show students the excitement and power of recombinant DNA technology. This laboratory experiment utilizes two plasmids of "Vibrio fischeri" in a transformation experiment. (LZ)

  8. Time-gated holographic imaging using photorefractive materials

    Hyde, Sam C. W.; Barry, Nicholas P.; Jones, Richard; Dainty, Christopher; French, Paul M. W.; Wechsler, Barry A.; Klein, Marvin B.

    1995-05-01

    We present results for real-time holographic imaging in the medical imaging window (approximately 650 to 1000 nm) using photorefractive media. By recording holograms of a USAF test chart using a Ti:sapphire laser, we have selected the coherent)mostly ballistic) light and formed an image through a scattering medium which simulates biological tissue. The advantage of this approach is that a 2D image may be acquired in 'real time', i.e. without scanning. This will permit electronic signal processing and averaging, which may be used to reduce the deleterious effects of speckle. If the holograom is written with ultrashort pulses, then time-gated imaging is possible. This may be used to provide depth information by recording the 'time of flight' of the image light. We have recorded such holograms with picosecond pulses and present time-gated image data.

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

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

    2013-10-01

    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

  10. Establishment of a bioluminescence model for microenvironmentally induced oral carcinogenesis with implications for screening bioengineered scaffolds.

    Suliman, Salwa; Parajuli, Himalaya; Sun, Yang; Johannessen, Anne Christine; Finne-Wistrand, Anna; McCormack, Emmet; Mustafa, Kamal Babikeir Eln; Costea, Daniela Elena

    2015-01-01

    Background: Microenvironmental cues play a major role in head and neck cancer. Biodegradable scaffolds used for bone regeneration might also act as stimulative cues for head and neck cancer. The purpose of this study was to establish an experimental model for precise and noninvasive evaluation of tumorigenic potential of microenvironmental cues in head and neck cancer. Methods: Bioluminescence was chosen to image tumor formation. Early neoplastic oral keratinocyte (DOK) cells were lucifera...

  11. A causal relation between bioluminescence and oxygen to quantify the cell niche

    Lambrechts, Dennis; Roeffaers, Maarten; Goossens, Karel; Hofkens, Johan; Vande Velde, Greetje; Van de Putte, Tom; Schrooten, Jan; Van Oosterwyck, Hans

    2014-01-01

    Bioluminescence imaging assays have become a widely integrated technique to quantify effectiveness of cell-based therapies by monitoring fate and survival of transplanted cells. To date these assays are still largely qualitative and often erroneous due to the complexity and dynamics of local micro-environments (niches) in which the cells reside. Here, we report, using a combined experimental and computational approach, on oxygen that besides being a critical niche component responsible for ce...

  12. Dual monitoring using 124I-FIAU and bioluminescence for HSV1-tk suicide gene therapy

    Herpes simplex virus type I thymidine kinase (HSV-tk) is the most common reporter gene and is used in cancer gene therapy with a prodrug nucleoside analog, ganciclovir (GCV). The aim of this study is to evaluate therapeutic efficacy of suicide gene therapy with 2'-fluoro-2'-deoxy-1-D-arabinofuranosyl-5-[124I] iodouracil (124I - FIAU) and bioluminescence in retrovirally HSV -tk and firefly luciferase transduced hepatoma model. The HSV -tk and firefly luciferase (Luc) was retrovirally transduced and expressed in MCA rat Morris hepatoma cells. Nude mice with subcutaneous tumors, MCA and MCA-TK-Luc, were subjected to GCV treatment (50mg/Kg/d intraperitoneally) for 5 day. PET imaging and biodistribution with (124I-FIAU) were performed at before and after initiation of therapy with GCV. Bioluminescent signal was also measured during GCV treatment. Before GCV treatment, no significant difference in tumor volume was found in tumors between MCA and MCA-TK-Luc. After GCV treatment, tumor volume of MCA-TK-Luc markedly reduced compared to that of MCA. In biodistribution study, 124I-FIAU uptake after GCV therapy significantly decreased compared with pretreatment levels (34.8 13.67 %ID/g vs 7.6 2.59 %ID/g) and bioluminescent signal was also significantly decreased compared with pretreatment levels. In small animal PET imaging, 124I-FIAU selectively localized in HSV -tk expressing tumor and the therapeutic efficacy of GCV treatment was evaluated by 124I-FIAU PET imaging. 124I-FIAU PET and bioluminescence imaging in HSV-tk suicide gene therapy were effective to evaluate the therapeutic response. 124I-FIAU may serve as an efficient and selective agent for monitoring of transduced HSV1-tk gene expression in vivo in clinical trials

  13. Establishment of a bioluminescence model for microenvironmentally induced oral carcinogenesis with implications for screening bioengineered scaffolds

    Suliman, Salwa; Parajuli, Himalaya; Sun, Yang(Department of Physics, Shanghai Jiao Tong University, Shanghai 200240, China); Johannessen, Anne Christine; Finne-Wistrand, Anna; McCormack, Emmet; Mustafa, Kamal; Costea, Daniela Elena

    2016-01-01

    Background. Microenvironmental cues play a major role in head and neck cancer. Biodegradable scaffolds used for bone regeneration might also act as stimulative cues for head and neck cancer. The purpose of this study was to establish an experimental model for precise and noninvasive evaluation of tumorigenic potential of microenvironmental cues in head and neck cancer. Methods. Bioluminescence was chosen to image tumor formation. Early neoplastic oral keratinocyte (DOK) cells were luciferase-...

  14. Bioluminescence ATP Monitoring for the Routine Assessment of Food Contact Surface Cleanliness in a University Canteen

    Andrea Osimani

    2014-10-01

    Full Text Available ATP bioluminescence monitoring and traditional microbiological analyses (viable counting of total mesophilic aerobes, coliforms and Escherichia coli were used to evaluate the effectiveness of Sanitation Standard Operating Procedures (SSOP at a university canteen which uses a HACCP-based approach. To that end, 10 cleaning control points (CPs, including food contact surfaces at risk of contamination from product residues or microbial growth, were analysed during an 8-month monitoring period. Arbitrary acceptability limits were set for both microbial loads and ATP bioluminescence readings. A highly significant correlation (r = 0.99 between the means of ATP bioluminescence readings and the viable counts of total mesophilic aerobes was seen, thus revealing a strong association of these parameters with the level of surface contamination. Among CPs, the raw meat and multi-purpose chopping boards showed the highest criticalities. Although ATP bioluminescence technology cannot substitute traditional microbiological analyses for the determination of microbial load on food contact surfaces, it has proved to be a powerful tool for the real time monitoring of surface cleanliness at mass catering plants, for verify the correct application of SSOP, and hence for their implementation/revision in the case of poor hygiene.

  15. Noctiluca sp. bioluminescence in response to the mechanical stimuli of a screw propeller

    Han Jing

    2015-01-01

    Full Text Available This work presents a new experiment method studying the Noctiluca sp. bioluminescence under the mechanical stimulation. It devoted to the study of the Noctiluca sp. bioluminescence triggered by the screw propeller’s mechanical stimuli in the tank. The size of the tank was 2*1*1m. The screw propeller is fixed on a shelf and the position relative to the tank was adjustable by moving the shelf. Two methods were carried out to control the running of the screw propeller. In the first scenario, the shelf was fixed in the center of the tank and the second scenario, the shelf moved from one side to the other in the tank. At the same time, the screw propeller was running with a certain velocity. The luminescent strength of Noctiluca sp. enhanced as the increase of the screw propeller’s running velocity. There were two obvious luminous areas nearby the screw propeller’s blades. The luminescent area was bigger in the second scenario. Thus, when navigational ship passing the sea area which filled with Noctiluca sp. or other luminescent halobios, it will stimulate the Noctiluca sp. or other luminescent halobios bioluminescence. The ship also can be detected using the bioluminescence.

  16. Image-Based Learning Approach Applied to Time Series Forecasting

    J. C. Chimal-Eguía; K. Ramírez-Amáro

    2012-01-01

    In this paper, a new learning approach based on time-series image information is presented. In order to implementthis new learning technique, a novel time-series input data representation is also defined. This input data representation is based on information obtained by image axis division into boxes. The difference between this new input data representation and the classical is that this technique is not time-dependent. This new information is implemented in the new Image-Based Learning A...

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

    Shinozaki, Yohei; Harada, Yasuhiro

    2014-06-01

    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

  18. Automated analysis of protein subcellular location in time series images

    Hu, Yanhua; Osuna-Highley, Elvira; Hua, Juchang; Nowicki, Theodore Scott; Stolz, Robert; McKayle, Camille; Murphy, Robert F.

    2010-01-01

    Motivation: Image analysis, machine learning and statistical modeling have become well established for the automatic recognition and comparison of the subcellular locations of proteins in microscope images. By using a comprehensive set of features describing static images, major subcellular patterns can be distinguished with near perfect accuracy. We now extend this work to time series images, which contain both spatial and temporal information. The goal is to use temporal features to improve...

  19. Real-Time Augmented Reality for Image-Guided Interventions

    Vogt, Sebastian

    2010-01-01

    The aim of this thesis is to design, develop, and study a new approach to interventional image guidance based on augmented reality visualization – the merging of real and virtual images. This work broadly reviews and categorizes all preliminary augmented reality prototype systems for interventional image guidance. A novel augmented reality setup is introduced and engineered for interventional guidance tasks, integrating pre-operative imaging, namely CT and MRI, as well as real-time ultrasound...

  20. Detection of ATP and NADH: A Bioluminescent Experience.

    Selig, Ted C.; And Others

    1984-01-01

    Described is a bioluminescent assay for adenosine triphosphate (ATP) and reduced nicotineamide-adenine dinucleotide (NADH) that meets the requirements of an undergraduate biochemistry laboratory course. The 3-hour experiment provides students with experience in bioluminescence and analytical biochemistry yet requires limited instrumentation,…

  1. Use of the liquid scintillation spectrometer in bioluminescence analysis

    This review covers publications concerning analytical bioluminescence which in the main have appeared between mid-1973 and mid-1976. Outlines of some new assays and techniques are given together with modifications of existing procedures. Comments are presented on the use of the liquid scintillation spectrometer and other equipment for measuring bioluminescence. New applications are detailed and discussed

  2. A REVIEW OF ENVIRONMENTAL APPLICATIONS OF BIOLUMINESCENCE MEASUREMENTS

    This review of the recent literature on environmental applications of bioluminescence systems will focus on in vivo and in vitro bioluminescence methods that have been utilized to elucidate properties of chemicals, toxic and mutagenic effects, and to estimate biomass. The unifyin...

  3. Analysis of time-varying psoriasis lesion image patterns

    Maletti, Gabriela Mariel; Ersbøll, Bjarne Kjær; Nielsen, Allan Aasbjerg;

    2004-01-01

    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.......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....

  4. Real-time hyperspectral imaging for food safety applications

    Multispectral imaging systems with selected bands can commonly be used for real-time applications of food processing. Recent research has demonstrated several image processing methods including binning, noise removal filter, and appropriate morphological analysis in real-time mode can remove most fa...

  5. Dinoflagellate bioluminescence in response to mechanical stimuli in water flows

    A. S. Cussatlegras

    2005-01-01

    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.

  6. Moment searching algorithm for bioluminescence tomography

    Ludong Jin; Yan Wu; Jie Tian; Heyu Huang; Xiaochao Qu

    2009-01-01

    To avoid the ill-posedness in the inverse problem of bioluminescence tomography, a moment searching algorithm fusing the finite element method (FEM) with the moment concept in theoretical mechanics is developed. In the algorithm, the source's information is mapped to the surface photon flux density by FEM, and the source's position is modified with the feedback through the algorithm of barycenter searching, which makes full use of the position information of the photon flux density on surface. The position is modified in every iterative step and will finally converge to the real source's value theoretically.

  7. Adaptive Real Time Imaging Synthesis Telescopes

    Wright, Melvyn

    2012-01-01

    The digital revolution is transforming astronomy from a data-starved to a data-submerged science. Instruments such as the Atacama Large Millimeter Array (ALMA), the Large Synoptic Survey Telescope (LSST), and the Square Kilometer Array (SKA) will measure their accumulated data in petabytes. The capacity to produce enormous volumes of data must be matched with the computing power to process that data and produce meaningful results. In addition to handling huge data rates, we need adaptive calibration and beamforming to handle atmospheric fluctuations and radio frequency interference, and to provide a user environment which makes the full power of large telescope arrays accessible to both expert and non-expert users. Delayed calibration and analysis limit the science which can be done. To make the best use of both telescope and human resources we must reduce the burden of data reduction. Our instrumentation comprises of a flexible correlator, beam former and imager with digital signal processing closely coupled...

  8. Real-time extended dynamic range imaging in shearography

    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

  9. Real-time digital x-ray subtraction imaging

    A method of producing visible difference images derived from an x-ray image of an anatomical subject is described. X-rays are directed through the subject, and the image is converted into television fields comprising trains of analog video signals. The analog signals are converted into digital signals, which are then integrated over a predetermined time corresponding to several television fields. Difference video signals are produced by performing a subtraction between the ongoing video signals and the corresponding integrated signals, and are converted into visible television difference images representing changes in the x-ray image

  10. Experimental ultrasound system for real-time synthetic imaging

    Jensen, Jørgen Arendt; Holm, Ole; Jensen, Lars Joost; Bendsen, Henrik; Pedersen, Henrik Møller; Salomonsen, Kent; Hansen, Johnny; Nikolov, Svetoslav

    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...... 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......-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...

  11. In Vivo Real-Time, Multicolor, Quantum Dot Lymphatic Imaging

    Kosaka, Nobuyuki; Ogawa, Mikako; Sato, Noriko; Choyke, Peter L.; Kobayashi, Hisataka

    2009-01-01

    The lymphatic network is complex and difficult to visualize in real-time in vivo. Moreover, the direction of flow within lymphatic networks is often unpredictable especially in areas with well-developed “watershed” or overlapping lymphatics. Herein, we report a method of in vivo real-time multicolor lymphatic imaging using cadmium–selenium quantum dots (Qdots) with a fluorescence imaging system that enables the simultaneous visualization of up to five distinct lymphatic basins in real-time. F...

  12. Algorithm for localized adaptive diffuse optical tomography and its application in bioluminescence tomography

    A reconstruction algorithm for diffuse optical tomography based on diffusion theory and finite element method is described. The algorithm reconstructs the optical properties in a permissible domain or region-of-interest to reduce the number of unknowns. The algorithm can be used to reconstruct optical properties for a segmented object (where a CT-scan or MRI is available) or a non-segmented object. For the latter, an adaptive segmentation algorithm merges contiguous regions with similar optical properties thereby reducing the number of unknowns. In calculating the Jacobian matrix the algorithm uses an efficient direct method so the required time is comparable to that needed for a single forward calculation. The reconstructed optical properties using segmented, non-segmented, and adaptively segmented 3D mouse anatomy (MOBY) are used to perform bioluminescence tomography (BLT) for two simulated internal sources. The BLT results suggest that the accuracy of reconstruction of total source power obtained without the segmentation provided by an auxiliary imaging method such as x-ray CT is comparable to that obtained when using perfect segmentation. (paper)

  13. CD4+ T cell effects on CD8+ T cell location defined using bioluminescence.

    Mitra Azadniv

    Full Text Available T lymphocytes of the CD8+ class are critical in delivering cytotoxic function and in controlling viral and intracellular infections. These cells are "helped" by T lymphocytes of the CD4+ class, which facilitate their activation, clonal expansion, full differentiation and the persistence of memory. In this study we investigated the impact of CD4+ T cells on the location of CD8+ T cells, using antibody-mediated CD4+ T cell depletion and imaging the antigen-driven redistribution of bioluminescent CD8+ T cells in living mice. We documented that CD4+ T cells influence the biodistribution of CD8+ T cells, favoring their localization to abdominal lymph nodes. Flow cytometric analysis revealed that this was associated with an increase in the expression of specific integrins. The presence of CD4+ T cells at the time of initial CD8+ T cell activation also influences their biodistribution in the memory phase. Based on these results, we propose the model that one of the functions of CD4+ T cell "help" is to program the homing potential of CD8+ T cells.

  14. Algorithm for localized adaptive diffuse optical tomography and its application in bioluminescence tomography

    Naser, Mohamed A.; Patterson, Michael S.; Wong, John W.

    2014-04-01

    A reconstruction algorithm for diffuse optical tomography based on diffusion theory and finite element method is described. The algorithm reconstructs the optical properties in a permissible domain or region-of-interest to reduce the number of unknowns. The algorithm can be used to reconstruct optical properties for a segmented object (where a CT-scan or MRI is available) or a non-segmented object. For the latter, an adaptive segmentation algorithm merges contiguous regions with similar optical properties thereby reducing the number of unknowns. In calculating the Jacobian matrix the algorithm uses an efficient direct method so the required time is comparable to that needed for a single forward calculation. The reconstructed optical properties using segmented, non-segmented, and adaptively segmented 3D mouse anatomy (MOBY) are used to perform bioluminescence tomography (BLT) for two simulated internal sources. The BLT results suggest that the accuracy of reconstruction of total source power obtained without the segmentation provided by an auxiliary imaging method such as x-ray CT is comparable to that obtained when using perfect segmentation.

  15. A bioluminescent assay for measuring glucose uptake.

    Valley, Michael P; Karassina, Natasha; Aoyama, Natsuyo; Carlson, Coby; Cali, James J; Vidugiriene, Jolanta

    2016-07-15

    Identifying activators and inhibitors of glucose uptake is critical for both diabetes management and anticancer therapy. To facilitate such studies, easy-to-use nonradioactive assays are desired. Here we describe a bioluminescent glucose uptake assay for measuring glucose transport in cells. The assay is based on the uptake of 2-deoxyglucose and the enzymatic detection of the 2-deoxyglucose-6-phosphate that accumulates. Uptake can be measured from a variety of cell types, it can be inhibited by known glucose transporter inhibitors, and the bioluminescent assay yields similar results when compared with the radioactive method. With HCT 116 cells, glucose uptake can be detected in as little as 5000 cells and remains linear up to 50,000 cells with signal-to-background values ranging from 5 to 45. The assay can be used to screen for glucose transporter inhibitors, or by multiplexing with viability readouts, changes in glucose uptake can be differentiated from overall effects on cell health. The assay also can provide a relevant end point for measuring insulin sensitivity. With adipocytes and myotubes, insulin-dependent increases in glucose uptake have been measured with 10- and 2-fold assay windows, respectively. Significant assay signals of 2-fold or more have also been measured with human induced pluripotent stem cell (iPSC)-derived cardiomyocytes and skeletal myoblasts. PMID:27130501

  16. THz time-domain spectroscopy imaging for mail inspection

    Zhang, Liquan; Wang, Zhongdong; Ma, Yanmei; Hao, Erjuan

    2011-08-01

    Acquiring messages from the mail but not destroying the envelope is a big challenge in the war of intelligence. If one can read the message of the mail when the envelope is closed, he will benefit from the message asymmetry and be on a good wicket in the competition. In this paper, we presented a transmitted imaging system using THz time-domain spectroscopy technology. We applied the system to image the mail inside an envelope by step-scanning imaging technology. The experimental results show that the THz spectroscopy can image the mail in an envelope. The words in the paper can be identified easily from the background. We also present the THz image of a metal blade in the envelope, in which we can see the metal blade clearly. The results show that it is feasible of THz Time-Domain Spectroscopy Imaging for mail inspection applications.

  17. Real-time digital x-ray subtraction imaging

    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

  18. Anisotropy signature in extended images from reverse-time migration

    Sava, Paul

    2012-11-04

    Reverse-time migration can accurately image complex geologic structures in anisotropic media. Extended images at selected locations in the earth, i.e. at common-image-point gathers (CIPs), carry enough information to characterize the angle-dependent illumination and to provide measurements for migration velocity analysis. Furthermore, inaccurate anisotropy leaves a distinctive signature in CIPs, which can be used to evaluate anisotropy through techniques similar to the ones used in conventional wavefield tomography.

  19. Bioluminogenic Imaging of AminopeptidaseN In Vitro and In Vivo.

    Wu, Wenxiao; Chen, Laizhong; Li, Jing; Du, Lupei; Li, Minyong

    2016-01-01

    Bioluminescence is a process that converts biochemical energy to visible light. Bioluminescence-based imaging technology has been widely applicable in the imaging of process envisioned in life sciences. As one of the most popular bioluminescence system, the firefly luciferin-luciferase system is exceptionally suitable for deep tissue imaging in living animals owing to its long wavelength emission light. Herein, we report the experimental detail of bioluminogenic imaging of aminopeptidase N activity both in vitro and in vivo. PMID:27424897

  20. Image reconstruction in PET using time of flight information

    Recent progresses in fast time coincidence technique have permitted the use of time of flight (TOF) information in positron Emission Tomography. We describe the basic concept of positron time of flight imaging and introduce new concepts in order to incorporate the TOF data in the reconstruction process. An algorithm to recover positron activity is then proposed. We describe the image reconstruction in the TTVO1 time of flight camera, the system architecture and the special purpose operators. The time of flight tomography offers large development possibilities and we look forward the new high resolution, high signal-to-noise TOF camera

  1. Fast magnetic resonance imaging technology for time-series image diagnosis

    To expand the range of applicability of magnetic resonance imaging (MRI) tests, the Toshiba Group is engaged in the development of a time-series imaging technology with the aim of achieving double the acquisition speed compared with the conventional parallel imaging (PI) method. Although the PI method, which is the principal method used in existing products, can provide acquired images with approximately two or three times the spatial and/or temporal resolution through the integration of multiple coils and image processing technologies, it is not sufficient for time-series image diagnosis. A time-series PI method to increase the acceleration factors of PI requires and additional time-series map scan for calibration, hindering the improvement of imaging speed. In order to achieve a breakthrough in this situation, we are developing a fast MRI technology that can reconstruct images using the time-series PI method without the need for an additional time-series map scan. The resulting enhancement of the imaging speed will be useful in various areas, such as the acquisition of cardiac images with higher resolution. (author)

  2. High time-resolution imaging with the MAMA detector systems

    Morgan, Jeffrey S.; Timothy, J. Gethyn; Smith, Andrew M.; Hill, Bob; Kasle, David B.

    1990-01-01

    Current uses of the MAMA detector which utilize the photon time-tagging capabilities of these detectors are reported. These applications currently include image stabilization by means of post-processing corrections of platform drift and speckle interferometry. The initial results of a sounding rocket experiment to obtain UV images of NGC 6240 and results from speckle interferometry of Neptune's moon Triton are presented.

  3. Spectrally resolved bioluminescence tomography with adaptive finite element analysis: methodology and simulation

    As a molecular imaging technique, bioluminescence tomography (BLT) with its highly sensitive detection and facile operation can significantly reveal molecular and cellular information in vivo at the whole-body small animal level. However, because of complex photon transportation in biological tissue and boundary detection data with high noise, bioluminescent sources in deeper positions generally cannot be localized. In our previous work, we used achromatic or monochromatic measurements and an a priori permissible source region strategy to develop a multilevel adaptive finite-element algorithm. In this paper, we propose a spectrally solved tomographic algorithm with a posteriori permissible source region selection. Multispectral measurements, and anatomical and optical information first deal with the nonuniqueness of BLT and constrain the possible solution of source reconstruction. The use of adaptive mesh refinement and permissible source region based on a posteriori measures not only avoids the dimension disaster arising from the multispectral measured data but also reduces the ill-posedness of BLT and therefore improves the reconstruction quality. Reconsideration of the optimization method and related modifications further enhance reconstruction robustness and efficiency. We also incorporate into the method some improvements for reducing computational burdens. Finally, using a whole-body virtual mouse phantom, we demonstrate the capability of the proposed BLT algorithm to reconstruct accurately bioluminescent sources in deeper positions. In terms of optical property errors and two sources of discernment in deeper positions, this BLT algorithm represents the unique predominance for BLT reconstruction

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

    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.

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

    Dominjon, A., E-mail: a.dominjon@ipnl.in2p3.fr [CNRS/IN2P3, Institut de Physique Nucleaire de Lyon, Villeurbanne F-69622 (France); Ageron, M. [CNRS/IN2P3, Centre de Physique des Particules de Marseille, Marseille, F-13288 (France); Barbier, R. [CNRS/IN2P3, Institut de Physique Nucleaire de Lyon, Villeurbanne F-69622 (France); Universite de Lyon, Universite Lyon 1, Lyon F-69003 (France); Billault, M.; Brunner, J. [CNRS/IN2P3, Centre de Physique des Particules de Marseille, Marseille, F-13288 (France); Cajgfinger, T. [CNRS/IN2P3, Institut de Physique Nucleaire de Lyon, Villeurbanne F-69622 (France); Universite de Lyon, Universite Lyon 1, Lyon F-69003 (France); Calabria, P. [CNRS/IN2P3, Institut de Physique Nucleaire de Lyon, Villeurbanne F-69622 (France); Chabanat, E. [CNRS/IN2P3, Institut de Physique Nucleaire de Lyon, Villeurbanne F-69622 (France); Universite de Lyon, Universite Lyon 1, Lyon F-69003 (France); Chaize, D.; Doan, Q.T.; Guerin, C.; Houles, J.; Vagneron, L. [CNRS/IN2P3, Institut de Physique Nucleaire de Lyon, Villeurbanne F-69622 (France)

    2012-12-11

    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.

  6. Construction of a bioluminescent reporter strain to detect polychlorinated biphenyls

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

    1998-12-01

    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.

  7. Real-time movie image enhancement in NMR

    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)

  8. Laminographic reconstruction from real-time radiographic images

    We report the application of digital laminography reconstruction methods to real-time radiographic (RTR) images. Multiple digital images were acquired with the part at several orientations. Several acquisition and reconstruction methods have been investigated and their effects on the depth resolution and signal-to-noise ratio of the reconstructed images are discussed. The standard method yields the best signal-to-noise but the worst depth separation; the extreme value method yields the best depth separation with a slight decrease in signal-to-noise; and the iterative method is a compromise between the two. Both the extreme value and iterative methods require care in properly normalizing the projection images

  9. Real time neutron image processing system in NRF

    The neutron radiography facility was installed at the neutron radiography beam tube of the HANARO research reactor. The NRF is used for the nondestructive test to inspect and evaluate the material defect and homogeneity by detecting the transmitted neutron image in the nuclear as well as non-nuclear industry. To analyze the dynamical neutron image effectively and efficiently, the real-time image processing system was developed in background subtraction, normalization, geometry correction and beam uniformity, contrast control, filtering. The image quality test and dimension measurements were performed for the neutron beam purity and sensitivity indication. The NRF beam condition represents the highest beam quality for neutron radiography.

  10. Dynamics of embryonic pancreas development using real-time imaging

    Puri, Sapna; Hebrok, Matthias

    2007-01-01

    Current knowledge about developmental processes in complex organisms has relied almost exclusively on analyses of fixed specimens. However, organ growth is highly dynamic, and visualization of such dynamic processes, e.g. real-time tracking of cell movement and tissue morphogenesis, is becoming increasingly important. Here, we use live imaging to investigate expansion of the embryonic pancreatic epithelium in mouse. Using time-lapse imaging of tissue explants in culture, fluorescently labeled...

  11. A short- time beltrami kernel for smoothing images and manifolds.

    Spira, Alon; Kimmel, Ron; Sochen, Nir

    2007-06-01

    We introduce a short-time kernel for the Beltrami image enhancing flow. The flow is implemented by "convolving" the image with a space dependent kernel in a similar fashion to the solution of the heat equation by a convolution with a Gaussian kernel. The kernel is appropriate for smoothing regular (flat) 2-D images, for smoothing images painted on manifolds, and for simultaneously smoothing images and the manifolds they are painted on. The kernel combines the geometry of the image and that of the manifold into one metric tensor, thus enabling a natural unified approach for the manipulation of both. Additionally, the derivation of the kernel gives a better geometrical understanding of the Beltrami flow and shows that the bilateral filter is a Euclidean approximation of it. On a practical level, the use of the kernel allows arbitrarily large time steps as opposed to the existing explicit numerical schemes for the Beltrami flow. In addition, the kernel works with equal ease on regular 2-D images and on images painted on parametric or triangulated manifolds. We demonstrate the denoising properties of the kernel by applying it to various types of images and manifolds. PMID:17547140

  12. Anisotropy signature in reverse-time migration extended images

    Sava, Paul C.

    2014-11-04

    Reverse-time migration can accurately image complex geologic structures in anisotropic media. Extended images at selected locations in the Earth, i.e., at common-image-point gathers, carry rich information to characterize the angle-dependent illumination and to provide measurements for migration velocity analysis. However, characterizing the anisotropy influence on such extended images is a challenge. Extended common-image-point gathers are cheap to evaluate since they sample the image at sparse locations indicated by the presence of strong reflectors. Such gathers are also sensitive to velocity error that manifests itself through moveout as a function of space and time lags. Furthermore, inaccurate anisotropy leaves a distinctive signature in common-image-point gathers, which can be used to evaluate anisotropy through techniques similar to the ones used in conventional wavefield tomography. It specifically admits a V-shaped residual moveout with the slope of the "V" flanks depending on the anisotropic parameter η regardless of the complexity of the velocity model. It reflects the fourth-order nature of the anisotropy influence on moveout as it manifests itself in this distinct signature in extended images after handling the velocity properly in the imaging process. Synthetic and real data observations support this assertion.

  13. Fast sensors for time-of-flight imaging applications.

    Vallance, Claire; Brouard, Mark; Lauer, Alexandra; Slater, Craig S; Halford, Edward; Winter, Benjamin; King, Simon J; Lee, Jason W L; Pooley, Daniel E; Sedgwick, Iain; Turchetta, Renato; Nomerotski, Andrei; John, Jaya John; Hill, Laura

    2014-01-14

    The development of sensors capable of detecting particles and radiation with both high time and high positional resolution is key to improving our understanding in many areas of science. Example applications of such sensors range from fundamental scattering studies of chemical reaction mechanisms through to imaging mass spectrometry of surfaces, neutron scattering studies aimed at probing the structure of materials, and time-resolved fluorescence measurements to elucidate the structure and function of biomolecules. In addition to improved throughput resulting from parallelisation of data collection - imaging of multiple different fragments in velocity-map imaging studies, for example - fast image sensors also offer a number of fundamentally new capabilities in areas such as coincidence detection. In this Perspective, we review recent developments in fast image sensor technology, provide examples of their implementation in a range of different experimental contexts, and discuss potential future developments and applications. PMID:24002354

  14. Real-time image fusion involving diagnostic ultrasound

    Ewertsen, Caroline; Săftoiu, Adrian; Gruionu, Lucian G;

    2013-01-01

    The aim of our article is to give an overview of the current and future possibilities of real-time image fusion involving ultrasound. We present a review of the existing English-language peer-reviewed literature assessing this technique, which covers technical solutions (for ultrasound and endosc......The aim of our article is to give an overview of the current and future possibilities of real-time image fusion involving ultrasound. We present a review of the existing English-language peer-reviewed literature assessing this technique, which covers technical solutions (for ultrasound...... and endoscopic ultrasound), image fusion in several anatomic regions, and electromagnetic needle tracking....

  15. Biocidal effects of silver and zinc oxide nanoparticles on the bioluminescent bacteria

    Taran, M. V.; Starodub, N. F.; Katsev, A. M.; Guidotti, M.; Khranovskyy, V. D.; Babanin, A. A.; Melnychuk, M. D.

    2013-11-01

    The effect of silver and zinc oxide nanoparticles in combination with alginate on bioluminescent Photobacterium leiognathi Sh1 bacteria was investigated. Silver nanoparticles were found to be more toxic than zinc oxide nanoparticles on bioluminescent bacteria. The nanoparticles and their ions released results in the same effect, however, it was absent in combination with alginate. The effective inhibiting concentration (EC50) for silver nanoparticles was found about 0.3 - 0.4 μg mL-1, which was up to two times larger then for zinc oxide nanoparticles. The absence of sodium chloride in the tested media prevented the formation of colloidal particles of larger size and the effective inhibition concentrations of metal derivatives were lower than in the presence of sodium chloride.

  16. Time-gated optical imaging through turbid media using stimulated Raman scattering: Studies on image contrast

    K Divakar Rao; H S Patel; B Jain; P K Gupta

    2005-02-01

    In this paper, we report the development of experimental set-up for timegated optical imaging through turbid media using stimulated Raman scattering. Our studies on the contrast of time-gated images show that for a given optical thickness, the image contrast is better for sample with lower scattering coefficient and higher physical thickness, and that the contrast improves with decreasing value of anisotropy parameters of the scatterers. These results are consistent with time-resolved Monte Carlo simulations.

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

    Dangendorf, Volker; Lauck, Ronald; Kaufmann, Frank; Barnstedt, Juergen; Breskin, Amos; Jagutzki, Ottmar; Kraemer, Michael; Vartsky, David

    2006-01-01

    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...

  18. Real-time transfer and display of radiography image

    The information process network of cobalt-60 container inspection system is a local area network based on PC. The system requires reliable transfer of radiography image between collection station and process station and the real-time display of radiography image on process station. Due to the very high data acquisition rate, in order to realize the real-time transfer and display of radiography image, 100 M Ethernet technology and network process communication technology are adopted in the system. Windows Sockets is the most common process communication technology up to now. Several kinds of process communication way under Windows Sockets technology are compared and tested. Finally the author realized 1 Mbyte/s' inerrant image transfer and real-time display with blocked datagram transfer technology

  19. Bioluminescent determination of free fatty acids.

    Kather, H; Wieland, E

    1984-08-01

    A simple, highly specific, and sensitive bioluminescent method for determination of free fatty acids in unextracted plasma or serum has been developed. The method is based on the activation of free fatty acids by acyl-CoA synthetase (EC 6.2.1.3). The pyrophosphate formed is used to phosphorylate fructose 6-phosphate in a reaction catalyzed by the enzyme pyrophosphate-fructose-6-phosphate phosphotransferase (EC 4.1.2.13). The triosephosphates produced from fructose 1,6-bisphosphate by aldolase are oxidized by NAD in the presence of arsenate to 3-phosphoglycerate. The NADH is detected via the bacterial NADH-linked luciferase system. Excellent agreement has been obtained by comparison with accepted methods. In addition, for the determination of serum free fatty acids, the method is particularly applicable for following lipolysis of isolated adipocytes. PMID:6486422

  20. Multimodal MR imaging of acute and subacute experimental traumatic brain injury: Time course and correlation with cerebral energy metabolites

    Traumatic brain injury (TBI) is one of the leading causes of death and permanent disability world-wide. The predominant cause of death after TBI is brain edema which can be quantified by non-invasive diffusion-weighted magnetic resonance imaging (DWI). To provide a better understanding of the early onset, time course, spatial development, and type of brain edema after TBI and to correlate MRI data and the cerebral energy state reflected by the metabolite adenosine triphosphate (ATP). The spontaneous development of lateral fluid percussion-induced TBI was investigated in the acute (6 h), subacute (48 h), and chronic (7 days) phase in rats by MRI of quantitative T2 and apparent diffusion coefficient (ADC) mapping as well as perfusion was combined with ATP-specific bioluminescence imaging and histology. An induced TBI led to moderate to mild brain damages, reflected by transient, pronounced development of vasogenic edema and perfusion reduction. Heterogeneous ADC patterns indicated a parallel, but mixed expression of vasogenic and cytotoxic edema. Cortical ATP levels were reduced in the acute and subacute phase by 13% and 27%, respectively, but were completely normalized at 7 days after injury. The partial ATP reduction was interpreted to be partially caused by a loss of neurons in parallel with transient dilution of the regional ATP concentration by pronounced vasogenic edema. The normalization of energy metabolism after 7 days was likely due to infiltrating glia and not to recovery. The MRI combined with metabolite measurement further improves the understanding and evaluation of brain damages after TBI

  1. Real-time imaging of ATP release induced by mechanical stretch in human airway smooth muscle cells.

    Takahara, Norihiro; Ito, Satoru; Furuya, Kishio; Naruse, Keiji; Aso, Hiromichi; Kondo, Masashi; Sokabe, Masahiro; Hasegawa, Yoshinori

    2014-12-01

    Airway smooth muscle (ASM) cells within the airway walls are continually exposed to mechanical stimuli, and exhibit various functions in response to these mechanical stresses. ATP acts as an extracellular mediator in the airway. Moreover, extracellular ATP is considered to play an important role in the pathophysiology of asthma and chronic obstructive pulmonary disease. However, it is not known whether ASM cells are cellular sources of ATP secretion in the airway. We therefore investigated whether mechanical stretch induces ATP release from ASM cells. Mechanical stretch was applied to primary human ASM cells cultured on a silicone chamber coated with type I collagen using a stretching apparatus. Concentrations of ATP in cell culture supernatants measured by luciferin-luciferase bioluminescence were significantly elevated by cyclic stretch (12 and 20% strain). We further visualized the stretch-induced ATP release from the cells in real time using a luminescence imaging system, while acquiring differential interference contrast cell images with infrared optics. Immediately after a single uniaxial stretch for 1 second, strong ATP signals were produced by a certain population of cells and spread to surrounding spaces. The cyclic stretch-induced ATP release was significantly reduced by inhibitors of Ca(2+)-dependent vesicular exocytosis, 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetraacetoxymethyl ester, monensin, N-ethylmaleimide, and bafilomycin. In contrast, the stretch-induced ATP release was not inhibited by a hemichannel blocker, carbenoxolone, or blockade of transient receptor potential vanilloid 4 by short interfering RNA transfection or ruthenium red. These findings reveal a novel property of ASM cells: mechanically induced ATP release may be a cellular source of ATP in the airway. PMID:24885163

  2. Dinoflagellate bioluminescence in response to mechanical stimuli in water flows

    A. S. Cussatlegras; P. Le Gal

    2005-01-01

    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 ...

  3. Dinoflagellate bioluminescence in response to mechanical stimuli in water flows

    A. S. Cussatlegras; Gal, P.

    2005-01-01

    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 d...

  4. Dinoflagellate bioluminescence in response to mechanical stimuli in water flows

    Cussatlegras, A. S.; Gal, P.

    2005-01-01

    International audience 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 performe...

  5. Shedding light on bioluminescence regulation in Vibrio fischeri

    Miyashiro, Tim; Ruby, Edward G.

    2012-01-01

    The bioluminescence emitted by the marine bacterium Vibrio fischeri is a particularly striking result of individual microbial cells coordinating a group behavior. The genes responsible for light production are principally regulated by the LuxR-LuxI quorum-sensing system. In addition to LuxR-LuxI, numerous other genetic elements and environmental conditions control bioluminescence production. Efforts to mathematically model the LuxR-LuxI system are providing insight into the dynamics of this a...

  6. Bioluminescence-Sensing Assay for Microbial Growth Recognition

    Heba Ramadan Eed; Abdel-Kader, Nora S.; Mahmoud Helmy El Tahan; Tianhong Dai; Rehab Amin

    2016-01-01

    The conventional methods for microbial viability quantification require cultivation and are laborious. There is consequently a widespread need for cultivation-free methods. The adenosine triphosphate (ATP) bioluminescence-sensing assay is considered an extremely effective biosensor; hence ATP is the energy currency of all living microbes and can be used as a rapid indicator of microbial viability. We developed an ATP bioluminescence-sensing assay to detect microbial viability. A biolumine...

  7. Image Segmentation in Liquid Argon Time Projection Chamber Detector

    Płoński, Piotr; Sulej, Robert; Zaremba, Krzysztof

    2015-01-01

    The Liquid Argon Time Projection Chamber (LAr-TPC) detectors provide excellent imaging and particle identification ability for studying neutrinos. An efficient and automatic reconstruction procedures are required to exploit potential of this imaging technology. Herein, a novel method for segmentation of images from LAr-TPC detectors is presented. The proposed approach computes a feature descriptor for each pixel in the image, which characterizes amplitude distribution in pixel and its neighbourhood. The supervised classifier is employed to distinguish between pixels representing particle's track and noise. The classifier is trained and evaluated on the hand-labeled dataset. The proposed approach can be a preprocessing step for reconstructing algorithms working directly on detector images.

  8. Precision cosmology with time delay lenses: high resolution imaging requirements

    Meng, Xiao-Lei; Agnello, Adriano; Auger, Matthew W; Liao, Kai; Marshall, Philip J

    2015-01-01

    Lens time delays are a powerful probe of cosmology, provided that the gravitational potential of the main deflector can be modeled with sufficient precision. Recent work has shown that this can be achieved by detailed modeling of the host galaxies of lensed quasars, which appear as "Einstein Rings" in high resolution images. We carry out a systematic exploration of the high resolution imaging required to exploit the thousands of lensed quasars that will be discovered by current and upcoming surveys with the next decade. Specifically, we simulate realistic lens systems as imaged by the Hubble Space Telescope (HST), James Webb Space Telescope (JWST), and ground based adaptive optics images taken with Keck or the Thirty Meter Telescope (TMT). We compare the performance of these pointed observations with that of images taken by the Euclid (VIS), Wide-Field Infrared Survey Telescope (WFIRST) and Large Synoptic Survey Telescope (LSST) surveys. We use as our metric the precision with which the slope $\\gamma'$ of the...

  9. Deep-sea bioluminescence blooms after dense water formation at the ocean surface.

    Christian Tamburini

    Full Text Available 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.

  10. Real-time airborne hyperspectral imaging of land mines

    Ivanco, Tyler; Achal, Steve; McFee, John E.; Anger, Cliff; Young, Jane

    2007-04-01

    DRDC Suffeld and Itres Research have jointly investigated the use of visible and infrared hyperspectral imaging (HSI) for surface and buried land mine detection since 1989. These studies have demonstrated reliable passive HSI detection of surface-laid mines, based on their reflectance spectra, from airborne and ground-based platforms. Commercial HSI instruments collect and store image data at aircraft speeds, but the data are analysed off- line. This is useful for humanitarian demining, but unacceptable for military countermine operations. We have developed a hardware and software system with algorithms that can process the raw hyperspectral data in real time to detect mines. The custom algorithms perform radiometric correction of the raw data, then classify pixels of the corrected data, referencing a spectral signature library. The classification results are stored and displayed in real time, that is, within a few frame times of the data acquisition. Such real-time mine detection was demonstrated for the first time from a slowly moving land vehicle in March 2000. This paper describes an improved system which can achieve real-time detection of mines from an airborne platform, with its commensurately higher data rates. The system is presently compatible with the Itres family of visible/near infrared, short wave infrared and thermal infrared pushbroom hyperspectral imagers and its broadband thermal infrared pushbroom imager. Experiments to detect mines from an airborne platform in real time were conducted at DRDC Suffield in November 2006. Surface-laid land mines were detected in real time from a slowly moving helicopter with generally good detection rates and low false alarm rates. To the authors' knowledge, this is the first time that land mines have been detected from an airborne platform in real time using hyperspectral imaging.

  11. Real-time oriented image analysis in microcirculatory research

    Pries, Axel R.; Eriksson, S. E.; Jepsen, H.

    1990-11-01

    A digital video image analysis system is presented which consists of a personal computer equipped with a real-time video digitizer and a graphic tablet controlled by a modular set of programs aimed at performing a number of real-time oriented measuring tasks in microcirculatory research. Such tasks comprize the continuous recording of vessel diameters flow velocities or light intensity profiles from video recordings obtained during intravital microscopy of the terminal vascular bed either in research animals or in human beings. Two outstanding features of the presented systems are (A) the spatial correlation module for velocity measurement and (B) the automatic background movement correction. A: The spatial correlation velocity measurement module combined with an asymmetric illumination or gating process for image generation allows measurement of flow velocities from video microscopic images up to 20 mm/sec. This is about 10 to 20 times faster than the maximum. velocities which can be measured using conventional video based techniques. B: The automatic background movement correction is designed to track translational movements of background image structures in a reference window in real time (with respect to the video system). The translational vector of the image background is then used to adjust the position of the individual measuring lines or windows used in the different application modules to their original position relative to the tissue structures which are investigated. Such an automatic real-time tracking system isvery often a

  12. Using bacterial bioluminescence to evaluate the impact of biofilm on porous media hydraulic properties.

    Bozorg, Ali; Gates, Ian D; Sen, Arindom

    2015-02-01

    Biofilm formation in natural and engineered porous systems can significantly impact hydrodynamics by reducing porosity and permeability. To better understand and characterize how biofilms influence hydrodynamic properties in porous systems, the genetically engineered bioluminescent bacterial strain Pseudomonas fluorescens HK44 was used to quantify microbial population characteristics and biofilm properties in a translucent porous medium. Power law relationships were found to exist between bacterial bioluminescence and cell density, fraction of void space occupied by biofilm (i.e. biofilm saturation), and hydraulic conductivity. The simultaneous evaluation of biofilm saturation and porous medium hydraulic conductivity in real time using a non-destructive approach enabled the construction of relative hydraulic conductivity curves. Such information can facilitate simulation studies related to biological activity in porous structures, and support the development of new models to describe the dynamic behavior of biofilm and fluid flow in porous media. The bioluminescence based approach described here will allow for improved understanding and control of industrially relevant processes such as biofiltration and bioremediation. PMID:25479429

  13. Time-Reversal Acoustics and Maximum-Entropy Imaging

    Berryman, J G

    2001-08-22

    Target location is a common problem in acoustical imaging using either passive or active data inversion. Time-reversal methods in acoustics have the important characteristic that they provide a means of determining the eigenfunctions and eigenvalues of the scattering operator for either of these problems. Each eigenfunction may often be approximately associated with an individual scatterer. The resulting decoupling of the scattered field from a collection of targets is a very useful aid to localizing the targets, and suggests a number of imaging and localization algorithms. Two of these are linear subspace methods and maximum-entropy imaging.

  14. The Real-Time Image Processing Technique Based on DSP

    QI Chang; CHEN Yue-hua; HUANG Tian-shu

    2005-01-01

    This paper proposes a novel real-time image processing technique based on digital singnal processor (DSP). At the aspect of wavelet transform(WT) algorithm, the technique uses algorithm of second generation wavelet transform-lifting scheme WT that has low calculation complexity property for the 2-D image data processing. Since the processing effect of lifting scheme WT for 1-D data is better than the effect of it for 2-D data obviously, this paper proposes a reformative processing method: Transform 2-D image data to 1-D data sequence by linearization method, then process the 1-D data sequence by algorithm of lifting scheme WT. The method changes the image convolution mode,which based on the cross filtering of rows and columns. At the aspect of hardware realization, the technique optimizes the program structure of DSP to exert the operation power with the in-chip memorizer of DSP. The experiment results show that the real-time image processing technique proposed in this paper can meet the real-time requirement of video-image transmitting in the video surveillance system of electric power. So the technique is a feasible and efficient DSP solution.

  15. Decomposing global light transport using time of flight imaging

    Wu, Di; O'Toole, Matthew; Velten, Andreas; Agrawal, Amit; Raskar, Ramesh

    2012-01-01

    Global light transport is composed of direct and indirect components. In this paper, we take the first steps toward analyzing light transport using high temporal resolution information via time of flight (ToF) images. The time profile at each pixel encodes complex interactions between the incident light and the scene geometry with spatially-varying material properties. We exploit the time profile to decompose light transport into its constituent direct, subsurface scattering, and interreflect...

  16. Real-time particle image velocimetry based on FPGA technology

    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.

  17. Compact wearable dual-mode imaging system for real-time fluorescence image-guided surgery

    Zhu, Nan; Huang, Chih-Yu; Mondal, Suman; Gao, Shengkui; Huang, Chongyuan; Gruev, Viktor; Achilefu, Samuel; Liang, Rongguang

    2015-09-01

    A wearable all-plastic imaging system for real-time fluorescence image-guided surgery is presented. The compact size of the system is especially suitable for applications in the operating room. The system consists of a dual-mode imaging system, see-through goggle, autofocusing, and auto-contrast tuning modules. The paper will discuss the system design and demonstrate the system performance.

  18. Real-Time Ellipsometry-Based Transmission Ultrasound Imaging

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

    2007-02-14

    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

  19. Magneto-optical system for high speed real time imaging

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

    2012-08-01

    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.

  20. New real-time image processing system for IRFPA

    WANG Bing-jian; LIU Shang-qian; CHENG Yu-bao

    2006-01-01

    Influenced by detectors' material,manufacturing technology etc,every detector in infrared focal plane array (IRFPA) will output different voltages even if their input radiation flux is the same.And this is called non-uniformity of IRFPA.At the same time,the high background temperature,low temperature difference between targets and background and the low responsivity of IRFPA result in low contrast of infrared images.So non-uniformity correction and image enhancement are important techniques for IRFPA imaging system.This paper proposes a new real-time infrared image processing system based on Field Programmable Gate Array(FPGA).The system implements non-uniformity correction,image enhancement and video synthesization etc.By using parallel architecture and pipeline technique,the system processing speed is as high as 50Mx12bits per second.It is appropriate greatly to a large IRFPA and a high frame frequency IRFPA imaging system.The system is miniatured in one FPGA.

  1. Dual monitoring using {sup 124}I-FIAU and bioluminescence for HSV1-tk suicide gene therapy

    Lee, T. S.; Kim, J. H.; Kwon, H. C. [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)] (and others)

    2007-07-01

    Herpes simplex virus type I thymidine kinase (HSV-tk) is the most common reporter gene and is used in cancer gene therapy with a prodrug nucleoside analog, ganciclovir (GCV). The aim of this study is to evaluate therapeutic efficacy of suicide gene therapy with 2'-fluoro-2'-deoxy-1-D-arabinofuranosyl-5-[{sup 124}I] iodouracil ({sup 124}I - FIAU) and bioluminescence in retrovirally HSV -tk and firefly luciferase transduced hepatoma model. The HSV -tk and firefly luciferase (Luc) was retrovirally transduced and expressed in MCA rat Morris hepatoma cells. Nude mice with subcutaneous tumors, MCA and MCA-TK-Luc, were subjected to GCV treatment (50mg/Kg/d intraperitoneally) for 5 day. PET imaging and biodistribution with ({sup 124}I-FIAU) were performed at before and after initiation of therapy with GCV. Bioluminescent signal was also measured during GCV treatment. Before GCV treatment, no significant difference in tumor volume was found in tumors between MCA and MCA-TK-Luc. After GCV treatment, tumor volume of MCA-TK-Luc markedly reduced compared to that of MCA. In biodistribution study, {sup 124}I-FIAU uptake after GCV therapy significantly decreased compared with pretreatment levels (34.8 13.67 %ID/g vs 7.6 2.59 %ID/g) and bioluminescent signal was also significantly decreased compared with pretreatment levels. In small animal PET imaging, {sup 124}I-FIAU selectively localized in HSV -tk expressing tumor and the therapeutic efficacy of GCV treatment was evaluated by {sup 124}I-FIAU PET imaging. {sup 124}I-FIAU PET and bioluminescence imaging in HSV-tk suicide gene therapy were effective to evaluate the therapeutic response. {sup 124}I-FIAU may serve as an efficient and selective agent for monitoring of transduced HSV1-tk gene expression in vivo in clinical trials.

  2. Brownian motion at fast time scales and thermal noise imaging

    Huang, Rongxin

    This dissertation presents experimental studies on Brownian motion at fast time scales, as well as our recent developments in Thermal Noise Imaging which uses thermal motions of microscopic particles for spatial imaging. As thermal motions become increasingly important in the studies of soft condensed matters, the study of Brownian motion is not only of fundamental scientific interest but also has practical applications. Optical tweezers with a fast position-sensitive detector provide high spatial and temporal resolution to study Brownian motion at fast time scales. A novel high bandwidth detector was developed with a temporal resolution of 30 ns and a spatial resolution of 1 A. With this high bandwidth detector, Brownian motion of a single particle confined in an optical trap was observed at the time scale of the ballistic regime. The hydrodynamic memory effect was fully studied with polystyrene particles of different sizes. We found that the mean square displacements of different sized polystyrene particles collapse into one master curve which is determined by the characteristic time scale of the fluid inertia effect. The particle's inertia effect was shown for particles of the same size but different densities. For the first time the velocity autocorrelation function for a single particle was shown. We found excellent agreement between our experiments and the hydrodynamic theories that take into account the fluid inertia effect. Brownian motion of a colloidal particle can be used to probe three-dimensional nano structures. This so-called thermal noise imaging (TNI) has been very successful in imaging polymer networks with a resolution of 10 nm. However, TNI is not efficient at micrometer scale scanning since a great portion of image acquisition time is wasted on large vacant volume within polymer networks. Therefore, we invented a method to improve the efficiency of large scale scanning by combining traditional point-to-point scanning to explore large vacant

  3. Time-lapse Raman imaging of osteoblast differentiation

    Hashimoto, Aya; Yamaguchi, Yoshinori; Chiu, Liang-Da; Morimoto, Chiaki; Fujita, Katsumasa; Takedachi, Masahide; Kawata, Satoshi; Murakami, Shinya; Tamiya, Eiichi

    2015-07-01

    Osteoblastic mineralization occurs during the early stages of bone formation. During this mineralization, hydroxyapatite (HA), a major component of bone, is synthesized, generating hard tissue. Many of the mechanisms driving biomineralization remain unclear because the traditional biochemical assays used to investigate them are destructive techniques incompatible with viable cells. To determine the temporal changes in mineralization-related biomolecules at mineralization spots, we performed time-lapse Raman imaging of mouse osteoblasts at a subcellular resolution throughout the mineralization process. Raman imaging enabled us to analyze the dynamics of the related biomolecules at mineralization spots throughout the entire process of mineralization. Here, we stimulated KUSA-A1 cells to differentiate into osteoblasts and conducted time-lapse Raman imaging on them every 4 hours for 24 hours, beginning 5 days after the stimulation. The HA and cytochrome c Raman bands were used as markers for osteoblastic mineralization and apoptosis. From the Raman images successfully acquired throughout the mineralization process, we found that β-carotene acts as a biomarker that indicates the initiation of osteoblastic mineralization. A fluctuation of cytochrome c concentration, which indicates cell apoptosis, was also observed during mineralization. We expect time-lapse Raman imaging to help us to further elucidate osteoblastic mineralization mechanisms that have previously been unobservable.

  4. A computational approach to real-time image processing for serial time-encoded amplified microscopy

    Oikawa, Minoru; Hiyama, Daisuke; Hirayama, Ryuji; Hasegawa, Satoki; Endo, Yutaka; Sugie, Takahisa; Tsumura, Norimichi; Kuroshima, Mai; Maki, Masanori; Okada, Genki; Lei, Cheng; Ozeki, Yasuyuki; Goda, Keisuke; Shimobaba, Tomoyoshi

    2016-03-01

    High-speed imaging is an indispensable technique, particularly for identifying or analyzing fast-moving objects. The serial time-encoded amplified microscopy (STEAM) technique was proposed to enable us to capture images with a frame rate 1,000 times faster than using conventional methods such as CCD (charge-coupled device) cameras. The application of this high-speed STEAM imaging technique to a real-time system, such as flow cytometry for a cell-sorting system, requires successively processing a large number of captured images with high throughput in real time. We are now developing a high-speed flow cytometer system including a STEAM camera. In this paper, we describe our approach to processing these large amounts of image data in real time. We use an analog-to-digital converter that has up to 7.0G samples/s and 8-bit resolution for capturing the output voltage signal that involves grayscale images from the STEAM camera. Therefore the direct data output from the STEAM camera generates 7.0G byte/s continuously. We provided a field-programmable gate array (FPGA) device as a digital signal pre-processor for image reconstruction and finding objects in a microfluidic channel with high data rates in real time. We also utilized graphics processing unit (GPU) devices for accelerating the calculation speed of identification of the reconstructed images. We built our prototype system, which including a STEAM camera, a FPGA device and a GPU device, and evaluated its performance in real-time identification of small particles (beads), as virtual biological cells, owing through a microfluidic channel.

  5. Established rheumatoid arthritis - new imaging modalities

    McQueen, Fiona M; Østergaard, Mikkel

    2007-01-01

    ) and single photon emission tomography (SPECT) can reveal actively metabolizing bone and the proliferation of synovial cells via radioactive labeling. Bioluminescence and fluorescence reflectance imaging are other approaches that allow imaging, and potentially the delivery of therapeutic agents, at a...

  6. Established rheumatoid arthritis - new imaging modalities

    McQueen, Fiona M; Østergaard, Mikkel

    2007-01-01

    ) and single photon emission tomography (SPECT) can reveal actively metabolizing bone and the proliferation of synovial cells via radioactive labeling. Bioluminescence and fluorescence reflectance imaging are other approaches that allow imaging, and potentially the delivery of therapeutic agents...

  7. Improving the Image Quality of Synthetic Transmit Aperture Ultrasound Images - Achieving Real-Time In-Vivo Imaging

    Gammelmark, Kim

    2004-01-01

    issues need to solved. The goal of this PhD study has been to find methods that can be used to overcome the above mentioned limitations, and hereby improve the image quality of STA imaging to a clinically desirable level, enabling real-time in-vivo STA imaging. The thesis investigates a new method to....... The thesis contains summaries of four journal articles and four corresponding conference publications, which comprise the primary contributions of the PhD. The first two papers give elaborated evaluations of TMS imaging for linear array and convex array imaging, respectively. The results, including...... presented in the third paper, based on a clinical trial with 7 healthy male volunteers. Real-time movie sequences of 3 seconds duration were acquired and analyzed by experienced medical doctors using blinded clinical evaluation. The results showed a statistically significant improvement in image quality of...

  8. A bioluminescent mouse model of proliferation to highlight early stages of pancreatic cancer: A suitable tool for preclinical studies.

    de Latouliere, Luisa; Manni, Isabella; Iacobini, Carla; Pugliese, Giuseppe; Grazi, Gian Luca; Perri, Pasquale; Cappello, Paola; Novelli, Franco; Menini, Stefano; Piaggio, Giulia

    2016-09-01

    Transgenic mouse models designed to recapitulate genetic and pathologic aspects of cancer are useful to study early stages of disease as well as its progression. Among several, two of the most sophisticated models for pancreatic ductal adenocarcinoma (PDAC) are the LSL-Kras(G12D/+);Pdx-1-Cre (KC) and LSL-Kras(G12D/+);LSL-Trp53(R172H/+);Pdx-1-Cre (KPC) mice, in which the Cre-recombinase regulated by a pancreas-specific promoter activates the expression of oncogenic Kras alone or in combination with a mutant p53, respectively. Non-invasive in vivo imaging offers a novel approach to preclinical studies introducing the possibility to investigate biological events in the spatio/temporal dimension. We recently developed a mouse model, MITO-Luc, engineered to express the luciferase reporter gene in cells undergoing active proliferation. In this model, proliferation events can be visualized non-invasively by bioluminescence imaging (BLI) in every body district in vivo. Here, we describe the development and characterization of MITO-Luc-KC- and -KPC mice. In these mice we have now the opportunity to follow PDAC evolution in the living animal in a time frame process. Moreover, by relating in vivo and ex vivo BLI and histopathological data we provide evidence that these mice could represents a suitable tool for pancreatic cancer preclinical studies. Our data also suggest that aberrant proliferation events take place early in pancreatic carcinogenesis, before tumour appearance. PMID:26704357

  9. Time-resolved neutron imaging at ANTARES cold neutron beamline

    Tremsin, A S; Tittelmeier, K; Schillinger, B; Schulz, M; Lerche, M; Feller, W B

    2015-01-01

    In non-destructive evaluation with X-rays light elements embedded in dense, heavy (or high-Z) matrices show little contrast and their structural details can hardly be revealed. Neutron radiography, on the other hand, provides a solution for those cases, in particular for hydrogenous materials, owing to the large neutron scattering cross section of hydrogen and uncorrelated dependency of neutron cross section on the atomic number. The majority of neutron imaging experiments at the present time is conducted with static objects mainly due to the limited flux intensity of neutron beamline facilities and sometimes due to the limitations of the detectors. However, some applications require the studies of dynamic phenomena and can now be conducted at several high intensity beamlines such as the recently rebuilt ANTARES beam line at the FRM-II reactor. In this paper we demonstrate the capabilities of time resolved imaging for repetitive processes, where different phases of the process can be imaged simultaneously and...

  10. Real-Time Air Pollutants Rendering based on Image Processing

    Demin Wang

    2011-11-01

    Full Text Available This paper presents a new method for realistic real-time rendering of air pollutants based on image processing. The air pollutants’ variable density can create many shapes of mist what can add a realistic environment to virtual scene. In order to achieve a realistic effect, we further enhance thus obtained air pollution data getting from monitor in spatial domain. In the proposed method we map the densities of air pollutants to different gray levels, and visualize them by blending those gray levels with background images. The proposed method can also visualize large-scale air pollution data from different viewpoints in real-time and provide the resulting image with any resolution theoretically, which is very important and favorable for the Internet transmission.

  11. A Simple Fusion Method for Image Time Series Based on the Estimation of Image Temporal Validity

    Mar Bisquert

    2015-01-01

    Full Text Available High-spatial-resolution satellites usually have the constraint of a low temporal frequency, which leads to long periods without information in cloudy areas. Furthermore, low-spatial-resolution satellites have higher revisit cycles. Combining information from high- and low- spatial-resolution satellites is thought a key factor for studies that require dense time series of high-resolution images, e.g., crop monitoring. There are several fusion methods in the bibliography, but they are time-consuming and complicated to implement. Moreover, the local evaluation of the fused images is rarely analyzed. In this paper, we present a simple and fast fusion method based on a weighted average of two input images (H and L, which are weighted by their temporal validity to the image to be fused. The method was applied to two years (2009–2010 of Landsat and MODIS (MODerate Imaging Spectroradiometer images that were acquired over a cropped area in Brazil. The fusion method was evaluated at global and local scales. The results show that the fused images reproduced reliable crop temporal profiles and correctly delineated the boundaries between two neighboring fields. The greatest advantages of the proposed method are the execution time and ease of use, which allow us to obtain a fused image in less than five minutes.

  12. Advances in Time-Resolved Tomographic Particle Image Velocimetry

    Lynch, K.P.

    2015-01-01

    This thesis details advanced developments in 3-D particle image velocimetry (PIV) based on the tomographic PIV technique, with an emphasis on time-resolved experiments. Tomographic PIV is a technique introduced in 2006 to measure the flow velocity in a three-dimensional volume. When measurements are performed at a rate high enough to sample the dynamical evolution of the flow, the measurement is considered time-resolved. The present work begins with a description of developments in tomographi...

  13. Time-of-flight imaging of invisibility cloaks

    Halimeh, Jad C.; Wegener, Martin

    2012-01-01

    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 t...

  14. Time-resolved fast neutron imaging: simulation of detector performance

    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

  15. Diffeomorphic image registration with automatic time-step adjustment

    Pai, Akshay Sadananda Uppinakudru; Klein, S.; Sommer, Stefan Horst; Darkner, Sune; Sporring, Jon; Nielsen, Mads

    In this paper, we propose an automated Euler's time-step adjustment scheme for diffeomorphic image registration using stationary velocity fields (SVFs). The proposed variational problem aims at bounding the inverse consistency error by adaptively adjusting the number of Euler's step required to...

  16. Time-resolved fast neutron imaging: simulation of detector performance

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

    2004-01-01

    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.

  17. Segmentation of Time-Lapse Images with Focus on Microscopic Images of Cells

    Soukup, Jindřich; Císař, P.; Šroubek, Filip

    Berlin: Springer-Verlag, 2013 - (Petrosino, A.), s. 71-80. (Lecture Notes in Computer Science. Image Processing , Computer Vision, Pattern Recognition, and Graphics. 8157). ISBN 978-3-642-41183-0. [International Conference on Image Analysis and Processing . Naples (IT), 11.09.2013-13.09.2013] R&D Projects: GA ČR GA13-29225S Grant ostatní: Grantová agentura UK(CZ) GAUK 914813/2013; GA MŠk(CZ) ED2.1.00/01.0024 Institutional support: RVO:67985556 Keywords : segmentation * time-lapse * microscopy imaging * phase constrast Subject RIV: JD - Computer Applications, Robotics http://library.utia.cas.cz/separaty/2013/ZOI/soukup-segmentation of time-lapse image s with focus on microscopic image s of cells.pdf

  18. A miniature real-time volumetric ultrasound imaging system

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

    2005-04-01

    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.

  19. Bubble masks for time-encoded imaging of fast neutrons.

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

    2013-09-01

    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.

  20. The global ultraviolet imager (GUVI) for the NASA TIMED mission

    The Global Ultraviolet Imager (GUVI) investigation is designed to provide quantitative observations and interpretation of the Earth's airglow and auroral emissions in support of the NASA Thermosphere, Ionosphere, Mesosphere, Energy and Dynamics (TIMED) mission. It will address TIMED objectives dealing with energetics, dynamics, and the specification of state variables. The instrument will provide multiple-wavelength, simultaneous ''monochromatic'' images of the far-ultraviolet emission (115 to 180 nm) using a scan mirror to sweep the instantaneous field of view of a spectrographic imager through an arc of up to 140 degree aligned perpendicular to the orbit plane of the spacecraft. The instantaneous field of view is 11.8 degree by 0.37 degree (adjustable) along the slit and perpendicular to the slit, respectively. The field of view is mapped to a two-dimensional image plane with up to 64 spatial pixels by 160 spectral pixels of spectral width 0.4 nm per pixel. Binning of pixels can be performed along both the spatial and spectral axes of the array to reduce the demands on the downlink telemetry. The f/3 Rowland circle scanning spectrographic imager is outfitted with a toroidal grating ruled at 1,200 grooves per millimeter. The fore-optics consist of a plane scanning mirror and an off-axis parabolic telescope. The detector is a photon-counting microchannel plate with a wedge and strip anode mounted in a sealed tube

  1. Modelling dinoflagellates as an approach to the seasonal forecasting of bioluminescence in the North Atlantic

    Marcinko, Charlotte L.J.; Martin, Adrian P.; Allen, John T.

    2014-01-01

    Bioluminescence within ocean surface waters is of significant interest because it can enhance the study of subsurface movement and organisms. Little is known about how bioluminescence potential (BPOT) varies spatially and temporally in the open ocean. However, light emitted from dinoflagellates often dominates the stimulated bioluminescence field. As a first step towards forecasting surface ocean bioluminescence in the open ocean, a simple ecological model is developed which simulates seasona...

  2. Vegetable seed radiosensitivity and kinetic analysis of super-weak bioluminescence

    Bioluminescence of several vegetable seeds induced by γ-rays was studied. The results show that positive relation exists between seeds bioluminescence and irradiation dose, which fits with equation Y=Y0eKD. The higher the K value is, the more intense the bioluminescence induced by γ-rays is. Significant differences among K values were found with different varieties. The bioluminescence and exterior measurement value of seed radiosensitivity showed good consistency

  3. Imaging the time sequence of latent electrostatic fingerprints

    Watson, P.; Prance, R. J.; Prance, H.; Beardsmore-Rust, S. T.

    2010-10-01

    Biometric identification for forensic investigations and security continues to depend on classic fingerprinting in many instances. Existing techniques rely on either visible deposits or hidden (latent) fingerprints resulting from the transfer of residues from the finger to the surface. However, one of the limitations of classic fingerprinting, for use as forensic evidence, is in determining a time sequence of events. It is extremely difficult to establish a timeline, from fingerprint evidence alone. We present the capability of a new technique which images the electrical charge deposited by tribocharging when a finger contacts an electrically insulated surface. The method is applicable to insulating surfaces and has been tested on PVC, PTFE, Acetate and PVDF sheets. The latent electrostatic charge pattern is detected using a novel, microscopic, electric potential sensor. The sensor is capable of imaging static charge distributions non-invasively, with no discharging effect on the sample. We present data showing the decay of the charge image with time, over a period up to 14 days. This capability has two major implications. First this technique does not suffer from ambiguities caused by a history of old fingerprints and second it has the potential to allow the time sequence of recent charge fingerprint images to be determined.

  4. Time required to stabilize thermographic images at rest

    Marins, João Carlos Bouzas; Moreira, Danilo Gomes; Cano, Sergio Piñonosa; Quintana, Manuel Sillero; Soares, Danusa Dias; Fernandes, Alex de Andrade; Silva, Fabrício Sousa da; Costa, Carlos Magno Amaral; Amorim, Paulo Roberto dos Santos

    2014-07-01

    Thermography for scientific research and practical purposes requires a series of procedures to obtain images that should be standardized; one of the most important is the time required for acclimatization in the controlled environment. Thus, the objective of this study was to identify the appropriate acclimatization time in rest to reach a thermal balance on young people skin. Forty-four subjects participated in the study, 18 men (22.3 ± 3.1 years) and 26 women (21.7 ± 2.5 years). Thermographic images were collected using a thermal imager (Fluke®), totaling 44 images over a period of 20 min. The skin temperature (TSK) was measured at the point of examination which included the 0 min, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20. The body regions of interest (ROI) analyzed included the hands, forearms, arms, thighs, legs, chest and abdomen. We used the Friedman test with post hoc Dunn's in order to establish the time at rest required to obtain a TSK balance and the Mann-Whitney test was used to compare age, BMI, body fat percentage and temperature variations between men and women, considering always a significance level of p body region of the abdomen obtained a significant variance (p body analysis it is recommended at least 10 min for both sexes.

  5. Implied Movement in Static Images Reveals Biological Timing Processing

    2015-01-01

    Visual perception is adapted toward a better understanding of our own movements than those of non-conspecifics. The present study determined whether time perception is affected by pictures of different species by considering the evolutionary scale. Static (“S”) and implied movement (“M”) images of a dog, cheetah, chimpanzee, and man were presented to undergraduate students. S and M images of the same species were presented in random order or one after the other (S-M or M-S) for two groups of ...

  6. Rapid detection (4 h) of methicillin-resistant Staphylococcus aureus by a bioluminescence method.

    Park, C. H.; Hixon, D L; McLaughlin, C M; Cook, J F

    1988-01-01

    A 4-h bioluminescence method for methicillin susceptibility determination was compared with reference methods. Of the Staphylococcus aureus strains tested, 80 were methicillin resistant, 180 were methicillin susceptible, and 10 were borderline susceptible. There was 100% correlation between bioluminescence and reference methods for methicillin-susceptible and methicillin-resistant strains. All borderline-susceptible strains were identified as methicillin resistant by bioluminescence.

  7. Propensity Scores for Prediction and Characterization of Bioluminescent Proteins from Sequences

    Huang, Hui-Ling

    2014-01-01

    Bioluminescent proteins (BLPs) are a class of proteins with various mechanisms of light emission such as bioluminescence and fluorescence from luminous organisms. While valuable for commercial and medical applications, identification of BLPs, including luciferases and fluorescent proteins (FPs), is rather challenging, owing to their high variety of protein sequences. Moreover, characterization of BLPs facilitates mutagenesis analysis to enhance bioluminescence and fluorescence. Therefore, thi...

  8. Novel bioluminescent coelenterazine derivatives with imidazopyrazinone C-6 extended substitution for Renilla luciferase.

    Jiang, Tianyu; Yang, Xiaofeng; Yang, Xingye; Yuan, Mingliang; Zhang, Tianchao; Zhang, Huateng; Li, Minyong

    2016-06-21

    Two series of novel coelenterazine analogues (alkynes and triazoles) with imidazopyrazinone C-6 extended substitution have been designed and synthesized successfully for the extension of bioluminescent substrates. After extensive evaluation, some compounds display excellent bioluminescence properties compared with DeepBlueC in cellulo, thus becoming potential molecules for bioluminescence techniques. PMID:27197767

  9. Time-resolved tomographic images of a relativistic electron beam

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

    1984-07-01

    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.

  10. Super-resolved imaging with ultimate time resolution

    Ashida, Yuto

    2015-01-01

    Precisely and accurately locating point objects is a long-standing common thread in science. Super-resolved imaging of single molecules has revolutionized our view of quasi-static nanostructures $\\it{in-vivo}$. A wide-field approach based on localizing individual fluorophores has emerged as a versatile method to surpass the standard resolution limit. In those techniques, the super-resolution is realized by sparse photoactivation and localization together with the statistical analysis based on point spread functions. Nevertheless, the slow temporal resolution of super-resolved imaging severely restricts the utility to the study of live-cell phenomena. Clearly, a major breakthrough to observe fast, nanoscale dynamics needs to be made. Here we present a super-resolved imaging method that achieves the theoretical-limit time resolution. By invoking information theory, we can achieve the robust localization of overlapped light emitters at an order of magnitude faster speed than the conventional super-resolution mic...

  11. Real-time digital X-ray subtraction imaging

    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)

  12. Cellular Neural Network for Real Time Image Processing

    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)

  13. A Colour Image Quantization Algorithm for Time-Constrained Applications

    Wattanapong KURDTHONGMEE

    2005-06-01

    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.

  14. Time-of-flight imaging of invisibility cloaks

    Halimeh, Jad C

    2011-01-01

    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 ...

  15. Automatic multimodal real-time tracking for image plane alignment in interventional Magnetic Resonance Imaging

    Interventional magnetic resonance imaging (MRI) aims at performing minimally invasive percutaneous interventions, such as tumor ablations and biopsies, under MRI guidance. During such interventions, the acquired MR image planes are typically aligned to the surgical instrument (needle) axis and to surrounding anatomical structures of interest in order to efficiently monitor the advancement in real-time of the instrument inside the patient's body. Object tracking inside the MRI is expected to facilitate and accelerate MR-guided interventions by allowing to automatically align the image planes to the surgical instrument. In this PhD thesis, an image-based work-flow is proposed and refined for automatic image plane alignment. An automatic tracking work-flow was developed, performing detection and tracking of a passive marker directly in clinical real-time images. This tracking work-flow is designed for fully automated image plane alignment, with minimization of tracking-dedicated time. Its main drawback is its inherent dependence on the slow clinical MRI update rate. First, the addition of motion estimation and prediction with a Kalman filter was investigated and improved the work-flow tracking performance. Second, a complementary optical sensor was used for multi-sensor tracking in order to decouple the tracking update rate from the MR image acquisition rate. Performance of the work-flow was evaluated with both computer simulations and experiments using an MR compatible test bed. Results show a high robustness of the multi-sensor tracking approach for dynamic image plane alignment, due to the combination of the individual strengths of each sensor. (author)

  16. Augmented reality based real-time subcutaneous vein imaging system.

    Ai, Danni; Yang, Jian; Fan, Jingfan; Zhao, Yitian; Song, Xianzheng; Shen, Jianbing; Shao, Ling; Wang, Yongtian

    2016-07-01

    A novel 3D reconstruction and fast imaging system for subcutaneous veins by augmented reality is presented. The study was performed to reduce the failure rate and time required in intravenous injection by providing augmented vein structures that back-project superimposed veins on the skin surface of the hand. Images of the subcutaneous vein are captured by two industrial cameras with extra reflective near-infrared lights. The veins are then segmented by a multiple-feature clustering method. Vein structures captured by the two cameras are matched and reconstructed based on the epipolar constraint and homographic property. The skin surface is reconstructed by active structured light with spatial encoding values and fusion displayed with the reconstructed vein. The vein and skin surface are both reconstructed in the 3D space. Results show that the structures can be precisely back-projected to the back of the hand for further augmented display and visualization. The overall system performance is evaluated in terms of vein segmentation, accuracy of vein matching, feature points distance error, duration times, accuracy of skin reconstruction, and augmented display. All experiments are validated with sets of real vein data. The imaging and augmented system produces good imaging and augmented reality results with high speed. PMID:27446690

  17. Real-time mid-infrared imaging of living microorganisms.

    Haase, Katharina; Kröger-Lui, Niels; Pucci, Annemarie; Schönhals, Arthur; Petrich, Wolfgang

    2016-01-01

    The speed and efficiency of quantum cascade laser-based mid-infrared microspectroscopy are demonstrated using two different model organisms as examples. For the slowly moving Amoeba proteus, a quantum cascade laser is tuned over the wavelength range of 7.6 µm to 8.6 µm (wavenumbers 1320 cm(-1) and 1160 cm(-1) , respectively). The recording of a hyperspectral image takes 11.3 s whereby an average signal-to-noise ratio of 29 is achieved. The limits of time resolution are tested by imaging the fast moving Caenorhabditis elegans at a discrete wavenumber of 1265 cm(-1) . Mid-infrared imaging is performed with the 640 × 480 pixel video graphics array (VGA) standard and at a full-frame time resolution of 0.02 s (i.e. well above the most common frame rate standards). An average signal-to-noise ratio of 16 is obtained. To the best of our knowledge, these findings constitute the first mid-infrared imaging of living organisms at VGA standard and video frame rate. PMID:26572683

  18. Timing and position response of a block detector for fast neutron time-of-flight imaging

    Our research effort seeks to improve the spatial and timing performance of a block detector made of a pixilated plastic scintillator (EJ-200), first demonstrated as part of Oak Ridge National Laboratory's Advanced Portable Neutron Imaging System. Improvement of the position and time response is necessary to achieve better resolution and contrast in the images of shielded special nuclear material. Time-of-flight is used to differentiate between gamma and different sources of neutrons (e.g., transmission and fission neutrons). Factors limiting the timing and position performance of the neutron detector have been revealed through simulations and measurements. Simulations have suggested that the degradation in the ability to resolve pixels in the neutron detector is due to those interactions occurring near the light guide. The energy deposition within the neutron detector is shown to affect position performance and imaging efficiency. This examination details how energy cuts improve the position performance and degrade the imaging efficiency. Measurements have shown the neutron detector to have a timing resolution of σ=238 ps. The majority of this timing uncertainty is from the depth-of-interaction (DOI) of the neutron which is confirmed by simulations and analytical calculations

  19. Nanoluciferase signal brightness using furimazine substrates opens bioluminescence resonance energy transfer to widefield microscopy.

    Kim, Jiho; Grailhe, Regis

    2016-08-01

    Fluorescence and bioluminescence resonance energy transfer (FRET, BRET) techniques are powerful tools for studying protein-protein interactions in cellular assays. In contrast to fluorescent proteins, chemiluminescent proteins do not require excitation light, known to trigger autofluorescence, phototoxicity, and photobleaching. Regrettably, low signal intensity of luciferase systems restricts their usage as they require specialized microscopes equipped with ultra low-light imaging cameras. In this study, we report that bioluminescence quantification in living cells using a standard widefield automated microscope dedicated to screening and high content analysis is possible with the newer luciferase systems, Nanoluciferase (Nluc). With such equipment, we showed that robust intramolecular BRET can be measured using a combination of Nluc and yellow fluorescent protein (YFP). Using the human Superoxide Dismutase 1 (SOD1) dimer model, we next validated that intermolecular BRET could be quantified at a single cell level. The enhanced signal brightness of Nluc enabling BRET imaging to widefield microscopy shows strong potential to open up single cell protein-protein interactions studies to a wider audience. © 2016 International Society for Advancement of Cytometry. PMID:27144967

  20. A fast reconstruction algorithm for bioluminescence tomography based on smoothed l0 norm regularization

    He, Xiaowei; Yu, Jingjing; Geng, Guohua; Guo, Hongbo

    2013-10-01

    As an important optical molecular imaging technique, bioluminescence tomography (BLT) offers an inexpensive and sensitive means for non-invasively imaging a variety of physiological and pathological activities at cellular and molecular levels in living small animals. The key problem of BLT is to recover the distribution of the internal bioluminescence sources from limited measurements on the surface. Considering the sparsity of the light source distribution, we directly formulate the inverse problem of BLT into an l0-norm minimization model and present a smoothed l0-norm (SL0) based reconstruction algorithm. By approximating the discontinuous l0 norm with a suitable continuous function, the SL0 norm method solves the problem of intractable computational load of the minimal l0 search as well as high sensitivity of l0-norm to noise. Numerical experiments on a mouse atlas demonstrate that the proposed SL0 norm based reconstruction method can obtain whole domain reconstruction without any a priori knowledge of the source permissible region, yielding almost the same reconstruction results to those of l1 norm methods.

  1. CMOS SPAD-based image sensor for single photon counting and time of flight imaging

    Dutton, Neale Arthur William

    2016-01-01

    The facility to capture the arrival of a single photon, is the fundamental limit to the detection of quantised electromagnetic radiation. An image sensor capable of capturing a picture with this ultimate optical and temporal precision is the pinnacle of photo-sensing. The creation of high spatial resolution, single photon sensitive, and time-resolved image sensors in complementary metal oxide semiconductor (CMOS) technology offers numerous benefits in a wide field of applications....

  2. Cardiac Time Intervals by Tissue Doppler Imaging M-Mode

    Biering-Sørensen, Tor; Mogelvang, Rasmus; de Knegt, Martina Chantal;

    2016-01-01

    PURPOSE: To define normal values of the cardiac time intervals obtained by tissue Doppler imaging (TDI) M-mode through the mitral valve (MV). Furthermore, to evaluate the association of the myocardial performance index (MPI) obtained by TDI M-mode (MPITDI) and the conventional method of obtaining...... MPI (MPIConv), with established echocardiographic and invasive measures of systolic and diastolic function. METHODS: In a large community based population study (n = 974), where all are free of any cardiovascular disease and cardiovascular risk factors, cardiac time intervals, including isovolumic...... relaxation time (IVRT), isovolumic contraction time (IVCT), and ejection time (ET) were obtained by TDI M-mode through the MV. IVCT/ET, IVRT/ET and the MPI ((IVRT+IVCT)/ET) were calculated. We also included a validation population (n = 44) of patients who underwent left heart catheterization and had the...

  3. Robust real time extraction of plane segments from time-of-flight camera images

    Dalbah, Yosef; Koltermann, Dirk; Wahl, Friedrich M.

    2014-04-01

    We present a method that extracts plane segments from images of a time-of-flight camera. Future driver assistance systems rely on an accurate description of the vehicle's environment. Time-of-flight cameras can be used for environment perception and for the reconstruction of the environment. Since most structures in urban environments are planar, extracted plane segments from single camera images can be used for the creation of a global map. We present a method for real time detection of planar surface structures from time-of-flight camera data. The concept is based on a planar surface segmentation that serves as the fundament for a subsequent global planar surface extraction. The evaluation demonstrates the ability of the described algorithm to detect planar surfaces form depth data of complex scenarios in real time. We compare our methods to state of the art planar surface extraction algorithms.

  4. Imaging cell biology in live animals: Ready for prime time

    Weigert, Roberto; Porat-Shliom, Natalie; Amornphimoltham, Panomwat

    2013-01-01

    Time-lapse fluorescence microscopy is one of the main tools used to image subcellular structures in living cells. Yet for decades it has been applied primarily to in vitro model systems. Thanks to the most recent advancements in intravital microscopy, this approach has finally been extended to live rodents. This represents a major breakthrough that will provide unprecedented new opportunities to study mammalian cell biology in vivo and has already provided new insight in the fields of neurobi...

  5. Time-resolved neutron imaging at ANTARES cold neutron beamline

    Tremsin, A.S.; Dangendorf, V.; Tittelmeier, K.; Schillinger, B.; Schulz, M.; Lerche, M.; Feller, W. B.

    2015-01-01

    In non-destructive evaluation with X-rays light elements embedded in dense, heavy (or high-Z) matrices show little contrast and their structural details can hardly be revealed. Neutron radiography, on the other hand, provides a solution for those cases, in particular for hydrogenous materials, owing to the large neutron scattering cross section of hydrogen and uncorrelated dependency of neutron cross section on the atomic number. The majority of neutron imaging experiments at the present time...

  6. Imaging gene expression in real-time using aptamers

    Shin, Ilchung [Iowa State Univ., Ames, IA (United States)

    2012-01-01

    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

  7. Imaging gene expression in real-time using aptamers

    Shin, Il Chung [Iowa State Univ., Ames, IA (United States)

    2011-01-01

    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

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

    Vânia da Silva Nunes-Halldorson; Norma Letícia Duran

    2003-01-01

    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...

  9. Change detection in a time series of polarimetric SAR images

    Skriver, Henning; Nielsen, Allan Aasbjerg; Conradsen, Knut

    a certain point can be used to detect at which points changes occur in the time series. [1] T. W. Anderson, An Introduction to Multivariate Statistical Analysis, John Wiley, New York, third edition, 2003. [2] K. Conradsen, A. A. Nielsen, J. Schou, and H. Skriver, “A test statistic in the complex...... to the complex Wishart distribution and demonstrate its application to change detection in truly multi-temporal, polarimetric SAR data. Results will be shown that demonstrate the difference between applying to time series of polarimetric SAR images, pairwise comparisons or the new omnibus test...

  10. Integration of image exposure time into a modified laser speckle imaging method

    Speckle-based methods have been developed to characterize tissue blood flow and perfusion. One such method, called modified laser speckle imaging (mLSI), enables computation of blood flow maps with relatively high spatial resolution. Although it is known that the sensitivity and noise in LSI measurements depend on image exposure time, a fundamental disadvantage of mLSI is that it does not take into account this parameter. In this work, we integrate the exposure time into the mLSI method and provide experimental support of our approach with measurements from an in vitro flow phantom.

  11. Time-resolved neutron imaging at ANTARES cold neutron beamline

    Tremsin, A. S.; Dangendorf, V.; Tittelmeier, K.; Schillinger, B.; Schulz, M.; Lerche, M.; Feller, W. B.

    2015-07-01

    In non-destructive evaluation with X-rays light elements embedded in dense, heavy (or high-Z) matrices show little contrast and their structural details can hardly be revealed. Neutron radiography, on the other hand, provides a solution for those cases, in particular for hydrogenous materials, owing to the large neutron scattering cross section of hydrogen and uncorrelated dependency of neutron cross section on the atomic number. The majority of neutron imaging experiments at the present time is conducted with static objects mainly due to the limited flux intensity of neutron beamline facilities and sometimes due to the limitations of the detectors. However, some applications require the studies of dynamic phenomena and can now be conducted at several high intensity beamlines such as the recently rebuilt ANTARES beam line at the FRM-II reactor. In this paper we demonstrate the capabilities of time resolved imaging for repetitive processes, where different phases of the process can be imaged simultaneously and integrated over multiple cycles. A fast MCP/Timepix neutron counting detector was used to image the water distribution within a model steam engine operating at 10 Hz frequency. Within neutron radiography for the future applications. The neutron spectrum of the ANTARES beamline as well as transmission spectra of a Fe sample were also measured with the Time Of Flight (TOF) technique in combination with a high resolution beam chopper. The energy resolution of our setup was found to be ~ 0.8% at 5 meV and ~ 1.7% at 25 meV. The background level (most likely gammas and epithermal/fast neutrons) of the ANTARES beamline was also measured in our experiments and found to be on the scale of 3% when no filters are installed in the beam. Online supplementary data available from stacks.iop.org/jinst/10/P07008/mmedia. The videos are given as supplementary material linked to the main article.

  12. Precision cosmology with time delay lenses: High resolution imaging requirements

    Meng, Xiao -Lei [Beijing Normal Univ., Beijing (China); Univ. of California, Santa Barbara, CA (United States); Treu, Tommaso [Univ. of California, Santa Barbara, CA (United States); Univ. of California, Los Angeles, CA (United States); Agnello, Adriano [Univ. of California, Santa Barbara, CA (United States); Univ. of California, Los Angeles, CA (United States); Auger, Matthew W. [Univ. of Cambridge, Cambridge (United Kingdom); Liao, Kai [Beijing Normal Univ., Beijing (China); Univ. of California, Santa Barbara, CA (United States); Univ. of California, Los Angeles, CA (United States); Marshall, Philip J. [Stanford Univ., Stanford, CA (United States)

    2015-09-28

    Lens time delays are a powerful probe of cosmology, provided that the gravitational potential of the main deflector can be modeled with sufficient precision. Recent work has shown that this can be achieved by detailed modeling of the host galaxies of lensed quasars, which appear as ``Einstein Rings'' in high resolution images. The distortion of these arcs and counter-arcs, as measured over a large number of pixels, provides tight constraints on the difference between the gravitational potential between the quasar image positions, and thus on cosmology in combination with the measured time delay. We carry out a systematic exploration of the high resolution imaging required to exploit the thousands of lensed quasars that will be discovered by current and upcoming surveys with the next decade. Specifically, we simulate realistic lens systems as imaged by the Hubble Space Telescope (HST), James Webb Space Telescope (JWST), and ground based adaptive optics images taken with Keck or the Thirty Meter Telescope (TMT). We compare the performance of these pointed observations with that of images taken by the Euclid (VIS), Wide-Field Infrared Survey Telescope (WFIRST) and Large Synoptic Survey Telescope (LSST) surveys. We use as our metric the precision with which the slope γ' of the total mass density profile ρtot∝ r–γ' for the main deflector can be measured. Ideally, we require that the statistical error on γ' be less than 0.02, such that it is subdominant to other sources of random and systematic uncertainties. We find that survey data will likely have sufficient depth and resolution to meet the target only for the brighter gravitational lens systems, comparable to those discovered by the SDSS survey. For fainter systems, that will be discovered by current and future surveys, targeted follow-up will be required. Furthermore, the exposure time required with upcoming facilitites such as JWST, the Keck Next Generation Adaptive

  13. Precision cosmology with time delay lenses: high resolution imaging requirements

    Meng, Xiao-Lei; Treu, Tommaso; Agnello, Adriano; Auger, Matthew W.; Liao, Kai; Marshall, Philip J.

    2015-09-01

    Lens time delays are a powerful probe of cosmology, provided that the gravitational potential of the main deflector can be modeled with sufficient precision. Recent work has shown that this can be achieved by detailed modeling of the host galaxies of lensed quasars, which appear as ``Einstein Rings'' in high resolution images. The distortion of these arcs and counter-arcs, as measured over a large number of pixels, provides tight constraints on the difference between the gravitational potential between the quasar image positions, and thus on cosmology in combination with the measured time delay. We carry out a systematic exploration of the high resolution imaging required to exploit the thousands of lensed quasars that will be discovered by current and upcoming surveys with the next decade. Specifically, we simulate realistic lens systems as imaged by the Hubble Space Telescope (HST), James Webb Space Telescope (JWST), and ground based adaptive optics images taken with Keck or the Thirty Meter Telescope (TMT). We compare the performance of these pointed observations with that of images taken by the Euclid (VIS), Wide-Field Infrared Survey Telescope (WFIRST) and Large Synoptic Survey Telescope (LSST) surveys. We use as our metric the precision with which the slope γ' of the total mass density profile ρtotpropto r-γ' for the main deflector can be measured. Ideally, we require that the statistical error on γ' be less than 0.02, such that it is subdominant to other sources of random and systematic uncertainties. We find that survey data will likely have sufficient depth and resolution to meet the target only for the brighter gravitational lens systems, comparable to those discovered by the SDSS survey. For fainter systems, that will be discovered by current and future surveys, targeted follow-up will be required. However, the exposure time required with upcoming facilitites such as JWST, the Keck Next Generation Adaptive Optics System, and TMT, will only be of

  14. Real-time Imaging Orientation Determination System to Verify Imaging Polarization Navigation Algorithm.

    Lu, Hao; Zhao, Kaichun; Wang, Xiaochu; You, Zheng; Huang, Kaoli

    2016-01-01

    Bio-inspired imaging polarization navigation which can provide navigation information and is capable of sensing polarization information has advantages of high-precision and anti-interference over polarization navigation sensors that use photodiodes. Although all types of imaging polarimeters exist, they may not qualify for the research on the imaging polarization navigation algorithm. To verify the algorithm, a real-time imaging orientation determination system was designed and implemented. Essential calibration procedures for the type of system that contained camera parameter calibration and the inconsistency of complementary metal oxide semiconductor calibration were discussed, designed, and implemented. Calibration results were used to undistort and rectify the multi-camera system. An orientation determination experiment was conducted. The results indicated that the system could acquire and compute the polarized skylight images throughout the calibrations and resolve orientation by the algorithm to verify in real-time. An orientation determination algorithm based on image processing was tested on the system. The performance and properties of the algorithm were evaluated. The rate of the algorithm was over 1 Hz, the error was over 0.313°, and the population standard deviation was 0.148° without any data filter. PMID:26805851

  15. Time-resolved spectral imaging: better photon economy, higher accuracy

    Fereidouni, Farzad; Reitsma, Keimpe; Blab, Gerhard A.; Gerritsen, Hans C.

    2015-03-01

    Lifetime and spectral imaging are complementary techniques that offer a non-invasive solution for monitoring metabolic processes, identifying biochemical compounds, and characterizing their interactions in biological tissues, among other tasks. Newly developed instruments that perform time-resolved spectral imaging can provide even more information and reach higher sensitivity than either modality alone. Here we report a multispectral lifetime imaging system based on a field-programmable gate array (FPGA), capable of operating at high photon count rates (12 MHz) per spectral detection channel, and with time resolution of 200 ps. We performed error analyses to investigate the effect of gate width and spectral-channel width on the accuracy of estimated lifetimes and spectral widths. Temporal and spectral phasors were used for analysis of recorded data, and we demonstrated blind un-mixing of the fluorescent components using information from both modalities. Fractional intensities, spectra, and decay curves of components were extracted without need for prior information. We further tested this approach with fluorescently doubly-labeled DNA, and demonstrated its suitability for accurately estimating FRET efficiency in the presence of either non-interacting or interacting donor molecules.

  16. Real-time Image Generation for Compressive Light Field Displays

    With the invention of integral imaging and parallax barriers in the beginning of the 20th century, glasses-free 3D displays have become feasible. Only today—more than a century later—glasses-free 3D displays are finally emerging in the consumer market. The technologies being employed in current-generation devices, however, are fundamentally the same as what was invented 100 years ago. With rapid advances in optical fabrication, digital processing power, and computational perception, a new generation of display technology is emerging: compressive displays exploring the co-design of optical elements and computational processing while taking particular characteristics of the human visual system into account. In this paper, we discuss real-time implementation strategies for emerging compressive light field displays. We consider displays composed of multiple stacked layers of light-attenuating or polarization-rotating layers, such as LCDs. The involved image generation requires iterative tomographic image synthesis. We demonstrate that, for the case of light field display, computed tomographic light field synthesis maps well to operations included in the standard graphics pipeline, facilitating efficient GPU-based implementations with real-time framerates.

  17. High-temperature real-time ultrasonic imaging

    Ultrasonic instrumentation capable of real-time imaging of materials at temperatures up to 2880C (5900F) 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

  18. Real time blood testing using quantitative phase imaging.

    Pham, Hoa V; Bhaduri, Basanta; Tangella, Krishnarao; Best-Popescu, Catherine; Popescu, Gabriel

    2013-01-01

    We demonstrate a real-time blood testing system that can provide remote diagnosis with minimal human intervention in economically challenged areas. Our instrument combines novel advances in label-free optical imaging with parallel computing. Specifically, we use quantitative phase imaging for extracting red blood cell morphology with nanoscale sensitivity and NVIDIA's CUDA programming language to perform real time cellular-level analysis. While the blood smear is translated through focus, our system is able to segment and analyze all the cells in the one megapixel field of view, at a rate of 40 frames/s. The variety of diagnostic parameters measured from each cell (e.g., surface area, sphericity, and minimum cylindrical diameter) are currently not available with current state of the art clinical instruments. In addition, we show that our instrument correctly recovers the red blood cell volume distribution, as evidenced by the excellent agreement with the cell counter results obtained on normal patients and those with microcytic and macrocytic anemia. The final data outputted by our instrument represent arrays of numbers associated with these morphological parameters and not images. Thus, the memory necessary to store these data is of the order of kilobytes, which allows for their remote transmission via, for example, the cellular network. We envision that such a system will dramatically increase access for blood testing and furthermore, may pave the way to digital hematology. PMID:23405194

  19. Capsule endoscopy: Improving transit time and image view

    Zvi Fireman; D Paz; Y Kopelman

    2005-01-01

    AIM: To evaluate the effect of various methods of small bowel preparation on the transit time and the quality of visualization of the entire small bowel mucosa.METHODS: Ninety-five patients underwent capsule endoscopy (CE) by easily swallowing the capsule. They were divided into three study groups according to the preparation used: group A (n = 26) by polyethylene glycol (PEG) liter or with sodium phosphate (SP) 12 h prior to the CE study; group B (n = 29) by erythromycin 1 h prior to the CE study; and group C (n = 40) without any preparation. Visualization ranged from good to satisfactory to poor.RESULTS: The gastric emptying time in the group prepared with erythromycin was shorter but without statistical significance and the small bowel transit time was unaffected. In elderly subjects prepared by PEG or SP, the gastric emptying time was significantly longer (163.7 min, P = 0.05). The transit times of the three sub-groups were not affected by gender or pathology.The grade of cleaning of the entire study group was 3.27±1.1. The erythromycin group presented significantly the worst quality of images (P = 0.05) compared to the other sub-groups. Age, gender, and pathology had no effect on the quality of the cleaning of the small bowel in the sub-groups. One (1.05%) case had no natural excretion.CONCLUSION: Erythromycin markedly reduces gastric emptying time, but has a negative effect on the quality of the image in the small bowel. The preparation of elderly subjects with PEG or SP has a negative effect on the small bowel transit time.

  20. Real-time dynamic imaging of virus distribution in vivo.

    Sean E Hofherr

    Full Text Available The distribution of viruses and gene therapy vectors is difficult to assess in a living organism. For instance, trafficking in murine models can usually only be assessed after sacrificing the animal for tissue sectioning or extraction. These assays are laborious requiring whole animal sectioning to ascertain tissue localization. They also obviate the ability to perform longitudinal or kinetic studies in one animal. To track viruses after systemic infection, we have labeled adenoviruses with a near-infrared (NIR fluorophore and imaged these after intravenous injection in mice. Imaging was able to track and quantitate virus particles entering the jugular vein simultaneous with injection, appearing in the heart within 500 milliseconds, distributing in the bloodstream and throughout the animal within 7 seconds, and that the bulk of virus distribution was essentially complete within 3 minutes. These data provide the first in vivo real-time tracking of the rapid initial events of systemic virus infection.

  1. Satellite image time series simulation for environmental monitoring

    Guo, Tao

    2014-11-01

    The performance of environmental monitoring heavily depends on the availability of consecutive observation data and it turns out an increasing demand in remote sensing community for satellite image data in the sufficient resolution with respect to both spatial and temporal requirements, which appear to be conflictive and hard to tune tradeoffs. Multiple constellations could be a solution if without concerning cost, and thus it is so far interesting but very challenging to develop a method which can simultaneously improve both spatial and temporal details. There are some research efforts to deal with the problem from various aspects, a type of approaches is to enhance the spatial resolution using techniques of super resolution, pan-sharpen etc. which can produce good visual effects, but mostly cannot preserve spectral signatures and result in losing analytical value. Another type is to fill temporal frequency gaps by adopting time interpolation, which actually doesn't increase informative context at all. In this paper we presented a novel method to generate satellite images in higher spatial and temporal details, which further enables satellite image time series simulation. Our method starts with a pair of high-low resolution data set, and then a spatial registration is done by introducing LDA model to map high and low resolution pixels correspondingly. Afterwards, temporal change information is captured through a comparison of low resolution time series data, and the temporal change is then projected onto high resolution data plane and assigned to each high resolution pixel referring the predefined temporal change patterns of each type of ground objects to generate a simulated high resolution data. A preliminary experiment shows that our method can simulate a high resolution data with a good accuracy. We consider the contribution of our method is to enable timely monitoring of temporal changes through analysis of low resolution images time series only, and usage of

  2. Time-resolved neutron imaging at ANTARES cold neutron beamline

    In non-destructive evaluation with X-rays light elements embedded in dense, heavy (or high-Z) matrices show little contrast and their structural details can hardly be revealed. Neutron radiography, on the other hand, provides a solution for those cases, in particular for hydrogenous materials, owing to the large neutron scattering cross section of hydrogen and uncorrelated dependency of neutron cross section on the atomic number. The majority of neutron imaging experiments at the present time is conducted with static objects mainly due to the limited flux intensity of neutron beamline facilities and sometimes due to the limitations of the detectors. However, some applications require the studies of dynamic phenomena and can now be conducted at several high intensity beamlines such as the recently rebuilt ANTARES beam line at the FRM-II reactor. In this paper we demonstrate the capabilities of time resolved imaging for repetitive processes, where different phases of the process can be imaged simultaneously and integrated over multiple cycles. A fast MCP/Timepix neutron counting detector was used to image the water distribution within a model steam engine operating at 10 Hz frequency. Within <10 minutes integration the amount of water was measured as a function of cycle time with a sub-mm spatial resolution, thereby demonstrating the capabilities of time-resolved neutron radiography for the future applications. The neutron spectrum of the ANTARES beamline as well as transmission spectra of a Fe sample were also measured with the Time Of Flight (TOF) technique in combination with a high resolution beam chopper. The energy resolution of our setup was found to be ∼ 0.8% at 5 meV and ∼ 1.7% at 25 meV. The background level (most likely gammas and epithermal/fast neutrons) of the ANTARES beamline was also measured in our experiments and found to be on the scale of 3% when no filters are installed in the beam. Online supplementary data available from stacks

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

    Fakhri, Georges El

    2012-01-01

    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 99mTc and 18F concentrations and illustrate the utility of such approaches in two sequential SPECT-PET/CT studies that include 99mTc-MAA/18F-NaF and 99mTc-Pentetate/18F-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

  4. The influence of SHFEMF on bioluminescence of V. Harveyi

    Exposure of bacteria V. harveyi grown on agar medium to 7 HHz electromagnetic field changes the intensity of their luminescence. It is suggested that the dynamics of the luminescence change reflects the adaptation processes in the microorganisms which accompany the electromao.netic field effect. The changes observed may be attributed to the temperature dependence of bioluminescence

  5. The mechanism of electronic excitation in the bacterial bioluminescent reaction

    The current state of the problem of formation of the electron-excited product in the chemiluminescent reaction that underlies the bacterial luminescence is analysed. Various schemes of chemical transformations capable of producing a bacterial bioluminescence emitter are presented. The problem of excitation of secondary emitters is considered; two possible mechanisms of their excitation are analysed.

  6. Red-Shifted Aequorin Variants Incorporating Non-Canonical Amino Acids: Applications in In Vivo Imaging

    Grinstead, Kristen M.; Rowe, Laura; Ensor, Charles M.; Joel, Smita; Daftarian, Pirouz; Dikici, Emre; Zingg, Jean-Marc; Daunert, Sylvia

    2016-01-01

    The increased importance of in vivo diagnostics has posed new demands for imaging technologies. In that regard, there is a need for imaging molecules capable of expanding the applications of current state-of-the-art imaging in vivo diagnostics. To that end, there is a desire for new reporter molecules capable of providing strong signals, are non-toxic, and can be tailored to diagnose or monitor the progression of a number of diseases. Aequorin is a non-toxic photoprotein that can be used as a sensitive marker for bioluminescence in vivo imaging. The sensitivity of aequorin is due to the fact that bioluminescence is a rare phenomenon in nature and, therefore, it does not suffer from autofluorescence, which contributes to background emission. Emission of bioluminescence in the blue-region of the spectrum by aequorin only occurs when calcium, and its luciferin coelenterazine, are bound to the protein and trigger a biochemical reaction that results in light generation. It is this reaction that endows aequorin with unique characteristics, making it ideally suited for a number of applications in bioanalysis and imaging. Herein we report the site-specific incorporation of non-canonical or non-natural amino acids and several coelenterazine analogues, resulting in a catalog of 72 cysteine-free, aequorin variants which expand the potential applications of these photoproteins by providing several red-shifted mutants better suited to use in vivo. In vivo studies in mouse models using the transparent tissue of the eye confirmed the activity of the aequorin variants incorporating L-4-iodophehylalanine and L-4-methoxyphenylalanine after injection into the eye and topical addition of coelenterazine. The signal also remained localized within the eye. This is the first time that aequorin variants incorporating non-canonical amino acids have shown to be active in vivo and useful as reporters in bioluminescence imaging. PMID:27367859

  7. Red-Shifted Aequorin Variants Incorporating Non-Canonical Amino Acids: Applications in In Vivo Imaging.

    Kristen M Grinstead

    Full Text Available The increased importance of in vivo diagnostics has posed new demands for imaging technologies. In that regard, there is a need for imaging molecules capable of expanding the applications of current state-of-the-art imaging in vivo diagnostics. To that end, there is a desire for new reporter molecules capable of providing strong signals, are non-toxic, and can be tailored to diagnose or monitor the progression of a number of diseases. Aequorin is a non-toxic photoprotein that can be used as a sensitive marker for bioluminescence in vivo imaging. The sensitivity of aequorin is due to the fact that bioluminescence is a rare phenomenon in nature and, therefore, it does not suffer from autofluorescence, which contributes to background emission. Emission of bioluminescence in the blue-region of the spectrum by aequorin only occurs when calcium, and its luciferin coelenterazine, are bound to the protein and trigger a biochemical reaction that results in light generation. It is this reaction that endows aequorin with unique characteristics, making it ideally suited for a number of applications in bioanalysis and imaging. Herein we report the site-specific incorporation of non-canonical or non-natural amino acids and several coelenterazine analogues, resulting in a catalog of 72 cysteine-free, aequorin variants which expand the potential applications of these photoproteins by providing several red-shifted mutants better suited to use in vivo. In vivo studies in mouse models using the transparent tissue of the eye confirmed the activity of the aequorin variants incorporating L-4-iodophehylalanine and L-4-methoxyphenylalanine after injection into the eye and topical addition of coelenterazine. The signal also remained localized within the eye. This is the first time that aequorin variants incorporating non-canonical amino acids have shown to be active in vivo and useful as reporters in bioluminescence imaging.

  8. Microsecond time-resolved 2D X-ray imaging

    A method is presented which allows to take two-dimensional X-ray images of repetitive processes with recording times in the sub-microsecond range. Various measurements have been performed with a recently introduced novel two-dimensional single photon counter which has been slightly modified in order to determine the exact arrival time of each detected photon. For this purpose a special clock signal is synchronized with the process and is digitized contemporaneously with each event. This technique can be applied even with rate limited detectors and low flux sources, since--unlike in conventional methods, where chopped beams or gated read out electronics are used--all photons are used for the image formation. For the measurements, rapidly moving mechanical systems and conventional X-ray sources have been used, reaching time resolutions of some 10 μs. The technique presented here opens a variety of new biological, medical and industrial applications which will be discussed. As a first application example, three dimensional tomographic reconstructions of rapidly rotating objects (4000 turns/min) are presented

  9. Detection of heterogeneous substrate distributions in tumors and spheroids by bioluminescence

    Heterogeneous cell populations within solid tumors often limit non-surgical tumor therapies. Partially, the biological variability among cancer cells in vivo is attributable to a non-uniform oxygenation and pH distribution as a consequence of spatial and temporal heterogeneities in the tumor microcirculation. In order to evaluate whether such inhomogeneities may also be found in the distribution of nutrients and metabolites, a method, originally developed for the determination of regional substrate distributions in brain tissue; has been applied to cryobiopsies of human tumor xenografts and of tumors in patients. In addition, this method has been adapted to multicellular tumor spheroids of human origin. The bioluminescence reactions are enzymatically linked to the substrate of interest. A cold cyrostat section of the frozen enzyme solution is laid upon a frozen cryostat section of a tumor or of a spheroid. Bioluminescence recorded by film exposure occurs upon thawing these sections. The exposed film is then evaluated by microdensitometry and by special image analysis. The regional distributions of glucose, lactate and ATP are obtained in relative units. The results show that all substances investigated exhibit large regional differences reflecting a great heterogeneity of the metabolic micromilieu in malignant tumors and even within tumor spheroids

  10. A novel real time imaging platform to quantify macrophage phagocytosis.

    Kapellos, Theodore S; Taylor, Lewis; Lee, Heyne; Cowley, Sally A; James, William S; Iqbal, Asif J; Greaves, David R

    2016-09-15

    Phagocytosis of pathogens, apoptotic cells and debris is a key feature of macrophage function in host defense and tissue homeostasis. Quantification of macrophage phagocytosis in vitro has traditionally been technically challenging. Here we report the optimization and validation of the IncuCyte ZOOM® real time imaging platform for macrophage phagocytosis based on pHrodo® pathogen bioparticles, which only fluoresce when localized in the acidic environment of the phagolysosome. Image analysis and fluorescence quantification were performed with the automated IncuCyte™ Basic Software. Titration of the bioparticle number showed that the system is more sensitive than a spectrofluorometer, as it can detect phagocytosis when using 20× less E. coli bioparticles. We exemplified the power of this real time imaging platform by studying phagocytosis of murine alveolar, bone marrow and peritoneal macrophages. We further demonstrate the ability of this platform to study modulation of the phagocytic process, as pharmacological inhibitors of phagocytosis suppressed bioparticle uptake in a concentration-dependent manner, whereas opsonins augmented phagocytosis. We also investigated the effects of macrophage polarization on E. coli phagocytosis. Bone marrow-derived macrophage (BMDM) priming with M2 stimuli, such as IL-4 and IL-10 resulted in higher engulfment of bioparticles in comparison with M1 polarization. Moreover, we demonstrated that tolerization of BMDMs with lipopolysaccharide (LPS) results in impaired E. coli bioparticle phagocytosis. This novel real time assay will enable researchers to quantify macrophage phagocytosis with a higher degree of accuracy and sensitivity and will allow investigation of limited populations of primary phagocytes in vitro. PMID:27475716

  11. Improved site contamination through time-lapse complex resistivity imaging

    Flores Orozco, Adrian; Kemna, Andreas; Cassiani, Giorgio; Binley, Andrew

    2016-04-01

    In the framework of the EU FP7 project ModelPROBE, time-lapse complex resistivity (CR) measurements were conducted at a test site close to Trecate (NW Italy). The objective was to investigate the capabilities of the CR imaging method to delineate the geometry and dynamics of subsurface hydrocarbon contaminant plume which resulted from a crude oil spill in 1994. To achieve this it is required to discriminate the electrical signal associated to static (i.e., lithology) from dynamic changes in the subsurface, with the latter associated to significant seasonal groundwater fluctuations. Previous studies have demonstrated the benefits of the CR method to gain information which is not accessible with common electrical resistivity tomography. However field applications are still rarely and neither the analysis of the data error for CR time-lapse measurements, nor the inversion itself haven not received enough attention. While the ultimate objective at the site is to characterize, here we address the discrimination of the lithological and hydrological controls on the IP response by considering data collected in an uncontaminated area of the site. In this study we demonstrate that an adequate error description of CR measurements provides images free of artifacts and quantitative superior than previous approaches. Based on this approach, differential images computed for time-lapse data exhibited anomalies well correlated with spatiotemporal changes correlated to seasonal fluctuations in the groundwater level. The proposed analysis may be useful in the characterization of fate and transport of hydrocarbon contaminants relevant for the site, which presents areas contaminated with crude oil.

  12. Real-time underwater image enhancement: An improved approach for imaging with AUV-150

    Jeet Banerjee; Ranjit Ray; Siva Ram Krishna Vadali; Sankar Nath Shome; Sambhunath Nandy

    2016-02-01

    An RGB YCbCr Processing method (RYPro) is proposed for underwater images commonly suffering from low contrast and poor color quality. The degradation in image quality may be attributed to absorption and backscattering of light by suspended underwater particles. Moreover, as the depth increases, different colors are absorbed by the surrounding medium depending on the wavelengths. In particular, blue/green color is dominant in the underwater ambience which is known as color cast. For further processing of the image, enhancement remains an essential preprocessing operation. Color equalization is a widely adopted approach for underwater image enhancement. Traditional methods normally involve blind color equalization for enhancing the image under test. In the present work, processing sequence of the proposed method includes noise removal using linear and non-linear filters followed by adaptive contrast correction in the RGB and YCbCr color planes. Performance of the proposed method is evaluated and compared with three golden methods, namely, Gray World (GW), White Patch (WP), Adobe Photoshop Equalization (APE) and a recently developed method entitled “Unsupervised Color Correction Method (UCM)”. In view of its simplicity and computational ease, the proposed method is recommended for real-time applications. Suitability of the proposed method is validated by real-time implementation during the testing of the Autonomous Underwater Vehicle (AUV-150) developed indigenously by CSIR-CMERI.

  13. Near infrared bioluminescence resonance energy transfer from firefly luciferase—quantum dot bionanoconjugates

    The bioluminescence resonance energy transfer (BRET) between firefly luciferase enzymes and semiconductive quantum dots (QDs) with near infrared emission is described. The QD were phase transferred to aqueous buffers using a histidine mediated phase transfer route, and incubated with a hexahistidine tagged, green emitting variant of firefly luciferase from Photinus pyralis (PPyGRTS). The PPyGRTS were bound to the QD interface via the hexahistidine tag, which effectively displaces the histidine layer and binds directly to the QD interfaces, allowing for short donor–acceptor distances (∼5.5 nm). Due to this, high BRET efficiency ratios of ∼5 were obtained. These PPyGRTS-QD bio-nano conjugates were characterized by transmission electron microscopy, thermal gravimetric analysis, Fourier transform infrared spectroscopy and BRET emission studies. The final optimized conjugate was easily observable by night vision imaging, demonstrating the potential of these materials in imaging and signaling/sensing applications. (paper)

  14. Near infrared bioluminescence resonance energy transfer from firefly luciferase—quantum dot bionanoconjugates

    Alam, Rabeka; Karam, Liliana M.; Doane, Tennyson L.; Zylstra, Joshua; Fontaine, Danielle M.; Branchini, Bruce R.; Maye, Mathew M.

    2014-12-01

    The bioluminescence resonance energy transfer (BRET) between firefly luciferase enzymes and semiconductive quantum dots (QDs) with near infrared emission is described. The QD were phase transferred to aqueous buffers using a histidine mediated phase transfer route, and incubated with a hexahistidine tagged, green emitting variant of firefly luciferase from Photinus pyralis (PPyGRTS). The PPyGRTS were bound to the QD interface via the hexahistidine tag, which effectively displaces the histidine layer and binds directly to the QD interfaces, allowing for short donor-acceptor distances (˜5.5 nm). Due to this, high BRET efficiency ratios of ˜5 were obtained. These PPyGRTS-QD bio-nano conjugates were characterized by transmission electron microscopy, thermal gravimetric analysis, Fourier transform infrared spectroscopy and BRET emission studies. The final optimized conjugate was easily observable by night vision imaging, demonstrating the potential of these materials in imaging and signaling/sensing applications.

  15. Polydiacetylene Liposomal Aequorin Bioluminescent Device for Detection of Hydrophobic Compounds.

    Yamamoto, Ryoko; Takegami, Shigehiko; Konishi, Atsuko; Horikawa, Hikari; Yonezawa, Sayumi; Kitade, Tatsuya

    2016-06-01

    In this study, a polydiacetylene liposomal aequorin bioluminescent device (PLABD) that functioned through control of the membrane transport of Ca(2+) ions was developed for detecting hydrophobic compounds. In the PLABD, aequorin was encapsulated in an internal water phase and a calcium ionophore (CI) was contained in a hydrophobic region. Membrane transport of Ca(2+) ions across the CI was suppressed by polymerization between diacetylene molecules. On addition of an analyte, the membrane transport of Ca(2+) ions across the CI increased, and Ca(2+) ions from the external water phase could diffuse into the internal water phase via the CI, which resulted in bioluminescence of the aequorin. Lidocaine, procaine, and procainamide were used as model compounds to test the validity of the detection mechanism of the PLABD. When each analyte was added to a suspension of the PLABD, bioluminescence from the aequorin in the PLABD was observed, and the level of this bioluminescence increased with increasing analyte concentration. There was a linear relationship between the logarithm of the analyte concentration and the bioluminescence for all analytes as follows: R = 0.89 from 10 nmol L(-1) to 10 mmol L(-1) for lidocaine, R = 0.66 from 10 nmol L(-1) to 100 μmol L(-1) for procaine, and R = 0.74 from 100 nmol L(-1) to 100 μmol L(-1) for procainamide. Compared to the traditional colorimetric method using polydiacetylene liposome, the PLABD was superior for both the sensitivity and dynamic range. Thus, PLABD is a valid, simple, and sensitive signal generator for detection of hydrophobic compounds that interact with PLABD membranes. PMID:27146598

  16. HOMOGENEOUS MULTISTAGE ARCHITECTURE FOR REAL-TIME IMAGE PROCESSING

    Hanen Chenini

    2012-12-01

    Full Text Available In this article, we present a new multistage architecture oriented to real-time complex processing applications. Given a set of rules, this proposed architecture allows the using of different communication links (point to point link, hardware router… to connect unlimited number of parallel computing elements (software processors to follow the increasing complexity of algorithms. In particular, this work brings out a parallel implementation of multihypothesis approach for road recognition application on the proposed Multiprocessor Systemon-Chip (MP-SoC architecture. This algorithm is usually the main part of the lane keeping applications. Experimental results using images of a real road scene are presented. Using a low cost FPGA-based System-on-Chip, our hardware architecture is able to detect and recognize the roadsides in a time limit of 60 mSec. Moreover, we demonstrate that our multistage architecture may be used to achieve good speed-up in solving automotive applications.

  17. Integrated CMOS photodetectors and signal processing for very low-level chemical sensing with the bioluminescent bioreporter integrated circuit

    Bolton, Eric K.; Sayler, Gary S.; Nivens, David E.; Rochelle, James M.; Ripp, Steven; Simpson, Michael L.

    2002-01-01

    We report an integrated CMOS microluminometer optimized for the detection of low-level bioluminescence as part of the bioluminescent bioreporter integrated circuit (BBIC). This microluminometer improves on previous devices through careful management of the sub-femtoampere currents, both signal and leakage, that flow in the front-end processing circuitry. In particular, the photodiode is operated with a reverse bias of only a few mV, requiring special attention to the reset circuitry of the current-to-frequency converter (CFC) that forms the front-end circuit. We report a sub-femtoampere leakage current and a minimum detectable signal (MDS) of 0.15 fA (1510 s integration time) using a room temperature 1.47 mm2 CMOS photodiode. This microluminometer can detect luminescence from as few as 5000 fully induced Pseudomonas fluorescens 5RL bacterial cells. c2002 Elsevier Science B.V. All rights reserved.

  18. Description of the larva of Alampoides alychnus (Kirsch, 1873, the first known species with bioluminescent immatures in Euplinthini (Elateridae, Agrypninae

    Simone Policena Rosa

    2013-01-01

    Full Text Available Mature larva of Alampoides alychnus (Kirsch is described and compared to known Pyrophorini immatures. Larvae were collected live in the soil of a region dominated by sugarcane plantation and gallery forest in Campo Novo dos Parecis, Mato Grosso, Brazil. They were maintained in laboratory and the pupal period lasted 14 days. This larva differs from other Pyrophorini larvae mainly by bioluminescent pattern: one pair of luminous spots on the mesonotum, and a longitudinal series of median spots on the metanotum and all abdominal segments. The morphology of larva and the bioluminescent pattern of larva and pupa are described for the first time to the genus and the tribe. The fact that adults show no trace of luminescence is emphasized.

  19. Advanced Imaging by Space-Time Deconvolution in Array GPR

    Savelyev, T. G.; van Tol, N. T.; Yarovoy, A. G.; Ligthart, L. P.

    Digital beamforming in array-based UWB radar delivers a high-resolution 3-D image of subsurface in GPR landmine detection while simultaneous data acquisition by elements of the array significantly increases the scanning speed. Such a GPR system with a single transmit antenna and a linear receive array has been developed in the Delft University of Technology. For online processing we propose an advanced imaging algorithm based on migration by regularized, parametric space-time deconvolution. The algorithm deconvolves a 3-D space-time array point spread function out of the data volume by means of FFT and inverse Wiener filter that is being controlled automatically with numerical criteria for stability and accuracy. The prior knowledge of GPR impulse response and ground impulse response is used to form a separate point spread function for each receiving antenna. The developed technique has been verified on experimental data for typical anti-personnel mines and compared with a classical migration by diffraction stacking.

  20. Satellite Image Time Series Decomposition Based on EEMD

    Yun-long Kong

    2015-11-01

    Full Text Available Satellite Image Time Series (SITS have recently been of great interest due to the emerging remote sensing capabilities for Earth observation. Trend and seasonal components are two crucial elements of SITS. In this paper, a novel framework of SITS decomposition based on Ensemble Empirical Mode Decomposition (EEMD is proposed. EEMD is achieved by sifting an ensemble of adaptive orthogonal components called Intrinsic Mode Functions (IMFs. EEMD is noise-assisted and overcomes the drawback of mode mixing in conventional Empirical Mode Decomposition (EMD. Inspired by these advantages, the aim of this work is to employ EEMD to decompose SITS into IMFs and to choose relevant IMFs for the separation of seasonal and trend components. In a series of simulations, IMFs extracted by EEMD achieved a clear representation with physical meaning. The experimental results of 16-day compositions of Moderate Resolution Imaging Spectroradiometer (MODIS, Normalized Difference Vegetation Index (NDVI, and Global Environment Monitoring Index (GEMI time series with disturbance illustrated the effectiveness and stability of the proposed approach to monitoring tasks, such as applications for the detection of abrupt changes.

  1. Time-Domain Fluorescence Lifetime Imaging Techniques Suitable for Solid-State Imaging Sensor Arrays

    Robert K. Henderson

    2012-05-01

    Full Text Available We have successfully demonstrated video-rate CMOS single-photon avalanche diode (SPAD-based cameras for fluorescence lifetime imaging microscopy (FLIM by applying innovative FLIM algorithms. We also review and compare several time-domain techniques and solid-state FLIM systems, and adapt the proposed algorithms for massive CMOS SPAD-based arrays and hardware implementations. The theoretical error equations are derived and their performances are demonstrated on the data obtained from 0.13 μm CMOS SPAD arrays and the multiple-decay data obtained from scanning PMT systems. In vivo two photon fluorescence lifetime imaging data of FITC-albumin labeled vasculature of a P22 rat carcinosarcoma (BD9 rat window chamber are used to test how different algorithms perform on bi-decay data. The proposed techniques are capable of producing lifetime images with enough contrast.

  2. Validation of constitutively expressed bioluminescent Pseudomonas aeruginosa as a rapid microbiological quantification tool.

    Shah, N; Naseby, D C

    2015-06-15

    Whole cell biosensors have been extensively used for monitoring toxicity and contamination of various compounds and xenobiotics in environmental biology and microbial ecology; their application in the pharmaceutical and cosmetics industries has been limited. According to several pharmacopoeias, pharmaceutical products must be tested for microbial activity using traditional viable count techniques; the use of whole cell microbial biosensors potentially provides an alternative, fast, and efficient method. However there is a lack of a validated bioluminescence method. Prototype whole cell microbial biosensors have already been developed in Pseudomonas aeruginosa ATCC 9027. Validation of the bioluminescent strains was performed in accordance with the pharmacopoeia, Parenteral Drug Association and International Organisation of Standardisation. These strains demonstrated that the bioluminescent method was accurate, precise and equivalent, as compared with plate counting at a range of 10(3)-10(7) CFU/mL. Percentage recoveries using the bioluminescent method were between 70% and 130% for all bioluminescent strains and therefore the bioluminescent method was accurate according to the criteria set in PDA technical report 33. The method was also more precise (relative standard deviation less than 15%) than the traditional plate counting method or the ATP bioluminescent method. The lower limit of detection was 10(3) CFU/mL. Two-way ANOVA showed no significant difference between the traditional plate counting and the novel bioluminescent method for all bioluminescent strains. The bioluminescent constructs passed/exceeded pharmacopoeia-specified criteria for range, limit of detection, accuracy, precision and equivalence. PMID:25618377

  3. BLProt: Prediction of bioluminescent proteins based on support vector machine and relieff feature selection

    Kandaswamy, Krishna Kumar

    2011-08-17

    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. 2011 Kandaswamy et al; licensee BioMed Central Ltd.

  4. Ultrasound contrast agent imaging: Real-time imaging of the superharmonics

    Peruzzini, D.; Viti, J. [MSD lab, Department of Information Engineering, Univ of Florence, Via S.Marta, 3, 50139 Firenze (Italy); Erasmus MC, ’s-Gravendijkwal 230, Faculty Building, Ee 2302, 3015 CE Rotterdam (Netherlands); Tortoli, P. [MSD lab, Department of Information Engineering, Univ of Florence, Via S.Marta, 3, 50139 Firenze (Italy); Verweij, M. D. [Acoustical Wavefield Imaging, ImPhys, Delft Univ Technology, van der Waalsweg 8, 2628 CH Delft (Netherlands); Jong, N. de; Vos, H. J., E-mail: h.vos@erasmusmc.nl [Erasmus MC, ’s-Gravendijkwal 230, Faculty Building, Ee 2302, 3015 CE Rotterdam (Netherlands); Acoustical Wavefield Imaging, ImPhys, Delft Univ Technology, van der Waalsweg 8, 2628 CH Delft (Netherlands)

    2015-10-28

    Currently, in medical ultrasound contrast agent (UCA) imaging the second harmonic scattering of the microbubbles is regularly used. This scattering is in competition with the signal that is caused by nonlinear wave propagation in tissue. It was reported that UCA imaging based on the third or higher harmonics, i.e. “superharmonic” imaging, shows better contrast. However, the superharmonic scattering has a lower signal level compared to e.g. second harmonic signals. This study investigates the contrast-to-tissue ratio (CTR) and signal to noise ratio (SNR) of superharmonic UCA scattering in a tissue/vessel mimicking phantom using a real-time clinical scanner. Numerical simulations were performed to estimate the level of harmonics generated by the microbubbles. Data were acquired with a custom built dual-frequency cardiac phased array probe. Fundamental real-time images were produced while beam formed radiofrequency (RF) data was stored for further offline processing. The phantom consisted of a cavity filled with UCA surrounded by tissue mimicking material. The acoustic pressure in the cavity of the phantom was 110 kPa (MI = 0.11) ensuring non-destructivity of UCA. After processing of the acquired data from the phantom, the UCA-filled cavity could be clearly observed in the images, while tissue signals were suppressed at or below the noise floor. The measured CTR values were 36 dB, >38 dB, and >32 dB, for the second, third, and fourth harmonic respectively, which were in agreement with those reported earlier for preliminary contrast superharmonic imaging. The single frame SNR values (in which ‘signal’ denotes the signal level from the UCA area) were 23 dB, 18 dB, and 11 dB, respectively. This indicates that noise, and not the tissue signal, is the limiting factor for the UCA detection when using the superharmonics in nondestructive mode.

  5. Ultrasound contrast agent imaging: Real-time imaging of the superharmonics

    Currently, in medical ultrasound contrast agent (UCA) imaging the second harmonic scattering of the microbubbles is regularly used. This scattering is in competition with the signal that is caused by nonlinear wave propagation in tissue. It was reported that UCA imaging based on the third or higher harmonics, i.e. “superharmonic” imaging, shows better contrast. However, the superharmonic scattering has a lower signal level compared to e.g. second harmonic signals. This study investigates the contrast-to-tissue ratio (CTR) and signal to noise ratio (SNR) of superharmonic UCA scattering in a tissue/vessel mimicking phantom using a real-time clinical scanner. Numerical simulations were performed to estimate the level of harmonics generated by the microbubbles. Data were acquired with a custom built dual-frequency cardiac phased array probe. Fundamental real-time images were produced while beam formed radiofrequency (RF) data was stored for further offline processing. The phantom consisted of a cavity filled with UCA surrounded by tissue mimicking material. The acoustic pressure in the cavity of the phantom was 110 kPa (MI = 0.11) ensuring non-destructivity of UCA. After processing of the acquired data from the phantom, the UCA-filled cavity could be clearly observed in the images, while tissue signals were suppressed at or below the noise floor. The measured CTR values were 36 dB, >38 dB, and >32 dB, for the second, third, and fourth harmonic respectively, which were in agreement with those reported earlier for preliminary contrast superharmonic imaging. The single frame SNR values (in which ‘signal’ denotes the signal level from the UCA area) were 23 dB, 18 dB, and 11 dB, respectively. This indicates that noise, and not the tissue signal, is the limiting factor for the UCA detection when using the superharmonics in nondestructive mode

  6. Time Lapse Electrical Imaging of Salt Affected Soil and Groundwater

    Hayley, Kevin

    This research was conducted to test the hypothesis that time-lapse electrical resistivity imaging (ERI) could be used as an effective tool for monitoring remediation of salt affected soil and groundwater. A time-lapse 3D ERI survey conducted over the span of 3 years was used to observe changes in an oilfield brine plume undergoing remediation using a tile drainage network. A theoretical model was modified to incorporate the results of an electrical conductivity (EC) temperature laboratory experiment and a linear EC temperature relationship was calibrated for the 0--25°C range. Using the linear approximation and theoretical model observed temperature and saturation variations could be accounted for in bulk EC measurements. ERI inversion parameters were selected to produce a model with the best agreement with collocated push tool conductivity logs, and the strongest correlation with core salinity measurements. The ERI models were transformed into salinity estimates in order to produce salt concentration maps and salt removal estimates. A temperature correction method was developed that acted on the collected electrical resistivity data, rather than on the inversion model. A data correction was sought in order to remove the effect of inversion smoothing from the temperature correction and to compensate for temperature prior to inversion opening the possibility of using time-lapse inversion techniques. First the data were inverted to produce a model, and then the model was converted to a standard temperature equivalent model using the previously developed EC temperature relationship. The difference between synthetic datasets produced using the inversion model and the standard temperature equivalent model was used as a data temperature correction factor and applied to the collected data. A simultaneous time-lapse inversion method was developed and compared to previously published methods. The simultaneous approach consisted of inverting the time-lapse datasets together

  7. A method for generating high resolution satellite image time series

    Guo, Tao

    2014-10-01

    There is an increasing demand for satellite remote sensing data with both high spatial and temporal resolution in many applications. But it still is a challenge to simultaneously improve spatial resolution and temporal frequency due to the technical limits of current satellite observation systems. To this end, much R&D efforts have been ongoing for years and lead to some successes roughly in two aspects, one includes super resolution, pan-sharpen etc. methods which can effectively enhance the spatial resolution and generate good visual effects, but hardly preserve spectral signatures and result in inadequate analytical value, on the other hand, time interpolation is a straight forward method to increase temporal frequency, however it increase little informative contents in fact. In this paper we presented a novel method to simulate high resolution time series data by combing low resolution time series data and a very small number of high resolution data only. Our method starts with a pair of high and low resolution data set, and then a spatial registration is done by introducing LDA model to map high and low resolution pixels correspondingly. Afterwards, temporal change information is captured through a comparison of low resolution time series data, and then projected onto the high resolution data plane and assigned to each high resolution pixel according to the predefined temporal change patterns of each type of ground objects. Finally the simulated high resolution data is generated. A preliminary experiment shows that our method can simulate a high resolution data with a reasonable accuracy. The contribution of our method is to enable timely monitoring of temporal changes through analysis of time sequence of low resolution images only, and usage of costly high resolution data can be reduces as much as possible, and it presents a highly effective way to build up an economically operational monitoring solution for agriculture, forest, land use investigation

  8. Time-gated FLIM microscope for corneal metabolic imaging

    Silva, Susana F.; Batista, Ana; Domingues, José Paulo; Quadrado, Maria João.; Morgado, António Miguel

    2016-03-01

    Detecting corneal cells metabolic alterations may prove a valuable tool in the early diagnosis of corneal diseases. Nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) are autofluorescent metabolic co-factors that allow the assessment of metabolic changes through non-invasive optical methods. These co-factors exhibit double-exponential fluorescence decays, with well-separated short and lifetime components, which are related to their protein-bound and free-states. Corneal metabolism can be assessed by measuring the relative contributions of these two components. For that purpose, we have developed a wide-field time-gated fluorescence lifetime microscope based on structured illumination and one-photon excitation to record FAD lifetime images from corneas. NADH imaging was not considered as its UV excitation peak is regarded as not safe for in vivo measurements. The microscope relies on a pulsed blue diode laser (λ=443 nm) as excitation source, an ultra-high speed gated image intensifier coupled to a CCD camera to acquire fluorescence signals and a Digital Micromirror Device (DMD) to implement the Structured Illumination technique. The system has a lateral resolution better than 2.4 μm, a field of view of 160 per 120 μm and an optical sectioning of 6.91 +/- 0.45 μm when used with a 40x, 0.75 NA, Water Immersion Objective. With this setup we were able to measure FAD contributions from ex-vivo chicken corneas collected from a local slaughterhouse..

  9. Real-time synthetic aperture ultrasonic scroll-imaging system and imaging experiment for nuclear power plant NDT

    The authors discuss their development of a real-time synthetic aperture imaging system which provides a cross sectional image of an object in real-time. They have already confirmed the performance of the system by imaging experiments using the normal-beam method. To confirm the wide applicability of the system, they have carried out imaging experiments with the angle-beam method. They considered four different sound paths for angle-beam SAFT imaging. After calculating sound path lengths, computer simulation and experiments were carried out. It is shown that there were two dominant paths for image reconstruction caused by the directivity of the transducer

  10. Image Corruption Detection in Diffusion Tensor Imaging for Post-Processing and Real-Time Monitoring

    Li, Yue; Shea, Steven M.; Lorenz, Christine H.; Jiang, Hangyi; Chou, Ming-Chung; Mori, Susumu

    2013-01-01

    Due to the high sensitivity of diffusion tensor imaging (DTI) to physiological motion, clinical DTI scans often suffer a significant amount of artifacts. Tensor-fitting-based, post-processing outlier rejection is often used to reduce the influence of motion artifacts. Although it is an effective approach, when there are multiple corrupted data, this method may no longer correctly identify and reject the corrupted data. In this paper, we introduce a new criterion called “corrected Inter-Slice Intensity Discontinuity” (cISID) to detect motion-induced artifacts. We compared the performance of algorithms using cISID and other existing methods with regard to artifact detection. The experimental results show that the integration of cISID into fitting-based methods significantly improves the retrospective detection performance at post-processing analysis. The performance of the cISID criterion, if used alone, was inferior to the fitting-based methods, but cISID could effectively identify severely corrupted images with a rapid calculation time. In the second part of this paper, an outlier rejection scheme was implemented on a scanner for real-time monitoring of image quality and reacquisition of the corrupted data. The real-time monitoring, based on cISID and followed by post-processing, fitting-based outlier rejection, could provide a robust environment for routine DTI studies. PMID:24204551

  11. Real Time Deconvolution of In-Vivo Ultrasound Images

    Jensen, Jørgen Arendt

    2013-01-01

    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...... 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...

  12. Time-resolved multiphoton imaging of basal cell carcinoma

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

    2007-02-01

    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.

  13. Results from laboratory tests of the two-dimensional Time-Encoded Imaging System.

    Marleau, Peter; Brennan, James S.; Brubaker, Erik; Gerling, Mark D; Le Galloudec, Nathalie Joelle

    2014-09-01

    A series of laboratory experiments were undertaken to demonstrate the feasibility of two dimensional time-encoded imaging. A prototype two-dimensional time encoded imaging system was designed and constructed. Results from imaging measurements of single and multiple point sources as well as extended source distributions are presented. Time encoded imaging has proven to be a simple method for achieving high resolution two-dimensional imaging with potential to be used in future arms control and treaty verification applications.

  14. Bioluminescent Ligand-Receptor Binding Assays for Protein or Peptide Hormones.

    Liu, Ya-Li; Guo, Zhan-Yun

    2016-01-01

    Bioluminescence has been widely used in biomedical research due to its high sensitivity, low background, and broad linear range. In recent studies, we applied bioluminescence to ligand-receptor binding assays for some protein or peptide hormones based on a newly developed small monomeric Nanoluciferase (NanoLuc) reporter that has the so far brightest bioluminescence. The conventional ligand-receptor binding assays rely on radioligands that have drawbacks, such as radioactive hazards and short shelf lives. In contrast, the novel bioluminescent binding assays use the NanoLuc-based protein or peptide tracers that are safe, stable, and ultrasensitive. Thus, the novel bioluminescent ligand-receptor binding assay would be applied to more and more protein or peptide hormones for ligand-receptor interaction studies in future. In the present article, we provided detailed protocols for setting up the novel bioluminescent ligand-receptor binding assays using two representative protein hormones as examples. PMID:27424896

  15. Modelling dinoflagellates as an approach to the seasonal forecasting of bioluminescence in the North Atlantic

    Marcinko, Charlotte L. J.; Martin, Adrian P.; Allen, John T.

    2014-11-01

    Bioluminescence within ocean surface waters is of significant interest because it can enhance the study of subsurface movement and organisms. Little is known about how bioluminescence potential (BPOT) varies spatially and temporally in the open ocean. However, light emitted from dinoflagellates often dominates the stimulated bioluminescence field. As a first step towards forecasting surface ocean bioluminescence in the open ocean, a simple ecological model is developed which simulates seasonal changes in dinoflagellate abundance. How forecasting seasonal changes in BPOT may be achieved through combining such a model with relationships derived from observations is discussed and an example is given. The study illustrates a potential new approach to forecasting BPOT through explicitly modelling the population dynamics of a prolific bioluminescent phylum. The model developed here offers a promising platform for the future operational forecasting of the broad temporal changes in bioluminescence within the North Atlantic. Such forecasting of seasonal patterns could provide valuable information for the targeting of scientific field campaigns.

  16. Time Reversal Reconstruction Algorithm Based on PSO Optimized SVM Interpolation for Photoacoustic Imaging

    Mingjian Sun

    2015-01-01

    Full Text Available Photoacoustic imaging is an innovative imaging technique to image biomedical tissues. The time reversal reconstruction algorithm in which a numerical model of the acoustic forward problem is run backwards in time is widely used. In the paper, a time reversal reconstruction algorithm based on particle swarm optimization (PSO optimized support vector machine (SVM interpolation method is proposed for photoacoustics imaging. Numerical results show that the reconstructed images of the proposed algorithm are more accurate than those of the nearest neighbor interpolation, linear interpolation, and cubic convolution interpolation based time reversal algorithm, which can provide higher imaging quality by using significantly fewer measurement positions or scanning times.

  17. Firefly Luciferase-Based Sequential Bioluminescence Resonance Energy Transfer (BRET)-Fluorescence Resonance Energy Transfer (FRET) Protease Assays.

    Branchini, Bruce

    2016-01-01

    We describe here the preparation of ratiometric luminescent probes that contain two well-separated emission peaks produced by a sequential bioluminescence resonance energy transfer (BRET)-fluorescence resonance energy transfer (FRET) process. The probes are single soluble fusion proteins consisting of a thermostable firefly luciferase variant that catalyzes yellow-green (560 nm maximum) bioluminescence and a red fluorescent protein covalently labeled with a near-Infrared fluorescent dye. The two proteins are connected by a decapeptide containing a protease recognition site specific for factor Xa, thrombin, or caspase 3. The rates of protease cleavage of the fusion protein substrates were monitored by recording emission spectra and plotting the change in peak ratios over time. Detection limits of 0.41 nM for caspase 3, 1.0 nM for thrombin, and 58 nM for factor Xa were realized with a scanning fluorometer. This method successfully employs an efficient sequential BRET-FRET energy transfer process based on firefly luciferase bioluminescence to assay physiologically important protease activities and should be generally applicable to the measurement of any endoprotease lacking accessible cysteine residues. PMID:27424898

  18. Imaging system for obtaining space- and time-resolved plasma images on TMX

    A Reticon 50 x 50 photodiode array camera has been placed on Livermore's Tandem Mirror Experiment to view a 56-cm diameter plasma source of visible, vacuum-ultraviolet, and x-ray photons. The compact camera views the source through a pinhole, filters, a fiber optic coupler, a microchannel plate intensifier (MCPI), and a reducer. The images are digitized (at 3.3 MHz) and stored in a large, high-speed memory that has a capacity of 45 images. A local LSI-11 microprocessor provides immediate processing and display of the data. The data are also stored on floppy disks that can be further processed on the large Livermore Computer System. The temporal resolution is limited by the fastest MCPI gate. The number of images recorded is determined by the read-out time of the Reticon camera (minimum 0.9 msec). The spatial resolution of approximately 1.4 cm is fixed by the geometry and the pinhole of 0.025 cm. Typical high-quality color representation of some plasma images are included

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

    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)

  20. L1/2 regularization based numerical method for effective reconstruction of bioluminescence tomography

    Even though bioluminescence tomography (BLT) exhibits significant potential and wide applications in macroscopic imaging of small animals in vivo, the inverse reconstruction is still a tough problem that has plagued researchers in a related area. The ill-posedness of inverse reconstruction arises from insufficient measurements and modeling errors, so that the inverse reconstruction cannot be solved directly. In this study, an l1/2 regularization based numerical method was developed for effective reconstruction of BLT. In the method, the inverse reconstruction of BLT was constrained into an l1/2 regularization problem, and then the weighted interior-point algorithm (WIPA) was applied to solve the problem through transforming it into obtaining the solution of a series of l1 regularizers. The feasibility and effectiveness of the proposed method were demonstrated with numerical simulations on a digital mouse. Stability verification experiments further illustrated the robustness of the proposed method for different levels of Gaussian noise

  1. Seasonal variation of deep-sea bioluminescence in the Ionian Sea

    The ICDeep (Image Intensified Charge Coupled Device for Deep sea research) profiler was used to measure the density of deep bioluminescent animals (BL) through the water column in the east, west and mid-Ionian Sea and in the Algerian Basin. A west to east decrease in BL density was found. Generalized additive modelling was used to investigate seasonal variation in the east and west Ionian Sea (NESTOR and NEMO neutrino telescope sites, respectively) from BL measurements in autumn 2008 and spring 2009. A significant seasonal effect was found in the west Ionian Sea (p<0.001), where a deep autumnal peak in BL density occurred between 500 and 2400 m. No significant seasonal variation in BL density was found in the east Ionian Sea (p=0.07). In both spring and autumn, significant differences in BL density were found through the water column between the east and west Ionian Sea (p<0.001).

  2. Seasonal variation of deep-sea bioluminescence in the Ionian Sea

    Craig, Jessica, E-mail: j.craig@abdn.ac.u [University of Aberdeen, Oceanlab, Main Street, Newburgh, Aberdeenshire, AB41 6AA (United Kingdom); Jamieson, Alan J.; Bagley, Philip M.; Priede, Imants G. [University of Aberdeen, Oceanlab, Main Street, Newburgh, Aberdeenshire, AB41 6AA (United Kingdom)

    2011-01-21

    The ICDeep (Image Intensified Charge Coupled Device for Deep sea research) profiler was used to measure the density of deep bioluminescent animals (BL) through the water column in the east, west and mid-Ionian Sea and in the Algerian Basin. A west to east decrease in BL density was found. Generalized additive modelling was used to investigate seasonal variation in the east and west Ionian Sea (NESTOR and NEMO neutrino telescope sites, respectively) from BL measurements in autumn 2008 and spring 2009. A significant seasonal effect was found in the west Ionian Sea (p<0.001), where a deep autumnal peak in BL density occurred between 500 and 2400 m. No significant seasonal variation in BL density was found in the east Ionian Sea (p=0.07). In both spring and autumn, significant differences in BL density were found through the water column between the east and west Ionian Sea (p<0.001).

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

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

    2011-01-01

    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...

  4. L1/2 regularization based numerical method for effective reconstruction of bioluminescence tomography

    Chen, Xueli; Yang, Defu; Zhang, Qitan; Liang, Jimin

    2014-05-01

    Even though bioluminescence tomography (BLT) exhibits significant potential and wide applications in macroscopic imaging of small animals in vivo, the inverse reconstruction is still a tough problem that has plagued researchers in a related area. The ill-posedness of inverse reconstruction arises from insufficient measurements and modeling errors, so that the inverse reconstruction cannot be solved directly. In this study, an l1/2 regularization based numerical method was developed for effective reconstruction of BLT. In the method, the inverse reconstruction of BLT was constrained into an l1/2 regularization problem, and then the weighted interior-point algorithm (WIPA) was applied to solve the problem through transforming it into obtaining the solution of a series of l1 regularizers. The feasibility and effectiveness of the proposed method were demonstrated with numerical simulations on a digital mouse. Stability verification experiments further illustrated the robustness of the proposed method for different levels of Gaussian noise.

  5. Bioluminescence determination of active caspase-3 in single apoptotic cells

    Lišková, Marcela; Klepárník, Karel; Matalová, Eva; Hegrová, Jitka; Přikryl, Jan; Švandová, Eva; Foret, František

    2013-01-01

    Roč. 34, č. 12 (2013), s. 1772-1777. ISSN 0173-0835 R&D Projects: GA ČR GAP206/11/2377 Grant ostatní: GA ČR(CZ) GAP502/12/1285 Institutional support: RVO:68081715 ; RVO:67985904 Keywords : apoptosis * bioluminescence * caspase-3 Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.161, year: 2013

  6. A Novel Bioluminescent Protease Assay Using Engineered Firefly Luciferase

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

    2008-01-01

    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...

  7. Bioluminescent bacteria as indicators of chemical contamination of coastal waters.

    Frischer, M E; Danforth, J M; Foy, T F; Juraske, R

    2005-01-01

    The ratio of bioluminescent to total bacteria (bioluminescent ratio, BLR) as an indicator of a variety of types of anthropogenic contamination of estuarine ecosystems was evaluated through a series of laboratory and field studies. Laboratory studies indicated that the BLR of natural bacterioplankton communities was proportionally reduced in the presence of a number of contaminants including diesel fuel and saltmarsh sediments co-contaminated with mercury and polychlorinated biphenyls (PCBs). Bioluminescent ratio inhibition was observed after short-term exposure to a contaminant suggesting a physiological rather than a population response of native microbial communities. Simulated eutrophication did not suppress the BLR. Field observations of the BLR were conducted weekly for a 2-yr period in the Skidaway River estuary, Georgia, USA. These observations revealed considerable seasonal variability associated with the BLR. Bioluminescent ratios were highest during the summer (25 +/- 15%), lower in the fall (6 +/- 5%) and spring (3 +/- 2%), and near zero during the winter. Although the BLR was not significantly correlated to salinity at a single site (Skidaway River estuary), the BLR was significantly correlated with salinity when sites within the same estuary system were compared (r2 = 0.93). Variation in BLR was not correlated to standard bacteriological indicators of water quality including total and fecal coliform bacteria. Comparison of the BLR from impacted and pristine estuarine sites during the fall suggested that anthropogenically impacted sites exhibited lower BLR than predicted from salinity versus BLR relationships developed in pristine systems. These observations suggest that the BLR could be used as a simple and reliable initial indicator of chemical contamination of estuarine systems resulting from human activity. PMID:15998855

  8. Detection of a bioluminescent milky sea from space

    Miller, Steven D.; Haddock, Steven H. D.; Elvidge, Christopher D.; Thomas F. Lee

    2005-01-01

    On many occasions over the centuries, mariners have reported witnessing surreal nocturnal displays where the surface of the sea produces an intense, uniform, and sustained glow that extends to the horizon in all directions. Although such emissions cannot be fully reconciled with the known features of any light-emitting organism, these so-called “milky seas” are hypothesized to be manifestations of unusually strong bioluminescence produced by colonies of bacteria in association with a microalg...

  9. Bioluminescence for USP sterility testing of pharmaceutical suspension products.

    Bussey, D M; K. Tsuji

    1986-01-01

    Bioluminescence measurement significantly improved the accuracy, sensitivity, precision, and reliability of the current visual endpoint determination for the USP sterility test and eliminated the day 7 transfer/dilution step required for testing suspension products. Thirteen strains of bacteria and fungi (representing potential contaminants in sterile products), three pharmaceutical suspension products, and four media were used in the experiment. No interference from suspension products was e...

  10. Forming rotated SAR images by real-time motion compensation.

    Doerry, Armin Walter

    2012-12-01

    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.

  11. Bioluminescence in the Ocean: Origins of Biological, Chemical, and Ecological Diversity

    Widder, E. A.

    2010-05-01

    From bacteria to fish, a remarkable variety of marine life depends on bioluminescence (the chemical generation of light) for finding food, attracting mates, and evading predators. Disparate biochemical systems and diverse phylogenetic distribution patterns of light-emitting organisms highlight the ecological benefits of bioluminescence, with biochemical and genetic analyses providing new insights into the mechanisms of its evolution. The origins and functions of some bioluminescent systems, however, remain obscure. Here, I review recent advances in understanding bioluminescence in the ocean and highlight future research efforts that will unite molecular details with ecological and evolutionary relationships.

  12. Real-time synthetic aperture imaging: opportunities and challenges

    Nikolov, Svetoslav; Tomov, Borislav Gueorguiev; Jensen, Jørgen Arendt

    2006-01-01

    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...

  13. Real-time sono-photoacoustic imaging of gold nanoemulsions

    Arnal, Bastien; Wei, Chen-Wei; Perez, Camilo; Lombardo, Michael; Pelivanov, Ivan M.; Pozzo, Danilo; O'Donnell, Matthew

    2015-03-01

    Phase transition contrast agents were first introduced in ultrasound (US) in the form of perfluorocarbon droplets. When their size is reduced to the nanoscale, surface tension dominates their stability and high pressure is required to vaporize them using long US emissions at high frequencies. Our group recently showed that nanoemulsion beads (100-300 nm) coated with gold nanopsheres could be used as non-linear contrast agents. Beads can be vaporized with light only, inducing stronger photoacoustic signals by increasing thermal expansion. A photoacoustic cavitation threshold study (US: 1.2 MHz, Laser 750 nm and 10-ns pulse) shows that the vaporization thresholds of NEB-GNS can be greatly reduced using simultaneous light and US excitations. The resulting signal is driven only by the pressure amplitude for a fluence higher than 2.4 mJ/cm2. At diagnostic exposures, it is possible to capture very high signals from the vaporized beads at concentrations reduced to 10 pM with optical absorption smaller than 0.01 cm-1. A real-time imaging mode selectively isolating vaporization signals was implemented on a Verasonics system. A linear US probe (L74, 3 MHz) launched short US bursts before light was emitted from the laser. Vaporization of NEB-GNS resulted in a persistent 30-dB signal enhancement compared to a dye with the same absorption. Specific vaporization signals were retrieved in phantom experiments with US scatterers. This technique, called sonophotoacoustics, has great potential for targeted molecular imaging and therapy using compact nanoprobes with potentially high-penetrability into tissue.

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

    Evangelos Zigouris; Dimitrios Markonis; Athanasios Kalantzopoulos

    2009-01-01

    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...

  15. Exploiting Molecular Biology by Time-Resolved Fluorescence Imaging

    Müller, Francis; Fattinger, Christof

    Many contemporary biological investigations rely on highly sensitive in vitro assays for the analysis of specific molecules in biological specimens, and the main part of these assays depends on high-sensitivity fluorescence detection techniques for the final readout. The analyzed molecules and molecular interactions in the specimen need to be detected in the presence of other highly abundant biomolecules, while the analyzed molecules themselves are only present at nano-, pico-, or even femtomolar concentration.A short scientific rationale of fluorescence is presented. It emphasizes the use of fluorescent labels for sensitive assays in life sciences and specifies the main properties of an ideal fluorophore. With fluorescence lifetimes in the microsecond range and fluorescence quantum yield of 0.4 some water soluble complexes of Ruthenium like modified Ru(sulfobathophenanthroline) complexes fulfill these properties. They are outstanding fluorescent labels for ultrasensitive assays as illustrated in two examples, in drug discovery and in point of care testing.We discuss the fundamentals and the state-of-the-art of the most sensitive time-gated fluorescence assays. We reflect on how the imaging devices currently employed for readout of these assays might evolve in the future. Many contemporary biological investigations rely on highly sensitive in vitro assays for the analysis of specific molecules in biological specimens, and the main part of these assays depends on high-sensitivity fluorescence detection techniques for the final readout. The analyzed molecules and molecular interactions in the specimen need to be detected in the presence of other highly abundant biomolecules, while the analyzed molecules themselves are only present at nano-, pico-, or even femtomolar concentration.A short scientific rationale of fluorescence is presented. It emphasizes the use of fluorescent labels for sensitive assays in life sciences and specifies the main properties of an ideal

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

    Ming-Fu Chen

    2014-02-01

    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

  17. Effects of acquisition time and reconstruction algorithm on image quality, quantitative parameters, and clinical interpretation of myocardial perfusion imaging

    Enevoldsen, Lotte Hahn; Menashi, Changez A K; Andersen, Ulrik B;

    2013-01-01

    Recently introduced iterative reconstruction algorithms with resolution recovery (RR) and noise-reduction technology seem promising for reducing scan time or radiation dose without loss of image quality. However, the relative effects of reduced acquisition time and reconstruction software have...

  18. High time-resolution sprite imaging: observations and implications

    Stenbaek-Nielsen, H C [Geophysical Institute, University of Alaska Fairbanks, 903 Koyukuk Drive, Fairbanks, Ak 99775-7320 (United States); McHarg, M G [Department of Physics, 2354 Fairchild Drive, Suite 2A31, US Air Force Academy, CO 80840 (United States)], E-mail: hnielsen@gi.alaska.edu, E-mail: matthew.mcharg@usafa.edu

    2008-12-07

    Sprites are large scale manifestations of electrical streamers triggered in the upper atmosphere by lightning in an underlying thunderstorm. Imaging of sprites at 10 000 frames per second has provided new insights into their spatial and temporal development. In this paper we discuss the experimental protocols that have been developed for performing high-speed observations of sprites and some new observations that have been obtained of relevance to laboratory experiments. Downward tendrils and upward branches, so characteristic in video recordings, are shown to be formed by very fast streamer heads with velocities up to half the speed of light. The streamer heads are spatially small, {approx}100 m or less, but very bright with emission rates up to {approx}10{sup 24} photons s{sup -1}. The sprite onset begins with a downward streamer. Then, in some sprites, at a little later time and from a lower altitude upward moving streamer heads may also appear. If there are no upward streamers the sprite would be classified as a 'C-sprite'; with both downward and upward streamers it would be a 'carrot sprite'. The optical emissions are primarily from the neutral molecular nitrogen first positive bands emitting in the near-infrared, but there are also blue emissions assumed to be from second positive bands of molecular nitrogen and from first negative bands of nitrogen ions. The streamer heads are observed at times to split into several streamer heads. This process appears to be more frequent in the core of larger sprites.

  19. Synchronization of circadian bioluminescence as a group-foraging strategy in cave glowworms.

    Maynard, Andrew J; Merritt, David J

    2013-07-01

    Flies of the genus Arachnocampa are sit-and-lure predators that use bioluminescence to attract flying prey to their silk webs. Some species are most common in rainforest habitat and others inhabit both caves and rainforest. We have studied the circadian regulation of bioluminescence in two species: one found in subtropical rainforest with no known cave populations and the other found in temperate rainforest with large populations in limestone caves. The rainforest species is typical of most nocturnal animals in that individuals are entrained by the light:dark (LD) cycle to be active at night; in this case, their propensity to bioluminesce is greatest at night. The dual-habitat species shows an opposite phase response to the same entrainment; its bioluminescence propensity rhythm is entrained by LD exposure to peak during the day. Nevertheless, in LD environments, individuals do not bioluminesce during the day because ambient light inhibits their bioluminescence (negative masking), pushing bioluminescence into the dark period. This unusual and unexpected phenomenon could be related to their association with caves and has been suggested to be an adaptation of the circadian system that promotes synchronization of a colony's output of bioluminescence. Here, we use controlled laboratory experiments to show that individuals do synchronize their bioluminescence rhythms when in visual contact with each other. Entrainment of the bioluminescence rhythm to the biological photophase causes colony-wide synchronization, creating a daily sinusoidal rhythm of the intensity of bioluminescence in the many thousands of individuals making up a colony. This synchronization could provide a group-foraging advantage, allowing the colony to glow most brightly when the prey are most likely to be active. PMID:23575257

  20. Cf-252 based neutron radiography using real-time image processing system

    For compact Cf-252 based neutron radiography, a real-time image processing system by particle counting technique has been developed. The electronic imaging system consists of a supersensitive imaging camera, a real-time corrector, a real-time binary converter, a real-time calculator for centroid, a display monitor and a computer. Three types of accumulated NR image; ordinary, binary and centroid images, can be observed during a measurement. Accumulated NR images were taken by the centroid mode, the binary mode and ordinary mode using of Cf-252 neutron source and those images were compared. The centroid mode presented the sharpest image and its statistical characteristics followed the Poisson distribution, while the ordinary mode showed the smoothest image as the averaging effect by particle bright spots with distributed brightness was most dominant. (author)

  1. Density resolutionary optimization of real time radiotherapy portal imagings

    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)

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

    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

    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

  3. Space-time encoding for high frame rate ultrasound imaging

    Misaridis, Thanssis; Jensen, Jørgen Arendt

    Frame rate in ultrasound imaging can be dramatically increased by using sparse synthetic transmit aperture (STA) beamforming techniques. The two main drawbacks of the method are the low signal-to-noise ratio (SNR) and the motion artifacts, that degrade the image quality. In this paper we propose ...

  4. Real-time progressive hyperspectral image processing endmember finding and anomaly detection

    Chang, Chein-I

    2016-01-01

    The book covers the most crucial parts of real-time hyperspectral image processing: causality and real-time capability. Recently, two new concepts of real time hyperspectral image processing, Progressive Hyperspectral Imaging (PHSI) and Recursive Hyperspectral Imaging (RHSI). Both of these can be used to design algorithms and also form an integral part of real time hyperpsectral image processing. This book focuses on progressive nature in algorithms on their real-time and causal processing implementation in two major applications, endmember finding and anomaly detection, both of which are fundamental tasks in hyperspectral imaging but generally not encountered in multispectral imaging. This book is written to particularly address PHSI in real time processing, while a book, Recursive Hyperspectral Sample and Band Processing: Algorithm Architecture and Implementation (Springer 2016) can be considered as its companion book. Includes preliminary background which is essential to those who work in hyperspectral ima...

  5. Collinear, two-color optical Kerr effect shutter for ultrafast time-resolved imaging

    Purwar, Harsh; Rozé, Claude; Sedarsky, David; Blaisot, Jean-Bernard

    2015-01-01

    Imaging with ultrashort exposure times is generally achieved with a crossed-beam geometry. In the usual arrangement, an off-axis gating pulse induces birefringence in a medium exhibiting a strong Kerr response (commonly carbon disulfide) which is followed by a polarizer aligned to fully attenuate the on-axis imaging beam. By properly timing the gate pulse, imaging light experiences a polarization change allowing time-dependent transmission through the polarizer to form an ultrashort image. The crossed-beam system is effective in generating short gate times, however, signal transmission through the system is complicated by the crossing angle of the gate and imaging beams. This work presents a robust ultrafast time-gated imaging scheme based on a combination of type-I frequency doubling and a collinear optical arrangement in carbon disulfide. We discuss spatial effects arising from crossed-beam Kerr gating, and examine the imaging spatial resolution and transmission timing affected by collinear activation of th...

  6. Accessible high performance computing solutions for near real-time image processing for time critical applications

    Bielski, Conrad; Lemoine, Guido; Syryczynski, Jacek

    2009-09-01

    High Performance Computing (HPC) hardware solutions such as grid computing and General Processing on a Graphics Processing Unit (GPGPU) are now accessible to users with general computing needs. Grid computing infrastructures in the form of computing clusters or blades are becoming common place and GPGPU solutions that leverage the processing power of the video card are quickly being integrated into personal workstations. Our interest in these HPC technologies stems from the need to produce near real-time maps from a combination of pre- and post-event satellite imagery in support of post-disaster management. Faster processing provides a twofold gain in this situation: 1. critical information can be provided faster and 2. more elaborate automated processing can be performed prior to providing the critical information. In our particular case, we test the use of the PANTEX index which is based on analysis of image textural measures extracted using anisotropic, rotation-invariant GLCM statistics. The use of this index, applied in a moving window, has been shown to successfully identify built-up areas in remotely sensed imagery. Built-up index image masks are important input to the structuring of damage assessment interpretation because they help optimise the workload. The performance of computing the PANTEX workflow is compared on two different HPC hardware architectures: (1) a blade server with 4 blades, each having dual quad-core CPUs and (2) a CUDA enabled GPU workstation. The reference platform is a dual CPU-quad core workstation and the PANTEX workflow total computing time is measured. Furthermore, as part of a qualitative evaluation, the differences in setting up and configuring various hardware solutions and the related software coding effort is presented.

  7. VISTA - A Constellation for Real Time Regional Imaging

    Meerman, Max; Boland, Lee; da Silva Curiel, Alex; Sweeting, Martin, , Sir

    2002-01-01

    The role of satellites in medium and high-resolution reconnaissance of the Earth's surface has been well demonstrated in recent years through missions such as Landsat, SPOT, IKONOS, ImageSat and Quickbird. The market for such data products is well served and likely to become more competitive with further very-high-resolution missions. Whereas commercial markets have concentrated on enhancing resolution, the small satellite sector has concentrated on reducing the cost of data products, and the development of systems providing niche services. One such EO requirement that can be well met by smaller satellites is the need for higher temporal resolution, as this typically requires a large number of satellites to operate as a constellation - thus far financially impractical using conventional EO satellites. Surrey is currently engaged in building its first constellation that will provide daily global coverage at moderate resolution (32-metre GSD and 600km swath) in three spectral bands. Targeted at providing timely quick-look data products for disaster mitigation and monitoring, the constellation comprises 7 satellites in a single orbital plane. Each satellite has a wide swath so that successive satellites progressively cover the entire globe in a single day. The Vista constellation takes this concept a step further, and is proposed for applications requiring near-continuous surveillance of regional activity. By introducing a multiple plane constellation of small Earth observation satellites, it is possible to monitor continuously selected regions anywhere on the globe. The paper describes the system trades and outlines the scope of the performance that could be obtained from such a system. A cost model illustrates that the balance between launch and space segment costs must be reached by considering suitable replacement strategies, and that the system is highly sensitive to requirement creep. Finally, it is shown that the use of cost effective, small satellites leads to

  8. Real-time single-molecule imaging of quantum interference.

    Juffmann, Thomas; Milic, Adriana; Müllneritsch, Michael; Asenbaum, Peter; Tsukernik, Alexander; Tüxen, Jens; Mayor, Marcel; Cheshnovsky, Ori; Arndt, Markus

    2012-05-01

    The observation of interference patterns in double-slit experiments with massive particles is generally regarded as the ultimate demonstration of the quantum nature of these objects. Such matter-wave interference has been observed for electrons, neutrons, atoms and molecules and, in contrast to classical physics, quantum interference can be observed when single particles arrive at the detector one by one. The build-up of such patterns in experiments with electrons has been described as the "most beautiful experiment in physics". Here, we show how a combination of nanofabrication and nano-imaging allows us to record the full two-dimensional build-up of quantum interference patterns in real time for phthalocyanine molecules and for derivatives of phthalocyanine molecules, which have masses of 514 AMU and 1,298 AMU respectively. A laser-controlled micro-evaporation source was used to produce a beam of molecules with the required intensity and coherence, and the gratings were machined in 10-nm-thick silicon nitride membranes to reduce the effect of van der Waals forces. Wide-field fluorescence microscopy detected the position of each molecule with an accuracy of 10 nm and revealed the build-up of a deterministic ensemble interference pattern from single molecules that arrived stochastically at the detector. In addition to providing this particularly clear demonstration of wave-particle duality, our approach could also be used to study larger molecules and explore the boundary between quantum and classical physics. PMID:22447163

  9. Using bioluminescent biosensors for hazard analysis and critical control point (HACCP) in wastewater control.

    Valat, C; Champiat, D; Degorce-Dumas, J R; Thomas, O

    2004-01-01

    Starting from a new approach for water pollution control and wastewater treatment plant management, the hazard analysis and critical control point (HACCP) quality concept, the interest for the development of new rapid and sensitive methods such as bioluminescence-based methods is evident. After an introduction of the HACCP procedure, a bibliographic study of the bioluminescence potentiality is presented and discussed. PMID:14979548

  10. Bacterial Bioluminescence: Spectral Study of the Emitters in the In Vivo Reaction

    Matheson, I.B.C.; Lee, J.; Muller, F.

    1981-01-01

    Transient fluorescent species are observed in the bioluminescent reactions of three reduced flavin mononucleotides with aliphatic aldehydes and oxygen, catalyzed by bacterial luciferase. In each case the fluorescence spectral distribution is similar to that of the bioluminescence but is readily dist

  11. Time-resolved computed tomography of the liver: retrospective, multi-phase image reconstruction derived from volumetric perfusion imaging

    To assess feasibility and image quality (IQ) of a new post-processing algorithm for retrospective extraction of an optimised multi-phase CT (time-resolved CT) of the liver from volumetric perfusion imaging. Sixteen patients underwent clinically indicated perfusion CT using 4D spiral mode of dual-source 128-slice CT. Three image sets were reconstructed: motion-corrected and noise-reduced (MCNR) images derived from 4D raw data; maximum and average intensity projections (time MIP/AVG) of the arterial/portal/portal-venous phases and all phases (total MIP/ AVG) derived from retrospective fusion of dedicated MCNR split series. Two readers assessed the IQ, detection rate and evaluation time; one reader assessed image noise and lesion-to-liver contrast. Time-resolved CT was feasible in all patients. Each post-processing step yielded a significant reduction of image noise and evaluation time, maintaining lesion-to-liver contrast. Time MIPs/AVGs showed the highest overall IQ without relevant motion artefacts and best depiction of arterial and portal/portal-venous phases respectively. Time MIPs demonstrated a significantly higher detection rate for arterialised liver lesions than total MIPs/AVGs and the raw data series. Time-resolved CT allows data from volumetric perfusion imaging to be condensed into an optimised multi-phase liver CT, yielding a superior IQ and higher detection rate for arterialised liver lesions than the raw data series. (orig.)

  12. Advances in photon counting for bioluminescence

    Ingle, Martin B.; Powell, Ralph

    1998-11-01

    Photon counting systems were originally developed for astronomy, initially by the astronomical community. However, a major application area is in the study of luminescent probes in living plants, fishes and cell cultures. For these applications, it has been necessary to develop camera system capability at very low light levels -- a few photons occasionally -- and also at reasonably high light levels to enable the systems to be focused and to collect quality images of the object under study. The paper presents new data on MTF at extremely low photon flux and conventional ICCD illumination, counting efficiency and dark noise as a function of temperature.

  13. Real Time Medical Image Consultation System Through Internet

    D. Durga Prasad

    2010-01-01

    Full Text Available Teleconsultation among doctors using a telemedicine system typically involves dealing with and sharing medical images of the patients. This paper describes a software tool written in Java which enables the participating doctors to view medical images such as blood slides, X-Ray, USG, ECG etc. online and even allows them to mark and/or zoom specific areas. It is a multi-party secure image communication system tool that can be used by doctors and medical consultants over the Internet.

  14. Using luciferase to image bacterial infections in mice.

    Chang, Mi Hee; Cirillo, Suat L G; Cirillo, Jeffrey D

    2011-01-01

    Imaging is a valuable technique that can be used to monitor biological processes. In particular, the presence of cancer cells, stem cells, specific immune cell types, viral pathogens, parasites and bacteria can be followed in real-time within living animals. Application of bioluminescence imaging to the study of pathogens has advantages as compared to conventional strategies for analysis of infections in animal models. Infections can be visualized within individual animals over time, without requiring euthanasia to determine the location and quantity of the pathogen. Optical imaging allows comprehensive examination of all tissues and organs, rather than sampling of sites previously known to be infected. In addition, the accuracy of inoculation into specific tissues can be directly determined prior to carrying forward animals that were unsuccessfully inoculated throughout the entire experiment. Variability between animals can be controlled for, since imaging allows each animal to be followed individually. Imaging has the potential to greatly reduce animal numbers needed because of the ability to obtain data from numerous time points without having to sample tissues to determine pathogen load. This protocol describes methods to visualize infections in live animals using bioluminescence imaging for recombinant strains of bacteria expressing luciferase. The click beetle (CBRLuc) and firefly luciferases (FFluc) utilize luciferin as a substrate. The light produced by both CBRluc and FFluc has a broad wavelength from 500 nm to 700 nm, making these luciferases excellent reporters for the optical imaging in living animal models. This is primarily because wavelengths of light greater than 600 nm are required to avoid absorption by hemoglobin and, thus, travel through mammalian tissue efficiently. Luciferase is genetically introduced into the bacteria to produce light signal. Mice are pulmonary inoculated with bioluminescent bacteria intratracheally to allow monitoring of

  15. Resources of Digital FIR Filters Hardware Implementation in FPGAs for Digital Image Processing in Real Time

    Peter Cvicela; Josef Huska; Peter Kulla

    2004-01-01

    The main image information content, from the human visual system viewing point, is focused into whole colorimetric and spatial informations. Because every image is result of some previous processes, the goal for all standard image processing methods is improvement colorimetric and spatial image parameters in relation maximum information content by the complicated and expensive systems for digital image processing in (quasi)real time [1] based on the flash signal (multi)processors. Some single...

  16. Rad Hard Imaging Array with Picosecond Timing Project

    National Aeronautics and Space Administration — For a wide range of remote sensing applications, there is a critical need to develop imaging arrays that simultaneously achieve high spatial resolution, high...

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

    Fakhri, Georges El

    2012-01-01

    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. T...

  18. Real-time intraoperative high-speed imaging during phacoemulsification.

    Srivastava, Samaresh; Vasavada, Abhay R; Vasavada, Vaishali A; Vasavada, Viraj A

    2012-09-01

    We describe the use of high-speed imaging during phacoemulsification in a clinical scenario. Images captured during surgery at high frame rates are converted into a slow-motion film to view and analyze various surgical steps. This technique highlights events that are not captured in a normal-speed video recording. It has obvious applications for understanding surgical techniques and technology. PMID:22841426

  19. Quantitative time-stretch imaging -- towards big-data bioassay

    Lau, Kam-seng; 劉金成

    2015-01-01

    The unrelenting growth in the field of optical bioimaging over the past half-century has been leading to a ubiquitously wide range of applications, from in-vitro single-cell analysis to large-scale in-vivo imaging. In light of the complexity and heterogeneity of the biological systems (from molecular, cellular to even the whole-organism level), development of new optical imaging/microscopy technologies should be focused on improving spatial resolution, temporal resolution and enlarging the qu...

  20. Monitoring Therapeutic Treatments against Burkholderia Infections Using Imaging Techniques

    Tiffany M. Mott

    2013-05-01

    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.