WorldWideScience

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. 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.

  3. 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

  4. Cellular bioluminescence imaging.

    Welsh, David K; Noguchi, Takako

    2012-08-01

    Bioluminescence imaging of live cells has recently been recognized as an important alternative to fluorescence imaging. Fluorescent probes are much brighter than bioluminescent probes (luciferase enzymes) and, therefore, provide much better spatial and temporal resolution and much better contrast for delineating cell structure. However, with bioluminescence imaging there is virtually no background or toxicity. As a result, bioluminescence can be superior to fluorescence for detecting and quantifying molecules and their interactions in living cells, particularly in long-term studies. Structurally diverse luciferases from beetle and marine species have been used for a wide variety of applications, including tracking cells in vivo, detecting protein-protein interactions, measuring levels of calcium and other signaling molecules, detecting protease activity, and reporting circadian clock gene expression. Such applications can be optimized by the use of brighter and variously colored luciferases, brighter microscope optics, and ultrasensitive, low-noise cameras. This article presents a review of how bioluminescence differs from fluorescence, its applications to cellular imaging, and available probes, optics, and detectors. It also gives practical suggestions for optimal bioluminescence imaging of single cells.

  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

    2017-01-01

    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. 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 enzyme with its substrate. Most imaging systems provide 2-dimensional (2D) information in rodents, showing the locations and intensity of light emitted from the animal in pseudo-color scaling. A 3-dimensional (3D) capability for bioluminescence imaging is now available, but is more expensive and less efficient; other disadvantages include the requirement for genetically encoded luciferase, the injection of the substrate to enable light emission, and the dependence of light signal on tissue depth. All of these problems make it unlikely that the method will be extended to human studies. However, in small animal models, bioluminescence imaging is now routinely applied to serially detect the location and burden of xenografted tumors, or identify and measure the number of immune or stem cells after an adoptive transfer. Bioluminescence imaging also makes it possible to track the relative amounts and locations of bacteria, viruses, and other pathogens over time. Specialized applications of bioluminescence also follow tissue-specific luciferase expression in transgenic mice, and monitor biological processes such as signaling or protein interactions in real time. In summary, bioluminescence imaging has become an important component of biomedical research that will continue in the future.

  10. 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

    2014-03-01

    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. As bioluminescence microscopy requires no excitation light, it lacks the photo-toxicity associated with fluorescence imaging and permits the long-term, nonlethal observation of living cells. Thus, bioluminescence microscopy would be a powerful tool in cellular biology that complements fluorescence microscopy.

  11. 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.

  12. Quantitative bioluminescence imaging of mouse tumor models.

    Tseng, Jen-Chieh; Kung, Andrew L

    2015-01-05

    Bioluminescence imaging (BLI) has become an essential technique for preclinical evaluation of anticancer therapeutics and provides sensitive and quantitative measurements of tumor burden in experimental cancer models. For light generation, a vector encoding firefly luciferase is introduced into human cancer cells that are grown as tumor xenografts in immunocompromised hosts, and the enzyme substrate luciferin is injected into the host. Alternatively, the reporter gene can be expressed in genetically engineered mouse models to determine the onset and progression of disease. In addition to expression of an ectopic luciferase enzyme, bioluminescence requires oxygen and ATP, thus only viable luciferase-expressing cells or tissues are capable of producing bioluminescence signals. Here, we summarize a BLI protocol that takes advantage of advances in hardware, especially the cooled charge-coupled device camera, to enable detection of bioluminescence in living animals with high sensitivity and a large dynamic range.

  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. In vivo cell tracking with bioluminescence imaging.

    Kim, Jung Eun; Kalimuthu, Senthilkumar; Ahn, Byeong-Cheol

    2015-03-01

    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.

  15. Using bioluminescence imaging in glioma research.

    Luwor, Rodney B; Stylli, Stanley S; Kaye, Andrew H

    2015-05-01

    Glioblastoma multiforme (GBM) is the most common malignant brain tumour and has the worst prognosis. Over the last decade, the use of bioluminescence imaging technology has rapidly become widespread to further understand the mechanisms that drive GBM development and progression. Pre-clinical evaluation and optimisation of therapeutic efficacy in GBM research has also utilised this simple non-invasive technology. Here we summarise recent advances made in glioma biology and therapeutic intervention using bioluminescence imaging. This review also describes the current knowledge regarding the use of luciferase-based reporters in examining the role of specific cancer signalling cascades that promote glioma progression.

  16. 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.

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

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

    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 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.

  18. Fluorescence and Bioluminescence Imaging of Orthotopic Brain Tumors in Mice.

    McKinnon, Emilie; Moore, Alfred; Dixit, Suraj; Zhu, Yun; Broome, Ann-Marie

    2017-01-01

    Optical imaging strategies, such as fluorescence and bioluminescence imaging, are non-invasive, in vivo whole body imaging techniques utilized to study cancer. Optical imaging is widely used in preclinical work because of its ease of use and cost-friendliness. It also provides the opportunity to study animals and biological responses longitudinally over time. Important considerations include depth of tissue penetration, photon scattering, absorption and the choice of light emitting probe, all of which affect the resolution (image quality and data information) and the signal to noise ratio of the image. We describe how to use bioluminescence and fluorescence imaging to track a chemotherapeutic delivery nanocarrier conjugated with a fluorophore to determine its localization in vivo.

  19. 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.

  20. 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.

  1. 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

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

    2008-05-15

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

  2. 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.

  3. A synthetic luciferin improves in vivo bioluminescence imaging of gene expression in cardiovascular brain regions.

    Simonyan, Hayk; Hurr, Chansol; Young, Colin N

    2016-10-01

    Bioluminescence imaging is an effective tool for in vivo investigation of molecular processes. We have demonstrated the applicability of bioluminescence imaging to spatiotemporally monitor gene expression in cardioregulatory brain nuclei during the development of cardiovascular disease, via incorporation of firefly luciferase into living animals, combined with exogenous d-luciferin substrate administration. Nevertheless, d-luciferin uptake into the brain tissue is low, which decreases the sensitivity of bioluminescence detection, particularly when considering small changes in gene expression in tiny central areas. Here, we tested the hypothesis that a synthetic luciferin, cyclic alkylaminoluciferin (CycLuc1), would be superior to d-luciferin for in vivo bioluminescence imaging in cardiovascular brain regions. Male C57B1/6 mice underwent targeted delivery of an adenovirus encoding the luciferase gene downstream of the CMV promoter to the subfornical organ (SFO) or paraventricular nucleus of hypothalamus (PVN), two crucial cardioregulatory neural regions. While bioluminescent signals could be obtained following d-luciferin injection (150 mg/kg), CycLuc1 administration resulted in a three- to fourfold greater bioluminescent emission from the SFO and PVN, at 10- to 20-fold lower substrate concentrations (7.5-15 mg/kg). This CycLuc1-mediated enhancement in bioluminescent emission was evident early following substrate administration (i.e., 6-10 min) and persisted for up to 1 h. When the exposure time was reduced from 60 s to 1,500 ms, minimal signal in the PVN was detectable with d-luciferin, whereas bioluminescent images could be reliably captured with CycLuc1. These findings demonstrate that bioluminescent imaging with the synthetic luciferin CycLuc1 provides an improved physiological genomics tool to investigate molecular events in discrete cardioregulatory brain nuclei.

  4. 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)

  5. Expanding the bioluminescent toolkit for in vivo imaging

    Paley, Miranda Amelia

    2014-01-01

    Bioluminescence imaging (BLI) is among the most dynamic imaging modalities for visualizing whole cells and gene expression patterns in vivo. This technique captures light emission from the luciferase-catalyzed oxidation of small molecule luciferins with highly sensitive CCD cameras. While powerful, current options for multiplexed BLI in mice are limited by the number of luciferase/luciferin pairs found in nature. Our lab aims to expand the bioluminescent toolkit by pairing mutant luciferases ...

  6. 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.

  7. 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.

    2015-01-01

    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. PMID:26516295

  8. Noninvasive bioluminescence imaging of dengue virus infection in the brain of A129 mice.

    Li, Xiao-Feng; Deng, Yong-Qiang; Zhao, Hui; Ye, Qing; Wang, Hong-Jiang; Li, Shi-Hua; Zhu, Shun-Ya; Shi, Pei-Yong; Qin, E-De; Zhang, Bo; Qin, Cheng-Feng

    2013-05-01

    Dengue virus (DENV) infection is one of the most important public health threats globally; however, no vaccines or effective antivirals are currently available. The bioluminescence imaging technique has emerged as a powerful tool for studies on viral pathogenesis in vitro and in vivo. In this study, using a recombinant DENV that stably expressed Renilla luciferase (Rluc-DENV), we used bioluminescence for imaging of DENV infection in the brain of A129 mice that lacked type I interferon receptors. Upon intracranial inoculation with Rluc-DENV, A129 mice developed typical neurological symptoms and rapidly succumbed to viral infection. Real-time bioluminescence intensity analysis revealed the replication kinetics of Rluc-DENV in the brain of A129 mice. Linear regression analyses showed a good correlation between photon flux and viral titers (R(2) = 0.9923). Finally, the bioluminescence model was validated using a known mouse monoclonal antibody, 2A10G6, and the therapeutic effects of this neutralizing antibody were readily monitored by live imaging in the same animal. The noninvasive bioluminescence imaging of DENV infection as described here shows distinct advantages over traditional animal models and provides a powerful tool for potential antiviral or vaccine assays against DENV infection in vivo.

  9. 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

  10. 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

  11. Advancing Molecular Therapies through In Vivo Bioluminescent Imaging

    Anton McCaffrey

    2003-04-01

    Full Text Available Effective development of therapeutics that target the molecular basis of disease is dependent on testing new therapeutic moieties and delivery strategies in animal models of human disease. Accelerating the analyses of these models and improving their predictive value through whole animal imaging methods, which provide data in real time and are sensitive to the subtle changes, are crucial for rapid advancement of these approaches. Modalities based on optics are rapid, sensitive, and accessible methods for in vivo analyses with relatively low instrumentation costs. In vivo bioluminescent imaging (BLI is one of these optically based imaging methods that enable rapid in vivo analyses of a variety of cellular and molecular events with extreme sensitivity. BLI is based on the use of light-emitting enzymes as internal biological light sources that can be detected externally as biological indicators. BLI has been used to test spatio-temporal expression patterns of both target and therapeutic genes in living laboratory animals where the contextual influences of whole biological systems are preserved. BLI has also been used to analyze gene delivery, immune cell therapies, and the in vivo efficacy of inhibitory RNAs. New tools for BLI are being developed that will offer greater flexibility in detection and analyses. BLI can be used to accelerate the evaluation of experimental therapeutic strategies and whole body imaging offers the opportunity of revealing the effects of novel approaches on key steps in disease processes.

  12. 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 facile method for monitoring the cytotoxic effects of a compound of interest. Mammalian cells stably expressing the full bacterial bioluminescence (luxCDABEfrp) gene cassette autonomously produce an optical signal that peaks at 490 nm without the addition of an expensive and possibly interfering luciferin substrate, excitation by an external energy source, or destruction of the sample that is traditionally performed during optical imaging procedures. This independence from external stimulation places the burden for maintaining the bioluminescent reaction solely on the cell, meaning that the resultant signal is only detected during active metabolism. This characteristic makes the lux-expressing cell line an excellent candidate for use as a biosentinel against cytotoxic effects because changes in bioluminescent production are indicative of adverse effects on cellular growth and metabolism. Similarly, the autonomous nature and lack of required sample destruction permits repeated imaging of the same sample in real-time throughout the period of toxicant exposure and can be performed across multiple samples using existing imaging equipment in an automated fashion. PMID:24193545

  13. In Vivo Bioluminescence Imaging for Longitudinal Monitoring of Inflammation in Animal Models of Uveitis

    Gutowski, Michal B.; Wilson, Leslie; Van Gelder, Russell N.; Pepple, Kathryn L.

    2017-01-01

    Purpose We develop a quantitative bioluminescence assay for in vivo longitudinal monitoring of inflammation in animal models of uveitis. Methods Three models of experimental uveitis were induced in C57BL/6 albino mice: primed mycobacterial uveitis (PMU), endotoxin-induced uveitis (EIU), and experimental autoimmune uveitis (EAU). Intraperitoneal injection of luminol sodium salt, which emits light when oxidized, provided the bioluminescence substrate. Bioluminescence images were captured by a PerkinElmer In Vivo Imaging System (IVIS) Spectrum and total bioluminescence was analyzed using Living Image software. Bioluminescence on day zero was compared to bioluminescence on the day of peak inflammation for each model. Longitudinal bioluminescence imaging was performed in EIU and EAU. Results In the presence of luminol, intraocular inflammation generates detectable bioluminescence in three mouse models of uveitis. Peak bioluminescence in inflamed PMU eyes (1.46 × 105 photons/second [p/s]) was significantly increased over baseline (1.47 × 104 p/s, P = 0.01). Peak bioluminescence in inflamed EIU eyes (3.18 × 104 p/s) also was significantly increased over baseline (1.09 × 104 p/s, P = 0.04), and returned to near baseline levels by 48 hours. In EAU, there was a nonsignificant increase in bioluminescence at peak inflammation. Conclusions In vivo bioluminescence may be used as a noninvasive, quantitative measure of intraocular inflammation in animal models of uveitis. Primed mycobacterial uveitis and EIU are both acute models with robust anterior inflammation and demonstrated significant changes in bioluminescence corresponding with peak inflammation. Experimental autoimmune uveitis is a more indolent posterior uveitis and generated a more modest bioluminescent signal. In vivo imaging system bioluminescence is a nonlethal, quantifiable assay that can be used for monitoring inflammation in animal models of uveitis. PMID:28278321

  14. Bioluminescence imaging: a shining future for cardiac regeneration

    Roura, Santiago; Gálvez-Montón, Carolina; Bayes-Genis, Antoni

    2013-01-01

    Advances in bioanalytical techniques have become crucial for both basic research and medical practice. One example, bioluminescence imaging (BLI), is based on the application of natural reactants with light-emitting capabilities (photoproteins and luciferases) isolated from a widespread group of organisms. The main challenges in cardiac regeneration remain unresolved, but a vast number of studies have harnessed BLI with the discovery of aequorin and green fluorescent proteins. First described...

  15. A human brainstem glioma xenograft model enabled for bioluminescence imaging

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

    2009-01-01

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

  16. Factors Influencing Quantification of in Vivo Bioluminescence Imaging: Application to Assessment of Pancreatic Islet Transplants

    John Virostko

    2004-10-01

    Full Text Available The aim of this study is to determine and characterize factors influencing in vivo bioluminescence imaging (BLI and apply them to the specific application of imaging transplanted pancreatic islets. Noninvasive quantitative assessment of transplanted pancreatic islets poses a formidable challenge. Murine pancreatic islets expressing firefly luciferase were transplanted under the renal capsule or into the portal vein of nonobese diabetic–severe combined immunodeficiency mice and the bioluminescence was quantified with a cooled charge coupled device camera and digital photon image analysis. The important, but often neglected, effects of wound healing, mouse positioning, and transplantation site on bioluminescence measurements were investigated by imaging a constant emission, isotropic light-emitting bead (λ = 600 implanted at the renal or hepatic site. The renal beads emitted nearly four times more light than hepatic beads with a smaller spot size, indicating that light absorption and scatter are greatly influenced by the transplant site and must be accounted for in BLI measurements. Detected luminescence decreased with increasing angle between the mouse surface normal and optical axis. By defining imaging parameters such as postsurgical effects, animal positioning, and light attenuation as a function of transplant site, this study develops BLI as a useful imaging modality for quantitative assessment of islets post-transplantation.

  17. Analysis of neurogenesis during experimental autoimmune encephalomyelitis reveals pitfalls of bioluminescence imaging.

    Ayzenberg, Ilya; Schlevogt, Sibylle; Metzdorf, Judith; Stahlke, Sarah; Pedreitturia, Xiomara; Hunfeld, Anika; Couillard-Despres, Sebastien; Kleiter, Ingo

    2015-01-01

    Bioluminescence imaging is a sensitive approach for longitudinal neuroimaging. Transgenic mice expressing luciferase under the promoter of doublecortin (DCX-luc), a specific marker of neuronal progenitor cells (NPC), allow monitoring of neurogenesis in living mice. Since the extent and time course of neurogenesis during autoimmune brain inflammation are controversial, we investigated neurogenesis in MOG-peptide induced experimental allergic encephalomyelitis (EAE) using DCX-luc reporter mice. We observed a marked, 2- to 4-fold increase of the bioluminescence signal intensity 10 days after EAE induction and a gradual decline 1-2 weeks thereafter. In contrast, immunostaining for DCX revealed no differences between EAE and control mice 2 and 4 weeks after immunization in zones of adult murine neurogenesis such as the dentate gyrus. Ex vivo bioluminescence imaging showed similar luciferase expression in brain homogenates of EAE and control animals. Apart from complete immunization including MOG-peptide also incomplete immunization with complete Freund´s adjuvant and pertussis toxin resulted in a rapid increase of the in vivo bioluminescence signal. Blood-brain barrier (BBB) leakage was demonstrated 10 days after both complete and incomplete immunization and might explain the increased bioluminescence signal in vivo. We conclude, that acute autoimmune inflammation in EAE does not alter neurogenesis, at least at the stage of DCX-expressing NPC. Effects of immunization on the BBB integrity must be considered when luciferase is used as a reporter within the CNS during the active stage of EAE. Models with stable CNS-restricted luciferase expression could serve as technically convenient way to evaluate BBB integrity in a longitudinal manner.

  18. Comparison between bioluminescence imaging technique and CFU count for the study of oropharyngeal candidiasis in mice.

    Gabrielli, Elena; Roselletti, Elena; Luciano, Eugenio; Sabbatini, Samuele; Mosci, Paolo; Pericolini, Eva

    2015-05-01

    We recently described a bioluminescence in vivo imaging technique, representing a powerful tool to test the real-time progression of oropharyngeal candidiasis, hence potentially useful to evaluate the efficacy of antifungal therapies. In this study, the in vivo imaging technique was compared with CFU measurement of target organs (tongue, esophagus and stomach) for monitoring and quantifying oropharyngeal candidiasis. We have correlated these two analytical methods at different times post-infection using engineered, luminescent Candida albicans in mice rendered susceptible to oral candidiasis by cortisone-acetate. Scatter plots, Pearson correlation and Student's t test were used to compare the methods. We observed that the bioluminescence in vivo imaging technique was more reliable than CFU counts in detecting early infection of, and its extent in, the oral cavity of the mouse. This was also evident following the introduction of a variable such as treatment with fluconazole. The results described in this study could validate the bioluminescence in vivo imaging technique as a method to monitor and quantify oropharyngeal candidiasis and to assess early discovery of active compounds in vivo.

  19. A novel luciferase fusion protein for highly sensitive optical imaging: from single-cell analysis to in vivo whole-body bioluminescence imaging.

    Mezzanotte, Laura; Blankevoort, Vicky; Löwik, Clemens W G M; Kaijzel, Eric L

    2014-09-01

    Fluorescence and bioluminescence imaging have different advantages and disadvantages depending on the application. Bioluminescence imaging is now the most sensitive optical technique for tracking cells, promoter activity studies, or for longitudinal in vivo preclinical studies. Far-red and near-infrared fluorescence imaging have the advantage of being suitable for both ex vivo and in vivo analysis and have translational potential, thanks to the availability of very sensitive imaging instrumentation. Here, we report the development and validation of a new luciferase fusion reporter generated by the fusion of the firefly luciferase Luc2 to the far-red fluorescent protein TurboFP635 by a 14-amino acid linker peptide. Expression of the fusion protein, named TurboLuc, was analyzed in human embryonic kidney cells, (HEK)-293 cells, via Western blot analysis, fluorescence microscopy, and in vivo optical imaging. The created fusion protein maintained the characteristics of the original bioluminescent and fluorescent protein and showed no toxicity when expressed in living cells. To assess the sensitivity of the reporter for in vivo imaging, transfected cells were subcutaneously injected in animals. Detection limits of cells were 5 × 10(3) and 5 × 10(4) cells for bioluminescent and fluorescent imaging, respectively. In addition, hydrodynamics-based in vivo gene delivery using a minicircle vector expressing TurboLuc allowed for the analysis of luminescent signals over time in deep tissue. Bioluminescence could be monitored for over 30 days in the liver of animals. In conclusion, TurboLuc combines the advantages of both bioluminescence and fluorescence and allows for highly sensitive optical imaging ranging from single-cell analysis to in vivo whole-body bioluminescence imaging.

  20. Assessing laser-tissue damage with bioluminescent imaging.

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

    2006-01-01

    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 (lambda=10.6 microm, 0.679 to 2.262 Wcm2, cw, unfocused Gaussian beam, omegaL=4.5 mm, 1 min exposure). The bioluminescence was monitored noninvasively with an IVIS 100 Bioluminescent Imaging System. BLI indicated that peak hsp70 expression occurs 4 to 12 h after exposure to thermal stress. A minimum irradiance of 0.679 Wcm2 activated the hsp70 response, and a higher irradiance of 2.262 Wcm2 was associated with a severe reduction in hsp70 response due to tissue ablation. Reverse transcription polymerase chain reaction demonstrated that hsp70 mRNA levels increased with prolonged heating exposures. Enzyme-linked immunosorbent protein assays confirmed that luciferase was an accurate surrogate for hsp70 intracellular protein levels. Hematoxylin

  1. 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

  2. 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.

  3. 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.

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

    Thomas Chuzel

    Full Text Available 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 has never been assessed on bacteria bioluminescent models. To this end, we investigated the effects of four anesthesia protocols on the bioluminescence flux in a central venous catheter murine model (SKH1-hr(hr mice infected by a bioluminescent S. aureus Xen36 strain. Bioluminescence imaging was performed on mice anesthetized by either ketamine/xylazine (with or without oxygen supplementation, or isoflurane carried with air or oxygen. Total flux emission was determined in vivo daily for 3 days and ex vivo at the end of the study together with a CFU counting of the biofilm in the catheter. Bioluminescence flux differences appear between the different anesthetic protocols. Using a ketamine/xylazine anesthesia (with air, bacteria detection was impossible since the bioluminescence signal remains in the background signal. Mice anesthetized with isoflurane and oxygen led to a signal significantly higher to the background all along the kinetics. The use of isoflurane in air presents a bioluminescence signal similar to the use of ketamine/xylazine with oxygen. These data highlight the importance of oxygen to improve bioluminescence flux by bacteria with isoflurane as well as with ketamine/xylazine anesthetics. As a conclusion, we recommend the use of isoflurane anesthetic with oxygen to increase the bioluminescence sensitivity in this kind of study.

  5. 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 has never been assessed on bacteria bioluminescent models. To this end, we investigated the effects of four anesthesia protocols on the bioluminescence flux in a central venous catheter murine model (SKH1-hr(hr) mice) infected by a bioluminescent S. aureus Xen36 strain. Bioluminescence imaging was performed on mice anesthetized by either ketamine/xylazine (with or without oxygen supplementation), or isoflurane carried with air or oxygen. Total flux emission was determined in vivo daily for 3 days and ex vivo at the end of the study together with a CFU counting of the biofilm in the catheter. Bioluminescence flux differences appear between the different anesthetic protocols. Using a ketamine/xylazine anesthesia (with air), bacteria detection was impossible since the bioluminescence signal remains in the background signal. Mice anesthetized with isoflurane and oxygen led to a signal significantly higher to the background all along the kinetics. The use of isoflurane in air presents a bioluminescence signal similar to the use of ketamine/xylazine with oxygen. These data highlight the importance of oxygen to improve bioluminescence flux by bacteria with isoflurane as well as with ketamine/xylazine anesthetics. As a conclusion, we recommend the use of isoflurane anesthetic with oxygen to increase the bioluminescence sensitivity in this kind of study.

  6. 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.

  7. 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.

  8. 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

  9. 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.

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

    Berger, Frank; Paulmurugan, Ramasamy; Bhaumik, Srabani; Gambhir, Sanjiv Sam

    2008-01-01

    Purpose 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 g

  11. In vivo bioluminescence imaging of Ca signalling in the brain of Drosophila.

    Martin, Jean-René; Rogers, Kelly L; Chagneau, Carine; Brûlet, Philippe

    2007-03-07

    Many different cells' signalling pathways are universally regulated by Ca(2+) concentration [Ca(2+)] rises that have highly variable amplitudes and kinetic properties. Optical imaging can provide the means to characterise both the temporal and spatial aspects of Ca(2+) signals involved in neurophysiological functions. New methods for in vivo imaging of Ca(2+) signalling in the brain of Drosophila are required for probing the different dynamic aspects of this system. In studies here, whole brain Ca(2+) imaging was performed on transgenic flies with targeted expression of the bioluminescent Ca(2+) reporter GFP-aequorin (GA) in different neural structures. A photon counting based technique was used to undertake continuous recordings of cytosolic [Ca(2+)] over hours. Time integrals for reconstructing images and analysis of the data were selected offline according to the signal intensity. This approach allowed a unique Ca(2+) response associated with cholinergic transmission to be identified by whole brain imaging of specific neural structures. Notably, [Ca(2+)] transients in the Mushroom Bodies (MBs) following nicotine stimulation were accompanied by a delayed secondary [Ca(2+)] rise (up to 15 min. later) in the MB lobes. The delayed response was sensitive to thapsigargin, suggesting a role for intra-cellular Ca(2+) stores. Moreover, it was reduced in dunce mutant flies, which are impaired in learning and memory. Bioluminescence imaging is therefore useful for studying Ca(2+) signalling pathways and for functional mapping of neurophysiological processes in the fly brain.

  12. In vivo Bioluminescence Imaging of Ca2+ Signalling in the Brain of Drosophila

    Chagneau, Carine; Brûlet, Philippe

    2007-01-01

    Many different cells' signalling pathways are universally regulated by Ca2+ concentration [Ca2+] rises that have highly variable amplitudes and kinetic properties. Optical imaging can provide the means to characterise both the temporal and spatial aspects of Ca2+ signals involved in neurophysiological functions. New methods for in vivo imaging of Ca2+ signalling in the brain of Drosophila are required for probing the different dynamic aspects of this system. In studies here, whole brain Ca2+ imaging was performed on transgenic flies with targeted expression of the bioluminescent Ca2+ reporter GFP-aequorin (GA) in different neural structures. A photon counting based technique was used to undertake continuous recordings of cytosolic [Ca2+] over hours. Time integrals for reconstructing images and analysis of the data were selected offline according to the signal intensity. This approach allowed a unique Ca2+ response associated with cholinergic transmission to be identified by whole brain imaging of specific neural structures. Notably, [Ca2+] transients in the Mushroom Bodies (MBs) following nicotine stimulation were accompanied by a delayed secondary [Ca2+] rise (up to 15 min. later) in the MB lobes. The delayed response was sensitive to thapsigargin, suggesting a role for intra-cellular Ca2+ stores. Moreover, it was reduced in dunce mutant flies, which are impaired in learning and memory. Bioluminescence imaging is therefore useful for studying Ca2+ signalling pathways and for functional mapping of neurophysiological processes in the fly brain. PMID:17342209

  13. In vivo bioluminescence imaging of Ca signalling in the brain of Drosophila.

    Jean-René Martin

    Full Text Available Many different cells' signalling pathways are universally regulated by Ca(2+ concentration [Ca(2+] rises that have highly variable amplitudes and kinetic properties. Optical imaging can provide the means to characterise both the temporal and spatial aspects of Ca(2+ signals involved in neurophysiological functions. New methods for in vivo imaging of Ca(2+ signalling in the brain of Drosophila are required for probing the different dynamic aspects of this system. In studies here, whole brain Ca(2+ imaging was performed on transgenic flies with targeted expression of the bioluminescent Ca(2+ reporter GFP-aequorin (GA in different neural structures. A photon counting based technique was used to undertake continuous recordings of cytosolic [Ca(2+] over hours. Time integrals for reconstructing images and analysis of the data were selected offline according to the signal intensity. This approach allowed a unique Ca(2+ response associated with cholinergic transmission to be identified by whole brain imaging of specific neural structures. Notably, [Ca(2+] transients in the Mushroom Bodies (MBs following nicotine stimulation were accompanied by a delayed secondary [Ca(2+] rise (up to 15 min. later in the MB lobes. The delayed response was sensitive to thapsigargin, suggesting a role for intra-cellular Ca(2+ stores. Moreover, it was reduced in dunce mutant flies, which are impaired in learning and memory. Bioluminescence imaging is therefore useful for studying Ca(2+ signalling pathways and for functional mapping of neurophysiological processes in the fly brain.

  14. Real-Time Bioluminescent Tracking of Cellular Population Dynamics

    Close, Dan; Xu, Tingling; Ripp, Steven; Sayler, Gary

    2015-01-01

    Cellular population dynamics are routinely monitored across many diverse fields for a variety of purposes. In general, these dynamics are assayed either through the direct counting of cellular aliquots followed by extrapolation to the total population size, or through the monitoring of signal intensity from any number of externally stimulated reporter proteins. While both viable methods, here we describe a novel technique that allows for the automated, non-destructive tracking of cellular population dynamics in real-time. This method, which relies on the detection of a continuous bioluminescent signal produced through expression of the bacterial luciferase gene cassette, provides a low cost, low time-intensive means for generating additional data compared to alternative methods. PMID:24166372

  15. Real-Time Bioluminescent Tracking of Cellular Population Dynamics

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

    2014-01-01

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

  16. 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.

  17. 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

  18. High resolution in vivo bioluminescent imaging for the study of bacterial tumour targeting.

    Michelle Cronin

    Full Text Available The ability to track microbes in real time in vivo is of enormous value for preclinical investigations in infectious disease or gene therapy research. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumours following systemic administration. Bioluminescent Imaging (BLI represents a powerful tool for use with bacteria engineered to express reporter genes such as lux. BLI is traditionally used as a 2D modality resulting in images that are limited in their ability to anatomically locate cell populations. Use of 3D diffuse optical tomography can localize the signals but still need to be combined with an anatomical imaging modality like micro-Computed Tomography (μCT for interpretation.In this study, the non-pathogenic commensal bacteria E. coli K-12 MG1655 and Bifidobacterium breve UCC2003, or Salmonella Typhimurium SL7207 each expressing the luxABCDE operon were intravenously (i.v. administered to mice bearing subcutaneous (s.c FLuc-expressing xenograft tumours. Bacterial lux signal was detected specifically in tumours of mice post i.v.-administration and bioluminescence correlated with the numbers of bacteria recovered from tissue. Through whole body imaging for both lux and FLuc, bacteria and tumour cells were co-localised. 3D BLI and μCT image analysis revealed a pattern of multiple clusters of bacteria within tumours. Investigation of spatial resolution of 3D optical imaging was supported by ex vivo histological analyses. In vivo imaging of orally-administered commensal bacteria in the gastrointestinal tract (GIT was also achieved using 3D BLI. This study demonstrates for the first time the potential to simultaneously image multiple BLI reporter genes three dimensionally in vivo using approaches that provide unique information on spatial locations.

  19. 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

    2016-06-14

    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.

  20. 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.

  1. 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.

  2. In vivo bioluminescence imaging of neurogenesis - the role of the blood brain barrier in an experimental model of Parkinson's disease.

    Fricke, Inga B; Schelhaas, Sonja; Zinnhardt, Bastian; Viel, Thomas; Hermann, Sven; Couillard-Després, Sébastien; Jacobs, Andreas H

    2017-02-13

    Bioluminescence imaging in transgenic mice expressing firefly luciferase in Doublecortin(+) (Dcx) neuroblasts might serve as a powerful tool to study the role of neurogenesis in models of brain injury and neurodegeneration using non-invasive, longitudinal in vivo imaging. Therefore, we aimed to use BLI in B6(Cg)-Tyrc-2J/J Dcx-Luc (Doublecortin-Luciferase, Dcx-Luc) mice to investigate its suitability to assess neurogenesis in a unilateral injection model of Parkinson's disease. We further aimed to assess the blood brain barrier leakage associated with the intranigral 6-OHDA injection to evaluate its impact on substrate delivery and bioluminescence signal intensity. Two weeks after lesion, we observed an increase in bioluminescence signal in the ipsilateral hippocampal region in both, 6-OHDA and vehicle injected Dcx-Luc mice. At the same time, no corresponding increase in Dcx(+) neuroblast numbers could be observed in the dentate gyrus of C57Bl6 mice. Blood brain barrier leakage was observed in the hippocampal region and in the degenerating substantia nigra of C57Bl6 mice in vivo using T1 weighted Magnetic Resonance Imaging with Gadovist(®) and ex vivo using Evans Blue Fluorescence Reflectance Imaging and mouse Immunoglobulin G staining. Our data suggests a BLI signal dependency on blood brain barrier permeability, underlining a major pitfall of substrate/tracer dependent imaging in invasive disease models.

  3. 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

  4. 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.

  5. 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

  6. 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.

  7. Direct Evidence of Mesenchymal Stem Cell Tropism for Tumor and Wounding Microenvironments using In Vivo Bioluminescence Imaging

    Kidd, Shannon; Spaeth, Erika; Dembinski, Jennifer L.; Dietrich, Martin; Watson, Keri; Klopp, Ann; Battula, Lokesh; Weil, Micheal; Andreeff, Michael; Marini, Frank C.

    2014-01-01

    Multipotent mesenchymal stromal/stem cells (MSC) have shown potential clinical utility. However, previous assessments of MSC behavior in recipients have relied on visual detection in host tissue following sacrifice, failing to monitor in vivo MSC dispersion in a single animal and limiting the number of variables that can be observed concurrently. In this study, we utilized noninvasive, in vivo bioluminescent imaging to determine conditions under which MSC selectively engraft in sites of inflammation. MSC modified to express firefly luciferase (MSC-ffLuc) were injected into healthy mice or mice bearing inflammatory insults, and MSC localization was followed with bioluminescent imaging. Inflammatory insults investigated included cutaneous needle-stick and surgical incision wounds, as well as xenogeneic and syngeneic tumors. We also compared tumor models in which MSC were intraveneously or intraperitoneally delivered. Our results demonstrate hMSC-ffLuc systemically delivered to non-tumor bearing animals initially reside in the lungs, then egress to the liver and spleen and decrease in signal over time. However, hMSC in wounded mice engraft and remain detectable only in injured sites. Similarly, in syngeneic and xenogeneic breast carcinoma-bearing mice, bioluminescent detection of systemically delivered MSC revealed persistent, specific co-localization with sites of tumor development. This pattern of tropism was also observed in an ovarian tumor model in which MSC were IP injected. In this study we have identified conditions under which MSC tropism and selective engraftment in sites of inflammation can be monitored by bioluminescent imaging over time. Importantly, these consistent findings were independent of tumor type, immunocompetence and route of MSC delivery. PMID:19650040

  8. Direct evidence of mesenchymal stem cell tropism for tumor and wounding microenvironments using in vivo bioluminescent imaging.

    Kidd, Shannon; Spaeth, Erika; Dembinski, Jennifer L; Dietrich, Martin; Watson, Keri; Klopp, Ann; Battula, Venkata Lokesh; Weil, Micheal; Andreeff, Michael; Marini, Frank C

    2009-10-01

    Multipotent mesenchymal stromal/stem cells (MSC) have shown potential clinical utility. However, previous assessments of MSC behavior in recipients have relied on visual detection in host tissue following sacrifice, failing to monitor in vivo MSC dispersion in a single animal and limiting the number of variables that can be observed concurrently. In this study, we used noninvasive, in vivo bioluminescent imaging to determine conditions under which MSC selectively engraft in sites of inflammation. MSC modified to express firefly luciferase (ffLuc-MSC) were injected into healthy mice or mice bearing inflammatory insults, and MSC localization was followed with bioluminescent imaging. The inflammatory insults investigated included cutaneous needle-stick and surgical incision wounds, as well as xenogeneic and syngeneic tumors. We also compared tumor models in which MSC were i.v. or i.p. delivered. Our results demonstrate that ffLuc-expressing human MSC (hMSC) systemically delivered to nontumor-bearing animals initially reside in the lungs, then egress to the liver and spleen, and decrease in signal over time. However, hMSC in wounded mice engraft and remain detectable only at injured sites. Similarly, in syngeneic and xenogeneic breast carcinoma-bearing mice, bioluminescent detection of systemically delivered MSC revealed persistent, specific colocalization with sites of tumor development. This pattern of tropism was also observed in an ovarian tumor model in which MSC were i.p. injected. In this study, we identified conditions under which MSC tropism and selective engraftment in sites of inflammation can be monitored by bioluminescent imaging over time. Importantly, these consistent findings were independent of tumor type, immunocompetence, and route of MSC delivery.

  9. 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.

  10. Global solution of the finite element shape-from-shading model with a bioluminescent molecular imaging application.

    Zhong, Jianghong; Tian, Jie; Yang, Xin; Qin, Chenghu

    2010-01-01

    Only a planar bioluminescence image acquired from an ordinary cooled charge-coupled device (CCD) array every time, how to re-establish the three-dimensional small animal shape and light intensity distribution on the surface has become urgent to be solved as a bottleneck of bioluminescence tomography (BLT) reconstruction. In this paper, a finite element algorithm to solve the Dirichlet type problem for the first order Hamilton-Jacobi equation related to the shape-fromshading model is adopted. The algorithm outputting the globally maximal solution of the above problem avoids cumbersome boundary conditions on the interfaces between light and shadows and the use of additional information on the surface. The results of the optimization method are satisfied. It demonstrates the feasibility and potential of the finite element shape-fromshading (FE-SFS) model for reconstructing the small animal surface that lays one of key foundations for a fast low-cost application of the BLT in the next future.

  11. A label-free bioluminescent sensor for real-time monitoring polynucleotide kinase activity.

    Du, Jiao; Xu, Qinfeng; Lu, Xiaoquan; Zhang, Chun-yang

    2014-08-19

    Polynucleotide kinase (PNK) plays a crucial role in maintaining the genomic stability of cells and is becoming a potential target in the radio-therapeutic treatment of cancers. The fluorescent method is usually used to measure the PNK activity, but it is impossible to obtain the real-time monitoring without the employment of the labeled DNA probes. Here, we report a label-free bioluminescent sensor for PNK activity assay through real-time monitoring of the phosphorylation-dependent DNA ligation reaction. In this bioluminescent sensor, two hairpin DNA probes with 5'-protruding terminal are designed as the phosphate acceptor, and the widely used phosphate donor of ATP is substituted by dCTP. In the absence of PNK, the ligation reaction cannot be triggered due to the lack of 5'-phosphoryl groups in the probes, and the background signal is negligible. With the addition of PNK, the phosphorylation-ligation reaction of the probes is initiated with the release of AMP, and the subsequent conversion of AMP to ATP leads to the generation of distinct bioluminescence signal. The PNK activity assay can be performed in real time by continuously monitoring the bioluminescence signal. This bioluminescent sensor is much simpler, label-free, cost-effective, and free from the autofluorescence interference of biological matrix, and can be further used for quantitative, kinetic, and inhibition assay.

  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. Dual-color bioluminescence imaging assay using green- and red-emitting beetle luciferases at subcellular resolution.

    Yasunaga, Mayu; Nakajima, Yoshihiro; Ohmiya, Yoshihiro

    2014-09-01

    Bioluminescence imaging is widely used to monitor cellular events, including gene expression in vivo and in vitro. Moreover, recent advances in luciferase technology have made possible imaging at the single-cell level. To improve the bioluminescence imaging system, we have developed a dual-color imaging system in which the green-emitting luciferase from a Brazilian click beetle (Emerald Luc, ELuc) and the red-emitting luciferase from a railroad worm (Stable Luciferase Red, SLR) were used as reporters, which were localized to the peroxisome and the nucleus, respectively. We clearly captured simultaneously the subcellular localization of ELuc in the peroxisome and SLR in the nucleus of a single cell using a high-magnification objective lens with 3-min exposure time without binning using a combination of optical filters. Furthermore, to apply this system to quantitative time-lapse imaging, the activation of nuclear factor triggered by tumor necrosis factor α was measured using nuclear-targeted SLR and peroxisome-targeted ELuc as the test and internal control reporters, respectively. We successfully quantified the kinetics of activation of nuclear factor κB using nuclear-targeted SLR and the transcriptional change of the internal control promoter using peroxisome-targeted ELuc simultaneously in a single cell, and showed that the activation kinetics, including activation rate and amplitude, differed among cells. The results demonstrated that this imaging system can visualize the subcellular localization of reporters and track the expressions of two genes simultaneously at subcellular resolution.

  14. Imaging of bubonic plague dynamics by in vivo tracking of bioluminescent Yersinia pestis.

    Toan Nham

    Full Text Available Yersinia pestis dissemination in a host is usually studied by enumerating bacteria in the tissues of animals sacrificed at different times. This laborious methodology gives only snapshots of the infection, as the infectious process is not synchronized. In this work we used in vivo bioluminescence imaging (BLI to follow Y. pestis dissemination during bubonic plague. We first demonstrated that Y. pestis CO92 transformed with pGEN-luxCDABE stably emitted bioluminescence in vitro and in vivo, while retaining full virulence. The light produced from live animals allowed to delineate the infected organs and correlated with bacterial loads, thus validating the BLI tool. We then showed that the first step of the infectious process is a bacterial multiplication at the injection site (linea alba, followed by a colonization of the draining inguinal lymph node(s, and subsequently of the ipsilateral axillary lymph node through a direct connection between the two nodes. A mild bacteremia and an effective filtering of the blood stream by the liver and spleen probably accounted for the early bacterial blood clearance and the simultaneous development of bacterial foci within these organs. The saturation of the filtering capacity of the spleen and liver subsequently led to terminal septicemia. Our results also indicate that secondary lymphoid tissues are the main targets of Y. pestis multiplication and that colonization of other organs occurs essentially at the terminal phase of the disease. Finally, our analysis reveals that the high variability in the kinetics of infection is attributable to the time the bacteria remain confined at the injection site. However, once Y. pestis has reached the draining lymph nodes, the disease progresses extremely rapidly, leading to the invasion of the entire body within two days and to death of the animals. This highlights the extraordinary capacity of Y. pestis to annihilate the host innate immune response.

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

    Wang, Yan; Zhang, Beilei; Liu, Wei; Dai, Yunpeng; Shi, Yaru; Zeng, Qi; Wang, Fu

    2016-04-19

    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.

  16. A single-cell bioluminescence imaging system for monitoring cellular gene expression in a plant body.

    Muranaka, Tomoaki; Kubota, Saya; Oyama, Tokitaka

    2013-12-01

    Gene expression is a fundamental cellular process and expression dynamics are of great interest in life science. We succeeded in monitoring cellular gene expression in a duckweed plant, Lemna gibba, using bioluminescent reporters. Using particle bombardment, epidermal and mesophyll cells were transfected with the luciferase gene (luc+) under the control of a constitutive [Cauliflower mosaic virus 35S (CaMV35S)] and a rhythmic [Arabidopsis thaliana CIRCADIAN CLOCK ASSOCIATED 1 (AtCCA1)] promoter. Bioluminescence images were captured using an EM-CCD (electron multiply charged couple device) camera. Luminescent spots of the transfected cells in the plant body were quantitatively measured at the single-cell level. Luminescence intensities varied over a 1,000-fold range among CaMV35S::luc+-transfected cells in the same plant body and showed a log-normal-like frequency distribution. We monitored cellular gene expression under light-dark conditions by capturing bioluminescence images every hour. Luminescence traces of ≥50 individual cells in a frond were successfully obtained in each monitoring procedure. Rhythmic and constitutive luminescence behaviors were observed in cells transfected with AtCCA1::luc+ and CaMV35S::luc+, respectively. Diurnal rhythms were observed in every AtCCA1::luc+-introduced cell with traceable luminescence, and slight differences were detected in their rhythmic waveforms. Thus the single-cell bioluminescence monitoring system was useful for the characterization of cellular gene expression in a plant body.

  17. 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.

  18. A dual-color far-red to near-infrared firefly luciferin analogue designed for multiparametric bioluminescence imaging.

    Jathoul, Amit P; Grounds, Helen; Anderson, James C; Pule, Martin A

    2014-11-24

    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 (Fluc). 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, exhibits pH dependent fluorescence spectra and emits bioluminescence of different colors with different engineered Fluc enzymes. Our analogue produces different far-red to nIR emission maxima up to λ(max)=706 nm with different Fluc mutants. This emission is the most red-shifted bioluminescence reported without using a resonance energy transfer acceptor. This improvement should allow tissues to be more effectively probed using multiparametric deep-tissue bioluminescence imaging.

  19. Monitoring the Response of Hyperbilirubinemia in the Mouse Brain by In Vivo Bioluminescence Imaging.

    Manni, Isabella; Di Rocco, Giuliana; Fusco, Salvatore; Leone, Lucia; Barbati, Saviana Antonella; Carapella, Carmine Maria; Grassi, Claudio; Piaggio, Giulia; Toietta, Gabriele

    2016-12-28

    Increased levels of unconjugated bilirubin are neurotoxic, but the mechanism leading to neurological damage has not been completely elucidated. Innovative strategies of investigation are needed to more precisely define this pathological process. By longitudinal in vivo bioluminescence imaging, we noninvasively visualized the brain response to hyperbilirubinemia in the MITO-Luc mouse, in which light emission is restricted to the regions of active cell proliferation. We assessed that acute hyperbilirubinemia promotes bioluminescence in the brain region, indicating an increment in the cell proliferation rate. Immunohistochemical detection in brain sections of cells positive for both luciferase and the microglial marker allograft inflammatory factor 1 suggests proliferation of microglial cells. In addition, we demonstrated that brain induction of bioluminescence was altered by pharmacological displacement of bilirubin from its albumin binding sites and by modulation of the blood-brain barrier permeability, all pivotal factors in the development of bilirubin-induced neurologic dysfunction. We also determined that treatment with minocycline, an antibiotic with anti-inflammatory and neuroprotective properties, or administration of bevacizumab, an anti-vascular endothelial growth factor antibody, blunts bilirubin-induced bioluminescence. Overall the study supports the use of the MITO-Luc mouse as a valuable tool for the rapid response monitoring of drugs aiming at preventing acute bilirubin-induced neurological dysfunction.

  20. 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.

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

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

    2010-01-01

    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 progressio...

  2. Molecular Origin of Color Variation in Firefly (Beetle) Bioluminescence: A Chemical Basis for Biological Imaging.

    Hirano, Takashi

    2016-01-01

    Firefly shows bioluminescence by "luciferin-luciferase" (L-L) reaction using luciferin, luciferase, ATP and O2. The chemical photon generation by an enzymatic reaction is widely utilized for analytical methods including biological imaging in the life science fields. To expand photondetecting analyses with firefly bioluminescence, it is important for users to understand the chemical basis of the L-L reaction. In particular, the emission color variation of the L-L reaction is one of the distinguishing characteristics for multicolor luciferase assay and in vivo imaging. From the viewpoint of fundamental chemistry, this review explains the recent progress in the studies on the molecular mechanism of emission color variation after showing the outline of the reaction mechanism of the whole L-L reaction. On the basis of the mechanism, the progresses in organic synthesis of luciferin analogs modulating their emission colors are also presented to support further developments of red/near infrared in vivo biological imaging utility of firefly bioluminescence.

  3. 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.

  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.

  5. In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin.

    Lark, Arianna R; Kitamoto, Toshihiro; Martin, Jean-René

    2016-01-08

    Functional in vivo imaging has become a powerful approach to study the function and physiology of brain cells and structures of interest. Recently a new method of Ca(2+)-imaging using the bioluminescent reporter GFP-aequorin (GA) has been developed. This new technique relies on the fusion of the GFP and aequorin genes, producing a molecule capable of binding calcium and - with the addition of its cofactor coelenterazine - emitting bright light that can be monitored through a photon collector. Transgenic lines carrying the GFP-aequorin gene have been generated for both mice and Drosophila. In Drosophila, the GFP-aequorin gene has been placed under the control of the GAL4/UAS binary expression system allowing for targeted expression and imaging within the brain. This method has subsequently been shown to be capable of detecting both inward Ca(2+)-transients and Ca(2+)-released from inner stores. Most importantly it allows for a greater duration in continuous recording, imaging at greater depths within the brain, and recording at high temporal resolutions (up to 8.3 msec). Here we present the basic method for using bioluminescent imaging to record and analyze Ca(2+)-activity within the mushroom bodies, a structure central to learning and memory in the fly brain.

  6. 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.

  7. 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.

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

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

    2011-01-01

    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. PMID:22346573

  9. Dynamic Modeling of Marine Bioluminescence and Night Time Leaving Radiance

    2012-09-30

    person shall be subject to a penalty for failing to comply with a collection of information if it does not display a currently valid OMB control ...in agreement with observations. In the model, the BL potential dynamics were controlled by advective and diffusive processes only, and as it was...Chapter in the Book: Subsea Optics and Imaging, accepted, to be published by Woodhead Publishing. Shulman, I., B. Penta, M. A. Moline, S. H. D

  10. 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.

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

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

    2013-05-17

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

  12. 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

  13. 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.

  14. Quantitative bioluminescence imaging--a method for the detection of metabolite distributions in frozen tissues

    Mueller-Klieser, Wolfgang; Walenta, Stefan; Schwickert, Georg

    1994-02-01

    A novel technique allows for measurement of metabolite distributions in tissue cryosections at a microscopic level using bioluminescence, single photon imaging, and computerized image analysis. Metabolites, such as ATP, glucose and lactate are registered in absolute concentration units, and the respective images can be correlated with each other and with histological structures by specific algorithms. One striking difference between malignant tumors and normal tissue is the pronounced heterogeneity of metabolite distributions in malignancies contrasted by rather homogeneous patterns obtained in many normal organs. The heterogeneous distribution of metabolites in solid tumors reflects the chaotic organization of the histological architecture and of the microvascular supply in cancerous tissue. Pixel-to-pixel comparison of metabolite distributions measured in cervix cancers of patients revealed a negative linear correlation between glucose and ATP concentrations at identical locations. In contrast, local lactate concentration was positively correlated with ATP.

  15. Bioluminescence imaging reveals dynamics of beta cell loss in the non-obese diabetic (NOD) mouse model.

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

    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 initial BLI at 10 weeks of age, whereas hyperglycemia did not ensue until mice were at least 16 weeks old. Mice that did not become diabetic maintained insulin secretion and had less of a decline in bioluminescence than mice that became diabetic. Bioluminescence measurements predicted a decline in beta cell mass prior to the onset of hyperglycemia and tracked beta cell loss. This model should be useful for investigating the fundamental processes underlying autoimmune diabetes and developing new therapies targeting beta cell protection and regeneration.

  16. 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

  17. The Drug Excipient Cyclodextrin Interacts With d-Luciferin and Interferes With Bioluminescence Imaging.

    Kumar, Jeyan S; Miller Jenkins, Lisa M; Gottesman, Michael M; Hall, Matthew D

    2016-01-01

    Cyclodextrins are well-characterized, barrel-shaped molecules that can solubilize organic small molecules in aqueous solution via host-guest interactions. As such, cyclodextrins are used as excipients for experimental therapeutics in vivo. We observed unanticipated modifications to bioluminescence imaging (BLI) signal intensity when 2-hydroxy-propyl-β-cyclodextrin (HPCD) was coinjected as an excipient. We hypothesized that HPCD bindsd-luciferin and interferes with the BLI signal. Using luciferase-expressing cell lines, we showed that HPCD lowers the BLI signal in a concentration-dependent manner. Flow cytometry revealed that HPCD resulted in reduced cellular accumulation ofd-luciferin, and mass spectrometry revealedd-luciferin HPCD species, confirming a direct interaction. In vivo imaging using a luciferase mouse model demonstrated that HPCD reduced luciferin-mediated BLI compared to luciferin alone. The implications of using HPCD as an excipient in BLI studies are discussed.

  18. Quantitative Imaging of D-2-Hydroxyglutarate in Selected Histological Tissue Areas by a Novel Bioluminescence Technique.

    Voelxen, Nadine F; Walenta, Stefan; Proescholdt, Martin; Dettmer, Katja; Pusch, Stefan; Mueller-Klieser, Wolfgang

    2016-01-01

    Patients with malignant gliomas have a poor prognosis with average survival of less than 1 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 that was discovered first in this tumor entity. D2HG is generated in large amounts due to various "gain-of-function" mutations in the isocitrate dehydrogenases IDH1 and IDH2. 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 (bioluminescence assay for determining D2HG in sections of snap-frozen tissue. The assay was verified independently by photometric tests and liquid chromatography/mass spectrometry. 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.

  19. 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

  20. In Vivo Follow-up of Brain Tumor Growth via Bioluminescence Imaging and Fluorescence Tomography

    Coralie Genevois

    2016-10-01

    Full Text Available Reporter gene-based strategies are widely used in experimental oncology. Bioluminescence imaging (BLI using the firefly luciferase (Fluc as a reporter gene and d-luciferin as a substrate is currently the most widely employed technique. The present paper compares the performances of BLI imaging with fluorescence imaging using the near infrared fluorescent protein (iRFP to monitor brain tumor growth in mice. Fluorescence imaging includes fluorescence reflectance imaging (FRI, fluorescence diffuse optical tomography (fDOT, and fluorescence molecular Imaging (FMT®. A U87 cell line was genetically modified for constitutive expression of both the encoding Fluc and iRFP reporter genes and assayed for cell, subcutaneous tumor and brain tumor imaging. On cultured cells, BLI was more sensitive than FRI; in vivo, tumors were first detected by BLI. Fluorescence of iRFP provided convenient tools such as flux cytometry, direct detection of the fluorescent protein on histological slices, and fluorescent tomography that allowed for 3D localization and absolute quantification of the fluorescent signal in brain tumors.

  1. Light Emission Requires Exposure to the Atmosphere in Ex Vivo Bioluminescence Imaging

    Yusuke Inoue

    2006-04-01

    Full Text Available The identification of organs bearing luciferase activity by in vivo bioluminescence imaging (BLI is often difficult, and ex vivo imaging of excised organs plays a complementary role. This study investigated the importance of exposure to the atmosphere in ex vivo BLI. Mice were inoculated with murine pro-B cell line Ba/F3 transduced with firefly luciferase and p190 BCR-ABL. They were killed following in vivo BLI, and whole-body imaging was done after death and then after intraperitoneal air injection. In addition, the right knee was exposed and imaged before and after the adjacent bones were cut. Extensive light signals were seen on in vivo imaging. The luminescence disappeared after the animal was killed, and air injection restored the light emission from the abdomen only, suggesting a critical role of atmospheric oxygen in luminescence after death. Although no substantial light signal at the right knee was seen before bone cutting, light emission was evident after cutting. In conclusion, in ex vivo BLI, light emission requires exposure to the atmosphere. Bone destruction is required to demonstrate luciferase activity in the bone marrow after death.

  2. Engraftment and bone mass are enhanced by PTHrP 1-34 in ectopically transplanted vertebrae (vossicle model) and can be non-invasively monitored with bioluminescence and fluorescence imaging.

    Hildreth, Blake Eason; Williams, Michelle M; Dembek, Katarzyna A; Hernon, Krista M; Rosol, Thomas J; Toribio, Ramiro E

    2015-12-01

    Evidence exists that parathyroid hormone-related protein (PTHrP) 1-34 may be more anabolic in bone than parathyroid hormone 1-34. While optical imaging is growing in popularity, scant information exists on the relationships between traditional bone imaging and histology and bioluminescence (BLI) and fluorescence (FLI) imaging. We aimed to evaluate the effects of PTHrP 1-34 on bone mass and determine if relationships existed between radiographic and histologic findings in bone and BLI and FLI indices. Vertebrae (vossicles) from mice coexpressing luciferase and green fluorescent protein were implanted subcutaneously into allogenic nude mice. Transplant recipients were treated daily with saline or PTHrP 1-34 for 4 weeks. BLI, FLI, radiography, histology, and µCT of the vossicles were performed over time. PTHrP 1-34 increased bioluminescence the most after 2 weeks, fluorescence at all time points, and decreased the time to peak bioluminescence at 4 weeks (P ≤ 0.027), the latter of which suggesting enhanced engraftment. PTHrP 1-34 maximized vertebral body volume at 4 weeks (P bioluminescence (r = 0.595; P = 0.019); (2) total fluorescence (r = 0.474; P = 0.074); and (3) max fluorescence (r = 0.587; P = 0.021). In conclusion, PTHrP 1-34 enhances engraftment and bone mass, which can be monitored non-invasively by BLI and FLI.

  3. 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

    2015-12-01

    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 Etests demonstrated increased bioluminescence at sub-MICs of different antibiotics. This study aimed to further evaluate the influence of antibiotic pressure on bioluminescence in S. aureus Xen29. Bioluminescence of S. aureus Xen29, grown planktonically in tryptone soy broth, was quantified in the absence and presence of different concentrations of vancomycin, ciprofloxacin, erythromycin or chloramphenicol and was related to expression of the luxA gene under antibiotic pressure measured using real-time PCR. In the absence of antibiotics, staphylococcal bioluminescence increased over time until a maximum after ca. 6h of growth, and subsequently decreased to the detection threshold after 24h of growth owing to reduced bacterial metabolic activity. Up to MICs of the antibiotics, bioluminescence increased according to a similar pattern up to 6h of growth, but after 24h bioluminescence was higher than in the absence of antibiotics. Contrary to expectations, bioluminescence per organism (CFU) after different growth periods in the absence and at MICs of different antibiotics decreased with increasing expression of luxA. Summarising, antibiotic pressure impacts the relation between CFU and bioluminescence. Under antibiotic pressure, bioluminescence is not controlled by luxA expression but by co-factors impacting the bacterial metabolic activity. This conclusion is of utmost importance when evaluating antibiotic efficacy in live animals using bioluminescent bacterial strains.

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

    Mueller-Klieser, W.; Walenta, S.; Paschen, W.; Kallinowski, F.; Vaupel, P.

    1988-08-03

    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.

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

    Juan Cao

    2016-01-01

    Full Text Available 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.

  6. 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.

  7. Multicolor bioluminescence boosts malaria research: quantitative dual-color assay and single-cell imaging in Plasmodium falciparum parasites.

    Cevenini, Luca; Camarda, Grazia; Michelini, Elisa; Siciliano, Giulia; Calabretta, Maria Maddalena; Bona, Roberta; Kumar, T R Santha; Cara, Andrea; Branchini, Bruce R; Fidock, David A; Roda, Aldo; Alano, Pietro

    2014-09-02

    New reliable and cost-effective antimalarial drug screening assays are urgently needed to identify drugs acting on different stages of the parasite Plasmodium falciparum, and particularly those responsible for human-to-mosquito transmission, that is, the P. falciparum gametocytes. Low Z' factors, narrow dynamic ranges, and/or extended assay times are commonly reported in current gametocyte assays measuring gametocyte-expressed fluorescent or luciferase reporters, endogenous ATP levels, activity of gametocyte enzymes, or redox-dependent dye fluorescence. We hereby report on a dual-luciferase gametocyte assay with immature and mature P. falciparum gametocyte stages expressing red and green-emitting luciferases from Pyrophorus plagiophthalamus under the control of the parasite sexual stage-specific pfs16 gene promoter. The assay was validated with reference antimalarial drugs and allowed to quantitatively and simultaneously measure stage-specific drug effects on parasites at different developmental stages. The optimized assay, requiring only 48 h incubation with drugs and using a cost-effective luminogenic substrate, significantly reduces assay cost and time in comparison to state-of-the-art analogous assays. The assay had a Z' factor of 0.71 ± 0.03, and it is suitable for implementation in 96- and 384-well microplate formats. Moreover, the use of a nonlysing D-luciferin substrate significantly improved the reliability of the assay and allowed one to perform, for the first time, P. falciparum bioluminescence imaging at single-cell level.

  8. Bioluminescent luciferase-modified magnetic nanoparticles as potential imaging agents for mammalian spermatozoa detection and tracking

    Background: Nanoparticles have emerged as key materials for developing applications in nanomedicine, nanobiotechnology, bioimaging and theranostics. Existing bioimaging technologies include bioluminescent resonance energy transfer-conjugated quantum dots (BRET-QDs). Despite the current use of BRET-Q...

  9. 106 ASSESSMENT OF ANTI-BACTERIAL EFFECTS OF PEGYLATED SILVER-COATED CARBON NANOTUBES ON CAUSATIVE BACTERIA OF BOVINE INFERTILITY USING BIOLUMINESCENCE IMAGING SYSTEM.

    Park, S; Chaudhari, A A; Pillai, S; Singh, S R; Willard, S T; Ryan, P L; Feugang, J M

    2016-01-01

    Pathogenic bacteria including Escherichia coli and Salmonella sp. are the major causative agents of endometritis and can cause infertility in livestock animals. Antibiotics are commonly used to terminate bacterial infections, but the development of bacterial antibiotic resistance is often encountered. Nanotechnology associated with silver nanoparticles has been highlighted as an alternative anti-bacterial agent, and pegylated silver-coated single-walled carbon nanotubes have high anti-bacterial effects and are non-toxic to human and murine cells in vitro. Here we verified whether a real-time bioluminescence monitoring system could be an alternative tool to assess anti-bacterial effects of nanotubes in a noninvasive approach. Escherichia coli and Salmonella sp. were transfected with plasmids containing constructs for luciferase enzyme (LuxCDABE) and substrate (luciferin) to create self-illuminating bioluminescent bacteria. Pathogens were grown in LB broth at 37°C, adjusted to 10(7) cfumL(-1), and placed in 96-well plates for treatments. Pegylated (pSWCNTs-Ag) and non-pegylated (SWCNTs-Ag) nanotubes were prepared and added to culture wells at various concentrations (31.25-125µgmL(-1)). The control group corresponded to bacteria without nanotubes (0µgmL(-1)). Anti-bacterial effects of nanotubes were determined every 10min until 1h, then every 30min up to 6h incubation through optical density (600nm) measurements and bioluminescence imaging (BLI) and quantification using an IVIS system. Optical density and BLI data were compared at each time-point using 2-way ANOVA, with PBioluminescence signals emitted by both bacteria stains appeared within 10min of incubation. Thereafter, control bacteria showed exponential growth that was detected as early as 25min post-incubation. Bioluminescence imaging revealed dose-dependent anti-bacterial activities of both pSWCNTs-Ag and SWCNTs-Ag on each E. coli and Salmonella sp. (P0.05); meanwhile, pSWCNTs-Ag nanotubes exhibited

  10. Bioluminescent, Nonlytic, Real-Time Cell Viability Assay and Use in Inhibitor Screening.

    Duellman, Sarah J; Zhou, Wenhui; Meisenheimer, Poncho; Vidugiris, Gediminas; Cali, James J; Gautam, Prson; Wennerberg, Krister; Vidugiriene, Jolanta

    2015-10-01

    Real-time continuous monitoring of cellular processes offers distinct advantages over traditional endpoint assays. A comprehensive representation of the changes occurring in live cells over the entire length of an experiment provides information about the biological status of the cell and informs decisions about the timing of treatments or the use of other functional endpoint assays. We describe a homogeneous, nonlytic, bioluminescent assay that measures cell viability in real time. This time-dependent measurement allowed us to monitor cell health for 72 h from the same test samples, distinguish differential cell growth, and investigate drug mechanism of action by analyzing time- and dose-dependent drug effects. The real-time measurements also allowed us to detect cell death immediately (>75% signal decrease within 15 min of digitonin addition), analyze drug potency versus efficacy, and identify cytostatic versus toxic drug effects. We screened an oncology compound library (Z' = 0.7) and identified compounds with varying activity at different time points (1.6% of the library showed activity within 3 h, whereas 35.4% showed a response by 47 h). The assay compared well with orthogonal endpoint cell viability assays and additionally provided data at multiple time points and the opportunity to multiplex assays on the same cells. To test the advantage of time-dependent measurements to direct optimal timing of downstream applications, we used the real-time cell viability assay to determine the ideal time to measure caspase activity by monitoring the onset of cell death and multiplexing a luminescent caspase activation assay on the same test samples.

  11. Mouse Reporter Strain for Noninvasive Bioluminescent Imaging of Cells that have Undergone Cre-Mediated Recombination

    Michal Safran

    2003-10-01

    Full Text Available Conditional alleles containing LoxP recombination sites, in conjunction with Cre recombinase delivered by a variety of means, allows for spatial and temporal control of gene expression in mouse models. Here we describe a mouse strain in which a luciferase (Luc cDNA, preceded by a LoxP-stop-LoxP (L-S-L cassette, was introduced into the ubiquitously expressed ROSA26 locus. Mouse embryo fibroblasts derived from this strain expressed luciferase after Cre-mediated recombination in vitro. ROSA26 L-S-L-Luc/+ mice expressed luciferase in a diffuse or liver-restricted pattern, as determined by noninvasive, bioluminescent imaging, when crossed to transgenic mice in which Cre was under the control of a zygotically expressed (EIIA-Cre, or a liver-restricted (albumin-Cre, promoter, respectively. Organ-specific luciferase expression was also seen after intraparenchymal administration of an adenovirus encoding Cre. The ROSA26 L-S-L-Luc/+ strain should be useful for characterizing Cre mouse strains and for following the fate of cells that have undergone Cre-mediated recombination in vivo.

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

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

    2014-01-01

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

  13. Continuous and real-time bioaerosol monitoring by combined aerosol-to-hydrosol sampling and ATP bioluminescence assay.

    Park, Ji-Woon; Kim, Hyeong Rae; Hwang, Jungho

    2016-10-19

    We present a methodology for continuous and real-time bioaerosol monitoring wherein an aerosol-to-hydrosol sampler is integrated with a bioluminescence detector. Laboratory test was conducted by supplying an air flow with entrained test bacteria (Staphylococcus epidermidis) to the inlet of the sampler. High voltage was applied between the discharge electrode and the ground electrode of the sampler to generate air ions by corona discharge. The bacterial aerosols were charged by the air ions and sampled in a flowing liquid containing both a cell lysis buffer and adenosine triphosphate (ATP) bioluminescence reagents. While the liquid was delivered to the bioluminescence detector, sampled bacteria were dissolved by the cell lysis buffer and ATP was extracted. The ATP was reacted with the ATP bioluminescence reagents, causing light to be emitted. When the concentration of bacteria in the aerosols was varied, the ATP bioluminescence signal in relative light units (RLUs) closely tracked the concentration in particles per unit air volume (# cm(-3)), as measured by an aerosol particle sizer. The total response time required for aerosol sampling and ATP bioluminescence detection increased from 30 s to 2 min for decreasing liquid sampling flow rate from 800 to 200 μLPM, respectively. However, lower concentration of S. epidermidis aerosols was able to be detected with lower liquid sampling flow rate (1 RLU corresponded to 6.5 # cm(-3) of S. epidermidis aerosols at 200 μLPM and 25.5 # cm(-3) at 800 μLPM). After obtaining all data sets of concentration of S. epidermidis aerosols and concentration of S. epidermidis particles collected in the flowing liquid, it was found that with our bioluminescence detector, 1 RLU corresponded to 1.8 × 10(5) (±0.2 × 10(5)) # mL(-1) of S. epidermidis in liquid. After the lab-test with S. epidermidis, our bioaerosol monitoring device was located in the lobby of a building. Air sampling was conducted continuously for 90

  14. 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.

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

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

    2014-02-15

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

  16. Monitoring Disease Progression and Therapeutic Response in a Disseminated Tumor Model for Non-Hodgkin Lymphoma by Bioluminescence Imaging

    Margarethe Köberle

    2015-07-01

    Full Text Available Xenograft tumor models are widely studied in cancer research. Our aim was to establish and apply a model for aggressive CD20-positive B-cell non-Hodgkin lymphomas, enabling us to monitor tumor growth and shrinkage in a noninvasive manner. By stably transfecting a luciferase expression vector, we created two bioluminescent human non-Hodgkin lymphoma cell lines, Jeko1(luci and OCI-Ly3(luci, that are CD20 positive, a prerequisite to studying rituximab, a chimeric anti-CD20 antibody. To investigate the therapy response in vivo, we established a disseminated xenograft tumor model injecting these cell lines in NOD/SCID mice. We observed a close correlation of bioluminescence intensity and tumor burden, allowing us to monitor therapy response in the living animal. Cyclophosphamide reduced tumor burden in mice injected with either cell line in a dose-dependent manner. Rituximab alone was effective in OCI-Ly3(luci-injected mice and acted additively in combination with cyclophosphamide. In contrast, it improved the therapeutic outcome of Jeko1(luci-injected mice only in combination with cyclophosphamide. We conclude that well-established bioluminescence imaging is a valuable tool in disseminated xenograft tumor models. Our model can be translated to other cell lines and used to examine new therapeutic agents and schedules.

  17. Bioluminescent imaging and histopathologic characterization of WEEV neuroinvasion in outbred CD-1 mice.

    Aaron T Phillips

    Full Text Available Western equine encephalitis virus (WEEV; Alphavirus is a mosquito-borne virus that can cause severe encephalitis in humans and equids. Previous studies have shown that intranasal infection of outbred CD-1 mice with the WEEV McMillan (McM strain result in high mortality within 4 days of infection. Here in vivo and ex vivo bioluminescence (BLM imaging was applied on mice intranasally infected with a recombinant McM virus expressing firefly luciferase (FLUC to track viral neuroinvasion by FLUC detection and determine any correlation between BLM and viral titer. Immunological markers of disease (MCP-1 and IP-10 were measured and compared to wild type virus infection. Histopathology was guided by corresponding BLM images, and showed that neuroinvasion occurred primarily through cranial nerves, mainly in the olfactory tract. Olfactory bulb neurons were initially infected with subsequent spread of the infection into different regions of the brain. WEEV distribution was confirmed by immunohistochemistry as having marked neuronal infection but very few infected glial cells. Axons displayed infection patterns consistent with viral dissemination along the neuronal axis. The trigeminal nerve served as an additional route of neuroinvasion showing significant FLUC expression within the brainstem. The recombinant virus WEEV.McM.FLUC had attenuated replication kinetics and induced a weaker immunological response than WEEV.McM but produced comparable pathologies. Immunohistochemistry staining for FLUC and WEEV antigen showed that transgene expression was present in all areas of the CNS where virus was observed. BLM provides a quantifiable measure of alphaviral neural disease progression and a method for evaluating antiviral strategies.

  18. ATP-Binding Cassette Transporters Modulate Both Coelenterazine- and D-Luciferin-Based Bioluminescence Imaging

    Ruimin Huang

    2011-05-01

    Full Text Available 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. To investigate the effect of ATP-binding cassette (ABC transporters on BLI readout, we generated click beetle (cLuc, firefly (fLuc, Renilla (rLuc, and Gaussia (gLuc luciferase HEK-293 reporter cells that overexpressed different ABC transporters (ABCB1, ABCC1, and ABCG2. In vitro studies showed a significant BLI intensity decrease in intact cells compared to cell lysates, when ABCG2 was overexpressed in HEK-293/cLuc, fLuc, and rLuc cells. Selective ABC transporter inhibitors were also applied. Inhibition of ABCG2 activity increased the BLI intensity more than two-fold in HEK-293/cLuc, fLuc, and rLuc cells; inhibition of ABCB1 elevated the BLI intensity two-fold only in HEK-293/rLuc cells. BLI of xenografts derived from HEK-293/ABC transporter/luciferase reporter cells confirmed the results of inhibitor treatment in vivo. These findings demonstrate that coelenterazine-based rLuc-BLI intensity can be modulated by ABCB1 and ABCG2. ABCG2 modulates d-luciferin-based BLI in a luciferase type–independent manner. Little ABC transporter effect on gLuc-BLI intensity is observed because a large fraction of gLuc is secreted. The expression level of ABC transporters is one key factor affecting BLI intensity, and this may be particularly important in luciferase-based applications in stem cell research.

  19. Stably luminescent Staphylococcus aureus clinical strains for use in bioluminescent imaging.

    Roger D Plaut

    Full Text Available In vivo bioluminescent imaging permits the visualization of bacteria in live animals, allowing researchers to monitor, both temporally and spatially, the progression of infection in each animal. We sought to engineer stably luminescent clinical strains of Staphylococcus aureus, with the goal of using such strains in mouse models. The gram-positive shuttle vector pMAD was used as the backbone for an integration plasmid. A chloramphenicol resistance gene, a modified lux operon from Photorhabdus luminescens, and approximately 650 bp of homology to the chromosome of the USA300 S. aureus strain NRS384 were added, generating plasmid pRP1195. Electroporation into strain RN4220 followed by temperature shift led to integration of pRP1195 into the chromosome. The integrated plasmid was transferred to clinical strains by phage transduction. Luminescent strains displayed no in vitro growth defects. Moreover, luminescence was stable in vitro after three rounds of subculture over 48 hours of growth in the absence of antibiotics. Mice were infected with a luminescent strain of NRS384 in skin and intravenous models. In a mouse skin model, luminescent bacteria were present in lesions that formed and cleared over the course of several days, and in an intravenous model, bacteria inoculated in the mouse tail vein were observed spreading to multiple tissues. No statistically significant difference in virulence was observed between NRS384 and the luminescent strain in either infection model. These preliminary data suggest that this luminescent USA300 strain is suitable for use in mouse models. Similar strains were engineered using other sequenced clinical strains. Because these strains are stably luminescent, they should prove useful in animal models of infection.

  20. 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.

  1. Theoretical Study of Dinoflagellate Bioluminescence.

    Wang, Ming-Yu; Liu, Ya-Jun

    2017-03-01

    Dinoflagellates are the most ubiquitous luminescent protists in the marine environment and have drawn much attention for their crucial roles in marine ecosystems. Dinoflagellate bioluminescence has been applied in underwater target detection. The luminescent system of dinoflagellates is a typical luciferin-luciferase one. However, the excited-state oxyluciferin is not the light emitter of dinoflagellate bioluminescence as in most luciferin-luciferase bioluminescent organisms. The oxyluciferin of bioluminescent dinoflagellates is not fluorescent, whereas its luciferin emits bright fluorescence with similar wavelength of the bioluminescence. What is the light emitter of dinoflagellate bioluminescence and what is the chemical process of the light emission like? These questions have not been answered by the limited experimental evidence so far. In this study, for the first time, the density functional calculation is employed to investigate the geometries and properties of luciferin and oxyluciferin of bioluminescent dinoflagellate. The calculated results agree with the experimental observations and indicate the luciferin or its analogue, rather than oxyluciferin, is the bioluminophore of dinoflagellate bioluminescence. A rough mechanism involving energy transfer is proposed for dinoflagellate bioluminescence.

  2. Non-invasive Bioluminescence Imaging of Myoblast-Mediated Hypoxia-Inducible Factor-1 Alpha Gene Transfer

    Gheysens, Olivier; Chen, Ian Y.; Rodriguez-Porcel, Martin; Chan, Carmel; Rasooly, Julia; Vaerenberg, Caroline; Paulmurugan, Ramasamy; Willmann, Juergen K.; Deroose, Christophe; Wu, Joseph; Gambhir, Sanjiv S.

    2011-01-01

    Purpose We tested a novel imaging strategy, in which both the survival of transplanted myoblasts and their therapeutic transgene expression, a recombinant hypoxia-inducible factor-1α (HIF-1α-VP2), can be monitored using firefly luciferase (fluc) and Renilla luciferase (hrl) bioluminescence reporter genes, respectively. Procedures The plasmid pUbi-hrl-pUbi-HIF-1α-VP2, which expresses both hrl and HIF-1α-VP2 using two ubiquitin promoters, was characterized in vitro. C2c12 myoblasts stably expressing fluc and transiently transfected with pUbi-hrl-pUbi-HIF-1α-VP2 were injected into the mouse hindlimb. Both hrl and fluc expression were monitored using bioluminescence imaging (BLI). Results Strong correlations existed between the expression of hRL and each of HIF-1α-VP2, VEGF, and PlGF (r2>0.83, r2>0.82, and r2>0.97, respectively). In vivo, both transplanted cells and HIF-1α-VP2 transgene expression were successfully imaged using BLI. Conclusions An objective evaluation of myoblast-mediated gene transfer in living mice can be performed by monitoring both the survival and the transgene expression of transplanted myoblasts using the techniques developed herein. PMID:21267661

  3. 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.

  4. 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

  5. Further assessment of Monkeypox Virus infection in Gambian pouched rats (Cricetomys gambianus) using in vivo bioluminescent imaging

    Falendysz, Elizabeth; Lopera, Juan G.; Faye Lorenzsonn,; Salzer, Johanna S.; Hutson, Christina L.; Doty, Jeffrey; Gallardo-Romero, Nadia; Carroll, Darin S.; Osorio, Jorge E.; Rocke, Tonie E.

    2015-01-01

    Monkeypox is a zoonosis clinically similar to smallpox in humans. Recent evidence has shown a potential risk of increased incidence in central Africa. Despite attempts to isolate the virus from wild rodents and other small mammals, no reservoir host has been identified. In 2003,Monkeypox virus (MPXV) was accidentally introduced into the U.S. via the pet trade and was associated with the Gambian pouched rat (Cricetomys gambianus). Therefore, we investigated the potential reservoir competence of the Gambian pouched rat for MPXV by utilizing a combination of in vivo and in vitro methods. We inoculated three animals by the intradermal route and three animals by the intranasal route, with one mock-infected control for each route. Bioluminescent imaging (BLI) was used to track replicating virus in infected animals and virological assays (e.g. real time PCR, cell culture) were used to determine viral load in blood, urine, ocular, nasal, oral, and rectal swabs. Intradermal inoculation resulted in clinical signs of monkeypox infection in two of three animals. One severely ill animal was euthanized and the other affected animal recovered. In contrast, intranasal inoculation resulted in subclinical infection in all three animals. All animals, regardless of apparent or inapparent infection, shed virus in oral and nasal secretions. Additionally, BLI identified viral replication in the skin without grossly visible lesions. These results suggest that Gambian pouched rats may play an important role in transmission of the virus to humans, as they are hunted for consumption and it is possible for MPXV-infected pouched rats to shed infectious virus without displaying overt clinical signs.

  6. An enhanced chimeric firefly luciferase-inspired enzyme for ATP detection and bioluminescence reporter and imaging applications.

    Branchini, Bruce R; Southworth, Tara L; Fontaine, Danielle M; Kohrt, Dawn; Talukder, Munya; Michelini, Elisa; Cevenini, Luca; Roda, Aldo; Grossel, Martha J

    2015-09-01

    Firefly luciferases, which emit visible light in a highly specific ATP-dependent process, have been adapted for a variety of applications, including gene reporter assays, whole-cell biosensor measurements, and in vivo imaging. We previously reported the approximately 2-fold enhanced activity and 1.4-fold greater bioluminescence quantum yield properties of a chimeric enzyme that contains the N-domain of Photinus pyralis luciferase joined to the C-domain of Luciola italica luciferase. Subsequently, we identified 5 amino acid changes based on L. italica that are the main determinants of the improved bioluminescence properties. Further engineering to enhance thermal and pH stability produced a novel luciferase called PLG2. We present here a systematic comparison of the spectral and physical properties of the new protein with P. pyralis luciferase and demonstrate the potential of PLG2 for use in assays based on the detection of femtomole levels of ATP. In addition, we compared the performance of a mammalian codon-optimized version of the cDNA for PLG2 with the luc2 gene in HEK293T cells. Using an optimized low-cost assay system, PLG2 activity can be monitored in mammalian cell lysates and living cells with 4.4-fold and approximately 3.0-fold greater sensitivity, respectively. PLG2 could be an improved alternative to Promega's luc2 for reporter and imaging applications.

  7. Multicolor Bioluminescence Obtained Using Firefly Luciferin.

    Kiyama, Masahiro; Saito, Ryohei; Iwano, Satoshi; Obata, Rika; Niwa, Haruki; Maki, Shojiro A

    2016-01-01

    Firefly bioluminescence is widely used in life science research as a useful analysis tool. For example, the adenosine-5`-triphosphate (ATP)-dependent enzymatic firefly bioluminescence reaction has long been utilized as a microbial monitoring tool. Rapid and sensitive firefly luciferin-luciferase combinations are used not only to measure cell viability but also for reporter-gene assays. Recently, bioluminescence was utilized as a noninvasive, real-time imaging tool for living subjects to monitor cells and biological events. However, the number of commercialized luciferase genes is limited and tissue-permeable near-infrared (NIR) region emitting light is required for in vivo imaging. In this review, recent studies describing synthetic luciferin analogues predicted to have red-shifted bioluminescence are summarized. Luciferase substrates emitting red, green, and blue light that were designed and developed in our laboratory are presented. The longest emission wavelength of the synthesized luciferin analogues was recorded at 675 nm, which is within the NIR region. This compound is now commercially available as "Aka Lumine®".

  8. Horse species symposium: a novel approach to monitoring pathogen progression during uterine and placental infection in the mare using bioluminescence imaging technology and lux-modified bacteria.

    Ryan, P L; Christiansen, D L; Hopper, R M; Walters, F K; Moulton, K; Curbelo, J; Greene, J M; Willard, S T

    2011-05-01

    Uterine and placental infections are the leading cause of abortion, stillbirth, and preterm delivery in the mare. Whereas uterine and placental infections in women have been studied extensively, a comprehensive examination of the pathogenic processes leading to this unsatisfactory pregnancy outcome in the mare has yet to be completed. Most information in the literature relating to late-term pregnancy loss in mares is based on retrospective studies of clinical cases submitted for necropsy. Here we report the development and application of a novel approach, whereby transgenically modified bacteria transformed with lux genes of Xenorhabdus luminescens or Photorhabdus luminescens origin and biophotonic imaging are utilized to better understand pathogen-induced preterm birth in late-term pregnant mares. This technology uses highly sensitive bioluminescence imaging camera systems to localize and monitor pathogen progression during tissue invasion by measuring the bioluminescent signatures emitted by the lux-modified pathogens. This method has an important advantage in that it allows for the potential tracking of pathogens in vivo in real time and over time, which was hitherto impossible. Although the application of this technology in domestic animals is in its infancy, investigators were successful in identifying the fetal lungs, sinuses, nares, urinary, and gastrointestinal systems as primary tissues for pathogen invasion after experimental infection of pregnant mares with lux-modified Escherichia coli. It is important that pathogens were not detected in other vital organs, such as the liver, brain, and cardiac system. Such precision in localizing sites of pathogen invasion provides potential application for this novel approach in the development of more targeted therapeutic interventions for pathogen-related diseases in the equine and other domestic species.

  9. 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

  10. Evaluation of impaired beta-cell function in nonobese-diabetic (NOD) mouse model using bioluminescence imaging.

    Sever, Dror; Eldor, Roy; Sadoun, Gadi; Amior, Livnat; Dubois, Daniele; Boitard, Christian; Aflalo, Claude; Melloul, Danielle

    2011-02-01

    Insulin-producing pancreatic β cells are functionally impaired or destroyed in diabetes mellitus. The onset of type 1 diabetes (T1D) represents the culmination of a prolonged prediabetic phase of immune-mediated β-cell destruction. To assess the in vivo metabolic status of these cells, we used the ATP-sensitive firefly luciferase bioluminescence imaging approach, as a noninvasive probe to monitor pathological alterations in β-cell function in the nonobese-diabetic (NOD) mouse model of T1D. Hence, we generated the ToIβ-NOD transgenic mice in which doxycycline-inducible luciferase gene is selectively expressed in β cells. A sharp reduction in bioluminescence emitted in vivo from β cells at the early stages, preceded by several weeks of a limited reduction in β-cell mass. Since this decline could be due to the ongoing inflammatory process occurring in vivo, we exposed control islets to inflammatory cytokines and observed a dramatic decrease in luciferase luminescence, which appears to be due in part to a decrease in protein levels and a drop in intracellular ATP levels. This is the first evidence that selective expression of the luciferase gene represents a sensitive method for noninvasive in vivo monitoring of early β-cell dysfunction, subtle metabolic changes, such as endogenous ATP levels, indicative of a pathological condition in a tissue at the cellular level.

  11. 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.

  12. 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.

  13. 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.

  14. Molecular phylogeny and node time estimation of bioluminescent Lantern Sharks (Elasmobranchii: Etmopteridae).

    Straube, Nicolas; Iglésias, Samuel P; Sellos, Daniel Y; Kriwet, Jürgen; Schliewen, Ulrich K

    2010-09-01

    Deep-sea Lantern Sharks (Etmopteridae) represent the most speciose family within Dogfish Sharks (Squaliformes). We compiled an extensive DNA dataset to estimate the first molecular phylogeny of the family and to provide node age estimates for the origin and diversification for this enigmatic group. Phylogenetic inferences yielded consistent and well supported hypotheses based on 4685bp of both nuclear (RAG1) and mitochondrial genes (COI, 12S-partial 16S, tRNAVal and tRNAPhe). The monophyletic family Etmopteridae originated in the early Paleocene around the C/T boundary, and split further into four morphologically distinct lineages supporting three of the four extant genera. The exception is Etmopterus which is paraphyletic with respect to Miroscyllium. Subsequent rapid radiation within Etmopterus in the Oligocene/early Miocene was accompanied by divergent evolution of bioluminescent flank markings which morphologically characterize the four lineages. Higher squaliform interrelationships could not be satisfactorily identified, but convergent evolution of bioluminescence in Dalatiidae and Etmopteridae is supported.

  15. The Influence of Hypoxia and pH on Bioluminescence Imaging of Luciferase-Transfected Tumor Cells and Xenografts

    Ashraf A. Khalil

    2013-01-01

    Full Text Available Bioluminescence imaging (BLI is a relatively new noninvasive technology used for quantitative assessment of tumor growth and therapeutic effect in living animal models. BLI involves the generation of light by luciferase-expressing cells following administration of the substrate luciferin in the presence of oxygen and ATP. In the present study, the effects of hypoxia, hypoperfusion, and pH on BLI signal (BLS intensity were evaluated in vitro using cultured cells and in vivo using a xenograft model in nude mice. The intensity of the BLS was significantly reduced in the presence of acute and chronic hypoxia. Changes in cell density, viability, and pH also affected BLS. Although BLI is a convenient non-invasive tool for tumor assessment, these factors should be considered when interpreting BLS intensity, especially in solid tumors that could be hypoxic due to rapid growth, inadequate blood supply, and/or treatment.

  16. 5-Fluorouracil induced intestinal mucositis via nuclear factor-κB activation by transcriptomic analysis and in vivo bioluminescence imaging.

    Chung-Ta Chang

    Full Text Available 5-Fluorouracil (5-FU is a commonly used drug for the treatment of malignant cancers. However, approximately 80% of patients undergoing 5-FU treatment suffer from gastrointestinal mucositis. The aim of this report was to identify the drug target for the 5-FU-induced intestinal mucositis. 5-FU-induced intestinal mucositis was established by intraperitoneally administering mice with 100 mg/kg 5-FU. Network analysis of gene expression profile and bioluminescent imaging were applied to identify the critical molecule associated with 5-FU-induced mucositis. Our data showed that 5-FU induced inflammation in the small intestine, characterized by the increased intestinal wall thickness and crypt length, the decreased villus height, and the increased myeloperoxidase activity in tissues and proinflammatory cytokine production in sera. Network analysis of 5-FU-affected genes by transcriptomic tool showed that the expression of genes was regulated by nuclear factor-κB (NF-κB, and NF-κB was the central molecule in the 5-FU-regulated biological network. NF-κB activity was activated by 5-FU in the intestine, which was judged by in vivo bioluminescence imaging and immunohistochemical staining. However, 5-aminosalicylic acid (5-ASA inhibited 5-FU-induced NF-κB activation and proinflammatory cytokine production. Moreover, 5-FU-induced histological changes were improved by 5-ASA. In conclusion, our findings suggested that NF-κB was the critical molecule associated with the pathogenesis of 5-FU-induced mucositis, and inhibition of NF-κB activity ameliorated the mucosal damage caused by 5-FU.

  17. Bioluminescence tracking of alginate micro-encapsulated cell transplants.

    Tiernan, Aubrey R; Sambanis, Athanassios

    2017-02-01

    Cell-based therapies to treat loss-of-function hormonal disorders such as diabetes and Parkinson's disease are routinely coupled with encapsulation strategies, but an understanding of when and why grafts fail in vivo is lacking. Consequently, investigators cannot clearly define the key factors that influence graft success. Although bioluminescence is a popular method to track the survival of free cells transplanted in preclinical models, little is known of the ability to use bioluminescence for real-time tracking of microencapsulated cells. Furthermore, the impact that dynamic imaging distances may have, due to freely-floating microcapsules in vivo, on cell survival monitoring is unknown. This work addresses these questions by applying bioluminescence to a pancreatic substitute based on microencapsulated cells. Recombinant insulin-secreting cells were transduced with a luciferase lentivirus and microencapsulated in Ba(2+) crosslinked alginate for in vitro and in vivo studies. In vitro quantitative bioluminescence monitoring was possible and viable microencapsulated cells were followed in real time under both normoxic and anoxic conditions. Although in vivo dispersion of freely-floating microcapsules in the peritoneal cavity limited the analysis to a qualitative bioluminescence evaluation, signals consistently four orders of magnitude above background were clear indicators of temporal cell survival. Strong agreement between in vivo and in vitro cell proliferation over time was discovered by making direct bioluminescence comparisons between explanted microcapsules and parallel in vitro cultures. Broader application of this bioluminescence approach to retrievable transplants, in supplement to currently used end-point physiological tests, could improve understanding and accelerate development of cell-based therapies for critical clinical applications. Copyright © 2014 John Wiley & Sons, Ltd.

  18. 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.

  19. 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.

  20. Lessons in microcapsule assembly from imaging delivery of a bioluminescent enzyme.

    Pavlov, Anton M; Sukhorukov, Gleb B; Gould, David J

    2013-03-11

    Layer-by-layer assembled microcapsules have potential applications as delivery and biosensing systems, which make them attractive tools for use in various aspects of nanomedicine. We examined the effect of microcapsule location on activity of the bioluminescent enzyme luciferase in both intact capsules and following cell uptake. In intact capsules, the rate of reaction of luciferase was greatest for luciferase in the outer layer and least in the core. Following cell uptake, luciferase in the outer layer was rapidly reactive, and a similar rate of reaction and activity was observed for luciferase placed in capsule interior (core). By contrast, there was minimal activity detected when microcapsules with luciferase sandwiched between polyelectrolytes in a middle layer were delivered to cells. This study informs us of the availability of bioactive molecules located in different positions within microcapsules and will enable better microcapsule construction in line with the intended application, particularly delivery of functional proteins to cells.

  1. 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.

  2. POTENTIAL OF INDUCED METABOLIC BIOLUMINESCENCE IMAGING TO UNCOVER METABOLIC EFFECTS OF ANTI-ANGIOGENIC THERAPY IN TUMORS

    Stefano eIndraccolo

    2016-02-01

    Full Text Available Tumor heterogeneity at the genetic level has been illustrated by a multitude of studies on the genomics of cancer, but whether tumors can be heterogeneous at the metabolic level is an issue which has been less systematically investigated so far. A burning related question is whether the metabolic features of tumors can change either following natural tumor progression (i.e. in primary tumors versus metastasis or therapeutic interventions. In this regard, recent findings by independent teams indicate that anti-angiogenic drugs cause metabolic perturbations in tumors as well as metabolic adaptations associated with increased malignancy. Induced metabolic bioluminescence imaging (imBI is an imaging technique which enables detection of key metabolites associated with glycolysis, including lactate, glucose, pyruvate and ATP in tumor sections. Signals captured by imBI can be used to visualize the topographic distribution of these metabolites and quantify their absolute amount. ImBI can be very useful for metabolic classification of tumors as well as to track metabolic changes in the glycolytic pathway associated with certain therapies. Imaging of the metabolic changes induced by anti-angiogenic drugs in tumors by imBI or other emerging technologies is a valuable tool to uncover molecular sensors engaged by metabolic stress and offers an opportunity to understand how metabolism-based approaches could improve cancer therapy.

  3. In Vivo Bioluminescence Imaging Validation of a Human Biopsy–Derived Orthotopic Mouse Model of Glioblastoma Multiforme

    Monika A. Jarzabek

    2013-05-01

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

  4. 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.

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

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

    2013-01-01

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

  6. 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.

  7. In vivo visualization and monitoring of viable neural stem cells using noninvasive bioluminescence imaging in the 6-hydroxydopamine-induced mouse model of Parkinson disease.

    Im, Hyung-Jun; Hwang, Do Won; Lee, Han Kyu; Jang, Jaeho; Lee, Song; Youn, Hyewon; Jin, Yeona; Kim, Seung U; Kim, E Edmund; Kim, Yong Sik; Lee, Dong Soo

    2013-06-01

    Transplantation of neural stem cells (NSCs) has been proposed as a treatment for Parkinson disease (PD). The aim of this study was to monitor the viability of transplanted NSCs expressing the enhanced luciferase gene in a mouse model of PD in vivo. The PD animal model was induced by unilateral injection of 6-hydroxydopamine (6-OHDA). The behavioral test using apomorphine-induced rotation and positron emission tomography with [18F]N-(3-fluoropropyl)-2'-carbomethoxy-3'-(4-iodophenyl)nortropane ([18F]FP-CIT) were conducted. HB1.F3 cells transduced with an enhanced firefly luciferase retroviral vector (F3-effLuc cells) were transplanted into the right striatum. In vivo bioluminescence imaging was repeated for 2 weeks. Four weeks after transplantation, [18F]FP-CIT PET and the rotation test were repeated. All 6-OHDA-injected mice showed markedly decreased [18F]FP-CIT uptake in the right striatum. Transplanted F3-effLuc cells were visualized on the right side of the brain in all mice by bioluminescence imaging. The bioluminescence intensity of the transplanted F3-effLuc cells gradually decreased until it was undetectable by 10 days. The behavioral test showed that stem cell transplantation attenuated the motor symptoms of PD. No significant change was found in [18F]FP-CIT imaging after cell transplantation. We successfully established an in vivo bioluminescence imaging system for the detection of transplanted NSCs in a mouse model of PD. NSC transplantation induced behavioral improvement in PD model mice.

  8. Real-time luminescence imaging of cellular ATP release.

    Furuya, Kishio; Sokabe, Masahiro; Grygorczyk, Ryszard

    2014-03-15

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

  9. 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.

  10. Measuring cytotoxicity by bioluminescence imaging outperforms the standard chromium-51 release assay.

    Mobin A Karimi

    Full Text Available The chromium-release assay developed in 1968 is still the most commonly used method to measure cytotoxicity by T cells and by natural killer cells. Target cells are loaded in vitro with radioactive chromium and lysis is determined by measuring chromium in the supernatant released by dying cells. Since then, alternative methods have been developed using different markers of target cell viability that do not involve radioactivity. Here, we compared and contrasted a bioluminescence (BLI-based cytotoxicity assay to the standard radioactive chromium-release assay using an identical set of effector cells and tumor target cells. For this, we stably transduced several human and murine tumor cell lines to express luciferase. When co-cultured with cytotoxic effector cells, highly reproducible decreases in BLI were seen in an effector to target cell dose-dependent manner. When compared to results obtained from the chromium release assay, the performance of the BLI-based assay was superior, because of its robustness, increased signal-to-noise ratio, and faster kinetics. The reduced/delayed detection of cytotoxicity by the chromium release method was attributable to the association of chromium with structural components of the cell, which are released quickly by detergent solubilization but not by hypotonic lysis. We conclude that the (BLI-based measurement of cytotoxicity offers a superior non-radioactive alternative to the chromium-release assay that is more robust and quicker to perform.

  11. Boosting bioluminescence neuroimaging: an optimized protocol for brain studies.

    Aswendt, Markus; Adamczak, Joanna; Couillard-Despres, Sebastien; Hoehn, Mathias

    2013-01-01

    Bioluminescence imaging is widely used for optical cell tracking approaches. However, reliable and quantitative bioluminescence of transplanted cells in the brain is highly challenging. In this study we established a new bioluminescence imaging protocol dedicated for neuroimaging, which increases sensitivity especially for noninvasive tracking of brain cell grafts. Different D-Luciferin concentrations (15, 150, 300 and 750 mg/kg), injection routes (i.v., i.p., s.c.), types of anesthesia (Isoflurane, Ketamine/Xylazine, Pentobarbital) and timing of injection were compared using DCX-Luc transgenic mice for brain specific bioluminescence. Luciferase kinetics was quantitatively evaluated for maximal photon emission, total photon emission and time-to-peak. Photon emission followed a D-Luciferin dose-dependent relation without saturation, but with delay in time-to-peak increasing for increasing concentrations. The comparison of intravenous, subcutaneous and intraperitoneal substrate injection reflects expected pharmacokinetics with fastest and highest photon emission for intravenous administration. Ketamine/Xylazine and Pentobarbital anesthesia showed no significant beneficial effect on maximal photon emission. However, a strong difference in outcome was observed by injecting the substrate pre Isoflurane anesthesia. This protocol optimization for brain specific bioluminescence imaging comprises injection of 300 mg/kg D-Luciferin pre Isoflurane anesthesia as an efficient and stable method with a signal gain of approx. 200% (compared to 150 mg/kg post Isoflurane). Gain in sensitivity by the novel imaging protocol was quantitatively assessed by signal-to-noise calculations of luciferase-expressing neural stem cells grafted into mouse brains (transplantation of 3,000-300,000 cells). The optimized imaging protocol lowered the detection limit from 6,000 to 3,000 cells by a gain in signal-to-noise ratio.

  12. Evaluation of bioluminescent imaging for noninvasive monitoring of colorectal cancer progression in the liver and its response to immunogene therapy

    Gonzalez-Aparicio Manuela

    2009-01-01

    Full Text Available Abstract Background Bioluminescent imaging (BLI is based on the detection of light emitted by living cells expressing a luciferase gene. Stable transfection of luciferase in cancer cells and their inoculation into permissive animals allows the noninvasive monitorization of tumor progression inside internal organs. We have applied this technology for the development of a murine model of colorectal cancer involving the liver, with the aim of improving the pre-clinical evaluation of new anticancer therapies. Results A murine colon cancer cell line stably transfected with the luciferase gene (MC38Luc1 retains tumorigenicity in immunocompetent C57BL/6 animals. Intrahepatic inoculation of MC38Luc1 causes progressive liver infiltration that can be monitored by BLI. Compared with ultrasonography (US, BLI is more sensitive, but accurate estimation of tumor mass is impaired in advanced stages. We applied BLI to evaluate the efficacy of an immunogene therapy approach based on the liver-specific expression of the proinflammatory cytokine interleukin-12 (IL-12. Individualized quantification of light emission was able to determine the extent and duration of antitumor responses and to predict long-term disease-free survival. Conclusion We show that BLI is a rapid, convenient and safe technique for the individual monitorization of tumor progression in the liver. Evaluation of experimental treatments with complex mechanisms of action such as immunotherapy is possible using this technology.

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

    Chun, Nicole A. L.; Murakami, Takashi

    2010-02-01

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

  14. 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.

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

    Jiang, Tianyu; Du, Lupei; Li, Minyong

    2016-04-01

    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.

  16. Time-Encoded Imagers.

    Marleau, Peter; Brubaker, Erik

    2014-11-01

    This report provides a short overview of the DNN R&D funded project, Time-Encoded Imagers. The project began in FY11 and concluded in FY14. The Project Description below provides the overall motivation and objectives for the project as well as a summary of programmatic direction. It is followed by a short description of each task and the resulting deliverables.

  17. 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

  18. 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

    2013-12-01

    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 regeneration and engraftment in freely moving adult zebrafish. We show that luciferase-based live imaging reliably estimates muscle quantity in an internal organ, the heart, and can longitudinally follow cardiac regeneration in individual animals after major injury. Furthermore, luciferase-based detection enables visualization and quantification of engraftment in live recipients of transplanted hematopoietic stem cell progeny, with advantages in sensitivity and gross spatial resolution over fluorescence detection. Our findings present a versatile resource for monitoring and dissecting vertebrate stem cell and regeneration biology.

  19. 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.

  20. 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

  1. Combining fluorescence and bioluminescence microscopy.

    Goda, Kazuhito; Hatta-Ohashi, Yoko; Akiyoshi, Ryutaro; Sugiyama, Takashi; Sakai, Ikuko; Takahashi, Takeo; Suzuki, Hirobumi

    2015-08-01

    Bioluminescence microscopy has revealed that gene expression in individual cells can respond differently to the same stimulus. To understand this phenomenon, it is important to sequentially observe the series of events from cellular signal transduction to gene expression regulated by specific transcription factors derived from signaling cascades in individual cells. However, these processes have been separately analyzed with fluorescence and bioluminescence microscopy. Furthermore, in culture medium, the background fluorescence of luciferin-a substrate of luciferase in promoter assays of gene expression in cultured cells-confounds the simultaneous observation of fluorescence and bioluminescence. Therefore, we optimized conditions for optical filter sets based on spectral properties and the luciferin concentration based on cell permeability for fluorescence observation combined with bioluminescence microscopy. An excitation and emission filter set (492-506 nm and 524-578 nm) was suitable for green fluorescent protein and yellow fluorescent protein imaging of cells, and >100 μM luciferin was acceptable in culture medium based on kinetic constants and the estimated intracellular concentration. Using these parameters, we present an example of sequential fluorescence and bioluminescence microscopic observation of signal transduction (translocation of protein kinase C alpha from the cytoplasm to the plasma membrane) coupled with activation of gene expression by nuclear factor of kappa light polypeptide B in individual cells and show that the gene expression response is not completely concordant with upstream signaling following stimulation with phorbol-12-myristate-13-acetate. Our technique is a powerful imaging tool for analysis of heterogeneous gene expression together with upstream signaling in live single cells.

  2. High Resolution In Vivo Bioluminescent Imaging for the Study of Bacterial Tumour Targeting

    Cronin, Michelle; Akin, Ali R.; Collins, Sara A.; Meganck, Jeff; Kim, Jae-Beom; Baban, Chwanrow K.; Joyce, Susan A.; van Dam, Gooitzen M.; Zhang, Ning; van Sinderen, Douwe; O'Sullivan, Gerald C.; Kasahara, Noriyuki; Gahan, Cormac G.; Francis, Kevin P.; Tangney, Mark

    2012-01-01

    The ability to track microbes in real time in vivo is of enormous value for preclinical investigations in infectious disease or gene therapy research. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumours following syste

  3. 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...

  4. 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.

  5. Autonomous Bioluminescent Expression of the Bacterial Luciferase Gene Cassette (lux) in a Mammalian Cell Line

    Close, Dan M.; Patterson, Stacey S.; Ripp, Steven; Baek, Seung J.; Sanseverino, John; Sayler, Gary S.

    2010-01-01

    Background 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. Methodology/Principal Findings 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 (FMNH2) 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. FMNH2 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. Conclusions/Significance 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. PMID:20805991

  6. 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.

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

    Wang Lihong V

    2004-05-01

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

  8. 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.

  9. 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.

  10. Detection of the onset of ischemia and carcinogenesis by hypoxia-inducible transcription factor-based in vivo bioluminescence imaging.

    Tetsuya Kadonosono

    Full Text Available An animal model for the early detection of common fatal diseases such as ischemic diseases and cancer is desirable for the development of new drugs and treatment strategies. Hypoxia-inducible factor 1 (HIF-1 is a transcription factor that regulates oxygen homeostasis and plays key roles in a number of diseases, including cancer. Here, we established transgenic (Tg mice that carry HRE/ODD-luciferase (HOL gene, which generates bioluminescence in an HIF-1-dependent manner and was successfully used in this study to monitor HIF-1 activity in ischemic tissues. To monitor carcinogenesis in vivo, we mated HOL mice with rasH2 Tg mice, which are highly sensitive to carcinogens and are used for short-term carcinogenicity assessments. After rasH2-HOL Tg mice were treated with N-methyl-N-nitrosourea, bioluminescence was detected noninvasively as early as 9 weeks in tissues that contained papillomas and malignant lesions. These results suggest that the Tg mouse lines we established hold significant potential for monitoring the early onset of both ischemia and carcinogenesis and that these lines will be useful for screening chemicals for carcinogenic potential.

  11. 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.

  12. 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.

    2016-01-01

    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

  13. 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.

  14. 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.

  15. Bioluminescence as a classroom tool for scientist volunteers.

    Hammer, M; Andrade, J D

    2000-01-01

    There is a great need for practicing scientists to volunteer their time and expertise in the K-12th grade science classroom. We have found that bioluminescence is a fun and exciting way to teach basic science concepts and is an excellent tool for the volunteering scientist. We have had very positive reactions from both teachers and students. The excitement of the students when they first see bioluminescence is contagious. Bioluminescent dinoflagellates are one of the easiest ways to introduce students to this fascinating topic. Many activities and experiments can be done using the bioluminescent dinoflagellates and many students and teachers could benefit from your knowledge and expertise. See you in the classroom.

  16. Ginger and Zingerone Ameliorate Lipopolysaccharide-Induced Acute Systemic Inflammation in Mice, Assessed by Nuclear Factor-κB Bioluminescent Imaging.

    Hsiang, Chien-Yun; Cheng, Hui-Man; Lo, Hsin-Yi; Li, Chia-Cheng; Chou, Pei-Chi; Lee, Yu-Chen; Ho, Tin-Yun

    2015-07-08

    Ginger is a commonly used spice in cooking. In this study, we comprehensively evaluated the anti-inflammatory activities of ginger and its component zingerone in lipopolysaccharide (LPS)-induced acute systemic inflammation in mice via nuclear factor-κB (NF-κB) bioluminescent imaging. Ginger and zingerone significantly suppressed LPS-induced NF-κB activities in cells in a dose-dependent manner, and the maximal inhibition (84.5% ± 3.5% and 96.2% ± 0.6%) was observed at 100 μg/mL ginger and zingerone, respectively. Moreover, dietary ginger and zingerone significantly reduced LPS-induced proinflammatory cytokine production in sera by 62.9% ± 18.2% and 81.3% ± 6.2%, respectively, and NF-κB bioluminescent signals in whole body by 26.9% ± 14.3% and 38.5% ± 6.2%, respectively. In addition, ginger and zingerone suppressed LPS-induced NF-κB-driven luminescent intensities in most organs, and the maximal inhibition by ginger and zingerone was observed in small intestine. Immunohistochemical staining further showed that ginger and zingerone decreased interleukin-1β (IL-1β)-, CD11b-, and p65-positive areas in jejunum. In conclusion, our findings suggested that ginger and zingerone were likely to be broad-spectrum anti-inflammatory agents in most organs that suppressed the activation of NF-κB, the production of IL-1β, and the infiltration of inflammatory cells in mice.

  17. Dual-Color Bioluminescence Imaging for Simultaneous Monitoring of the Intestinal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice.

    Daniel, Catherine; Poiret, Sabine; Dennin, Véronique; Boutillier, Denise; Lacorre, Delphine Armelle; Foligné, Benoit; Pot, Bruno

    2015-08-15

    Lactic acid bacteria are found in the gastrointestinal tract of mammals and have received tremendous attention due to their health-promoting properties. We report the development of two dual-color luciferase-producing Lactobacillus (Lb.) plantarum and Lactococcus (Lc.) lactis strains for noninvasive simultaneous tracking in the mouse gastrointestinal tract. We previously described the functional expression of the red luciferase mutant (CBRluc) from Pyrophorus plagiophthalamus in Lb. plantarum NCIMB8826 and Lc. lactis MG1363 (C. Daniel, S. Poiret, V. Dennin, D. Boutillier, and B. Pot, Appl Environ Microbiol 79:1086-1094, 2013, http://dx.doi.org/10.1128/AEM.03221-12). In this study, we determined that CBRluc is a better-performing luciferase for in vivo localization of both lactic acid bacteria after oral administration than the green click beetle luciferase mutant construct developed in this study. We further established the possibility to simultaneously detect red- and green-emitting lactic acid bacteria by dual-wavelength bioluminescence imaging in combination with spectral unmixing. The difference in spectra of light emission by the red and green click beetle luciferase mutants and dual bioluminescence detection allowed in vitro and in vivo quantification of the red and green emitted signals; thus, it allowed us to monitor the dynamics and fate of the two bacterial populations simultaneously. Persistence and viability of both strains simultaneously administered to mice in different ratios was studied in vivo in anesthetized mice and ex vivo in mouse feces. The application of dual-luciferase-labeled bacteria has considerable potential to simultaneously study the interactions and potential competitions of different targeted bacteria and their hosts.

  18. 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.

  19. Let there be bioluminescence: development of a biophotonic imaging platform for in situ analyses of oral biofilms in animal models.

    Merritt, Justin; Senpuku, Hidenobu; Kreth, Jens

    2016-01-01

    In the current study, we describe a novel biophotonic imaging-based reporter system that is particularly useful for the study of virulence in polymicrobial infections and interspecies interactions within animal models. A suite of luciferase enzymes was compared using three early colonizing species of the human oral flora (Streptococcus mutans, Streptococcus gordonii and Streptococcus sanguinis) to determine the utility of the different reporters for multiplexed imaging studies in vivo. Using the multiplex approach, we were able to track individual species within a dual-species oral infection model in mice with both temporal and spatial resolution. We also demonstrate how biophotonic imaging of multiplexed luciferase reporters could be adapted for real-time quantification of bacterial gene expression in situ. By creating an inducible dual-luciferase expressing reporter strain of S. mutans, we were able to exogenously control and measure expression of nlmAB (encoding the bacteriocin mutacin IV) within mice to assess its importance for the persistence ability of S. mutans in the oral cavity. The imaging system described in the current study circumvents many of the inherent limitations of current animal model systems, which should now make it feasible to test hypotheses that were previously impractical to model.

  20. Gombrich on image and time

    Kristóf Nyíri

    2009-12-01

    Full Text Available There is a very close, indeed intrinsic, connection between the notions of image and time. Images are incomplete unless they are moving ones – unless, that is, they happen in time. On the other hand, time cannot be conceptualized except by metaphors, and so ultimately by images, of movement in space. That only the moving image is a full-fledged one is a fact that was fully recognized and articulated by Ernst Gombrich. Also, Gombrich entertained, and argued for, a rich and well-balanced view of the relationships between pictorial and verbal representation. An antidote to the unholy influence of Goodman, Gombrich deserves to be rediscovered as the figure whose work is ideally suited to providing a founding paradigm for a truly successful philosophy of images.

  1. 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

  2. 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

  3. 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.

  4. Bioluminescence patterns among North American Armillaria species.

    Mihail, Jeanne D

    2015-06-01

    Bioluminescence is widely recognized among white-spored species of Basidiomycota. Most reports of fungal bioluminescence are based upon visual light perception. When instruments such as photomultipliers have been used to measure fungal luminescence, more taxa have been discovered to produce light, albeit at a range of magnitudes. The present studies were undertaken to determine the prevalence of bioluminescence among North American Armillaria species. Consistent, constitutive bioluminescence was detected for the first time for mycelia of Armillaria calvescens, Armillaria cepistipes, Armillaria gemina, Armillaria nabsnona, and Armillaria sinapina and confirmed for mycelia of Armillaria gallica, Armillaria mellea, Armillaria ostoyae, and Armillaria tabescens. Emission spectra of mycelia representing all species had maximum intensity in the range 515-525 nm confirming that emitted light was the result of bioluminescence rather than chemiluminescence. Time series analysis of 1000 consecutive luminescence measurements revealed a highly significant departure from random variation. Mycelial luminescence of eight species exhibited significant, stable shifts in magnitude in response to a series of mechanical disturbance treatments, providing one mechanism for generating observed luminescence variation.

  5. A radar image time series

    Leberl, F.; Fuchs, H.; Ford, J. P.

    1981-01-01

    A set of ten side-looking radar images of a mining area in Arizona that were aquired over a period of 14 yr are studied to demonstrate the photogrammetric differential-rectification technique applied to radar images and to examine changes that occurred in the area over time. Five of the images are rectified by using ground control points and a digital height model taken from a map. Residual coordinate errors in ground control are reduced from several hundred meters in all cases to + or - 19 to 70 m. The contents of the radar images are compared with a Landsat image and with aerial photographs. Effects of radar system parameters on radar images are briefly reviewed.

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

    Bang-Wen Xie

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

  7. Thoughts on the diversity of convergent evolution of bioluminescence on earth

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

    2012-10-01

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

  8. 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.

  9. Ratiometric bioluminescence indicators for monitoring cyclic adenosine 3',5'-monophosphate in live cells based on luciferase-fragment complementation.

    Takeuchi, Masaki; Nagaoka, Yasutaka; Yamada, Toshimichi; Takakura, Hideo; Ozawa, Takeaki

    2010-11-15

    Bioluminescent indicators for cyclic 3',5'-monophosphate AMP (cAMP) are powerful tools for noninvasive detection with high sensitivity. However, the absolute photon counts are affected substantially by adenosine 5'-triphosphate (ATP) and d-luciferin concentrations, limiting temporal analysis in live cells. This report describes a genetically encoded bioluminescent indicator for detecting intracellular cAMP based on complementation of split fragments of two-color luciferase mutants originated from click beetles. A cAMP binding domain of protein kinase A was connected with an engineered carboxy-terminal fragment of luciferase, of which ends were connected with amino-terminal fragments of green luciferase and red luciferase. We demonstrated that the ratio of green to red bioluminescence intensities was less influenced by the changes of ATP and d-luciferin concentrations. We also showed an applicability of the bioluminescent indicator for time-course and quantitative assessments of intracellular cAMP in living cells and mice. The bioluminescent indicator will enable quantitative analysis and imaging of spatiotemporal dynamics of cAMP in opaque and autofluorescent living subjects.

  10. 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

  11. 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.

  12. Bioluminescent bioreporter integrated circuit

    Simpson, Michael L. (Knoxville, TN); Sayler, Gary S. (Blaine, TN); Paulus, Michael J. (Knoxville, TN)

    2000-01-01

    Disclosed are monolithic bioelectronic devices comprising a bioreporter and an OASIC. These bioluminescent bioreporter integrated circuit are useful in detecting substances such as pollutants, explosives, and heavy-metals residing in inhospitable areas such as groundwater, industrial process vessels, and battlefields. Also disclosed are methods and apparatus for environmental pollutant detection, oil exploration, drug discovery, industrial process control, and hazardous chemical monitoring.

  13. 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.

  14. Bioluminescent bioreporter sensing of foodborne toxins

    Fraley, Amanda C.; Ripp, Steven; Sayler, Gary S.

    2004-06-01

    Histamine is the primary etiological agent in the foodborne disease scombrotoxicosis, one of the most common food toxicities related to fish consumption. Procedures for detecting histamine in fish products are available, but are often too expensive or too complex for routine use. As an alternative, a bacterial bioluminescent bioreporter has been constructed to develop a biosensor system that autonomously responds to low levels of histamine. The bioreporter contains a promoterless Photorhabdus luminescens lux operon (luxCDABE) fused with the Vibrio anguillarum angR regulatory gene promoter of the anguibactin biosynthetic operon. The bioreporter emitted 1.46 times more bioluminescence than background, 30 minutes after the addition of 100mM histamine. However, specificity was not optimal, as this biosensor generated significant bioluminescence in the presence of L-proline and L-histidine. As a means towards improving histamine specificity, the promoter region of a histamine oxidase gene from Arthrobacter globiformis was cloned upstream of the promotorless lux operon from Photorhabdus luminescens. This recently constructed whole-cell, lux-based bioluminescent bioreporter is currently being tested for optimal performance in the presence of histamine in order to provide a rapid, simple, and inexpensive model sensor for the detection of foodborne toxins.

  15. 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

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

    Zinn Kurt R

    2009-08-01

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

  17. Sensitive Dual Color in vivo Bioluminescence Imaging Using a New Red Codon Optimized Firefly Luciferase and a Green Click Beetle Luciferase

    2011-04-01

    expression of the bacterial luciferase gene cassette ( lux ) in a mammalian cell line. PLoS One 5: e12441. 6. Cheong WF, Prahl SA, Welch AJ (1990) A...Chemistry, Connecticut College, New London, Connecticut, United States of America Abstract Background: Despite a plethora of bioluminescent reporter genes ...challenging. This is partly attributable to the lack of optimization of cell reporter gene expression as well as too much spectral overlap of the color- coupled

  18. Smartphone-based low light detection for bioluminescence application

    Kim, Huisung; Jung, Youngkee; Doh, Iyll-Joon; Lozano-Mahecha, Roxana Andrea; Applegate, Bruce; Bae, Euiwon

    2017-01-01

    We report a smartphone-based device and associated imaging-processing algorithm to maximize the sensitivity of standard smartphone cameras, that can detect the presence of single-digit pW of radiant flux intensity. The proposed hardware and software, called bioluminescent-based analyte quantitation by smartphone (BAQS), provides an opportunity for onsite analysis and quantitation of luminescent signals from biological and non-biological sensing elements which emit photons in response to an analyte. A simple cradle that houses the smartphone, sample tube, and collection lens supports the measuring platform, while noise reduction by ensemble averaging simultaneously lowers the background and enhances the signal from emitted photons. Five different types of smartphones, both Android and iOS devices, were tested, and the top two candidates were used to evaluate luminescence from the bioluminescent reporter Pseudomonas fluorescens M3A. The best results were achieved by OnePlus One (android), which was able to detect luminescence from ~106 CFU/mL of the bio-reporter, which corresponds to ~107 photons/s with 180 seconds of integration time.

  19. Smartphone-based low light detection for bioluminescence application

    Kim, Huisung; Jung, Youngkee; Doh, Iyll-Joon; Lozano-Mahecha, Roxana Andrea; Applegate, Bruce; Bae, Euiwon

    2017-01-01

    We report a smartphone-based device and associated imaging-processing algorithm to maximize the sensitivity of standard smartphone cameras, that can detect the presence of single-digit pW of radiant flux intensity. The proposed hardware and software, called bioluminescent-based analyte quantitation by smartphone (BAQS), provides an opportunity for onsite analysis and quantitation of luminescent signals from biological and non-biological sensing elements which emit photons in response to an analyte. A simple cradle that houses the smartphone, sample tube, and collection lens supports the measuring platform, while noise reduction by ensemble averaging simultaneously lowers the background and enhances the signal from emitted photons. Five different types of smartphones, both Android and iOS devices, were tested, and the top two candidates were used to evaluate luminescence from the bioluminescent reporter Pseudomonas fluorescens M3A. The best results were achieved by OnePlus One (android), which was able to detect luminescence from ~106 CFU/mL of the bio-reporter, which corresponds to ~107 photons/s with 180 seconds of integration time. PMID:28067287

  20. 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.

  1. Bioluminescent assay for human lymphocyte blast transformation.

    Bulanova, E G; Budagyan, V M; Romanova, N A; Brovko LYu; Ugarova, N N

    1995-05-01

    One of the basic tests of in vitro evaluation of immune cell functional activity is a proliferative response of lymphocytes on the action of external stimuli such as mitogenic lectines, antigens, etc. We compared two methods used to assess the lymphocyte functional status. (1) [3H]thymidine incorporation and (2) bioluminescence for determination of intracellular ATP in blast cells. Comparison has been done for healthy donors and patients with proven low immunological status. The proposed bioluminescent method for evaluation of the proliferative response was shown to be sensitive enough for diagnostic purposes. This method allows one to process a large number of samples at the same time and correlates highly with the radionuclide test use hazardous radioactive materials.

  2. 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.

  3. Filtrage et déconvolution en imagerie de bioluminescence chez le petit animal

    Akkoul, Smaïl

    2010-01-01

    This thesis is devoted to the 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 ...

  4. Phage-amplified bioluminescent bioreporters for the detection of foodborne pathogens

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

    2004-06-01

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

  5. Bioluminescence Potential Modeling and Forecasting

    2013-05-22

    bioluminescence in the wakes of ships, breaking waves, around the bodies of rapidly moving fish and mammals , and from simple agitation of the water with one’s hand...history of brilliant displays of bioluminescence in the wakes of ships, breaking waves, around the bodies of rapidly moving fish and mammals , and from...during the earlier stages of upwelling development. Later, the observed deep offshore BL potential maximum disappeared and became a shallower and much

  6. 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.

  7. 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...

  8. Magnified time-domain ghost imaging

    Ryczkowski, Piotr; Barbier, Margaux; Friberg, Ari T.; Dudley, John M.; Genty, Goëry

    2017-04-01

    Ghost imaging allows the imaging of an object without directly seeing this object. Originally demonstrated in the spatial domain, it was recently shown that ghost imaging can be transposed into the time domain to detect ultrafast signals, even in the presence of distortion. We propose and experimentally demonstrate a temporal ghost imaging scheme which generates a 5× magnified ghost image of an ultrafast waveform. Inspired by shadow imaging in the spatial domain and building on the dispersive Fourier transform of an incoherent supercontinuum in an optical fiber, the approach overcomes the resolution limit of standard time-domain ghost imaging generally imposed by the detectors speed. The method can be scaled up to higher magnification factors using longer fiber lengths and light source with shorter duration.

  9. Ex vivo bioluminescence detection of alcelaphine herpesvirus 1 infection during malignant catarrhal fever.

    Dewals, Benjamin; Myster, Françoise; Palmeira, Leonor; Gillet, Laurent; Ackermann, Mathias; Vanderplasschen, Alain

    2011-07-01

    Alcelaphine herpesvirus 1 (AlHV-1), carried by wildebeest asymptomatically, causes malignant catarrhal fever (WD-MCF) when cross-species transmitted to a variety of susceptible species of the Artiodactyla order. Experimentally, WD-MCF can be reproduced in rabbits. WD-MCF is described as a combination of lymphoproliferation and degenerative lesions in virtually all organs and is caused by unknown mechanisms. Recently, we demonstrated that WD-MCF is associated with the proliferation of CD8(+) cells supporting a latent type of infection in lymphoid tissues. Here, we investigated the macroscopic distribution of AlHV-1 infection using ex vivo bioluminescence imaging in rabbit to determine whether it correlates with the distribution of lesions in lymphoid and nonlymphoid organs. To reach that goal, a recombinant AlHV-1 strain was produced by insertion of a luciferase expression cassette (luc) in an intergenic region. In vitro, the reconstituted AlHV-1 luc(+) strain replicated comparably to the parental strain, and luciferase activity was detected by bioluminescence imaging. In vivo, rabbits infected with the AlHV-1 luc(+) strain developed WD-MCF comparably to rabbits infected with the parental wild-type strain, with hyperthermia and increases of both CD8(+) T cell frequencies and viral genomic charge over time in peripheral blood mononuclear cells and in lymph nodes at time of euthanasia. Bioluminescent imaging revealed that AlHV-1 infection could be detected ex vivo in lymphoid organs but also in lung, liver, and kidney during WD-MCF, demonstrating that AlHV-1 infection is prevalent in tissue lesions. Finally, we show that the infiltrating mononuclear leukocytes in nonlymphoid organs are mainly CD8(+) T cells and that latency is predominant during WD-MCF.

  10. Measuring IL-1β Processing by Bioluminescence Sensors I: Using a Bioluminescence Resonance Energy Transfer Biosensor.

    Compan, Vincent; Pelegrín, Pablo

    2016-01-01

    IL-1β processing is one of the hallmarks of inflammasome activation and drives the initiation of the inflammatory response. For decades, Western blot or ELISA have been extensively used to study this inflammatory event. Here, we describe the use of a bioluminescence resonance energy transfer (BRET) biosensor to monitor IL-1β processing in real time and in living macrophages either using a plate reader or a microscope.

  11. Space-Time Imaging Systems

    2009-02-20

    vorticity layer along the surface of the wing, when the excitation is at a subharmonic of the inherent instability frequency, i.e., fo/2. This pattern...34a Instantaneous Yorticity Time-a’ ·era ged S treamli.nes Figure 1: Small perturbations (one-half degree) of angle-of - attack at the subharmonic of

  12. 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.

  13. 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.

  14. 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

  15. Application of ATP-based bioluminescence for bioaerosol quantification: effect of sampling method.

    Han, Taewon; Wren, Melody; DuBois, Kelsey; Therkorn, Jennifer; Mainelis, Gediminas

    2015-12-01

    An adenosine triphosphate (ATP)-based bioluminescence has potential to offer a quick and affordable method for quantifying bioaerosol samples. Here we report on our investigation into how different bioaerosol aerosolization parameters and sampling methods affect bioluminescence output per bacterium, and implications of that effect for bioaerosol research. Bacillus atrophaeus and Pseudomonas fluorescens bacteria were aerosolized by using a Collison nebulizer (BGI Inc., Waltham, MA) with a glass or polycarbonate jar and then collected for 15 and 60 min with: (1) Button Aerosol Sampler (SKC Inc., Eighty Four, PA) with polycarbonate, PTFE, and cellulose nitrate filters, (2) BioSampler (SKC Inc.) with 5 and 20 mL of collection liquid, and (3) our newly developed Electrostatic Precipitator with Superhydrophobic Surface (EPSS). For all aerosolization and sampling parameters we compared the ATP bioluminescence output per bacterium relative to that before aerosolization and sampling. In addition, we also determined the ATP reagent storage and preparation conditions that that do not affect the bioluminescence signal intensity. Our results show that aerosolization by a Collison nebulizer with a polycarbonate jar yields higher bioluminescence output per bacterium compared to the glass jar. Interestingly enough, the bioluminescence output by P. fluorescens increased substantially after its aerosolization compared to the fresh liquid suspension. For both test microorganisms, the bioluminescence intensity per bacterium after sampling was significantly lower than that before sampling suggesting negative effect of sampling stress on bioluminescence output. The decrease in bioluminescence intensity was more pronounces for longer sampling times and significantly and substantially depended on the sampling method. Among the investigated method, the EPSS was the least injurious for both microorganisms and sampling times. While the ATP-based bioluminescence offers a quick bioaerosol

  16. Autofluorescence imaging device for real-time detection and tracking of pathogenic bacteria in a mouse skin wound model: preclinical feasibility studies

    Wu, Yichao Charlie; Kulbatski, Iris; Medeiros, Philip J.; Maeda, Azusa; Bu, Jiachuan; Xu, Lizhen; Chen, Yonghong; DaCosta, Ralph S.

    2014-08-01

    Bacterial infection significantly impedes wound healing. Clinical diagnosis of wound infections is subjective and suboptimal, in part because bacteria are invisible to the naked eye during clinical examination. Moreover, bacterial infection can be present in asymptomatic patients, leading to missed opportunities for diagnosis and treatment. We developed a prototype handheld autofluorescence (AF) imaging device (Portable Real-time Optical Detection, Identification and Guidance for Intervention-PRODIGI) to noninvasively visualize and measure bacterial load in wounds in real time. We conducted preclinical pilot studies in an established nude mouse skin wound model inoculated with bioluminescent Staphylococcus aureus bacteria. We tested the feasibility of longitudinal AF imaging for in vivo visualization of bacterial load in skin wounds, validated by bioluminescence imaging. We showed that bacteria (S. aureus), occult to standard examination, can be visualized in wounds using PRODIGI. We also detected quantitative changes in wound bacterial load over time based on the antibiotic treatment and the correlation of bacterial AF intensity with bacterial load. AF imaging of wounds offers a safe, noninvasive method for visualizing the presence, location, and extent of bacteria as well as measuring relative changes in bacterial load in wounds in real time.

  17. Let there be bioluminescence – Development of a biophotonic imaging platform for in situ analyses of oral biofilms in animal models

    Merritt, Justin; Senpuku, Hidenobu; Kreth, Jens

    2016-01-01

    Summary In the current study, we describe a novel biophotonic imaging-based reporter system that is particularly useful for the study of virulence in polymicrobial infections and interspecies interactions within animal models. A suite of luciferase enzymes was compared using three early colonizing species of the human oral flora (Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis) to determine the utility of the different reporters for multiplexed imaging studies in vivo. Using the multiplex approach, we were able to track individual species within a dual species oral infection model in mice with both temporal and spatial resolution. We also demonstrate how biophotonic imaging of multiplexed luciferase reporters could be adapted for real-time quantification of bacterial gene expression in situ. By creating an inducible dual-luciferase expressing reporter strain of S. mutans, we were able to exogenously control and measure expression of nlmAB (encoding the bacteriocin mutacin IV) within mice to assess its importance for the persistence ability of S. mutans in the oral cavity. The imaging system described in the current study circumvents many of the inherent limitations of current animal model systems, which should now make it feasible to test hypotheses that were previously impractical to model. PMID:26119252

  18. Exploiting in vitro and in vivo bioluminescence for the implementation of the three Rs principle (replacement, reduction, and refinement) in drug discovery.

    Michelini, Elisa; Cevenini, Luca; Calabretta, Maria Maddalena; Calabria, Donato; Roda, Aldo

    2014-09-01

    Bioluminescence-based analytical tools are suitable for high-throughput and high-content screening assays, finding widespread application in several fields related to the drug discovery process. Cell-based bioluminescence assays, because of their peculiar advantages of predictability, possibility of automation, multiplexing, and miniaturization, seem the most appealing tool for the high demands of the early stages of drug screening. Reporter gene technology and the bioluminescence resonance energy transfer principle are widely used, and receptor binding studies of new agonists/antagonists for a variety of human receptors expressed in different cell lines can be performed. Moreover, bioluminescence can be used for in vitro and in vivo real-time monitoring of pathophysiological processes within living cells and small animals. New luciferases and substrates have recently arrived on the market, further expanding the spectrum of applications. A new generation of probes are also emerging that promise to revolutionize the preclinical imaging market. This formidable toolbox is demonstrated to facilitate the implementation of the three Rs principle in the early drug discovery process, in compliance with ethical and responsible research to reduce cost and improve the reliability and predictability of results.

  19. Electromagnetic Time Reversal Imaging: Analysis and Experimentation

    2010-04-26

    Biomedical Imaging: From Nano to Macro, ISBI󈧌, Paris, Friance, May 14-17, 2008 [6] Y. Jin, J. M. F. Moura, N. O’Donoughue, "Adaptive Time Reversal...Zhu, and Q. He, “Breast cancer detection by time reversal imaging,” 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro...target (a galvanized steel sheet) is surrounded by a large amount of PVC rods. Our experiments showed that the collected EM data in frequency and

  20. Coronary magnetic resonance vein imaging: imaging contrast, sequence, and timing.

    Nezafat, Reza; Han, Yuchi; Peters, Dana C; Herzka, Daniel A; Wylie, John V; Goddu, Beth; Kissinger, Kraig K; Yeon, Susan B; Zimetbaum, Peter J; Manning, Warren J

    2007-12-01

    Recently, there has been increased interest in imaging the coronary vein anatomy to guide interventional cardiovascular procedures such as cardiac resynchronization therapy (CRT), a device therapy for congestive heart failure (CHF). With CRT the lateral wall of the left ventricle is electrically paced using a transvenous coronary sinus lead or surgically placed epicardial lead. Proper transvenous lead placement is facilitated by the knowledge of the coronary vein anatomy. Cardiovascular MR (CMR) has the potential to image the coronary veins. In this study we propose and test CMR techniques and protocols for imaging the coronary venous anatomy. Three aspects of design of imaging sequence were studied: magnetization preparation schemes (T(2) preparation and magnetization transfer), imaging sequences (gradient-echo (GRE) and steady-state free precession (SSFP)), and imaging time during the cardiac cycle. Numerical and in vivo studies both in healthy and CHF subjects were performed to optimize and demonstrate the utility of CMR for coronary vein imaging. Magnetization transfer was superior to T(2) preparation for contrast enhancement. Both GRE and SSFP were viable imaging sequences, although GRE provided more robust results with better contrast. Imaging during the end-systolic quiescent period was preferable as it coincided with the maximum size of the coronary veins.

  1. 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

  2. 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.

  3. TOWARD REAL TIME UAVS’ IMAGE MOSAICKING

    S. Mehrdad

    2016-06-01

    Full Text Available 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.

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

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

    2014-01-01

    Background Expression of autonomous bioluminescence from human cells was previously reported to be impossible, suggesting that all bioluminescent-based mammalian reporter systems must therefore require application of a potentially influential chemical substrate. While this was disproven when the bacterial luciferase (lux) cassette was demonstrated to function in a human cell, its expression required multiple genetic constructs, was functional in only a single cell type, and generated a significantly reduced signal compared to substrate-requiring systems. Here we investigate the use of a humanized, viral 2A-linked lux genetic architecture for the efficient introduction of an autobioluminescent phenotype across a variety of human cell lines. Methodology/Principal Findings The lux cassette was codon optimized and assembled into a synthetic human expression operon using viral 2A elements as linker regions. Human kidney, breast cancer, and colorectal cancer cell lines were both transiently and stably transfected with the humanized operon and the resulting autobioluminescent phenotype was evaluated using common imaging instrumentation. Autobioluminescent cells were screened for cytotoxic effects resulting from lux expression and their utility as bioreporters was evaluated through the demonstration of repeated monitoring of single populations over a prolonged period using both a modified E-SCREEN assay for estrogen detection and a classical cytotoxic compound detection assay for the antibiotic Zeocin. Furthermore, the use of self-directed bioluminescent initiation in response to target detection was assessed to determine its amenability towards deployment as fully autonomous sensors. In all cases, bioluminescent measurements were supported with traditional genetic and transcriptomic evaluations. Conclusions/Significance Our results demonstrate that the viral 2A-linked, humanized lux genetic architecture successfully produced autobioluminescent phenotypes in all cell lines

  5. 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 proje...

  6. Bioluminescence in the ghost fungus Omphalotus nidiformis does not attract potential spore dispersing insects.

    Weinstein, Philip; Delean, Steven; Wood, Tom; Austin, Andrew D

    2016-12-01

    Bioluminescence has been known from fungi since ancient times, but little work has been done to establish its potential role. There is evidence that some bioluminescent fungi differentially attract potential spore-dispersing insects, and we aimed to establish if this was the case for the ghost fungus, Omphalotus nidiformis (Agaricales,Marasmiaceae), a widespread Australian temperate zone species. We examined three corroborative lines of evidence: circadian rhythmicity of bioluminescence; field-recorded insect abundance at the time of basidiome production; and attractiveness of glowing fungi to flying insects. Basidiomes glowed continuously day and night, and were present in winter (June-July) when insect abundance was low. To assess attractiveness, we deployed sticky-traps in open woodland in the absence of light pollution, in Treatment (baited with fresh bioluminescent O. nidiformis) and Control pairs, for 480 trap-hours on moonless nights. There was no statistical difference in mean insect abundance between Treatment and Control traps (mean 0.33 and 0.54 individuals per trap night, respectively). To interpret these results, we provide a brief review of competing hypotheses for fungal bioluminescence, and conclude that for some fungi, bioluminescence may be an incidental by-product of metabolism rather than conferring any selective advantage. It is possible that the role of bioluminescence differs among evolutionary lineages of fungi and/or with attributes of their growth environments that could affect spore dispersal, such as wind and insect abundance.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. Evaluation of an improved bioluminescence assay for the detection of bacteria in soy milk.

    Shinozaki, Yohei; Sato, Jun; Igarashi, Toshinori; Suzuki, Shigeya; Nishimoto, Kazunori; Harada, Yasuhiro

    2013-01-01

    Because soy milk is nutrient rich and nearly neutral in pH, it favors the growth of microbial contaminants. To ensure that soy milk meets food-safety standards, it must be pasteurized and have its sterility confirmed. ATP bioluminescence assay has become a widely accepted means of detecting food microorganisms. However, the high background bioluminescence intensity of soy milk has rendered it unsuitable for ATP analysis. Here, we tested the efficacy of an improved pre-treated bioluminescence assay on soy milk. By comparing background bioluminescence intensities obtained by the conventional and improved methods, we demonstrated that our method significantly reduces soy milk background bioluminescence. The dose-response curve of the assay was tested with serial dilutions of Bacillus sp. culture. An extremely strong log-linear relation between the bioluminescence intensity relative light units and colony formation units CFU/ml emerged for the tested strain. The detection limit of the assay was estimated as 5.2×10(3) CFU/ml from the dose-response curve and an imposed signal limit was three times the background level. The results showed that contaminated samples could be easily detected within 24 h using our improved bioluminescence assay.

  12. 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.

  13. 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.

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

    Lu, Yujie; Douraghy, Ali; Machado, Hidevaldo B.; Stout, David; Tian, Jie; Herschman, Harvey; Chatziioannou, Arion F.

    2009-11-01

    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.

  15. Cloaking and imaging at the same time

    Wu, Qiannan; Xu, Yadong; Li, Hui; Chen, Huanyang

    2013-02-01

    In this letter, we propose a conceptual device to perform good imaging with positive refraction. At the same time, this device is an isotropic omnidirectional cloak with a perfect electric conductor hiding region and shows versatile illusion optical effects. Numerical simulations are performed to verify the functionalities.

  16. Bioluminescence assay for cell viability.

    Lomakina, G Yu; Modestova, Yu A; Ugarova, N N

    2015-06-01

    Theoretical aspects of the adenosine triphosphate bioluminescence assay based on the use of the firefly luciferin-luciferase system are considered, as well as its application for assessing cell viability in microbiology, sanitation, medicine, and ecology. Various approaches for the analysis of individual or mixed cultures of microorganisms are presented, and capabilities of the method for investigation of biological processes in live cells including necrosis, apoptosis, as well as for investigation of the dynamics of metabolism are described.

  17. 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

  18. Real-Time Imaging of Quantum Entanglement

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

    2013-01-01

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

  19. The influence of hypoxia on bioluminescence in luciferase-transfected gliosarcoma tumor cells in vitro.

    Moriyama, Eduardo H; Niedre, Mark J; Jarvi, Mark T; Mocanu, Joseph D; Moriyama, Yumi; Subarsky, Patrick; Li, Buhong; Lilge, Lothar D; Wilson, Brian C

    2008-06-01

    Firefly luciferase catalyzes the emission of light from luciferin in the presence of oxygen and adenosine triphosphate. This bioluminescence is commonly employed in imaging mode to monitor tumor growth and treatment responses in vivo. A potential concern is that, since solid tumors are often hypoxic, either constitutively and/or as a result of treatment, the oxygen available for the bioluminescence reaction could be reduced to limiting levels, leading to underestimation of the actual number of luciferase-labeled cells during in vivo experiments. We present studies of the oxygen dependence of bioluminescence in vitro in rat 9 L gliosarcoma cells tagged with the firefly luciferase gene (9L(luc)). We demonstrate that the bioluminescence signal decreases at pO(2) ATP due to the reduction of mitochondrial membrane potential. Hence, the data suggest that the decrease of intracellular ATP level in vitro is the limiting factor for bioluminescence reaction and so is responsible for the reduction of bioluminescence signal in 9L(luc) cells in acute hypoxia, rather than luciferase expression or oxygen itself.

  20. 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.

  1. Polynomial-time solutions to image segmentation

    Asano, Tetsuo [Osaka Electro-Communication Univ., Neyagawa (Japan); Chen, D.Z. [Notre Dame, South Bend, IN (United States); Katoh, Naoki [Kobe Univ. of Commerce (Japan)

    1996-12-31

    Separating an object in an image from its background is a central problem (called segmentation) in pattern recognition and computer vision. In this paper, we study the complexity of the segmentation problem, assuming that the object forms a connected region in an intensity image. We show that the optimization problem of separating a connected region in an n-pixel grid is NP-hard under the interclass variance, a criterion that is used in discriminant analysis. More importantly, we consider the basic case in which the object is separated by two x-monotone curves (i.e., the object itself is x-monotone), and present polynomial-time algorithms for computing exact and approximate optimal segmentation. Our main algorithm for exact optimal segmentation by two x-monotone curves runs in O(n{sup 2}) time; this algorithm is based on several techniques such as a parametric optimization formulation, a hand-probing algorithm for the convex hull of an unknown point set, and dynamic programming using fast matrix searching. Our efficient approximation scheme obtains an {epsilon}-approximate solution in O({epsilon}{sup -1} n log L) time, where {epsilon} is any fixed constant with 1 > {epsilon} > 0, and L is the total sum of the absolute values of brightness levels of the image.

  2. 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...

  3. 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.

  4. Combining biofunctional magnetic nanoparticles and ATP bioluminescence for rapid detection of Escherichia coli.

    Cheng, Yuxiao; Liu, Yajun; Huang, Jingjing; Li, Kang; Zhang, Wen; Xian, Yuezhong; Jin, Litong

    2009-02-15

    A rapid, specific and sensitive method for assay of Escherichia coli (E. coli) using biofunctional magnetic nanoparticles (BMNPs) in combination with adenosine triphosphate (ATP) bioluminescence was proposed. The BMNPs were fabricated by immobilizing a specific anti-E. coli antibody on the surface of amine-functionalized magnetic nanoparticles (about 20nm in diameter), and then was applied to capture the target bacteria E. coli from samples. The BMNPs exhibited high capture efficiency to E. coli. Transmission electron microscope (TEM) images showed that the BMNPs were bound to the surface of entire E. coli cells. The target bacteria became magnetic so that could be isolated easily from the sample solution by employing an external magnetic field. The concentration of E. coli captured by the BMNPs was then detected by an ATP bioluminescence method. The optimization of ATP measurement was carried out to improve the detection sensitivity. The proposed method was applied to detect the E. coli inoculated into pasteurized milk with low detection limit (20 cfu/mL) and short detection time (about 1h).

  5. 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.

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

    Golberg, Karina; Elbaz, Amit; McNeil, Ronald; Kushmaro, Ariel; Geddes, Chris D.; Marks, Robert S.

    2014-12-01

    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.

  7. Doppler time-of-flight imaging

    Heide, Felix

    2015-07-30

    Over the last few years, depth cameras have become increasingly popular for a range of applications, including human-computer interaction and gaming, augmented reality, machine vision, and medical imaging. Many of the commercially-available devices use the time-of-flight principle, where active illumination is temporally coded and analyzed on the camera to estimate a per-pixel depth map of the scene. In this paper, we propose a fundamentally new imaging modality for all time-of-flight (ToF) cameras: per-pixel velocity measurement. The proposed technique exploits the Doppler effect of objects in motion, which shifts the temporal frequency of the illumination before it reaches the camera. Using carefully coded illumination and modulation frequencies of the ToF camera, object velocities directly map to measured pixel intensities. We show that a slight modification of our imaging system allows for color, depth, and velocity information to be captured simultaneously. Combining the optical flow computed on the RGB frames with the measured metric axial velocity allows us to further estimate the full 3D metric velocity field of the scene. We believe that the proposed technique has applications in many computer graphics and vision problems, for example motion tracking, segmentation, recognition, and motion deblurring.

  8. Interactive Real-time Magnetic Resonance Imaging

    Brix, Lau

    seeks to implement and assess existing reconstruction algorithms using multi-processors of modern graphics cards and many-core computer processors and to cover some of the potential clinical applications which might benefit from using an interactive real-time MRI system. First an off......-line, but interactive, slice alignment tool was used to support the notion that 3D blood flow quantification in the heart possesses the ability to obtain curves and volumes which are not statistical different from standard 2D flow. Secondly, the feasibility of an interactive real-time MRI system was exploited...... with regard to optimal sampling strategy for detecting motion in four different anatomies on two different MRI scanner brands. A fully implemented interactive real-time MRI system was exploited in a group of healthy fetuses and proved its eligibility as an alternative diagnostic tool for fetal imaging...

  9. 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.

  10. 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 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. PMID:18256736

  11. The Chemical Basis of Fungal Bioluminescence.

    Purtov, Konstantin V; Petushkov, Valentin N; Baranov, Mikhail S; Mineev, Konstantin S; Rodionova, Natalja S; Kaskova, Zinaida M; Tsarkova, Aleksandra S; Petunin, Alexei I; Bondar, Vladimir S; Rodicheva, Emma K; Medvedeva, Svetlana E; Oba, Yuichi; Oba, Yumiko; Arseniev, Alexander S; Lukyanov, Sergey; Gitelson, Josef I; Yampolsky, Ilia V

    2015-07-06

    Many species of fungi naturally produce light, a phenomenon known as bioluminescence, however, the fungal substrates used in the chemical reactions that produce light have not been reported. We identified the fungal compound luciferin 3-hydroxyhispidin, which is biosynthesized by oxidation of the precursor hispidin, a known fungal and plant secondary metabolite. The fungal luciferin does not share structural similarity with the other eight known luciferins. Furthermore, it was shown that 3-hydroxyhispidin leads to bioluminescence in extracts from four diverse genera of luminous fungi, thus suggesting a common biochemical mechanism for fungal bioluminescence.

  12. QM/MM study on the light emitters of aequorin chemiluminescence, bioluminescence, and fluorescence: a general understanding of the bioluminescence of several marine organisms.

    Chen, Shu-Feng; Ferré, Nicolas; Liu, Ya-Jun

    2013-06-24

    Aequorea victoria is a type of jellyfish that is known by its famous protein, green fluorescent protein (GFP), which has been widely used as a probe in many fields. Aequorea has another important protein, aequorin, which is one of the members of the EF-hand calcium-binding protein family. Aequorin has been used for intracellular calcium measurements for three decades, but its bioluminescence mechanism remains largely unknown. One of the important reasons is the lack of clear and reliable knowledge about the light emitters, which are complex. Several neutral and anionic forms exist in chemiexcited, bioluminescent, and fluorescent states and are connected with the H-bond network of the binding cavity in the protein. We first theoretically investigated aequorin chemiluminescence, bioluminescence, and fluorescence in real proteins by performing hybrid quantum mechanics and molecular mechanics methods combined with a molecular dynamics method. For the first time, this study reported the origin and clear differences in the chemiluminescence, bioluminescence and fluorescence of aequorin, which is important for understanding the bioluminescence not only of jellyfish, but also of many other marine organisms (that have the same coelenterazine caved in different coelenterazine-type luciferases).

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. The Repetitive Detection of Toluene with Bioluminescence Bioreporter Pseudomonas putida TVA8 Encapsulated in Silica Hydrogel on an Optical Fiber

    Gabriela Kuncová

    2016-06-01

    Full Text Available Living cells of the lux-based bioluminescent bioreporter Pseudomonas putida TVA8 were encapsulated in a silica hydrogel attached to the distal wider end of a tapered quartz fiber. Bioluminescence of immobilized cells was induced with toluene at high (26.5 mg/L and low (5.3 mg/L concentrations. Initial bioluminescence maxima were achieved after >12 h. One week after immobilization, a biofilm-like layer of cells had formed on the surface of the silica gel. This resulted in shorter response times and more intensive bioluminescence maxima that appeared as rapidly as 2 h after toluene induction. Considerable second bioluminescence maxima were observed after inductions with 26.5 mg toluene/L. The second and third week after immobilization the biosensor repetitively and semiquantitatively detected toluene in buffered medium. Due to silica gel dissolution and biofilm detachment, the bioluminescent signal was decreasing 20–32 days after immobilization and completely extinguished after 32 days. The reproducible formation of a surface cell layer on the wider end of the tapered optical fiber can be translated to various whole cell bioluminescent biosensor devices and may serve as a platform for in-situ sensors.

  19. 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.

  20. 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.

  1. Use of Bioluminescence Markers To Detect Pseudomonas spp. in the Rhizosphere

    de Weger, Letty A.; Dunbar, Paul; Mahafee, Walter F.; Lugtenberg, Ben J. J.; Sayler, Gary S.

    1991-01-01

    The use of bioluminescence as a sensitive marker for detection of Pseudomonas spp. in the rhizosphere was investigated. Continuous expression of the luxCDABE genes, required for bioluminescence, was not detectable in the rhizosphere. However, when either a naphthalene-inducible luxCDABE construct or a constitutive luxAB construct (coding only for the luciferase) was introduced into the Pseudomonas cells, light emission could be initiated just prior to measurement by the addition of naphthalene or the substrate for luciferase, n-decyl aldehyde, respectively. These Pseudomonas cells could successfully be detected in the rhizosphere by using autophotography or optical fiber light measurement techniques. Detection required the presence of 103 to 104 CFU/cm of root, showing that the bioluminescence technique is at least 1,000-fold more sensitive than β-galactosidase-based systems. Images PMID:16348610

  2. Bioluminescence in Dinoflagellates: Evidence that the Adaptive Value of Bioluminescence in Dinoflagellates is Concentration Dependent.

    Hanley, Karen A; Widder, Edith A

    2017-03-01

    Three major hypotheses have been proposed to explain why dinoflagellate bioluminescence deters copepod grazing: startle response, aposematic warning, and burglar alarm. These hypotheses propose dinoflagellate bioluminescence (A) startles predatory copepods, (B) warns potential predators of toxicity, and (C) draws the attention of higher order visual predators to the copepod's location. While the burglar alarm is the most commonly accepted hypothesis, it requires a high concentration of bioluminescent dinoflagellates to be effective, meaning the bioluminescence selective advantage at lower, more commonly observed, dinoflagellate concentrations may result from another function (e.g. startle response or aposematic warning). Therefore, a series of experiments was conducted to evaluate copepod grazing (Acartia tonsa) on bioluminescent dinoflagellates (during bioluminescent and nonbioluminescent phases, corresponding to night and day, respectively) at different concentrations (10, 1000, and 3000 cells mL(-1) ), on toxic (Pyrodinium bahamense var. bahamense) and nontoxic (Lingulodinium polyedrum) bioluminescent dinoflagellates, and in the presence of nonluminescent diatoms (Thalassiosira eccentrica). Changes in copepod ingestion rates, clearance rates, and feeding preferences as a result of these experimental factors, particularly during the mixed trails with nonluminescent diatoms, indicate there is a concentration threshold at which the burglar alarm becomes effective and below which dinoflagellate bioluminescence functions as an aposematic warning.

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

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

    1999-01-01

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

  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. 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.

  6. Real-time in vivo photoacoustic and ultrasound imaging

    Kolkman, Roy G.M.; Brands, Peter J.; Steenbergen, Wiendelt; Leeuwen, van Ton G.

    2008-01-01

    A real-time photoacoustic imaging system is designed and built. This system is based on a commercially available ultrasound imaging system. It can achieve a frame rate of 8 frames/sec. Vasculature in the hand of a human volunteer is imaged, and the resulting photoacoustic image is combined with the

  7. Time-resolved spectroscopy and imaging

    Chance, Britton

    1995-05-01

    In response to the conference organizer's request, I am presenting a summary of the current status of medical optical imaging and spectroscopy. This is a topic which is advancing rapidly and on which there have been a number of conferences recently, and yet there has not been presented an overview of the field and some idea of what the advantages and disadvantages of the photon diffusion technology may be. Thus, this paper emphasizes diffusion waves for spectroscopy and imaging deep within the tissue and, at the same time, for providing specificity information of both absorption and scattering. In achieving this goal, I will not be able to cite all of the advantages of technologies that view the superficial layers of skin, retina, etc., on the one hand, nor those that measure the photons that have been scattered minimally on the transit between input and output. One of the main reasons for this is that specificity of the optical methods requires all of the information available: absorption and scattering of intrinsic signals naturally in the tissue, and of extrinsic signal due to contrast agents that have been artificially lodged in strategic tissue volumes. Since this paper is essentially the transcript of a lecture, it is not proposed as a topic review and does not contain full-scale bibliographic references, some of which may be found in a recent review elsewhere. This paper highlights what we all might accomplish in order to bring to bear with maximal effectiveness the optical method for altering the outcome of medical problems. I have not emphasized the mathematics of photon diffusion so well represented by the papers of this symposium volume. The achievable goals of the optical methods are to speed detection, improve diagnosis, guide therapy, and what appears in the minds of most, contribute to the improvement of medical economics. In order to fulfill these objectives, we will in the end have to demonstrate by lengthy and expensive clinical studies that the

  8. High-sensitivity real-time imaging of dual protein-protein interactions in living subjects using multicolor luciferases.

    Naoki Hida

    Full Text Available Networks of protein-protein interactions play key roles in numerous important biological processes in living subjects. An effective methodology to assess protein-protein interactions in living cells of interest is protein-fragment complement assay (PCA. Particularly the assays using fluorescent proteins are powerful techniques, but they do not directly track interactions because of its irreversibility or the time for chromophore formation. By contrast, PCAs using bioluminescent proteins can overcome these drawbacks. We herein describe an imaging method for real-time analysis of protein-protein interactions using multicolor luciferases with different spectral characteristics. The sensitivity and signal-to-background ratio were improved considerably by developing a carboxy-terminal fragment engineered from a click beetle luciferase. We demonstrate its utility in spatiotemporal characterization of Smad1-Smad4 and Smad2-Smad4 interactions in early developing stages of a single living Xenopus laevis embryo. We also describe the value of this method by application of specific protein-protein interactions in cell cultures and living mice. This technique supports quantitative analyses and imaging of versatile protein-protein interactions with a selective luminescence wavelength in opaque or strongly auto-fluorescent living subjects.

  9. Is it time for cardiac innervation imaging?

    Knuuti, J. [Turku Univ., Turku (Finland) Turku PET Center; Sipola, P. [Kuopio Univ., Kuopio (Finland)

    2005-03-01

    The autonomic nervous system plays an important role in the regulation of cardiac function and the regional distribution of cardiac nerve terminals can be visualized using scintigraphic techniques. The most commonly used tracer is iodine-123-metaiodobenzylguanidine (MIBG) but C-11-hydroxyephedrine has also been used with PET. When imaging with MIBG, the ratio of heart-to-mediastinal counts is used as an index of tracer uptake, and regional distribution is also assessed from tomographic images. The rate of clearance of the tracer can also be measured and indicates the function of the adrenergic system. Innervation imaging has been applied in patients with susceptibility to arrythmias, coronary artery disease, hypertrophic and dilated cardiomyopathy and anthracycline induced cardiotoxicity. Abnormal adrenergic innervation or function appear to exist in many pathophysiological conditions indicating that sympathetic neurons are very susceptible to damage. Abnormal findings in innervation imaging also appear to have significant prognostic value especially in patients with cardiomyopathy. Recently, it has also been shown that innervation imaging can monitor drug-induced changes in cardiac adrenergic activity. Although innervation imaging holds great promise for clinical use, the method has not received wider clinical acceptance. Larger randomized studies are required to confirm the value of innervation imaging in various specific indications.

  10. Novel in vivo imaging analysis of an inner ear drug delivery system in mice: comparison of inner ear drug concentrations over time after transtympanic and systemic injections.

    Sho Kanzaki

    Full Text Available OBJECTIVE: Systemic steroid injections are used to treat idiopathic sudden-onset sensorineural hearing loss (ISSHL and some inner ear disorders. Recent studies show that transtympanic (TT steroid injections are effective for treating ISSHL. As in vivo monitoring of drug delivery dynamics for inner ear is lacking, its time course and dispersion of drugs is unknown. Here, we used a new in vivo imaging system to monitor drug delivery in live mice and to compare drug concentrations over time after TT and systemic injections. METHODS: Luciferin delivered into the inner ears of GFAP-Luc transgenic mice reacted with luciferase in GFAP-expressing cells in the cochlear spiral ganglion, resulting in photon bioluminescence. We used the Xenogen IVIS® imaging system to measure how long photons continued to be emitted in the inner ear after TT or systemic injections of luciferin, and then compared the associated drug dynamics. RESULTS: The response to TT and IP injections differed significantly. Photons were detected five minutes after TT injection, peaking at ~20 minutes. By contrast, photons were first detected 30 minutes after i.p. injection. TT and i.p. drug delivery time differed considerably. With TT injections, photons were detected earlier than with IP injections. Photon bioluminescence also disappeared sooner. Delivery time varied with TT injections. CONCLUSIONS: We speculate that the drug might enter the Eustachian tube from the middle ear. We conclude that inner-ear drug concentration can be maintained longer if the two injection routes are combined. As the size of luciferin differs from that of therapeutics like dexamethasone, combining drugs with luciferin may advance our understanding of in vivo drug delivery dynamics in the inner ear.

  11. Bioluminescent bioreporter assays for targeted detection of chemical and biological agents

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

    2008-04-01

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

  12. 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.

  13. Immobilized Bioluminescent Reagents in Flow Injection Analysis.

    Nabi, Abdul

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

  14. Automated Real-Time Conjunctival Microvasculature Image Stabilization.

    Felder, Anthony E; Mercurio, Cesare; Wanek, Justin; Ansari, Rashid; Shahidi, Mahnaz

    2016-07-01

    The bulbar conjunctiva is a thin, vascularized membrane covering the sclera of the eye. Non-invasive imaging techniques have been utilized to assess the conjunctival vasculature as a means of studying microcirculatory hemodynamics. However, eye motion often confounds quantification of these hemodynamic properties. In the current study, we present a novel optical imaging system for automated stabilization of conjunctival microvasculature images by real-time eye motion tracking and realignment of the optical path. The ability of the system to stabilize conjunctival images acquired over time by reducing image displacements and maintaining the imaging area was demonstrated.

  15. Monitoring of environmental pollutants by bioluminescent bacteria.

    Girotti, Stefano; Ferri, Elida Nora; Fumo, Maria Grazia; Maiolini, Elisabetta

    2008-02-04

    This review deals with the applications of bioluminescent bacteria to the environmental analyses, published during the years 2000-2007. The ecotoxicological assessment, by bioassays, of the environmental risks and the luminescent approaches are reported. The review includes a brief introduction to the characteristics and applications of bioassays, a description of the characteristics and applications of natural bioluminescent bacteria (BLB), and a collection of the main applications to organic and inorganic pollutants. The light-emitting genetically modified bacteria applications, as well as the bioluminescent immobilized systems and biosensors are outlined. Considerations about commercially available BLB and BLB catalogues are also reported. Most of the environmental applications, here mentioned, of luminescent organisms are on wastewater, seawater, surface and ground water, tap water, soil and sediments, air. Comparison to other bioindicators and bioassay has been also made. Various tables have been inserted, to make easier to take a rapid glance at all possible references concerning the topic of specific interest.

  16. Detection of bacteria with bioluminescent reporter bacteriophage.

    Klumpp, Jochen; Loessner, Martin J

    2014-01-01

    Bacteriophages are viruses that exclusively infect bacteria. They are ideally suited for the development of highly specific diagnostic assay systems. Bioluminescent reporter bacteriophages are designed and constructed by integration of a luciferase gene in the virus genome. Relying on the host specificity of the phage, the system enables rapid, sensitive, and specific detection of bacterial pathogens. A bioluminescent reporter phage assay is superior to any other molecular detection method, because gene expression and light emission are dependent on an active metabolism of the bacterial cell, and only viable cells will yield a signal. In this chapter we introduce the concept of creating reporter phages, discuss their advantages and disadvantages, and illustrate the advances made in developing such systems for different Gram-negative and Gram-positive pathogens. The application of bioluminescent reporter phages for the detection of foodborne pathogens is emphasized.

  17. The CUBLAS and CULA based GPU acceleration of adaptive finite element framework for bioluminescence tomography.

    Zhang, Bo; Yang, Xiang; Yang, Fei; Yang, Xin; Qin, Chenghu; Han, Dong; Ma, Xibo; Liu, Kai; Tian, Jie

    2010-09-13

    In molecular imaging (MI), especially the optical molecular imaging, bioluminescence tomography (BLT) emerges as an effective imaging modality for small animal imaging. The finite element methods (FEMs), especially the adaptive finite element (AFE) framework, play an important role in BLT. The processing speed of the FEMs and the AFE framework still needs to be improved, although the multi-thread CPU technology and the multi CPU technology have already been applied. In this paper, we for the first time introduce a new kind of acceleration technology to accelerate the AFE framework for BLT, using the graphics processing unit (GPU). Besides the processing speed, the GPU technology can get a balance between the cost and performance. The CUBLAS and CULA are two main important and powerful libraries for programming on NVIDIA GPUs. With the help of CUBLAS and CULA, it is easy to code on NVIDIA GPU and there is no need to worry about the details about the hardware environment of a specific GPU. The numerical experiments are designed to show the necessity, effect and application of the proposed CUBLAS and CULA based GPU acceleration. From the results of the experiments, we can reach the conclusion that the proposed CUBLAS and CULA based GPU acceleration method can improve the processing speed of the AFE framework very much while getting a balance between cost and performance.

  18. 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.

  19. Dual-time-point Imaging and Delayed-time-point Fluorodeoxyglucose-PET/Computed Tomography Imaging in Various Clinical Settings.

    Houshmand, Sina; Salavati, Ali; Segtnan, Eivind Antonsen; Grupe, Peter; Høilund-Carlsen, Poul Flemming; Alavi, Abass

    2016-01-01

    The techniques of dual-time-point imaging (DTPI) and delayed-time-point imaging, which are mostly being used for distinction between inflammatory and malignant diseases, has increased the specificity of fluorodeoxyglucose (FDG)-PET for diagnosis and prognosis of certain diseases. A gradually increasing trend of FDG uptake over time has been shown in malignant cells, and a decreasing or constant trend has been shown in inflammatory/infectious processes. Tumor heterogeneity can be assessed by using early and delayed imaging because differences between primary versus metastatic sites become more detectable compared with single time points. This article discusses the applications of DTPI and delayed-time-point imaging.

  20. Cloaking and imaging at the same time

    Wu, Qiannan; Chen, Huanyang

    2012-01-01

    In this letter, we propose a conceptual device to perform subwavelength imaging with positive refraction. The key to this proposal is that a drain is no longer a must for some cases. What's more, this device is an isotropic omnidirectional cloak with a perfect electric conductor hiding region and shows versatile illusion optical effects. Numerical simulations are performed to verify the functionalities.

  1. 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.

  2. Numerical modeling of the dynamic response of a bioluminescent bacterial biosensor.

    Affi, Mahmoud; Solliec, Camille; Legentilhomme, Patrick; Comiti, Jacques; Legrand, Jack; Jouanneau, Sulivan; Thouand, Gérald

    2016-12-01

    Water quality and water management are worldwide issues. The analysis of pollutants and in particular, heavy metals, is generally conducted by sensitive but expensive physicochemical methods. Other alternative methods of analysis, such as microbial biosensors, have been developed for their potential simplicity and expected moderate cost. Using a biosensor for a long time generates many changes in the growth of the immobilized bacteria and consequently alters the robustness of the detection. This work simulated the operation of a biosensor for the long-term detection of cadmium and improved our understanding of the bioluminescence reaction dynamics of bioreporter bacteria inside an agarose matrix. The choice of the numerical tools is justified by the difficulty to measure experimentally in every condition the biosensor functioning during a long time (several days). The numerical simulation of a biomass profile is made by coupling the diffusion equation and the consumption/reaction of the nutrients by the bacteria. The numerical results show very good agreement with the experimental profiles. The growth model verified that the bacterial growth is conditioned by both the diffusion and the consumption of the nutrients. Thus, there is a high bacterial density in the first millimeter of the immobilization matrix. The growth model has been very useful for the development of the bioluminescence model inside the gel and shows that a concentration of oxygen greater than or equal to 22 % of saturation is required to maintain a significant level of bioluminescence. A continuous feeding of nutrients during the process of detection of cadmium leads to a biofilm which reduces the diffusion of nutrients and restricts the presence of oxygen from the first layer of the agarose (1 mm) and affects the intensity of the bioluminescent reaction. The main advantage of this work is to link experimental works with numerical models of growth and bioluminescence in order to provide a

  3. In Vivo Real Time Volumetric Synthetic Aperture Ultrasound Imaging

    Bouzari, Hamed; Rasmussen, Morten Fischer; Brandt, Andreas Hjelm;

    2015-01-01

    Synthetic aperture (SA) imaging can be used to achieve real-time volumetric ultrasound imaging using 2-D array transducers. The sensitivity of SA imaging is improved by maximizing the acoustic output, but one must consider the limitations of an ultrasound system, both technical and biological...

  4. Real-time video-image analysis

    Eskenazi, R.; Rayfield, M. J.; Yakimovsky, Y.

    1979-01-01

    Digitizer and storage system allow rapid random access to video data by computer. RAPID (random-access picture digitizer) uses two commercially-available, charge-injection, solid-state TV cameras as sensors. It can continuously update its memory with each frame of video signal, or it can hold given frame in memory. In either mode, it generates composite video output signal representing digitized image in memory.

  5. 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.

  6. 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....

  7. Bioluminescent bioreporter integrated circuit detection methods

    Simpson, Michael L.; Paulus, Michael J.; Sayler, Gary S.; Applegate, Bruce M.; Ripp, Steven A.

    2005-06-14

    Disclosed are monolithic bioelectronic devices comprising a bioreporter and an OASIC. These bioluminescent bioreporter integrated circuit are useful in detecting substances such as pollutants, explosives, and heavy-metals residing in inhospitable areas such as groundwater, industrial process vessels, and battlefields. Also disclosed are methods and apparatus for detection of particular analytes, including ammonia and estrogen compounds.

  8. 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.

  9. Bubble stimulation efficiency of dinoflagellate bioluminescence.

    Deane, Grant B; Stokes, M Dale; Latz, Michael I

    2016-02-01

    Dinoflagellate bioluminescence, a common source of bioluminescence in coastal waters, is stimulated by flow agitation. Although bubbles are anecdotally known to be stimulatory, the process has never been experimentally investigated. This study quantified the flash response of the bioluminescent dinoflagellate Lingulodinium polyedrum to stimulation by bubbles rising through still seawater. Cells were stimulated by isolated bubbles of 0.3-3 mm radii rising at their terminal velocity, and also by bubble clouds containing bubbles of 0.06-10 mm radii for different air flow rates. Stimulation efficiency, the proportion of cells producing a flash within the volume of water swept out by a rising bubble, decreased with decreasing bubble radius for radii less than approximately 1 mm. Bubbles smaller than a critical radius in the range 0.275-0.325 mm did not stimulate a flash response. The fraction of cells stimulated by bubble clouds was proportional to the volume of air in the bubble cloud, with lower stimulation levels observed for clouds with smaller bubbles. An empirical model for bubble cloud stimulation based on the isolated bubble observations successfully reproduced the observed stimulation by bubble clouds for low air flow rates. High air flow rates stimulated more light emission than expected, presumably because of additional fluid shear stress associated with collective buoyancy effects generated by the high air fraction bubble cloud. These results are relevant to bioluminescence stimulation by bubbles in two-phase flows, such as in ship wakes, breaking waves, and sparged bioreactors.

  10. Bioluminophore and Flavin Mononucleotide Fluorescence Quenching of Bacterial Bioluminescence-A Theoretical Study.

    Luo, Yanling; Liu, Ya-Jun

    2016-11-02

    Bacterial bioluminescence with continuous glow has been applied to the fields of environmental toxin monitoring, drug screening, and in vivo imaging. Nonetheless, the chemical form of the bacterial bioluminophore is still a bone of contention. Flavin mononucleotide (FMN), one of the light-emitting products, and 4a-hydroxy-5-hydro flavin mononucleotide (HFOH), an intermediate of the chemical reactions, have both been assumed candidates for the light emitter because they have similar molecular structures and fluorescence wavelengths. The latter is preferred in experiments and was assigned in our previous density functional study. HFOH displays weak fluorescence in solutions, but exhibits strong bioluminescence in the bacterial luciferase. FMN shows the opposite behavior; its fluorescence is quenched when it is bound to the luciferase. This is the first example of flavin fluorescence quenching observed in bioluminescent systems and is merely an observation, both the quenching mechanism and quencher are still unclear. Based on theoretical analysis of high-level quantum mechanics (QM), combined QM and molecular mechanics (QM/MM), and molecular dynamics (MD), this paper confirms that HFOH in its first singlet excited state is the bioluminophore of bacterial bioluminescence. More importantly, the computational results indicate that Tyr110 in the luciferase quenches the FMN fluorescence via an electron-transfer mechanism.

  11. Amplitude metrics for cellular circadian bioluminescence reporters.

    St John, Peter C; Taylor, Stephanie R; Abel, John H; Doyle, Francis J

    2014-12-01

    Bioluminescence rhythms from cellular reporters have become the most common method used to quantify oscillations in circadian gene expression. These experimental systems can reveal phase and amplitude change resulting from circadian disturbances, and can be used in conjunction with mathematical models to lend further insight into the mechanistic basis of clock amplitude regulation. However, bioluminescence experiments track the mean output from thousands of noisy, uncoupled oscillators, obscuring the direct effect of a given stimulus on the genetic regulatory network. In many cases, it is unclear whether changes in amplitude are due to individual changes in gene expression level or to a change in coherence of the population. Although such systems can be modeled using explicit stochastic simulations, these models are computationally cumbersome and limit analytical insight into the mechanisms of amplitude change. We therefore develop theoretical and computational tools to approximate the mean expression level in large populations of noninteracting oscillators, and further define computationally efficient amplitude response calculations to describe phase-dependent amplitude change. At the single-cell level, a mechanistic nonlinear ordinary differential equation model is used to calculate the transient response of each cell to a perturbation, whereas population-level dynamics are captured by coupling this detailed model to a phase density function. Our analysis reveals that amplitude changes mediated at either the individual-cell or the population level can be distinguished in tissue-level bioluminescence data without the need for single-cell measurements. We demonstrate the effectiveness of the method by modeling experimental bioluminescence profiles of light-sensitive fibroblasts, reconciling the conclusions of two seemingly contradictory studies. This modeling framework allows a direct comparison between in vitro bioluminescence experiments and in silico ordinary

  12. 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...

  13. Dual-time-point Imaging and Delayed-time-point Fluorodeoxyglucose-PET/Computed Tomography Imaging in Various Clinical Settings

    Houshmand, Sina; Salavati, Ali; Antonsen Segtnan, Eivind;

    2016-01-01

    The techniques of dual-time-point imaging (DTPI) and delayed-time-point imaging, which are mostly being used for distinction between inflammatory and malignant diseases, has increased the specificity of fluorodeoxyglucose (FDG)-PET for diagnosis and prognosis of certain diseases. A gradually...

  14. Efficient hybrid method for time reversal superresolution imaging

    Xiaohua Wang,Wei Gao,; Bingzhong Wang

    2015-01-01

    An efficient hybrid time reversal (TR) imaging method based on signal subspace and noise subspace is proposed for electromagnetic superresolution detecting and imaging. First, the locations of targets are estimated by the transmitting-mode decom-position of the TR operator (DORT) method employing the signal subspace. Then, the TR multiple signal classification (TR-MUSIC) method employing the noise subspace is used in the estimated target area to get the superresolution imaging of targets. Two examples with homogeneous and inhomogeneous background mediums are considered, respectively. The results show that the proposed hybrid method has advantages in CPU time and memory cost because of the combination of rough and fine imaging.

  15. Image-driven pharmacokinetics: nanomedicine concentration across space and time.

    Brill, Dab A; MacKay, J Andrew

    2015-01-01

    Clinical pharmacokinetics (PK) primarily measures the concentration of drugs in the blood. For nanomedicines it may be more relevant to determine concentration within a target tissue. The emerging field of image-driven PK, which utilizes clinically accepted molecular imaging technology, empirically and noninvasively, measures concentration in multiple tissues. Image-driven PK represents the intersection of PK and biodistribution, combining to provide models of concentration across space and time. Image-driven PK can be used both as a research tool and in the clinic. This review explores the history of pharmacokinetics, technologies used in molecular imaging (especially positron emission tomography) and research using image-driven pharmacokinetic analysis. When standardized, image-driven PK may have significant implications in preclinical development as well as clinical optimization of targeted nanomedicines.

  16. 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.

  17. Image Space and Time Interpolation for Video Navigation

    2011-01-01

    English: The aim of image-based video navigation is essentially to achieve a continuous change in the viewpoint without the need of a complete camera coverage of the space of interest. By making use of image interpolation, the need for video hardware can be reduced drastically by replacing them, in the desired viewpoints, with virtual video cameras. In this work, based on previously published approaches, an algorithm for time and space image interpolation is developed with a video application...

  18. Time-resolved imaging of latent fingerprints with nanosecond resolution

    Seah, L. K.; Dinish, U. S.; Ong, S. K.; Chao, Z. X.; Murukeshan, V. M.

    2004-07-01

    Imaging of latent fingerprints using time-resolved (TR) method offers a broader platform to eliminate the unwanted background emission. In this paper, a novel TR imaging technique is demonstrated and implemented, which facilitates the detection of latent fingerprints with nanosecond resolution. Simulated experiments were carried out with two overlapping fingerprints treated with two fluorescent powders having different lifetimes in nanosecond range. The dependence of the fluorescence emission intensity in nanosecond resolution of TR imaging is also revealed.

  19. Ultrafast quantitative time-stretch imaging flow cytometry of phytoplankton

    Lai, Queenie T. K.; Lau, Andy K. S.; Tang, Anson H. L.; Wong, Kenneth K. Y.; Tsia, Kevin K.

    2016-03-01

    Comprehensive quantification of phytoplankton abundance, sizes and other parameters, e.g. biomasses, has been an important, yet daunting task in aquatic sciences and biofuel research. It is primarily because of the lack of effective tool to image and thus accurately profile individual microalgae in a large population. The phytoplankton species are highly diversified and heterogeneous in terms of their sizes and the richness in morphological complexity. This fact makes time-stretch imaging, a new ultrafast real-time optical imaging technology, particularly suitable for ultralarge-scale taxonomic classification of phytoplankton together with quantitative image recognition and analysis. We here demonstrate quantitative imaging flow cytometry of single phytoplankton based on quantitative asymmetric-detection time-stretch optical microscopy (Q-ATOM) - a new time-stretch imaging modality for label-free quantitative phase imaging without interferometric implementations. Sharing the similar concept of Schlieren imaging, Q-ATOM accesses multiple phase-gradient contrasts of each single phytoplankton, from which the quantitative phase profile is computed. We employ such system to capture, at an imaging line-scan rate of 11.6 MHz, high-resolution images of two phytoplankton populations (scenedesmus and chlamydomonas) in ultrafast microfluidic flow (3 m/s). We further perform quantitative taxonomic screening analysis enabled by this technique. More importantly, the system can also generate quantitative phase images of single phytoplankton. This is especially useful for label-free quantification of biomasses (e.g. lipid droplets) of the particular species of interest - an important task adopted in biofuel applications. Combining machine learning for automated classification, Q-ATOM could be an attractive platform for continuous and real-time ultralarge-scale single-phytoplankton analysis.

  20. Luciferin Amides Enable in Vivo Bioluminescence Detection of Endogenous Fatty Acid Amide Hydrolase Activity.

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

    2015-07-15

    Firefly luciferase is homologous to fatty acyl-CoA synthetases. We hypothesized that the firefly luciferase substrate d-luciferin and its analogs are fatty acid mimics that are ideally suited to probe the chemistry of enzymes that release fatty acid products. Here, we synthesized luciferin amides and found that these molecules are hydrolyzed to substrates for firefly luciferase by the enzyme fatty acid amide hydrolase (FAAH). In the presence of luciferase, these molecules enable highly sensitive and selective bioluminescent detection of FAAH activity in vitro, in live cells, and in vivo. The potency and tissue distribution of FAAH inhibitors can be imaged in live mice, and luciferin amides serve as exemplary reagents for greatly improved bioluminescence imaging in FAAH-expressing tissues such as the brain.

  1. Bioluminescence as an ecological factor during high Arctic polar night

    Cronin, Heather A.; Cohen, Jonathan H.; Berge, Jørgen; Johnsen, Geir; Moline, Mark A.

    2016-11-01

    Bioluminescence commonly influences pelagic trophic interactions at mesopelagic depths. Here we characterize a vertical gradient in structure of a generally low species diversity bioluminescent community at shallower epipelagic depths during the polar night period in a high Arctic fjord with in situ bathyphotometric sampling. Bioluminescence potential of the community increased with depth to a peak at 80 m. Community composition changed over this range, with an ecotone at 20–40 m where a dinoflagellate-dominated community transitioned to dominance by the copepod Metridia longa. Coincident at this depth was bioluminescence exceeding atmospheric light in the ambient pelagic photon budget, which we term the bioluminescence compensation depth. Collectively, we show a winter bioluminescent community in the high Arctic with vertical structure linked to attenuation of atmospheric light, which has the potential to influence pelagic ecology during the light-limited polar night.

  2. Bioluminescence-Sensing Assay for Microbial Growth Recognition

    Heba Ramadan Eed

    2016-01-01

    Full Text Available 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 bioluminescent recombinant E. coli strain was used with luciferase extracted from transformed bacteria. Results showed that there is a direct correlation between the bioluminescence intensity of the ATP bioluminescence-sensing assay and the microbial viability. Bacterial counts from food samples were detected using the developed sensing assay and validated by the traditional plate-counting method. Compared with the plate-counting method, ATP bioluminescence-sensing assay is a more rapid and efficient approach for detecting microbial viability.

  3. 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 endoscopic ultrasound), image fusion in several anatomic regions, and electromagnetic needle tracking....

  4. Spatiotemporal expression of heme oxygenase-1 detected by in vivo bioluminescence after hepatic ischemia in HO-1/luc mice

    Su, Huawei; van Dam, Gooitzen M.; Buis, Carlijn I.; Visser, Dorien S.; Hesselink, Jan Willem; Schuurs, Theo A.; Leuvenink, Henri G. D.; Contag, Christopher H.; Porte, Robert J.

    2006-01-01

    Upregulation of heme oxygenase-1 (HO-1) has been proposed as a critical mechanism protecting against cellular stress during liver transplantation, providing a potential target for new therapeutic interventions. We investigated the feasibility of in vivo bioluminescence imaging (BLI) to noninvasively

  5. 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.

  6. Experimental ultrasound system for real-time synthetic imaging

    1999-01-01

    Digital signal processing is being employed more and more in modern ultrasound scanners. This has made it possible to do dynamic receive focusing for each sample and implement other advanced imaging methods. The processing, however, has to be very fast and cost-effective at the same time. Dedicated chips are used in order to do real time processing. This often makes it difficult to implement radically different imaging strategies on one platform and makes the scanners less accessible for rese...

  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. 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.

  9. 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.

  10. Bioluminescence ATP monitoring for the routine assessment of food contact surface cleanliness in a university canteen.

    Osimani, Andrea; Garofalo, Cristiana; Clementi, Francesca; Tavoletti, Stefano; Aquilanti, Lucia

    2014-10-17

    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.

  11. Sequences for real-time magnetic particle imaging

    Weber Matthias

    2015-09-01

    Full Text Available Magnetic Particle Imaging (MPI is a new imaging modality with the potential to be a new medical tool for angiographic diagnostics. It is capable of visualizing the spatial distribution of super-paramagnetic nanoparticles in high temporal and spatial resolution. Furthermore, the new spatial encoding scheme of a field free line (FFL promises a ten-fold higher sensitivity. So far, all know imaging devices featuring this new technique feature slow data acquisition and thus, are far away from real-time imaging capability. An actual real-time approach requires a complex field generator and an application of currents with very precise amplitude and phase. Here, we present the first implementation and calibration of a dynamic FFL field sequence enabling the acquisition of 50 MPI images per second in a mouse sized scanner.

  12. Time multiplexed pinhole array based lensless three-dimensional imager

    Schwarz, Ariel; Wang, Jingang; Shemer, Amir; Zalevsky, Zeev; Javidi, Bahram

    2016-06-01

    We present an overview of multi variable coded aperture (MVCA) for lensless three-dimensional integral imaging (3D II) systems. The new configuration is based on a time multiplexing method using a variable pinholes array design. The system provides higher resolution 3D images with improved light intensity and signal to noise ratio as compared to single pinhole system. The MVCA 3D II system configuration can be designed to achieve high light intensity for practical use as micro lenslets arrays. This new configuration preserves the advantages of pinhole optics while solving the resolution limitation problem and the long exposure time of such systems. The three dimensional images are obtained with improved resolution, signal to noise ratio and sensitivity efficiency. This integral imaging lensless system is characterized by large depth of focus, simplicity and low cost. In this paper we present numerical simulations as well as experimental results that validate the proposed lensless imaging configuration.

  13. Infrared Image Real-time Enhancement Based on DSP

    DAI Shao-sheng; YUAN Xiang-hui; XUE Lian

    2004-01-01

    Since image real- time processing requires vast amount of computation and high- speed hardware,it is difficult to be implemented with general microcomputer system. In order to solve the problem,a powerful digital signal processing (DSP) hardware system is proposed,which is able to meet needs of image real-time processing. There are many approaches to enhance infrared image. But only histogram equalization is discussed because it is the most common and effective way. On the basis of histogram equalization principle, the specific procedures implemented in DSP are shown. At last the experimental results are given.

  14. Time-of-flight flow imaging using NMR remote detection

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

    2005-05-05

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

  15. 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.

  16. Implied Movement in Static Images Reveals Biological Timing Processing

    Francisco Carlos Nather

    2015-08-01

    Full Text Available 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 participants. Movement, Velocity, and Arousal semantic scales were used to characterize some properties of the images. Implied movement affected time perception, in which M images were overestimated. The results are discussed in terms of visual motion perception related to biological timing processing that could be established early in terms of the adaptation of humankind to the environment.

  17. Experimental ultrasound system for real-time synthetic imaging

    Jensen, Jørgen Arendt; Holm, Ole; Jensen, Lars Joost

    1999-01-01

    Digital signal processing is being employed more and more in modern ultrasound scanners. This has made it possible to do dynamic receive focusing for each sample and implement other advanced imaging methods. The processing, however, has to be very fast and cost-effective at the same time. Dedicated...... chips are used in order to do real time processing. This often makes it difficult to implement radically different imaging strategies on one platform and makes the scanners less accessible for research purposes. Here flexibility is the prime concern, and the storage of data from all transducer elements......-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...

  18. Coherent temporal imaging with analog time-bandwidth compression

    Asghari, Mohammad H

    2013-01-01

    We introduce the concept of coherent temporal imaging and its combination with the anamorphic stretch transform. The new system can measure both temporal profile of fast waveforms as well as their spectrum in real time and at high-throughput. We show that the combination of coherent detection and warped time-frequency mapping also performs time-bandwidth compression. By reducing the temporal width without sacrificing spectral resolution, it addresses the Big Data problem in real time instruments. The proposed method is the first application of the recently demonstrated Anamorphic Stretch Transform to temporal imaging. Using this method narrow spectral features beyond the spectrometer resolution can be captured. At the same time the output bandwidth and hence the record length is minimized. Coherent detection allows the temporal imaging and dispersive Fourier transform systems to operate in the traditional far field as well as in near field regimes.

  19. Quasi-real-time fluorescence imaging with lifetime dependent contrast

    Jiang, Pei-Chi; Grundfest, Warren S.; Stafsudd, Oscar M.

    2011-08-01

    Conventional fluorescence lifetime imaging requires complicated algorithms to extract lifetimes of fluorophores and acquisition of multiple data points at progressively longer delay times to characterize tissues. To address diminishing signal-to-noise ratios at these progressively longer time delays, we report a time-resolved fluorescence imaging method, normalized fluorescence yield imaging that does not require the extraction of lifetimes. The concept is to extract the ``contrast'' instead of the lifetime value of the fluorophores by using simple mathematical algorithms. This process converts differences in decay times directly to different intensities. The technique was verified experimentally using a gated iCCD camera and an ultraviolet light-emitting diode light source. It was shown that this method can distinguish between chemical dyes (Fluorescein and Rhodamine-B) and biomedical samples, such as powders of elastin and collagen. Good contrast was obtained between fluorophores that varied by less than 6% in lifetime. Additionally, it was shown that long gate times up to 16 ns achieve good contrast depending upon the samples to be studied. These results support the feasibility of time-resolved fluorescence imaging without lifetime extraction, which has a potential clinical role in noninvasive real-time imaging.

  20. Time-resolved multispectral imaging of combustion reaction

    Huot, Alexandrine; Gagnon, Marc-André; Jahjah, Karl-Alexandre; Tremblay, Pierre; Savary, Simon; Farley, Vincent; Lagueux, Philippe; Guyot, Éric; Chamberland, Martin; Marcotte, Fréderick

    2015-05-01

    Thermal infrared imaging is a field of science that evolves rapidly. Scientists have used for years the simplest tool: thermal broadband cameras. This allows to perform target characterization in both the longwave (LWIR) and midwave (MWIR) infrared spectral range. Infrared thermal imaging is used for a wide range of applications, especially in the combustion domain. For example, it can be used to follow combustion reactions, in order to characterize the injection and the ignition in a combustion chamber or even to observe gases produced by a flare or smokestack. Most combustion gases such as carbon dioxide (CO2) selectively absorb/emit infrared radiation at discrete energies, i.e. over a very narrow spectral range. Therefore, temperatures derived from broadband imaging are not reliable without prior knowledge about spectral emissivity. This information is not directly available from broadband images. However, spectral information is available using spectral filters. In this work, combustion analysis was carried out using Telops MS-IR MW camera which allows multispectral imaging at a high frame rate. A motorized filter wheel allowing synchronized acquisitions on eight (8) different channels was used to provide time-resolved multispectral imaging of combustion products of a candle in which black powder has been burnt to create a burst. It was then possible to estimate the temperature by modeling spectral profile derived from information obtained with the different spectral filters. Comparison with temperatures obtained using conventional broadband imaging illustrates the benefits of time-resolved multispectral imaging for the characterization of combustion processes.

  1. Time-resolved multispectral imaging of combustion reactions

    Huot, Alexandrine; Gagnon, Marc-André; Jahjah, Karl-Alexandre; Tremblay, Pierre; Savary, Simon; Farley, Vincent; Lagueux, Philippe; Guyot, Éric; Chamberland, Martin; Marcotte, Frédérick

    2015-10-01

    Thermal infrared imaging is a field of science that evolves rapidly. Scientists have used for years the simplest tool: thermal broadband cameras. These allow to perform target characterization in both the longwave (LWIR) and midwave (MWIR) infrared spectral range. Infrared thermal imaging is used for a wide range of applications, especially in the combustion domain. For example, it can be used to follow combustion reactions, in order to characterize the injection and the ignition in a combustion chamber or even to observe gases produced by a flare or smokestack. Most combustion gases, such as carbon dioxide (CO2), selectively absorb/emit infrared radiation at discrete energies, i.e. over a very narrow spectral range. Therefore, temperatures derived from broadband imaging are not reliable without prior knowledge of spectral emissivity. This information is not directly available from broadband images. However, spectral information is available using spectral filters. In this work, combustion analysis was carried out using a Telops MS-IR MW camera, which allows multispectral imaging at a high frame rate. A motorized filter wheel allowing synchronized acquisitions on eight (8) different channels was used to provide time-resolved multispectral imaging of combustion products of a candle in which black powder has been burnt to create a burst. It was then possible to estimate the temperature by modeling spectral profiles derived from information obtained with the different spectral filters. Comparison with temperatures obtained using conventional broadband imaging illustrates the benefits of time-resolved multispectral imaging for the characterization of combustion processes.

  2. 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.

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

    J. C. Chimal-Eguía

    2012-06-01

    Full Text Available 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 datarepresentation is based on information obtained by image axis division into boxes. The difference between this newinput data representation and the classical is that this technique is not time-dependent. This new information isimplemented in the new Image-Based Learning Approach (IBLA and by means of a probabilistic mechanism thislearning technique is applied to the interesting problem of time series forecasting. The experimental results indicatethat by using the methodology proposed in this article, it is possible to obtain better results than with the classicaltechniques such as artificial neuronal networks and support vector machines.

  4. 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.

  5. In Vivo Real Time Volumetric Synthetic Aperture Ultrasound Imaging

    Bouzari, Hamed; Rasmussen, Morten Fischer; Brandt, Andreas Hjelm

    2015-01-01

    . This paper investigates the in vivo applicability and sensitivity of volumetric SA imaging. Utilizing the transmit events to generate a set of virtual point sources, a frame rate of 25 Hz for a 90° x 90° field-of-view was achieved. Data were obtained using a 3.5 MHz 32 x 32 elements 2-D phased array......Synthetic aperture (SA) imaging can be used to achieve real-time volumetric ultrasound imaging using 2-D array transducers. The sensitivity of SA imaging is improved by maximizing the acoustic output, but one must consider the limitations of an ultrasound system, both technical and biological...... transducer connected to the experimental scanner (SARUS). Proper scaling is applied to the excitation signal such that intensity levels are in compliance with the U.S. Food and Drug Administration regulations for in vivo ultrasound imaging. The measured Mechanical Index and spatial-peak- temporal...

  6. 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...

  7. Programmable Real-time Clinical Photoacoustic and Ultrasound Imaging System.

    Kim, Jeesu; Park, Sara; Jung, Yuhan; Chang, Sunyeob; Park, Jinyong; Zhang, Yumiao; Lovell, Jonathan F; Kim, Chulhong

    2016-10-12

    Photoacoustic imaging has attracted interest for its capacity to capture functional spectral information with high spatial and temporal resolution in biological tissues. Several photoacoustic imaging systems have been commercialized recently, but they are variously limited by non-clinically relevant designs, immobility, single anatomical utility (e.g., breast only), or non-programmable interfaces. Here, we present a real-time clinical photoacoustic and ultrasound imaging system which consists of an FDA-approved clinical ultrasound system integrated with a portable laser. The system is completely programmable, has an intuitive user interface, and can be adapted for different applications by switching handheld imaging probes with various transducer types. The customizable photoacoustic and ultrasound imaging system is intended to meet the diverse needs of medical researchers performing both clinical and preclinical photoacoustic studies.

  8. Pixel timing correction in time-lapsed calcium imaging using point scanning microscopy.

    Boiroux, Dimitri; Oke, Yoshihiko; Miwakeichi, Fumikazu; Oku, Yoshitaka

    2014-11-30

    In point scanning imaging, data are acquired by sequentially scanning each pixel of a predetermined area. This way of scanning leads to time delays between pixels, especially for lower scanning speed or large scanned areas. Therefore, experiments are often performed at lower framerates in order to ensure a sufficient signal-to-noise ratio, even though framerates above 30 frames per second are technically feasible. For these framerates, we suggest that it becomes crucial to correct the time delay between image pixels prior to analyses. In this paper, we apply temporal interpolation (or pixel timing correction) for calcium imaging in two-photon microscopy as an example of fluorescence imaging. We present and compare three interpolation methods (linear, Lanczos and cubic B-spline). We test these methods on a simulated network of coupled bursting neurons at different framerates. In this network, we introduce a time delay to simulate a scanning by point scanning microscopy. We also assess these methods on actual microscopic calcium imaging movies recorded at usual framerates. Our numerical results suggest that point scanning microscopy imaging introduces statistically significant time delays between image pixels at low frequency. However, we demonstrate that pixel timing correction compensates for these time delays, regardless of the used interpolation method.

  9. Real-time chemiluminescence imaging using nano-lantern probes.

    Arai, Yoshiyuki; Nagai, Takeharu

    2014-01-01

    Chemiluminescence imaging can be performed without excitation light sources at various spatial levels ranging from a single cell to the whole body. Thus far, chemiluminescence imaging has been primarily performed with long exposure times because of weak signals, resulting in low temporal resolution. Recently, the brightest-known chemiluminescent proteins--Nano-lantern and Nano-lantern-based functional indicators--have been developed. Nano-lantern probes break the limitation of temporal resolution and enable chemiluminescence imaging of living samples such as cells, plants, and small animals at video rates. This unit describes one protocol for observation of a freely moving unshaved mouse transplanted with Nano-lantern-expressing tumor cells, and another for compatible use of optogenetic tools and a Nano-lantern calcium indicator. Both protocols utilize the synchronization of illumination and camera acquisition sessions, thereby enabling real-time chemiluminescence imaging.

  10. Mathematical methods in time series analysis and digital image processing

    Kurths, J; Maass, P; Timmer, J

    2008-01-01

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

  11. Two-dimensional random arrays for real time volumetric imaging

    Davidsen, Richard E.; Jensen, Jørgen Arendt; Smith, Stephen W.

    1994-01-01

    Two-dimensional arrays are necessary for a variety of ultrasonic imaging techniques, including elevation focusing, 2-D phase aberration correction, and real time volumetric imaging. In order to reduce system cost and complexity, sparse 2-D arrays have been considered with element geometries...... real time volumetric imaging system, which employs a wide transmit beam and receive mode parallel processing to increase image frame rate. Depth-of-field comparisons were made from simulated on-axis and off-axis beamplots at ranges from 30 to 160 mm for both coaxial and offset transmit and receive...... selected ad hoc, by algorithm, or by random process. Two random sparse array geometries and a sparse array with a Mills cross receive pattern were simulated and compared to a fully sampled aperture with the same overall dimensions. The sparse arrays were designed to the constraints of the Duke University...

  12. 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.

  13. Investigations on time stability of passive THz imaging

    Kowalski, Marcin; Palka, Norbert; Zyczkowski, Marek; Szustakowski, Mieczyslaw

    2014-10-01

    Terahertz radiation is within the frequency range from 100 GHz to 10THz. This radiation has specific characteristics in terms of imaging. The radiation is harmless to the human body because the energy transferred by electromagnetic waves in this range of frequencies are very small thus there is no ionization of matter. The development of imaging devices and exploration of new spectral bands is a chance to introduce new equipment for assuring public safety. It has been proved that objects hidden under clothing can be detected and visualized using terahertz (THz) cameras. However, passive THz cameras still offer too low image resolution for objects recognition. In order to determine the properties of terahertz imaging for detection of hidden objects several aspects need to be considered. Taking into account the fact that the image captured by the terahertz camera reflects the spatial distribution of the relative temperature of the observed objects, the effect of the measurement time on the imaging capabilities should be examined. A very important aspect is the influence of the type (material composition) of coating material, as well as the type of an object hidden under clothing (size and material). The purpose of the studies is to investigate the time stability of passive THz imaging on 250 GHz for detection of concealed objects. In the article, we present the measurement setup, the measurement methodology as well as the initial results of measurements with various types of clothing and test objects.

  14. 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

    2016-01-01

    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. V...

  15. 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.

  16. Real-time functional near-infrared imager

    Luo, Qingming; Zeng, Shaoqun; Chance, Britton; Nioka, Shoko

    1998-08-01

    A real time functional Near-InfraRed Imager (fNIRI) was presented in this paper. We developed a continuous wave (cw) light imaging probe which includes 9 light sources and 4 pairs detectors (each pair has one 850 nm filtered detector and one 760 nm filtered detector). There are 16 measurement sections and total detection area is 9 cm X 4 cm. The detector- source uses 2.5 cm spacing. The light sources are controlled by a computer and the signals from the detectors are converted and processed in real time by the computer. The user-friendly software was programmed with Visual C++ language. Relative changes of oxy-Hb, Hb, and total blood concentration in 16 channels and the corresponding images combined by 16 channels could be displayed in real time on computer. With this cw imaging probe, we have measured motor function in motor cortex area, visual function in occipital area, and cognitive activity in frontal forehead area of the human brain when the subjects are stimulated by moving fingers, viewing a flashing light and doing an analogy test, respectively. The experimental results show that the cw imaging probe can be used for functional images of brain activity, based upon changes of oxygenation and blood volume due to the stimulus.

  17. 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.

  18. 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...

  19. 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,…

  20. Single-cell bioluminescence and GFP in biofilm research

    Palmer, R.J. Jr, Sayler, G., White, D.C. [Tennessee Univ., Knoxville, TN (United States), Ctr. Env. Biotech; Phiefer, C. [Oak Ridge National Lab., TN (United States), Environmental Sciences Div.

    1996-12-31

    Using flow cells and a combination of microscopy techniques, we can unequivocally identify single bacterial cells that express bioluminescent and fluorescent bioreporters. We have shown that, for attached cells, bioluminescence output within a bacterial strain can vary greatly from cell to cell.

  1. Microcirculation monitoring with real time spatial frequency domain imaging

    Chen, Xinlin; Cao, Zili; Lin, Weihao; Zhu, Danfeng; Zhu, Xiuwei; Zeng, Bixin; Xu, M.

    2017-01-01

    We present a spatial frequency domain imaging (SFDI) study of local hemodynamics in the forearm of healthy volunteers performing paced breathing. Real time Single Snapshot Multiple Frequency Demodulation - Spatial Frequency Domain Imaging (SSMD-SFDI) was used to map the optical properties of the subsurface of the forearm continuously. The oscillations of the concentrations of deoxy- and oxyhemoglobin at the subsurface of the forearm induced by paced breathing are found to be close to out-of-phase, attributed to the dominance of the blood flow modulation by paced breathing. The properties of local microcirculation including the blood transit times through capillaries and venules are extracted by fitting to Simplified Hemodynamics Model. Our preliminary results suggest that the real time SSMD-SFDI platform may serve as one effective imaging modality for microcirculation monitoring.

  2. D City Transformations by Time Series of Aerial Images

    Adami, A.

    2015-02-01

    Recent photogrammetric applications, based on dense image matching algorithms, allow to use not only images acquired by digital cameras, amateur or not, but also to recover the vast heritage of analogue photographs. This possibility opens up many possibilities in the use and enhancement of existing photos heritage. The research of the original figuration of old buildings, the virtual reconstruction of disappeared architectures and the study of urban development are some of the application areas that exploit the great cultural heritage of photography. Nevertheless there are some restrictions in the use of historical images for automatic reconstruction of buildings such as image quality, availability of camera parameters and ineffective geometry of image acquisition. These constrains are very hard to solve and it is difficult to discover good dataset in the case of terrestrial close range photogrammetry for the above reasons. Even the photographic archives of museums and superintendence, while retaining a wealth of documentation, have no dataset for a dense image matching approach. Compared to the vast collection of historical photos, the class of aerial photos meets both criteria stated above. In this paper historical aerial photographs are used with dense image matching algorithms to realize 3d models of a city in different years. The models can be used to study the urban development of the city and its changes through time. The application relates to the city centre of Verona, for which some time series of aerial photographs have been retrieved. The models obtained in this way allowed, right away, to observe the urban development of the city, the places of expansion and new urban areas. But a more interesting aspect emerged from the analytical comparison between models. The difference, as the Euclidean distance, between two models gives information about new buildings or demolitions. As considering accuracy it is necessary point out that the quality of final

  3. 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.

  4. 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.

  5. Single photon imaging and timing array sensor apparatus and method

    Smith, R. Clayton

    2003-06-24

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

  6. 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.

  7. Reverse time migration based on normalized wavefield decomposition imaging condition

    YU Jianglong; HAN Liguo; ZHOU Yan; ZHANG Yongsheng

    2016-01-01

    With the increasing complexity of prospecting objectives,reverse time migration (RTM)has attracted more and more attention due to its outstanding imaging quality.RTMis based on two-way wave equation,so it can avoid the limits of angle in traditional one-way wave equation migration,image reverse branch,prism waves and multi-reflected wave precisely and obtain accurate dynamic information.However,the huge demands for storage and computation as well as low frequency noises restrict its wide application.The normalized cross-correlation ima-ging conditions based on wave field decomposition are derived from traditional cross-correlation imaging condition, and it can eliminate the low-frequency noises effectively and improve the imaging resolution.The practical proce-dure includes separating source and receiver wave field into one-way components respectively,and conducting cross-correlation imaging condition to the post-separated wave field.In this way,the resolution and precision of the imaging result will be promoted greatly.

  8. IMPLEMENTATION OF IMAGE PROCESSING IN REAL TIME CAR PARKING SYSTEM

    SAYANTI BANERJEE,

    2011-02-01

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

  9. A Robust Time Efficient Watermarking Technique for Stereo Images

    M. A. Abdou

    2015-01-01

    Full Text Available Stereoscopic and multiview imaging techniques are used for reproducing a natural or real world scene. However, the fact that more information is displayed requires supporting technologies to ensure the storage and transmission of the sequences. Beyond these supports comes watermarking as a desirable alternative solution for copyright protection of stereo images and videos. This paper introduces a watermarking method applied to stereo images in wavelet domain. This method uses a particle swarm optimization (PSO evolutionary computation method. The aim is to solve computational complexity problems as well as satisfy an execution time that complies with normal PCs or smart phones processors. Robustness against image attacks is tested, and results are shown.

  10. Time Domain Terahertz (T-Ray) Subsurface and Structural Imaging

    Zimdars, David; White, Jeffrey S.; Stuk, G.; Chernovsky, A.; Fichter, G.; Sucha, G.; Williamson, S.

    2007-03-01

    The technology, methods, and examples of high speed time domain terahertz (T-Ray) imaging non-destructive examination (NDE) for 2 and 3 dimensional structural and material content characterization are discussed. T-Ray imaging can be utilized for non-contact transmission and/or monostatic reflection inspection of non-conductive materials such as plastics, foam, composites, ceramics, paper, wood and glass. Example subsurface homeland security images of concealed items in baggage and on personnel are shown. We tabulate attenuation and penetration characteristics through a selection of building materials, and demonstrate the ability of T-ray instrumentation to sub-surface image building structures such as wall framing and interior wiring and conduits.

  11. 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.

  12. 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.

  13. Detection of dichloromethane with a bioluminescent (lux) bacterial bioreporter.

    Lopes, Nicholas; Hawkins, Shawn A; Jegier, Patricia; Menn, Fu-Min; Sayler, Gary S; Ripp, Steven

    2012-01-01

    The focus of this research effort was to develop an autonomous, inducible, lux-based bioluminescent bioreporter for the real-time detection of dichloromethane. Dichloromethane (DCM), also known as methylene chloride, is a volatile organic compound and one of the most commonly used halogenated solvents in the U.S., with applications ranging from grease and paint stripping to aerosol propellants and pharmaceutical tablet coatings. Predictably, it is released into the environment where it contaminates air and water resources. Due to its classification as a probable human carcinogen, hepatic toxin, and central nervous system effector, DCM must be carefully monitored and controlled. Methods for DCM detection usually rely on analytical techniques such as solid-phase microextraction (SPME) and capillary gas chromatography or photoacoustic environmental monitors, all of which require trained personnel and/or expensive equipment. To complement conventional monitoring practices, we have created a bioreporter for the self-directed detection of DCM by taking advantage of the evolutionary adaptation of bacteria to recognize and metabolize chemical agents. This bioreporter, Methylobacterium extorquens DCM( lux ), was engineered to contain a bioluminescent luxCDABE gene cassette derived from Photorhabdus luminescens fused downstream to the dcm dehalogenase operon, which causes the organism to generate visible light when exposed to DCM. We have demonstrated detection limits down to 1.0 ppm under vapor phase exposures and 0.1 ppm under liquid phase exposures with response times of 2.3 and 1.3 h, respectively, and with specificity towards DCM under relevant industrial environmental monitoring conditions.

  14. Space, time, error, and power optimization of image compression transforms

    Schmalz, Mark S.; Ritter, Gerhard X.; Caimi, Frank M.

    2000-11-01

    The implementation of an image compression transform on one or more small, embedded processors typically involves stringent constraints on power consumption and form factor. Traditional methods of optimizing compression algorithm performance typically emphasize joint minimization of space and time complexity, often without significant consideration of arithmetic accuracy or power consumption. However, small autonomous imaging platforms typically require joint optimization of space, time, error (or accuracy), and power (STEP) parameters, which the authors call STEP optimization. In response to implementational constraints on space and power consumption, the authors have developed systems and techniques for STEP optimization that are based on recent research in VLSI circuit design, as well as extensive previous work in system optimization. Building on the authors' previous research in embedded processors as well as adaptive or reconfigurable computing, it is possible to produce system-independent STEP optimization that can be customized for a given set of system-specific constraints. This approach is particularly useful when algorithms for image and signal processing (ISP) computer vision (CV), or automated target recognition (ATR), expressed in a machine- independent notation, are mapped to one or more heterogeneous processors (e.g., digital signal processors or DSPs, SIMD mesh processors, or reconfigurable logic). Following a theoretical summary, this paper illustrates various STEP optimization techniques via case studies, for example, real-time compression of underwater imagery on board an autonomous vehicle. Optimization algorithms are taken from the literature, and error profiling/analysis methodologies developed in the authors' previous research are employed. This yields a more rigorous basis for the simulation and evaluation of compression algorithms on a wide variety of hardware models. In this study, image algebra is employed as the notation of choice

  15. 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.

  16. 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.

  17. 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.

  18. 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...

  19. Diffeomorphic image registration with automatic time-step adjustment

    Pai, Akshay Sadananda Uppinakudru; Klein, S.; Sommer, Stefan Horst;

    2015-01-01

    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...

  20. The Targets/IMage Energy (TIME) 1.0 Model

    Vries B de; Wijngaart RA van den; MNV

    1996-01-01

    Documentation of the five submodels of Targets/IMage Energy (TIME) 1.0 model are presented. Energy Demand, Liquid Fuel (LF), Gaseous Fuel (GF), Solid Fuel (SF) and Electric Power Generation (EPG) are described in detail. Some results of the model calibration for the world 1900-1990 are presented as

  1. 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

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

    Brubaker, Erik; Brennan, James S.; Marleau, Peter; Nowack, Aaron B.; Steele, John T.; 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 inducedtypically, 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 gapsbubblespropagate 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.

  3. 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.

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

    Lv Yujie [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, PO Box 2728, Beijing 100080 (China); Tian Jie [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, PO Box 2728, Beijing 100080 (China); Cong Wenxiang [Division of Biomedical Imaging, VT-WFU School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 (United States); Wang Ge [Division of Biomedical Imaging, VT-WFU School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 (United States); Yang Wei [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, PO Box 2728, Beijing 100080 (China); Qin Chenghu [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, PO Box 2728, Beijing 100080 (China); Xu Min [Medical Image Processing Group, Institute of Automation, Chinese Academy of Sciences, PO Box 2728, Beijing 100080 (China)

    2007-08-07

    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.

  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...

  6. 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...

  7. Heterologous Matrix Metalloproteinase Gene Promoter Activity Allows In Vivo Real-time Imaging of Bleomycin-Induced Lung Fibrosis in Transiently Transgenized Mice

    Stellari, Fabio Franco; Ruscitti, Francesca; Pompilio, Daniela; Ravanetti, Francesca; Tebaldi, Giulia; Macchi, Francesca; Verna, Andrea Elizabeth; Villetti, Gino; Donofrio, Gaetano

    2017-01-01

    Idiopathic pulmonary fibrosis is a very common interstitial lung disease derived from chronic inflammatory insults, characterized by massive scar tissue deposition that causes the progressive loss of lung function and subsequent death for respiratory failure. Bleomycin is used as the standard agent to induce experimental pulmonary fibrosis in animal models for the study of its pathogenesis. However, to visualize the establishment of lung fibrosis after treatment, the animal sacrifice is necessary. Thus, the aim of this study was to avoid this limitation by using an innovative approach based on a double bleomycin treatment protocol, along with the in vivo images analysis of bleomycin-treated mice. A reporter gene construct, containing the luciferase open reading frame under the matrix metalloproteinase-1 promoter control region, was tested on double bleomycin-treated mice to investigate, in real time, the correlation between bleomycin treatment, inflammation, tissue remodeling and fibrosis. Bioluminescence emitted by the lungs of bleomycin-treated mice, corroborated by fluorescent molecular tomography, successfully allowed real time monitoring of fibrosis establishment. The reporter gene technology experienced in this work could represent an advanced functional approach for real time non-invasive assessment of disease evolution during therapy, in a reliable and translational living animal model. PMID:28298912

  8. 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.

  9. 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 ...

  10. 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 < 0.05. Results showed that women had significantly higher temperature variations than men (p < 0.01) along the time. In men, only the body region of the abdomen obtained a significant variance (p < 0.05) on the analyzed period, both in the anterior and posterior part. In women, the anterior abdomen and thighs, and the posterior part of the hands, forearms and abdomen showed significant differences (p < 0.05). Based on our results, it can be concluded that the time in rest condition required reaching a TSK balance in young men and women is variable, but for whole body analysis it is recommended at least 10 min for both sexes.

  11. 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.

  12. Time reversal imaging for sensor networks with optimal compensation in time.

    Derveaux, Grégoire; Papanicolaou, George; Tsogka, Chrysoula

    2007-04-01

    Using extensive numerical simulations, several distributed sensor imaging algorithms for localized damage in a structure are analyzed. Given a configuration of ultrasonic transducers, a full response matrix for the healthy structure is assumed known. It is used as a basis for comparison with the response matrix that is recorded when there is damage. Numerical simulations are done with the wave equation in two dimensions. The healthy structure contains many scatterers. The aim is to image point-like defects with several regularly distributed sensors. Because of the complexity of the environment, the recorded traces have a lot of delay spread and travel time migration does not work so well. Instead, the traces are back propagated numerically assuming that there is some knowledge of the background. Since the time at which the back propagated field will focus on the defects is unknown, the Shannon entropy or the bounded variation norm of the image is computed and the time where it is minimal is picked. This imaging method performs well because it produces a tight image near the location of the defects at the time of refocusing. When there are several defects, the singular value decomposition of the response matrix is also carried out.

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

    Laubach, M. A.; Hayward, J. P.; Zhang, X.; Cates, J. W.

    2014-11-01

    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.

  14. Lightweight distributed computing for intraoperative real-time image guidance

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

    2012-02-01

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

  15. Inertial bioluminescence rhythms at the Capo Passero (KM3NeT-Italia) site, Central Mediterranean Sea

    Aguzzi, J.; Fanelli, E.; Ciuffardi, T.; Schirone, A.; Craig, J.; Aiello, S.; Ameli, F.; Anghinolfi, M.; Barbarino, G.; Barbarito, E.; Beverini, N.; Biagi, S.; Biagioni, A.; Bouhadef, B.; Bozza, C.; Cacopardo, G.; Calamai, M.; Calì, C.; Capone, A.; Caruso, F.; Cecchini, S.; Ceres, A.; Chiarusi, T.; Circella, M.; Cocimano, R.; Coniglione, R.; Costa, M.; Cuttone, G.; D’Amato, C.; D’Amico, A.; De Bonis, G.; De Luca, V.; Deniskina, N.; Distefano, C.; Di Mauro, L. S.; Fermani, P.; Ferrara, G.; Flaminio, V.; Fusco, L. A.; Garufi, F.; Giordano, V.; Gmerk, A.; Grasso, R.; Grella, G.; Hugon, C.; Imbesi, M.; Kulikovskiy, V.; Larosa, G.; Lattuada, D.; Leismüller, K. P.; Leonora, E.; Litrico, P.; Lonardo, A.; Longhitano, F.; Presti, D. Lo; Maccioni, E.; Margiotta, A.; Marinelli, A.; Martini, A.; Masullo, R.; Mele, R.; Migliozzi, P.; Migneco, E.; Miraglia, A.; Mollo, C. M.; Mongelli, M.; Morganti, M.; Musico, P.; Musumeci, M.; Nicolau, C. A.; Orlando, A.; Orzelli, A.; Papaleo, R.; Pellegrino, C.; Pellegriti, M. G.; Perrina, C.; Piattelli, P.; Poma, E.; Pulvirenti, S.; Raffaelli, F.; Randazzo, N.; Riccobene, G.; Rovelli, A.; Sanguineti, M.; Sapienza, P.; Sciacca, V.; Sgura, I.; Simeone, F.; Sipala, V.; Speziale, F.; Spitaleri, A.; Spurio, M.; Stellacci, S. M.; Taiuti, M.; Terreni, G.; Trasatti, L.; Trovato, A.; Versari, F.; Vicini, P.; Viola, S.; Vivolo, D.

    2017-01-01

    In the deep sea, the sense of time is dependent on geophysical fluctuations, such as internal tides and atmospheric-related inertial currents, rather than day-night rhythms. Deep-sea neutrino telescopes instrumented with light detecting Photo-Multiplier Tubes (PMT) can be used to describe the synchronization of bioluminescent activity of abyssopelagic organisms with hydrodynamic cycles. PMT readings at 8 different depths (from 3069 to 3349 m) of the NEMO Phase 2 prototype, deployed offshore Capo Passero (Sicily) at the KM3NeT-Italia site, were used to characterize rhythmic bioluminescence patterns in June 2013, in response to water mass movements. We found a significant (p bioluminescence signal, corresponding to inertial fluctuations. Waveform and Fourier analyses of PMT data and tower orientation were carried out to identify phases (i.e. the timing of peaks) by subdividing time series on the length of detected inertial periodicity. A phase overlap between rhythms and cycles suggests a mechanical stimulation of bioluminescence, as organisms carried by currents collide with the telescope infrastructure, resulting in the emission of light. A bathymetric shift in PMT phases indicated that organisms travelled in discontinuous deep-sea undular vortices consisting of chains of inertially pulsating mesoscale cyclones/anticyclones, which to date remain poorly known. PMID:28332561

  16. APPLICATION OF MVP IN REAL-TIME IMAGE PROCESSING

    2000-01-01

    MVP is a digital signal processor, which is of MIMD structure and fit for multimedia application. MVP has several processors in it, and its operation is characteristic of parallelism and pipeline; therefore, real-time signal processing can be done on it. This paper presents the image processing system based on MVP, explains the principles of parallel task assignment and hardware pipeline design, and gives out the example of target tracking and edge detection.

  17. 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

  18. 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

  19. Transient imaging for real-time tracking around a corner

    Klein, Jonathan; Laurenzis, Martin; Hullin, Matthias

    2016-10-01

    Non-line-of-sight imaging is a fascinating emerging area of research and expected to have an impact in numerous application fields including civilian and military sensing. Performance of human perception and situational awareness can be extended by the sensing of shapes and movement around a corner in future scenarios. Rather than seeing through obstacles directly, non-line-of-sight imaging relies on analyzing indirect reflections of light that traveled around the obstacle. In previous work, transient imaging was established as the key mechanic to enable the extraction of useful information from such reflections. So far, a number of different approaches based on transient imaging have been proposed, with back projection being the most prominent one. Different hardware setups were used for the acquisition of the required data, however all of them have severe drawbacks such as limited image quality, long capture time or very high prices. In this paper we propose the analysis of synthetic transient renderings to gain more insights into the transient light transport. With this simulated data, we are no longer bound to the imperfect data of real systems and gain more flexibility and control over the analysis. In a second part, we use the insights of our analysis to formulate a novel reconstruction algorithm. It uses an adapted light simulation to formulate an inverse problem which is solved in an analysis-by-synthesis fashion. Through rigorous optimization of the reconstruction, it then becomes possible to track known objects outside the line of side in real time. Due to the forward formulation of the light transport, the algorithm is easily expandable to more general scenarios or different hardware setups. We therefore expect it to become a viable alternative to the classic back projection approach in the future.

  20. 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...... the MPITDI and MPIConv measured. RESULTS: IVRT, IVRT/ET and MPI all increased significantly with increasing age in both genders (pcardiac function. MPITDI...

  1. Integration of image exposure time into a modified laser speckle imaging method

    RamIrez-San-Juan, J C; Salazar-Hermenegildo, N; Ramos-Garcia, R; Munoz-Lopez, J [Optics Department, INAOE, Puebla (Mexico); Huang, Y C [Department of Electrical Engineering and Computer Science, University of California, Irvine, CA (United States); Choi, B, E-mail: jcram@inaoep.m [Beckman Laser Institute and Medical Clinic, University of California, Irvine, CA (United States)

    2010-11-21

    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.

  2. Bioluminescence-activated deep-tissue photodynamic therapy of cancer.

    Kim, Yi Rang; Kim, Seonghoon; Choi, Jin Woo; Choi, Sung Yong; Lee, Sang-Hee; Kim, Homin; Hahn, Sei Kwang; Koh, Gou Young; Yun, Seok Hyun

    2015-01-01

    Optical energy can trigger a variety of photochemical processes useful for therapies. Owing to the shallow penetration of light in tissues, however, the clinical applications of light-activated therapies have been limited. Bioluminescence resonant energy transfer (BRET) may provide a new way of inducing photochemical activation. Here, we show that efficient bioluminescence energy-induced photodynamic therapy (PDT) of macroscopic tumors and metastases in deep tissue. For monolayer cell culture in vitro incubated with Chlorin e6, BRET energy of about 1 nJ per cell generated as strong cytotoxicity as red laser light irradiation at 2.2 mW/cm(2) for 180 s. Regional delivery of bioluminescence agents via draining lymphatic vessels killed tumor cells spread to the sentinel and secondary lymph nodes, reduced distant metastases in the lung and improved animal survival. Our results show the promising potential of novel bioluminescence-activated PDT.

  3. Real-time Imaging Orientation Determination System to Verify Imaging Polarization Navigation Algorithm

    Hao Lu

    2016-01-01

    Full Text Available 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.

  4. On noise in time-delay integration CMOS image sensors

    Levski, Deyan; Choubey, Bhaskar

    2016-05-01

    Time delay integration sensors are of increasing interest in CMOS processes owing to their low cost, power and ability to integrate with other circuit readout blocks. This paper presents an analysis of the noise contributors in current day CMOS Time-Delay-Integration image sensors with various readout architectures. An analysis of charge versus voltage domain readout modes is presented, followed by a noise classification of the existing Analog Accumulator Readout (AAR) and Digital Accumulator Readout (DAR) schemes for TDI imaging. The analysis and classification of existing readout schemes include, pipelined charge transfer, buffered direct injection, voltage as well as current-mode analog accumulators and all-digital accumulator techniques. Time-Delay-Integration imaging modes in CMOS processes typically use an N-number of readout steps, equivalent to the number of TDI pixel stages. In CMOS TDI sensors, where voltage domain readout is used, the requirements over speed and noise of the ADC readout chain are increased due to accumulation of the dominant voltage readout and ADC noise with every stage N. Until this day, the latter is the primary reason for a leap-back of CMOS TDI sensors as compared to their CCD counterparts. Moreover, most commercial CMOS TDI implementations are still based on a charge-domain readout, mimicking a CCD-like operation mode. Thus, having a good understanding of each noise contributor in the signal chain, as well as its magnitude in different readout architectures, is vital for the design of future generation low-noise CMOS TDI image sensors based on a voltage domain readout. This paper gives a quantitative classification of all major noise sources for all popular implementations in the literature.

  5. Bioluminescence-Sensing Assay for Microbial Growth Recognition

    Heba Ramadan Eed; Nora S. Abdel-Kader; 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...

  6. 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

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

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

    2013-01-01

    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.

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

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

    2013-01-01

    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. PMID:23874425

  9. Use of Saccharomyces cerevisiae BLYES Expressing Bacterial Bioluminescence for Rapid, Sensitive Detection of Estrogenic Compounds

    Sanseverino, John; Gupta, Rakesh K.; Layton, Alice C.; Patterson, Stacey S.; Ripp, Steven A.; Saidak, Leslie; Simpson, Michael L.; Schultz, T. Wayne; Sayler, Gary S.

    2005-01-01

    An estrogen-inducible bacterial lux-based bioluminescent reporter was developed in Saccharomyces cerevisiae for applications in chemical sensing and environmental assessment of estrogen disruptor activity. The strain, designated S. cerevisiae BLYES, was constructed by inserting tandem estrogen response elements between divergent yeast promoters GPD and ADH1 on pUTK401 (formerly pUA12B7) that constitutively express luxA and luxB to create pUTK407. Cotransformation of this plasmid with a second plasmid (pUTK404) containing the genes required for aldehyde synthesis (luxCDE) and FMN reduction (frp) yielded a bioluminescent bioreporter responsive to estrogen-disrupting compounds. For validation purposes, results with strain BLYES were compared to the colorimetric-based estrogenic assay that uses the yeast lacZ reporter strain (YES). Strains BLYES and YES were exposed to 17β-estradiol over the concentration range of 1.2 × 10−8 through 5.6 × 10−12 M. Calculated 50% effective concentration values from the colorimetric and bioluminescence assays (n = 7) were similar at (4.4 ± 1.1) × 10−10 and (2.4 ± 1.0) × 10−10 M, respectively. The lower and upper limits of detection for each assay were also similar and were approximately 4.5 × 10−11 to 2.8 × 10−9 M. Bioluminescence was observed in as little as 1 h and reached its maximum in 6 h. In comparison, the YES assay required a minimum of 3 days for results. Strain BLYES fills the niche for rapid, high-throughput screening of estrogenic compounds and has the ability to be used for remote, near-real-time monitoring of estrogen-disrupting chemicals in the environment. PMID:16085836

  10. 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.

  11. BIOLUMINESCENCE: TEACHING BIOCHEMISTRY BEYOND THE UNIVERSITY WALLS

    Ana Paula Jesus de Almeida

    2016-11-01

    Full Text Available INTRODUCTION: The use of video in teaching and learning processes provides a challenging environment, able to stimulate the intellect and facilitate understanding in life science studies. Videos can be of extraordinary importance in education and dissemination of knowledge, contributing to greater learning, but is rarely used and exploited properly, especially for teaching biochemistry. Biochemistry is considered complex because it involves many molecular structures and processes, especially considering the number of events and molecules involved in the metabolism. OBJECTIVES: This study aimed to introduce biochemistry for the students of basic education using the theme "Light, Science and Life" in a playful and fun way. MATERIALS AND METHODS: A video about bioluminescence was designed and prepared aiming to use it as a support for learning biochemistry by students of basic education of public schools located in Salvador, Bahia. In order to prepare the video, undergraduate students initially revised the literature in order to acquire proper knowledge, and along with their teacher advisor worked the elaboration of texts, textbook and questionnaire and applied at school. DISCUSSION AND RESULTS: Analysis the qualitative results of the experiment on the preparation and use of the video about "Bioluminescence" focused mainly on the content of biochemistry linked to theme Light, Science and Life, and demonstrated the importance of such work in the teaching-learning process. The dynamics used allowed greater interaction between students and teacher, and the teaching of biochemistry in a fun way beyond the university walls. CONCLUSION: The teaching through recreational resources, e.g. videos and other educational strategies that foster learning should be encouraged from basic education, always bearing in order to transmit through these teaching methods the main concepts covered in biochemistry.

  12. Real time blood testing using quantitative phase imaging.

    Hoa V Pham

    Full Text Available 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.

  13. Real-time Image Generation for Compressive Light Field Displays

    Wetzstein, G.; Lanman, D.; Hirsch, M.; Raskar, R.

    2013-02-01

    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.

  14. Time reversal for ultrasonic transcranial surgery and echographic imaging

    Tanter, Mickael; Aubry, Jean-Francois; Vignon, Francois; Fink, Mathias

    2005-09-01

    High-intensity focused ultrasound (HIFU) is able to induce non-invasively controlled and selective destruction of tissues by focusing ultrasonic beams within organs, analogous to a magnifying glass that concentrates enough sunlight to burn a hole in paper. The brain is an attractive organ in which to perform ultrasonic tissue ablation, but such an application has been hampered by the strong defocusing effect of the skull bone. Our group has been involved in this topic for several years, providing proofs of concept and proposing technological solutions to this problem. Thanks to a high-power time-reversal mirror, presented here are in vivo thermal lesions induced through the skull of 12 sheep. Thermal lesions were confirmed by T2-weighted magnetic resonance post-treatment images and histological examination. These results provide striking evidence that noninvasive ultrasound brain surgery is feasible. A recent approach for high-resolution brain ultrasonic imaging will also be discussed with a skull aberration correction technique based on twin arrays technology. The correction of transcranial ultrasonic images is implemented on a new generation of time-reversal mirrors relying on a fully programmable transmit and receive beamformer.

  15. Stimulation of bioluminescence in Noctiluca sp. using controlled temperature changes.

    Han, Jing; Li, GuiJuan; Liu, HuanYing; Hu, HaoHao; Zhang, XueGang

    2013-01-01

    Bioluminescence induced by multifarious stimuli has long been observed and is remains under investigation because of its great complexity. In particular, the exact mechanism underlying bioluminescence is not yet fully understood. This work presents a new experimental method for studying Noctiluca sp. bioluminescence under temperature change stimulation. It is a study of Noctiluca sp. bioluminescence using controlled temperature changes in a tank. A characteristic of this experiment is the large volume of water used (1 m(3) in a tank of 2 × 1 × 1 m). Temperature changes were controlled by two methods. In the first, a flask filled with hot water was introduced into the tank and in the second, a water heater was used in the tank. Temperature changes were recorded using sensors. Noctiluca sp. bioluminescence was recorded using a Canon 5D Mark II and this allowed the characteristics of Noctiluca sp. bioluminescence under temperature change stimulation to be monitored.

  16. 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.

  17. Comparison of human optimized bacterial luciferase, firefly luciferase, and green fluorescent protein for continuous imaging of cell culture and animal models

    Close, Dan M.; Hahn, Ruth E.; Patterson, Stacey S.; Baek, Seung J.; Ripp, Steven A.; Sayler, Gary S.

    2011-01-01

    Bioluminescent and fluorescent reporter systems have enabled the rapid and continued growth of the optical imaging field over the last two decades. Of particular interest has been noninvasive signal detection from mammalian tissues under both cell culture and whole animal settings. Here we report on the advantages and limitations of imaging using a recently introduced bacterial luciferase (lux) reporter system engineered for increased bioluminescent expression in the mammalian cellular environment. Comparison with the bioluminescent firefly luciferase (Luc) system and green fluorescent protein system under cell culture conditions demonstrated a reduced average radiance, but maintained a more constant level of bioluminescent output without the need for substrate addition or exogenous excitation to elicit the production of signal. Comparison with the Luc system following subcutaneous and intraperitoneal injection into nude mice hosts demonstrated the ability to obtain similar detection patterns with in vitro experiments at cell population sizes above 2.5 × 104 cells but at the cost of increasing overall image integration time. PMID:21529093

  18. 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.

  19. Real-time functional near-infrared imager and imaging of human brain activity

    Luo, Qingming; Zeng, Shaoqun; Gong, Hui; Chen, Weiguo; Zhang, Zhi; Chance, Britton

    1999-02-01

    A real time functional near infrared imager (fNIRI) was introduced. The imager was controlled by a computer and the signals from the detectors were converted and processed in real time. A user-friendly software was programmed with Visual C++ language. Relative changes of oxy - Hb, Hb, and total blood concentration in 16 channels and the corresponding images can be displayed in real time on the computer. The imager was used as a real time monitor in psychological tests to record the response of the frontal cortex of human subjects. In mental work and pattern recognition tests, we recorded oxygen consumption and blood flow changes of volunteers' frontal cortex. The psychological results showed that the lower part of the left frontal gyres had intensive relation to pattern recognition and has definite boundaries. However, the mental work involved more zones of the frontal gyres and may be a complex conceptual model. The results also suggested that the human brain has an precise and complicated adjustability. The oxygen supplement in the stimulated area increased as the neuron stimulation.

  20. 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.

  1. 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.

  2. Automated Hierarchical Time Gain Compensation for In Vivo Ultrasound Imaging

    Moshavegh, Ramin; Hemmsen, Martin Christian; Martins, Bo

    2015-01-01

    Time gain compensation (TGC) is essential to ensure the optimal image quality of the clinical ultrasound scans. When large fluid collections are present within the scan plane, the attenuation distribution is changed drastically and TGC compensation becomes challenging. This paper presents...... tissue and the ultrasound signal strength. The proposed algorithm was applied to a set of 44 in vivo abdominal movie sequences each containing 15 frames. Matching pairs of in vivo sequences, unprocessed and processed with the proposed AHTGC were visualized side by side and evaluated by two radiologists...

  3. In vivo real-time cavitation imaging in moving organs

    Arnal, B.; Baranger, J.; Demene, C.; Tanter, M.; Pernot, M.

    2017-02-01

    The stochastic nature of cavitation implies visualization of the cavitation cloud in real-time and in a discriminative manner for the safe use of focused ultrasound therapy. This visualization is sometimes possible with standard echography, but it strongly depends on the quality of the scanner, and is hindered by difficulty in discriminating from highly reflecting tissue signals in different organs. A specific approach would then permit clear validation of the cavitation position and activity. Detecting signals from a specific source with high sensitivity is a major problem in ultrasound imaging. Based on plane or diverging wave sonications, ultrafast ultrasonic imaging dramatically increases temporal resolution, and the larger amount of acquired data permits increased sensitivity in Doppler imaging. Here, we investigate a spatiotemporal singular value decomposition of ultrafast radiofrequency data to discriminate bubble clouds from tissue based on their different spatiotemporal motion and echogenicity during histotripsy. We introduce an automation to determine the parameters of this filtering. This method clearly outperforms standard temporal filtering techniques with a bubble to tissue contrast of at least 20 dB in vitro in a moving phantom and in vivo in porcine liver.

  4. Real-time image processing for particle tracking velocimetry

    Kreizer, Mark; Ratner, David; Liberzon, Alex

    2010-01-01

    We present a novel high-speed particle tracking velocimetry (PTV) experimental system. Its novelty is due to the FPGA-based, real-time image processing "on camera". Instead of an image, the camera transfers to the computer using a network card, only the relevant information of the identified flow tracers. Therefore, the system is ideal for the remote particle tracking systems in research and industrial applications, while the camera can be controlled and data can be transferred over any high-bandwidth network. We present the hardware and the open source software aspects of the PTV experiments. The tracking results of the new experimental system has been compared to the flow visualization and particle image velocimetry measurements. The canonical flow in the central cross section of a a cubic cavity (1:1:1 aspect ratio) in our lid-driven cavity apparatus is used for validation purposes. The downstream secondary eddy (DSE) is the sensitive portion of this flow and its size was measured with increasing Reynolds number (via increasing belt velocity). The size of DSE estimated from the flow visualization, PIV and compressed PTV is shown to agree within the experimental uncertainty of the methods applied.

  5. Optical imaging system-based real-time image saliency extraction method

    Zhao, Jufeng; Gao, Xiumin; Chen, Yueting; Feng, Huajun

    2015-04-01

    Saliency extraction has become a popular topic in imaging science. One of the challenges in image saliency extraction is to detect the saliency content efficiently with a full-resolution saliency map. Traditional methods only involve computer calculation and thus result in limitations in computational speed. An optical imaging system-based visual saliency extraction method is developed to solve this problem. The optical system is built by effectively implementing an optical Fourier process with a Fourier lens to form two frequency planes for further operation. The proposed method combines optical components and computer calculations and mainly relies on frequency selection with precise pinholes on the frequency planes to efficiently produce a saliency map. Comparison shows that the method is suitable for extracting salient information and operates in real time to generate a full-resolution saliency map with good boundaries.

  6. 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.

  7. Ultrasound contrast agent imaging: Real-time imaging of the superharmonics

    Peruzzini, D.; Viti, J.; Tortoli, P.; Verweij, M. D.; de Jong, N.; Vos, H. J.

    2015-10-01

    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.

  8. Real-time windowing in imaging radar using FPGA technique

    Ponomaryov, Volodymyr I.; Escamilla-Hernandez, Enrique

    2005-02-01

    The imaging radar uses the high frequency electromagnetic waves reflected from different objects for estimating of its parameters. Pulse compression is a standard signal processing technique used to minimize the peak transmission power and to maximize SNR, and to get a better resolution. Usually the pulse compression can be achieved using a matched filter. The level of the side-lobes in the imaging radar can be reduced using the special weighting function processing. There are very known different weighting functions: Hamming, Hanning, Blackman, Chebyshev, Blackman-Harris, Kaiser-Bessel, etc., widely used in the signal processing applications. Field Programmable Gate Arrays (FPGAs) offers great benefits like instantaneous implementation, dynamic reconfiguration, design, and field programmability. This reconfiguration makes FPGAs a better solution over custom-made integrated circuits. This work aims at demonstrating a reasonably flexible implementation of FM-linear signal and pulse compression using Matlab, Simulink, and System Generator. Employing FPGA and mentioned software we have proposed the pulse compression design on FPGA using classical and novel windows technique to reduce the side-lobes level. This permits increasing the detection ability of the small or nearly placed targets in imaging radar. The advantage of FPGA that can do parallelism in real time processing permits to realize the proposed algorithms. The paper also presents the experimental results of proposed windowing procedure in the marine radar with such the parameters: signal is linear FM (Chirp); frequency deviation DF is 9.375MHz; the pulse width T is 3.2μs taps number in the matched filter is 800 taps; sampling frequency 253.125*106 MHz. It has been realized the reducing of side-lobes levels in real time permitting better resolution of the small targets.

  9. 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.

  10. 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.

  11. Observations and Measurements of Planktonic Bioluminescence in and Around a Milky Sea

    1988-03-01

    produced by plankton subjected to mechanical stimulation) can be observed from breaking wave crests and swimming shoals of fish . The Arabian Sea is...identification: growth at 4 ’C, growth at 35 ’C, ainylase, lipase , gelatinase. growth on maltose, cellobiose, gluconate, BIOLUMINESCENCE IN MILKY SEA 57...neofluar oil -immersion objective. BIOLUMINESCENCE MEASUREMENTS Surface-water bioluminescence Surface-water bioluminescence was measured continuously during

  12. 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.

  13. 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

  14. A Double-Threshold Technique for Fast Time-Correspondence Imaging

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

    2013-01-01

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

  15. 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..

  16. Seasonal Changes of Bioluminescence in Photosynthetic and Heterotrophic Dinoflagellates at San Clemente Island

    2012-02-01

    2 Seasonal Changes of Bioluminescence in Photosynthetic and Heterotrophic Dinoflagellates at San Clemente Island David Lapota Space and Naval...Warfare Systems Center, Pacific USA 1. Introduction A significant portion of bioluminescence in all oceans is produced by dinoflagellates . Numerous...studies have documented the ubiquitous distribution of bioluminescent dinoflagellates in near surface waters (Seliger et al., 1961; Yentsch and Laird

  17. 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.

  18. Monitoring of historical frescoes by timed infrared imaging analysis

    Cadelano, G.; Bison, P.; Bortolin, A.; Ferrarini, G.; Peron, F.; Girotto, M.; Volinia, M.

    2015-03-01

    The subflorescence and efflorescence phenomena are widely acknowledged as the major causes of permanent damage to fresco wall paintings. They are related to the occurrence of cycles of dry/wet conditions inside the walls. Therefore, it is essential to identify the presence of water on the decorated surfaces and inside the walls. Nondestructive testing in industrial applications have confirmed that active infrared thermography with continuous timed images acquisition can improve the outcomes of thermal analysis aimed to moisture identification. In spite of that, in cultural heritage investigations these techniques have not been yet used extensively on a regular basis. This paper illustrates an application of these principles in order to evaluate the decay of fresco mural paintings in a medieval chapel located in North-West of Italy. One important feature of this study is the use of a robotic system called aIRview that can be utilized to automatically acquire and process thermal images. Multiple accurate thermal views of the inside walls of the building have been produced in a survey that lasted several days. Signal processing algorithms based on Fast Fourier Transform analysis have been applied to the acquired data in order to formulate trustworthy hypotheses about the deterioration mechanisms.

  19. Picosecond time-resolved imaging using SPAD cameras

    Gariepy, Genevieve; Leach, Jonathan; Warburton, Ryan; Chan, Susan; Henderson, Robert; Faccio, Daniele

    2016-10-01

    The recent development of 2D arrays of single-photon avalanche diodes (SPAD) has driven the development of applications based on the ability to capture light in motion. Such arrays are composed typically of 32x32 SPAD detectors, each having the ability to detect single photons and measure their time of arrival with a resolution of about 100 ps. Thanks to the single-photon sensitivity and the high temporal resolution of these detectors, it is now possible to image light as it is travelling on a centimetre scale. This opens the door for the direct observation and study of dynamics evolving over picoseconds and nanoseconds timescales such as laser propagation in air, laser-induced plasma and laser propagation in optical fibres. Another interesting application enabled by the ability to image light in motion is the detection of objects hidden from view, based on the recording of scattered waves originating from objects hidden by an obstacle. Similarly to LIDAR systems, the temporal information acquired at every pixel of a SPAD array, combined with the spatial information it provides, allows to pinpoint the position of an object located outside the line-of-sight of the detector. A non-line-of-sight tracking can be a valuable asset in many scenarios, including for search and rescue mission and safer autonomous driving.

  20. Detection of DNA adducts by bioluminescence

    Xu, Shunqing; Tan, Xianglin; Yao, Qunfeng; He, Min; Zhou, Yikai; Chen, Jian

    2001-09-01

    Luminescent assay for detection ATP is very sensitive with limitation of 10-17 moles. ATP using styrene oxide as a model carcinogen we currently apply a luminescence technique to detect the very low levels of carcinogen-DNA adducts in vitro and in vivo. The bioluminescent assay of DNA adducts entails three consecutive steps: digestion of modified DNA to adducted dinucleoside monophosphate and normal nucleotide are hydrolyzed to nucleosides (N) by nuclease P1 and prostatic acid phosphomonesterase (PAP); incorporation of (gamma) -P of ATP into normal nucleoside(N); detection of consumption of ATP by luminescence. This assay does not require separate manipulation because of the selective property of nuclease P1. One fmol of carcinogen- DNA adducts was detected by luminescent assay. A good correlation between results of luminescent assay and 32P-postlabeling procedures has been observed. We detect 1 adduct in 108 nucleotides for 10(mu) g DNA sample. The procedures of luminescent method is very simple and low- cost. IT appears applicable to the ultra sensitive detection of low levels of DNA adducts without radioactive isotope.

  1. Photon-Counting Arrays for Time-Resolved Imaging

    I. Michel Antolovic

    2016-06-01

    Full Text Available The paper presents a camera comprising 512 × 128 pixels capable of single-photon detection and gating with a maximum frame rate of 156 kfps. The photon capture is performed through a gated single-photon avalanche diode that generates a digital pulse upon photon detection and through a digital one-bit counter. Gray levels are obtained through multiple counting and accumulation, while time-resolved imaging is achieved through a 4-ns gating window controlled with subnanosecond accuracy by a field-programmable gate array. The sensor, which is equipped with microlenses to enhance its effective fill factor, was electro-optically characterized in terms of sensitivity and uniformity. Several examples of capture of fast events are shown to demonstrate the suitability of the approach.

  2. A look at some systemic properties of self-bioluminescent emission

    Creath, Katherine

    2008-08-01

    Self-bioluminescent emission (SBE) is a type of biological chemiluminescence where photons are emitted as part of chemical reactions occurring during metabolic processes. This emission is also known as biophoton emission, ultraweak photon emission and ultraweak bioluminescence. This paper outlines research over the past century on some systemic properties of SBE as measured with biological detectors, photomultiplier detectors and ultra-sensitive imaging arrays. There is an apparent consensus in the literature that emission in the deep blue and ultraviolet (150-450nm) is related to DNA / RNA processes while emission in the red and near infrared (600-1000nm) is related to mitochondria and oxidative metabolisms involving reactive oxygen species, singlet oxygen and free radicals in plant, animal and human cells along with chlorophyll fluorescent decay in plants. Additionally, there are trends showing that healthy, unstressed and uninjured samples have less emission than samples that are unhealthy, stressed or injured. Mechanisms producing this emission can be narrowed down by isolating the wavelength region of interest and waiting for short-term fluorescence to decay leaving the ultraweak long-term metabolic emission. Examples of imaging this emission in healthy versus unhealthy, stressed versus unstressed, and injured versus uninjured plant parts are shown. Further discussion poses questions still to be answered related to properties such as coherence, photon statistics, and methodological means of isolating mechanisms.

  3. 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.

  4. Time-lapse seismic imaging of the Reykjanes geothermal reservoir

    Weemstra, Cornelis; Obermann, Anne; Blanck, Hanna; Verdel, Arie; Paap, Bob; Árni Guðnason, Egill; Páll Hersir, Gylfi; Jousset, Philippe; Sigurðsson, Ómar

    2016-04-01

    We report on the results obtained from a dense seismic deployment over a geothermal reservoir. The reservoir has been producing continuously for almost a decade and is located on the tip of the Reykjanes peninsula, SW Iceland. The seismic stations on top of the reservoir have continuously recorded the ambient seismic wavefield between April 2014 and September 2015. The density of the seismic network makes the data well suited for time-lapse seismic imaging of the reservoir. To that end we compute time-lapse responses through the application of seismic interferometry. These interferometric lapse responses are obtained by simple crosscorrelation of the seismic noise recorded by the different seismic stations. We subsequently evaluate the temporal variation of the coda of these crosscorrelations. The term coda refers to the later arriving, multiple scattered waves. The multiple scattering implies that these waves have sampled the subsurface very densely and hence become highly sensitive to tiny mechanical and structural changes in that subsurface. This sensitivity allows one, in principle at least, to monitor the geothermal reservoir. Preliminary results indeed suggest a relation between the temporal variation of the coda waves and the reservoir. Ultimately, this method may lead to a means to monitor a geothermal reservoir in both space and time.

  5. 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.

  6. Total variation regularization for bioluminescence tomography with the split Bregman method.

    Feng, Jinchao; Qin, Chenghu; Jia, Kebin; Zhu, Shouping; Liu, Kai; Han, Dong; Yang, Xin; Gao, Quansheng; Tian, Jie

    2012-07-01

    Regularization methods have been broadly applied to bioluminescence tomography (BLT) to obtain stable solutions, including l2 and l1 regularizations. However, l2 regularization can oversmooth reconstructed images and l1 regularization may sparsify the source distribution, which degrades image quality. In this paper, the use of total variation (TV) regularization in BLT is investigated. Since a nonnegativity constraint can lead to improved image quality, the nonnegative constraint should be considered in BLT. However, TV regularization with a nonnegativity constraint is extremely difficult to solve due to its nondifferentiability and nonlinearity. The aim of this work is to validate the split Bregman method to minimize the TV regularization problem with a nonnegativity constraint for BLT. The performance of split Bregman-resolved TV (SBRTV) based BLT reconstruction algorithm was verified with numerical and in vivo experiments. Experimental results demonstrate that the SBRTV regularization can provide better regularization quality over l2 and l1 regularizations.

  7. Non-Invasive in vivo Imaging in Small Animal Research

    V. Koo

    2006-01-01

    Full Text Available Non-invasive real time in vivo molecular imaging in small animal models has become the essential bridge between in vitro data and their translation into clinical applications. The tremendous development and technological progress, such as tumour modelling, monitoring of tumour growth and detection of metastasis, has facilitated translational drug development. This has added to our knowledge on carcinogenesis. The modalities that are commonly used include Magnetic Resonance Imaging (MRI, Computed Tomography (CT, Positron Emission Tomography (PET, bioluminescence imaging, fluorescence imaging and multi-modality imaging systems. The ability to obtain multiple images longitudinally provides reliable information whilst reducing animal numbers. As yet there is no one modality that is ideal for all experimental studies. This review outlines the instrumentation available together with corresponding applications reported in the literature with particular emphasis on cancer research. Advantages and limitations to current imaging technology are discussed and the issues concerning small animal care during imaging are highlighted.

  8. Time-resolved image analysis for turbulent flows

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

    2013-01-01

    Classical Particle Image Velocimetry (PIV) uses two representations of the particle image distribution to determine the displacement of the particle image pattern by spatial cross-correlation. The accuracy and the robustness are however limited by the fact that only two representations at t and t +Δ

  9. High-resolution imaging with a real-time synthetic aperture ultrasound system: a phantom study

    Huang, Lianjie; Labyed, Yassin; Simonetti, Francesco; Williamson, Michael; Rosenberg, Robert; Heintz, Philip; Sandoval, Daniel

    2011-03-01

    It is difficult for ultrasound to image small targets such as breast microcalcifications. Synthetic aperture ultrasound imaging has recently developed as a promising tool to improve the capabilities of medical ultrasound. We use two different tissueequivalent phantoms to study the imaging capabilities of a real-time synthetic aperture ultrasound system for imaging small targets. The InnerVision ultrasound system DAS009 is an investigational system for real-time synthetic aperture ultrasound imaging. We use the system to image the two phantoms, and compare the images with those obtained from clinical scanners Acuson Sequoia 512 and Siemens S2000. Our results show that synthetic aperture ultrasound imaging produces images with higher resolution and less image artifacts than Acuson Sequoia 512 and Siemens S2000. In addition, we study the effects of sound speed on synthetic aperture ultrasound imaging and demonstrate that an accurate sound speed is very important for imaging small targets.

  10. Multiple repetition time balanced steady-state free precession imaging.

    Cukur, Tolga; Nishimura, Dwight G

    2009-07-01

    Although balanced steady-state free precession (bSSFP) imaging yields high signal-to-noise ratio (SNR) efficiency, the bright lipid signal is often undesirable. The bSSFP spectrum can be shaped to suppress the fat signal with scan-efficient alternating repetition time (ATR) bSSFP. However, the level of suppression is limited, and the pass-band is narrow due to its nonuniform shape. A multiple repetition time (TR) bSSFP scheme is proposed that creates a broad stop-band with a scan efficiency comparable with ATR-SSFP. Furthermore, the pass-band signal uniformity is improved, resulting in fewer shading/banding artifacts. When data acquisition occurs in more than a single TR within the multiple-TR period, the echoes can be combined to significantly improve the level of suppression. The signal characteristics of the proposed technique were compared with bSSFP and ATR-SSFP. The multiple-TR method generates identical contrast to bSSFP, and achieves up to an order of magnitude higher stop-band suppression than ATR-SSFP. In vivo studies at 1.5 T and 3 T demonstrate the superior fat-suppression performance of multiple-TR bSSFP.

  11. Real-time imaging of renin release in vitro.

    Peti-Peterdi, János; Fintha, Attila; Fuson, Amanda L; Tousson, Albert; Chow, Robert H

    2004-08-01

    Renin release from juxtaglomerular granular cells is considered the rate-limiting step in activation of the renin-angiotensin system that helps to maintain body salt and water balance. Available assays to measure renin release are complex, indirect, and work with significant internal errors. To directly visualize and study the dynamics of both the release and tissue activity of renin, we isolated and perfused afferent arterioles with attached glomeruli dissected from rabbit kidneys and used multiphoton fluorescence imaging. Acidotropic fluorophores, such as quinacrine and LysoTrackers, clearly and selectively labeled renin granules. Immunohistochemistry of mouse kidney with a specific renin antibody and quinacrine staining colocalized renin granules and quinacrine fluorescence. A low-salt diet for 1 wk caused an approximately fivefold increase in the number of both individual granules and renin-positive granular cells. Time-lapse imaging showed no signs of granule trafficking or any movement, only the dimming and disappearance of fluorescence from individual renin granules within 1 s in response to 100 microM isoproterenol. There appeared to be a quantal release of the granular contents; i.e., an all-or-none phenomenon. Using As4.1 cells, a granular cell line, we observed further classic signs of granule exocytosis, the emptying of granule content associated with a flash of quinacrine fluorescence. Using a fluorescence resonance energy transfer-based, 5-(2-aminoethylamino)naphthalene-1-sulfonic acid (EDANS)-conjugated renin substrate in the bath, an increase in EDANS fluorescence (renin activity) was observed around granular cells in response to isoproterenol. Fluorescence microscopy is an excellent tool for the further study of the mechanism, regulation, and dynamics of renin release.

  12. High Dynamic Range Image Based on Multiple Exposure Time Synthetization

    Yoshifumi Shimodaira

    2007-03-01

    Full Text Available High dynamic range of illumination may cause serious distortions and otherproblems in viewing and further processing of digital images. In this paper a new tonereproduction preprocessing algorithm is introduced which may help in developing hardly ornon-viewable features and content of the images. The method is based on the synthetizationof multiple exposure images from which the dense part, i.e. regions having the maximumlevel of detail are included in the output image. The resulted high quality HDR image makeseasier the information extraction and effectively supports the further processing of theimage.

  13. Bioluminescence as the Basis for the Detection of Trichothecenes

    1986-03-17

    screened for their ability to quench bioluminescence were obtained through the courtesy of Dr. Lou Carson, of the Toxicology Division of the Food and...34 Recent Adv. Phytochem . 9, 167 (1974). 13. Lyman, J. and Fleming, R.H., "Composition of Seawater," J. Mar. Res. 3, 134 (1940). 14. Mayer, C.F., "Endemic...DIELDRIN Cl CI Cl~c C 1 2I• HEPTACHLOR EPOXIDE OCTACHLOR EPOXIDE "Fig. 11 - Pesticides screened for ability to quench bioluminescence Ir £ d, PF K.I IR 10 R 125 - I ’S * N 586 9 -q

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

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

    2011-07-01

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

  15. Imaging tooth enamel using zero echo time (ZTE) magnetic resonance imaging

    Rychert, Kevin M.; Zhu, Gang; Kmiec, Maciej M.; Nemani, Venkata K.; Williams, Benjamin B.; Flood, Ann B.; Swartz, Harold M.; Gimi, Barjor

    2015-03-01

    In an event where many thousands of people may have been exposed to levels of radiation that are sufficient to cause the acute radiation syndrome, we need technology that can estimate the absorbed dose on an individual basis for triage and meaningful medical decision making. Such dose estimates may be achieved using in vivo electron paramagnetic resonance (EPR) tooth biodosimetry, which measures the number of persistent free radicals that are generated in tooth enamel following irradiation. However, the accuracy of dose estimates may be impacted by individual variations in teeth, especially the amount and distribution of enamel in the inhomogeneous sensitive volume of the resonator used to detect the radicals. In order to study the relationship between interpersonal variations in enamel and EPR-based dose estimates, it is desirable to estimate these parameters nondestructively and without adding radiation to the teeth. Magnetic Resonance Imaging (MRI) is capable of acquiring structural and biochemical information without imparting additional radiation, which may be beneficial for many EPR dosimetry studies. However, the extremely short T2 relaxation time in tooth structures precludes tooth imaging using conventional MRI methods. Therefore, we used zero echo time (ZTE) MRI to image teeth ex vivo to assess enamel volumes and spatial distributions. Using these data in combination with the data on the distribution of the transverse radio frequency magnetic field from electromagnetic simulations, we then can identify possible sources of variations in radiation-induced signals detectable by EPR. Unlike conventional MRI, ZTE applies spatial encoding gradients during the RF excitation pulse, thereby facilitating signal acquisition almost immediately after excitation, minimizing signal loss from short T2 relaxation times. ZTE successfully provided volumetric measures of tooth enamel that may be related to variations that impact EPR dosimetry and facilitate the development

  16. All-passive pixel super-resolution of time-stretch imaging

    Chan, Antony C. S.; Ng, Ho-Cheung; Bogaraju, Sharat C. V.; Hayden K. H. So; Lam, Edmund Y.; Tsia, Kevin K.

    2016-01-01

    Based on image encoding in a serial-temporal format, optical time-stretch imaging entails a stringent requirement of state-of-the- art fast data acquisition unit in order to preserve high image resolution at an ultrahigh frame rate --- hampering the widespread utilities of such technology. Here, we propose a pixel super-resolution (pixel-SR) technique tailored for time-stretch imaging that preserves pixel resolution at a relaxed sampling rate. It harnesses the subpixel shifts between image fr...

  17. High throughput web inspection system using time-stretch real-time imaging

    Kim, Chanju

    Photonic time-stretch is a novel technology that enables capturing of fast, rare and non-repetitive events. Therefore, it operates in real-time with ability to record over long period of time while having fine temporal resolution. The powerful property of photonic time-stretch has already been employed in various fields of application such as analog-to-digital conversion, spectroscopy, laser scanner and microscopy. Further expanding the scope, we fully exploit the time-stretch technology to demonstrate a high throughput web inspection system. Web inspection, namely surface inspection is a nondestructive evaluation method which is crucial for semiconductor wafer and thin film production. We successfully report a dark-field web inspection system with line scan speed of 90.9 MHz which is up to 1000 times faster than conventional inspection instruments. The manufacturing of high quality semiconductor wafer and thin film may directly benefit from this technology as it can easily locate defects with area of less than 10 microm x 10 microm where it allows maximum web flow speed of 1.8 km/s. The thesis provides an overview of our web inspection technique, followed by description of the photonic time-stretch technique which is the keystone in our system. A detailed explanation of each component is covered to provide quantitative understanding of the system. Finally, imaging results from a hard-disk sample and flexible films are presented along with performance analysis of the system. This project was the first application of time-stretch to industrial inspection, and was conducted under financial support and with close involvement by Hitachi, Ltd.

  18. Exploiting NanoLuc luciferase for smartphone-based bioluminescence cell biosensor for (anti)-inflammatory activity and toxicity.

    Cevenini, Luca; Calabretta, Maria Maddalena; Lopreside, Antonia; Tarantino, Giuseppe; Tassoni, Annalisa; Ferri, Maura; Roda, Aldo; Michelini, Elisa

    2016-12-01

    The availability of smartphones with high-performance digital image sensors and processing power has completely reshaped the landscape of point-of-need analysis. Thanks to the high maturity level of reporter gene technology and the availability of several bioluminescent proteins with improved features, we were able to develop a bioluminescence smartphone-based biosensing platform exploiting the highly sensitive NanoLuc luciferase as reporter. A 3D-printed smartphone-integrated cell biosensor based on genetically engineered Hek293T cells was developed. Quantitative assessment of (anti)-inflammatory activity and toxicity of liquid samples was performed with a simple and rapid add-and-measure procedure. White grape pomace extracts, known to contain several bioactive compounds, were analyzed, confirming the suitability of the smartphone biosensing platform for analysis of untreated complex biological matrices. Such approach could meet the needs of small medium enterprises lacking fully equipped laboratories for first-level safety tests and rapid screening of new bioactive products. Graphical abstract Smartphone-based bioluminescence cell biosensor.

  19. Design and manufacture of imaging time-of-propagation optics

    Albrecht, Mike; Fast, James; Schwartz, Alan

    2016-09-01

    There are several challenges associated with the design and manufacture of the optics required for the imaging time-of- propagation detector constructed for the Belle II particle physics experiment. This detector uses Cherenkov light radiated in quartz bars to identify subatomic particles: pions, kaons, and protons. The optics are physically large (125 cm x 45 cm x 2 cm bars and 45 cm x 10 cm x 5 cm prisms), all surfaces are optically polished, and there is very little allowance for chamfers or surface defects. In addition to the optical challenges, there are several logistical and handling challenges associated with measuring, assembling, cleaning, packaging, and shipping these delicate precision optics. This paper describes a collaborative effort between Pacific Northwest National Laboratory, the University of Cincinnati, and ZYGO Corporation for the design and manufacture of 48 fused silica optics (30 bars and 18 prisms) for the iTOP Detector. Details of the iTOP detector design that drove the challenging optical requirements are provided, along with material selection considerations. Since the optics are so large, precise, and delicate, special care had to be given to the selection of a manufacturing process capable of achieving the challenging optical and surface defect requirements on such large and high-aspect-ratio (66:1) components. A brief update on the current status and performance of these optics is also provided.

  20. Synaptic signals: time travelling through the brain in the neuro-image

    Pisters, P.

    2011-01-01

    This essay presents some thoughts on schizoanalysis and visual culture around the proposition that cinema survives in the digital age as a type of image that, after the movement-image and the time-image, could be called the neuro-image. By considering clinical schizophrenia as ‘degree zero’ of schiz

  1. A combination of NADHP and hispidin is not essential for bioluminescence in luminous fungal living gills of Mycena chlorophos.

    Teranishi, Katsunori

    2017-01-05

    The chemical mechanisms underlying visible bioluminescence in the fungus Mycena chlorophos are not clear. A combination of dihydronicotinamide adenine dinucleotide phosphate (NADPH) and hispidin, which has been reported to increase the intensity of in vitro luminescence in crude cold-water extracts prepared from the bioluminescent fruiting bodies of M. chlorophos, exhibited potential bioluminescence activation in the early bioluminescence stages, in which the bioluminescence was ultra-weak, for living gills and luminescence activation for non-bioluminescent gills, which was collapsed by freezing and subsequent thawing, at all bioluminescence stages. These abilities were not evident in considerably bioluminescent gills. These abilities were blocked by trans-4-hydroxycinnamic acid and trans-3,4-dihydroxycinnamic acid, which were identified as in vivo bioluminescence-activating components. Original bioluminescence and bioluminescence produced from the addition of trans-4-hydroxycinnamic acid and trans-3,4-dihydroxycinnamic acid in living gills were almost completely inhibited by 10 mM NaN3 , whereas the luminescence produced form the combination of NADPH and hispidin in thawed non-bioluminescent and living gills at the early weak bioluminescence stages was not inhibited by 10 mM NaN3 . Thus, the combination of NADPH and hispidin plays different roles in luminescence systems compared with essential bioluminescence systems, and the combination of NADPH and hispidin was not essential for visible bioluminescence in living gills.

  2. 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.

  3. Evaluation of On- and Off-Line Bioluminescence Tomography System for Focal Irradiation Guidance.

    Zhang, Bin; Wong, John W; Iordachita, Iulian I; Reyes, Juvenal; Nugent, Katriana; Tran, Phuoc T; Tuttle, Stephen W; Koumenis, Constantinos; Wang, Ken Kang-Hsin

    2016-12-01

    In response to the limitations of computed tomography (CT) and cone-beam CT (CBCT) in irradiation guidance, especially for soft-tissue targets without the use of contrast agents, our group developed a solution that implemented bioluminescence tomography (BLT) as the image-guidance modality for preclinical radiation research. However, adding such a system to existing small animal irradiators is no small task. A potential solution is to utilize an off-line BLT system in close proximity to the irradiator, with stable and effective animal transport between the two systems. In this study, we investigated the localization accuracy of an off-line BLT system when used for the small animal radiation research platform (SARRP) and compared the results with those of an on-line system. The CBCT was equipped on both the off-line BLT system and the SARRP, with a distance of 5 m between them. To evaluate the setup error during animal transport between the two systems, the mice underwent CBCT imaging on the SARRP and were then transported to the off-line system for a second CBCT imaging session. The normalized intensity difference of the two images and the corresponding histogram and correlation were computed to evaluate if the transport process perturbed animal positioning. Strong correlation (correlation coefficients >0.95) between the SARRP and the off-line mouse CBCT was observed. The offset of the implanted light source center can be maintained within 0.2 mm during transport. To compare the target localization accuracy using the on-line SARRP BLT and the off-line system, a self-illuminated bioluminescent source was implanted in the abdomen of anesthetized mice. In addition to the application for dose calculation, CBCT imaging was also employed to generate the mesh grid of the imaged mouse for BLT reconstruction. Two scenarios were devised and compared, which involved localization of the luminescence source based on either: 1. on-line SARRP bioluminescence image and CBCT; or 2

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

    Gammelmark, Kim

    2004-01-01

    of convex array TMS imaging compared to conventional convex array imaging. Only minor motion artifacts causing subtle image brightness fluctuations were reported in TMS imaging, which did not depreciate the diagnostic value of the images. The influence of tissue motion and a method for two...

  5. Space-time encoding for high frame rate ultrasound imaging

    Misaridis, Thanssis; Jensen, Jørgen Arendt

    2002-01-01

    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...

  6. Real-time synthetic aperture imaging: opportunities and challenges

    Nikolov, Svetoslav; Tomov, Borislav Gueorguiev; Jensen, Jørgen Arendt

    2006-01-01

    in the region of interest it is possible to beamform the signals along a desired path, thus, improving the estimation of blood flow. The transmission of coded excitations makes it possible to achieve higher contrast and larger penetration depth compared to "conventional" scanners. This paper presents......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 3D. This parametric description makes it possible to quickly change the image geometry during scanning, thus enabling adaptive imaging and precise flow estimation. The paper addresses problems such as large bandwidth and computational load and gives the solutions that have been adopted...

  7. Upgrading bioluminescent bacterial bioreporter performance by splitting the lux operon.

    Yagur-Kroll, Sharon; Belkin, Shimshon

    2011-05-01

    Bioluminescent bacterial bioreporters harbor a fusion of bacterial bioluminescence genes (luxCDABE), acting as the reporting element, to a stress-response promoter, serving as the sensing element. Upon exposure to conditions that activate the promoter, such as an environmental stress or the presence of an inducing chemical, the promoter::reporter fusion generates a dose-dependent bioluminescent signal. In order to improve bioluminescent bioreporter performance we have split the luxCDABE genes of Photorhabdus luminescens into two smaller functional units: luxAB, that encode for the luciferase enzyme, which catalyzes the luminescence reaction, and luxCDE that encode for the enzymatic complex responsible for synthesis of the reaction's substrate, a long-chain aldehyde. The expression of each subunit was put under the control of either an inducible stress-responsive promoter or a synthetic constitutive promoter, and different combinations of the two units were tested for their response to selected chemicals in Escherichia coli. In all cases tested, the split combinations proved to be superior to the native luxCDABE configuration, suggesting an improved efficiency in the transcription and/or translation of two small gene units instead of a larger one with the same genes. The best combination was that of an inducible luxAB and a constitutive luxCDE, indicating that aldehyde availability is limited when the five genes are expressed together in E. coli, and demonstrating that improved biosensor performance may be achieved by rearrangement of the lux operon genes.

  8. Unanimous Model for Describing the Fast Bioluminescence Kinetics of Ca

    Eremeeva, Elena V.; Bartsev, Sergey I.; Berkel, van Willem J.H.; Vysotski, Eugene S.

    2017-01-01

    Upon binding their metal ion cofactors, Ca2+-regulated photoproteins display a rapid increase of light signal, which reaches its peak within milliseconds. In the present study, we investigate bioluminescence kinetics of the entire photoprotein family. All five recombinant hydromedusan Ca2+-regulated

  9. Monitoring Bloom Dynamics of a Common Coastal Bioluminescent Ctenophore

    2010-09-30

    watershed run-off and discharge of submarine ground-water can profoundly impact growth conditions of bioluminescent plankton on very short space and...changes in marine ecosystems (Kane, 2009). Gelatinous zooplankton, such as Mnemiopsis sp., feed on mesozooplankton, with copepods being their main food

  10. Feasibility Study for a Compact, Multi-Purpose Bioluminescence Detector

    1998-09-30

    which is likely to be primarily dinoflagellates, ostracods and gelatinous zooplankton, however a faster pump speed would be desirable in a profiling...and defense sectors by providing a valuable tool for biological oceanographers interested in plankton distribution patterns as well as providing...quantifying, tracking and identifying bioluminescent plankton . IEEE J. Oceanic Engineering. Widder, E.A. (1997) In situ video recordings of

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

    Fang, W.

    1994-01-01

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

  12. MO-AB-BRA-02: A Novel Scatter Imaging Modality for Real-Time Image Guidance During Lung SBRT

    Redler, G; Bernard, D; Templeton, A; Chu, J [Rush University Medical Center, Chicago, IL (United States); Nair, C Kumaran [University of Chicago, Chicago, IL (United States); Turian, J [Rush University Medical Center, Chicago, IL (United States); Rush Radiosurgery LLC, Chicago, IL (United States)

    2015-06-15

    Purpose: A novel scatter imaging modality is developed and its feasibility for image-guided radiation therapy (IGRT) during stereotactic body radiation therapy (SBRT) for lung cancer patients is assessed using analytic and Monte Carlo models as well as experimental testing. Methods: During treatment, incident radiation interacts and scatters from within the patient. The presented methodology forms an image of patient anatomy from the scattered radiation for real-time localization of the treatment target. A radiographic flat panel-based pinhole camera provides spatial information regarding the origin of detected scattered radiation. An analytical model is developed, which provides a mathematical formalism for describing the scatter imaging system. Experimental scatter images are acquired by irradiating an object using a Varian TrueBeam accelerator. The differentiation between tissue types is investigated by imaging simple objects of known compositions (water, lung, and cortical bone equivalent). A lung tumor phantom, simulating materials and geometry encountered during lung SBRT treatments, is fabricated and imaged to investigate image quality for various quantities of delivered radiation. Monte Carlo N-Particle (MCNP) code is used for validation and testing by simulating scatter image formation using the experimental pinhole camera setup. Results: Analytical calculations, MCNP simulations, and experimental results when imaging the water, lung, and cortical bone equivalent objects show close agreement, thus validating the proposed models and demonstrating that scatter imaging differentiates these materials well. Lung tumor phantom images have sufficient contrast-to-noise ratio (CNR) to clearly distinguish tumor from surrounding lung tissue. CNR=4.1 and CNR=29.1 for 10MU and 5000MU images (equivalent to 0.5 and 250 second images), respectively. Conclusion: Lung SBRT provides favorable treatment outcomes, but depends on accurate target localization. A comprehensive

  13. Real-time computer-generated integral imaging and 3D image calibration for augmented reality surgical navigation.

    Wang, Junchen; Suenaga, Hideyuki; Liao, Hongen; Hoshi, Kazuto; Yang, Liangjing; Kobayashi, Etsuko; Sakuma, Ichiro

    2015-03-01

    Autostereoscopic 3D image overlay for augmented reality (AR) based surgical navigation has been studied and reported many times. For the purpose of surgical overlay, the 3D image is expected to have the same geometric shape as the original organ, and can be transformed to a specified location for image overlay. However, how to generate a 3D image with high geometric fidelity and quantitative evaluation of 3D image's geometric accuracy have not been addressed. This paper proposes a graphics processing unit (GPU) based computer-generated integral imaging pipeline for real-time autostereoscopic 3D display, and an automatic closed-loop 3D image calibration paradigm for displaying undistorted 3D images. Based on the proposed methods, a novel AR device for 3D image surgical overlay is presented, which mainly consists of a 3D display, an AR window, a stereo camera for 3D measurement, and a workstation for information processing. The evaluation on the 3D image rendering performance with 2560×1600 elemental image resolution shows the rendering speeds of 50-60 frames per second (fps) for surface models, and 5-8 fps for large medical volumes. The evaluation of the undistorted 3D image after the calibration yields sub-millimeter geometric accuracy. A phantom experiment simulating oral and maxillofacial surgery was also performed to evaluate the proposed AR overlay device in terms of the image registration accuracy, 3D image overlay accuracy, and the visual effects of the overlay. The experimental results show satisfactory image registration and image overlay accuracy, and confirm the system usability.

  14. A novel lux operon in the cryptically bioluminescent fish pathogen Vibrio salmonicida is associated with virulence.

    Nelson, Eric J; Tunsjø, Hege S; Fidopiastis, Pat M; Sørum, Henning; Ruby, Edward G

    2007-03-01

    The cold-water-fish pathogen Vibrio salmonicida expresses a functional bacterial luciferase but produces insufficient levels of its aliphatic-aldehyde substrate to be detectably luminous in culture. Our goals were to (i) better explain this cryptic bioluminescence phenotype through molecular characterization of the lux operon and (ii) test whether the bioluminescence gene cluster is associated with virulence. Cloning and sequencing of the V. salmonicida lux operon revealed that homologs of all of the genes required for luminescence are present: luxAB (luciferase) and luxCDE (aliphatic-aldehyde synthesis). The arrangement and sequence of these structural lux genes are conserved compared to those in related species of luminous bacteria. However, V. salmonicida strains have a novel arrangement and number of homologs of the luxR and luxI quorum-sensing regulatory genes. Reverse transcriptase PCR analysis suggests that this novel arrangement of quorum-sensing genes generates antisense transcripts that may be responsible for the reduced production of bioluminescence. In addition, infection with a strain in which the luxA gene was mutated resulted in a marked delay in mortality among Atlantic salmon relative to infection with the wild-type parent in single-strain challenge experiments. In mixed-strain competition between the luxA mutant and the wild type, the mutant was attenuated up to 50-fold. It remains unclear whether the attenuation results from a direct loss of luciferase or a polar disturbance elsewhere in the lux operon. Nevertheless, these findings document for the first time an association between a mutation in a structural lux gene and virulence, as well as provide a new molecular system to study Vibrio pathogenesis in a natural host.

  15. Comparison between Bioluminescent Imaging and Small-animal PET-CT in Xenotransplantation Tumor Model with Luc-expressing Cell%荧光素酶标的人肝癌细胞裸鼠异种移植瘤模型的生物发光成像和PET-CT成像比较

    李小颖; 高凯; 董伟; 张连峰

    2011-01-01

    Objective To establish hepatic carcinoma mouse model with human BEL-7402 cells expressing luciferase and to compare bioluminescence imaging with that by small-animal PET-CT.Method The vector containing luciferase gene was constructed and transfected into human BEL-7402 liver tumor cell line and selected with G418 to obtain stable Luc-expressing clones.5 × l05 cells, constitutively expressing luciferase, were inoculated to liver of BALB/cA-nu through hepatic portal vein for assessment of tumor growth with the whole-body optical imaging system and small-animal PET-CT.Result The stable Luc-expressing BEL-7402 cell lines were abtained, which could grow to tumor mass after implantation.There was high uptake of 18 F-FDG at the edge of liver with small-animal PET-CT.Conclusion Luc-labeled BEL-7402 cell lines and a mouse tumor model based on the cell lines are established, the whole-body optical imaging system combined with small-animal PET-CT provides a new and reliable technology for the in situ tumor model, which is a new tool to further study the mechanism of metastasis and drug discovery.%目的 利用荧光素酶基因标记的人肝癌细胞株BEL-7402建立裸鼠肝原位移植模型,及小鼠肝原位移植模型的生物发光和小动物PET-CT成像的比较.方法 构建表达荧光素酶基因的真核表达载体并将其转人人肝癌细胞BEL-7402,经梯度浓度G418筛选获得稳定表达荧光素酶基因的细胞克隆并扩大培养.BALB/cA-nu裸鼠肝门静脉接种5×10(5),个发光细胞使其成瘤,活体荧光成像和小动物PET-CT成像系统观察肿瘤的生长情况.结果 获得了稳定表达Luc的人肝癌细胞株,将其接种到裸鼠体内,活体荧光成像系统观察发现能够成瘤,小动物PET-CT影像观察发现小鼠肝脏边缘对18F-FDG有高摄取区域.结论 利用荧光素酶基因标记的人肝癌细胞BEL7402成功建立了原位肝癌裸鼠模型,小动物活体成像结合小动物PET-CT技术为原位肿瘤模

  16. 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...

  17. The reaction process of firefly bioluminescence triggered by photolysis of caged-ATP.

    Kageyama, Takeshi; Tanaka, Masatoshi; Sekiya, Takao; Ohno, Shin-Ya; Wada, Naohisa

    2011-01-01

    The reaction process of firefly bioluminescence was studied by photolyzing caged-ATP to adenosine triphosphate (ATP) within 100 ms. The intensity of luminescence increases markedly to reach a maximum within 1 s, maintains almost the same intensity up to 5 s and then decays monotonically. The rise γ(1) and decay γ(2) rate constants were determined to be about 5 s(-1) and 1 × 10(-2) s(-1), respectively, so as to phenomenologically fit the time course. A second luminescence peak appears after around 350 s. The dependence of the rate constants on the concentrations of reactants and a viscous reagent revealed that two kinds of reaction contribute the observed time course: (1) an intrinsic reaction by ATP photolyzed from caged-ATP that is already trapped in luciferase; and (2) a diffusion-controlled reaction by free ATP in the buffer solution outside luciferase. Numerical analysis based on reaction kinetics related γ(1) and γ(2) to the rate constants of a three-step reaction model, and accurately described the effects of concentration of reactants and a viscous reagent on the time courses of bioluminescence. Thus, it has been clearly concluded that the binding mode of caged-ATP at the catalytic center of luciferase is very different from that of ATP.

  18. Time-resolved computed tomography of the liver: retrospective, multi-phase image reconstruction derived from volumetric perfusion imaging

    Fischer, Michael A.; Kartalis, Nikolaos; Aspelin, Peter; Albiin, Nils; Brismar, Torkel B. [Karolinska University Hospital, Division of Medical Imaging and Technology, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm (Sweden); Leidner, Bertil; Svensson, Anders [Karolinska University Hospital, Division of Medical Imaging and Technology, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm (Sweden); Karolinska University Hospital Huddinge, Department of Radiology, Stockholm (Sweden)

    2014-01-15

    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.)

  19. 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.

  20. Landscape in Cinema: Images to Think Time through Space

    Filipa Rosário

    2017-01-01

    Full Text Available This text is an introduction to the special issue on ‘Landscape and Cinema’ published by Aniki. Portuguese Journal of the Moving Image. It is composed of a brief state of the art, which anchors this work within the bibliography on the subject, and a synthetic description of the discourses developed by the six papers included in the issue: the first one addresses the historical and aesthetic evolution of the background in the Russian and Soviet cinema of the first decades of the twentieth century; the second discusses the representation of the Portuguese landscape in Manuel Guimarães’ films; the third identifies a few cinematic techniques that work as conventions of psychological landscape film, such as juxtaposition and superimposition; the fourth interprets the meaning of Brazilian urban landscapes showed in the feature film A Vida Provisória (Maurício Gomes Leite, 1968; the fifth deals with the corporeal and conceptual relations between body, memory and landscape through a sensory reading of the films Ten Skies (James Benning, 2004 and Sin Peso (Cao Guimarães, 2007; and finally, the sixth undertakes a comparative analysis of three South American non-fiction works that show several waterscapes as places of memory: El botón de nácar (Patricio Guzmán, 2015, Los durmientes (Enrique Ramírez, 2015 and Las aguas del olvido (Jonathan Perel, 2013. The aim of this preliminary text is therefore to introduce the main contents of the issue and some of its conclusions. In this sense, it is necessary to emphasise the open and polysemic nature of landscape, which works, inside and outside cinema, as a palimpsest in which multiple meanings come together. For this reason, those films that depict, create and intervene in landscape allow to perceive, among other things, the overlapping of historical and subjective times that takes place in certain spaces.

  1. 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.

  2. Real-time digital signal processing for live electro-optic imaging.

    Sasagawa, Kiyotaka; Kanno, Atsushi; Tsuchiya, Masahiro

    2009-08-31

    We present an imaging system that enables real-time magnitude and phase detection of modulated signals and its application to a Live Electro-optic Imaging (LEI) system, which realizes instantaneous visualization of RF electric fields. The real-time acquisition of magnitude and phase images of a modulated optical signal at 5 kHz is demonstrated by imaging with a Si-based high-speed CMOS image sensor and real-time signal processing with a digital signal processor. In the LEI system, RF electric fields are probed with light via an electro-optic crystal plate and downconverted to an intermediate frequency by parallel optical heterodyning, which can be detected with the image sensor. The artifacts caused by the optics and the image sensor characteristics are corrected by image processing. As examples, we demonstrate real-time visualization of electric fields from RF circuits.

  3. 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.

  4. 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...

  5. Analysis, Modeling and Dynamic Optimization of 3D Time-of-Flight Imaging Systems

    Schmidt, Mirko

    2011-01-01

    The present thesis is concerned with the optimization of 3D Time-of-Flight (ToF) imaging systems. These novel cameras determine range images by actively illuminating a scene and measuring the time until the backscattered light is detected. Depth maps are constructed from multiple raw images. Usually two of such raw images are acquired simultaneously using special correlating sensors. This thesis covers four main contributions: A physical sensor model is presented which enables the analysis a...

  6. Real-time portal imaging devices operating on high-pressure gaseous electronic principles

    Giakos, George C.; Richardson, Donna B.; Ghotra, P.; Pillai, Bindu; Seetharaman, Lakshmi; Passalaqua, Anthony M.; DiBianca, Frank A.; Endorf, Robert J.; Devidas, Sreenivas

    1995-05-01

    A novel real-time portal imaging scanning detector, based on high-pressure gaseous electronics principles and operating up to 60 atmospheres, is presented and the predicted performance of this detector is analyzed. The idea is to utilize high pressure gaseous electronics imaging detectors operating in the saturation regime, aimed at improving image performance characteristics in real time portal imaging. As a result, beam localization errors are controlled, identified and corrected accurately and the patient radiotherapy treatment becomes more effective.

  7. 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.

  8. 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.

  9. Cine viability magnetic resonance imaging of the heart without increased scan time.

    Hassanein, Azza S; Khalifa, Ayman M; Ibrahim, El-Sayed H

    2016-02-01

    Cardiac magnetic resonance imaging (MRI) provides information about myocardial morphology, function, and viability from cine, tagged, and late gadolinium enhancement (LGE) images, respectively. While the cine and tagged images are acquired in a time-resolved fashion, the LGE images are acquired at a single timeframe. The purpose of this work is to develop a method for generating cine LGE images without additional scan time. The motion field is extracted from the tagged images, and is then used to guide the deformation of the infarcted region from the acquired LGE image at the acquired timeframe to any other timeframe. Major techniques for motion estimation, including harmonic phase (HARP) and optical flow analysis, are tested in this work for motion estimation. The proposed method is tested on numerical phantom and images from four human subjects. The generated cine LGE images showed both viability and wall motion information in the same set of images without additional scan time or image misregistration problems. The band-pass optical flow analysis resulted in the most accurate motion estimation compared to other methods, especially HARP, which fails to track points at the myocardial boundary. Infarct transmurality from the generated images showed good agreement with myocardial strain, and wall thickening showed good agreement with that measured from conventional cine images. In conclusion, the developed technique allows for generating cine LGE images that enable simultaneous display of wall motion and viability information. The generated images could be useful for estimating myocardial contractility reserve and for treatment prognosis.

  10. Toxicological study of pesticides in air and precipitations of Paris by means of a bioluminescence method.

    Trajkovska, S; Mbaye, M; Gaye Seye, M D; Aaron, J J; Chevreuil, M; Blanchoud, H

    2009-06-01

    A detailed toxicological study on several pesticides, including chlorothalonil, cyprodynil, dichlobénil, pendimethaline, trifluraline, and alpha-endosulfan, present at trace levels in air and total atmospheric precipitations of Paris is presented. The pesticides contained in the atmospheric samples, collected during sampling campaigns in February-March 2007, are identified and quantified by a high-performance liquid chromatographic (HPLC)-UV detection method. The toxicity measurements are performed by means of the Microtox bioluminescence method, based on the evaluation of the bioluminescence inhibition of the Vibrio fischeri marine bacteria at two exposure times to the pesticide solutions. The specific toxicity, corresponding to the particular toxicity of the compound under study and represented by the EC(50) parameter, is determined for these pesticides. Also, the global toxicity, which is the toxicity of all micro-pollutants present in the sample under study, is estimated for the extracts of air and atmospheric precipitation (rainwater) samples. The specific toxicities strongly vary with the nature of the pesticide, the EC(50) parameter values being comprised between 0.17 and 0.83 mg/mL and 0.15 and 0.66 mg/mL, respectively, for exposure times of 5 and 15 min. The importance of the atmospheric samples' global toxicity and the respective contribution of the toxic potency of the various pesticides contained in these samples are discussed.

  11. Antimicrobial properties of cyclodextrin-antiseptics-complexes determined by microplate laser nephelometry and ATP bioluminescence assay.

    Finger, Susanne; Wiegand, Cornelia; Buschmann, Hans-Jürgen; Hipler, Uta-Christina

    2012-10-15

    Antimicrobial effects of substances can be determined with different methods that measure distinct parameters. Thus, a comparison of the results obtained can be difficult. In this study, two in vitro methods were employed to determine concentration and time dependent effects of cyclodextrin (CD)-complexes with the antiseptics chlorhexidine diacetate (CHX), iodine (IOD) and polihexanide (PHMB) on Candida albicans and Malassezia pachydermatis. Using both, microplate laser nephelometry and the ATP bioluminescence assay, it could be shown that CD-antiseptics-complexes tested exhibited significant antifungal effects with the exception of γ-CD-CHX in the case of C. albicans. Microplate laser nephelometry (MLN) is an optical method and enables a quantitative determination of particle concentrations in solution. By means of this method, microbial growth under influence of potential antimicrobial substances can be monitored over a prolonged time period. In addition, the antimicrobial activity was analyzed by measurement of the microbial adenosine triphosphate (ATP) content with a bioluminescent assay. The luminescent signal is directly proportional to the amount of ATP, and thus, a linear function of the number of living microbial cells present. Both methods were compared according to the half maximal inhibitory concentration (IC(50)) calculated and the statistical evaluation of Pearson's correlation coefficient (r). In summary, it could be demonstrated that both methods yield similar results although they differ in the parameter.

  12. Real-Time Bioluminescent Tracking of Cellular Population Dynamics

    Close, Dan; Xu, Tingling; Ripp, Steven; Sayler, Gary

    2014-01-01

    Cellular population dynamics are routinely monitored across many diverse fields for a variety of purposes. In general, these dynamics are assayed either through the direct counting of cellular aliquots followed by extrapolation to the total population size, or through the monitoring of signal intensity from any number of externally stimulated reporter proteins. While both viable methods, here we describe a novel technique that allows for the automated, non-destructive tracking of cellular pop...

  13. Contribution of Reflection Terahertz Time Domain-Imaging (THz-TDI) to Imaging Analysis of Artworks

    Dandolo, Corinna Ludovica Koch; Fukunaga, Kaori; Kohzuma, Y.;

    Different kind s of artefacts (easel painting, panel paintings and Asian lacquerwares) have been scanned by THz - TDI and results have been compared with those obtained by others standard imaging techniques (x-ray radiography, cross sectional imaging, technical photography) ....

  14. Real-time maximum a-posteriori image reconstruction for fluorescence microscopy

    Anwar A. Jabbar

    2015-08-01

    Full Text Available Rapid reconstruction of multidimensional image is crucial for enabling real-time 3D fluorescence imaging. This becomes a key factor for imaging rapidly occurring events in the cellular environment. To facilitate real-time imaging, we have developed a graphics processing unit (GPU based real-time maximum a-posteriori (MAP image reconstruction system. The parallel processing capability of GPU device that consists of a large number of tiny processing cores and the adaptability of image reconstruction algorithm to parallel processing (that employ multiple independent computing modules called threads results in high temporal resolution. Moreover, the proposed quadratic potential based MAP algorithm effectively deconvolves the images as well as suppresses the noise. The multi-node multi-threaded GPU and the Compute Unified Device Architecture (CUDA efficiently execute the iterative image reconstruction algorithm that is ≈200-fold faster (for large dataset when compared to existing CPU based systems.

  15. Real-time maximum a-posteriori image reconstruction for fluorescence microscopy

    Jabbar, Anwar A.; Dilipkumar, Shilpa; C K, Rasmi; Rajan, K.; Mondal, Partha P.

    2015-08-01

    Rapid reconstruction of multidimensional image is crucial for enabling real-time 3D fluorescence imaging. This becomes a key factor for imaging rapidly occurring events in the cellular environment. To facilitate real-time imaging, we have developed a graphics processing unit (GPU) based real-time maximum a-posteriori (MAP) image reconstruction system. The parallel processing capability of GPU device that consists of a large number of tiny processing cores and the adaptability of image reconstruction algorithm to parallel processing (that employ multiple independent computing modules called threads) results in high temporal resolution. Moreover, the proposed quadratic potential based MAP algorithm effectively deconvolves the images as well as suppresses the noise. The multi-node multi-threaded GPU and the Compute Unified Device Architecture (CUDA) efficiently execute the iterative image reconstruction algorithm that is ≈200-fold faster (for large dataset) when compared to existing CPU based systems.

  16. A symmetrical image encryption scheme in wavelet and time domain

    Luo, Yuling; Du, Minghui; Liu, Junxiu

    2015-02-01

    There has been an increasing concern for effective storages and secure transactions of multimedia information over the Internet. Then a great variety of encryption schemes have been proposed to ensure the information security while transmitting, but most of current approaches are designed to diffuse the data only in spatial domain which result in reducing storage efficiency. A lightweight image encryption strategy based on chaos is proposed in this paper. The encryption process is designed in transform domain. The original image is decomposed into approximation and detail components using integer wavelet transform (IWT); then as the more important component of the image, the approximation coefficients are diffused by secret keys generated from a spatiotemporal chaotic system followed by inverse IWT to construct the diffused image; finally a plain permutation is performed for diffusion image by the Logistic mapping in order to reduce the correlation between adjacent pixels further. Experimental results and performance analysis demonstrate the proposed scheme is an efficient, secure and robust encryption mechanism and it realizes effective coding compression to satisfy desirable storage.

  17. Image-driven cardiac left ventricle segmentation for the evaluation of multiview fused real-time 3-dimensional echocardiography images.

    Rajpoot, Kashif; Noble, J Alison; Grau, Vicente; Szmigielski, Cezary; Becher, Harald

    2009-01-01

    Real-time 3-dimensional echocardiography (RT3DE) permits the acquisition and visualization of the beating heart in 3D. Despite a number of efforts to automate the left ventricle (LV) delineation from RT3DE images, this remains a challenging problem due to the poor nature of the acquired images usually containing missing anatomical information and high speckle noise. Recently, there have been efforts to improve image quality and anatomical definition by acquiring multiple single-view RT3DE images with small probe movements and fusing them together after alignment. In this work, we evaluate the quality of the multiview fused images using an image-driven semiautomatic LV segmentation method. The segmentation method is based on an edge-driven level set framework, where the edges are extracted using a local-phase inspired feature detector for low-contrast echocardiography boundaries. This totally image-driven segmentation method is applied for the evaluation of end-diastolic (ED) and end-systolic (ES) single-view and multiview fused images. Experiments were conducted on 17 cases and the results show that multiview fused images have better image segmentation quality, but large failures were observed on ED (88.2%) and ES (58.8%) single-view images.

  18. Visual Images of Subjective Perception of Time in a Literary Text

    Nesterik, Ella V.; Issina, Gaukhar I.; Pecherskikh, Taliya F.; Belikova, Oxana V.

    2016-01-01

    The article is devoted to the subjective perception of time, or psychological time, as a text category and a literary image. It focuses on the visual images that are characteristic of different types of literary time--accelerated, decelerated and frozen (vanished). The research is based on the assumption that the category of subjective perception…

  19. 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.

  20. Visualization of in Vivo Hydrogen Sulfide Production by a Bioluminescence Probe in Cancer Cells and Nude Mice.

    Tian, Xiaodong; Li, Zhiyan; Lau, Choiwan; Lu, Jianzhong

    2015-11-17

    Hydrogen sulfide (H2S) has emerged as an exciting endogenous gasotransmitter in addition to nitric oxide and carbon monoxide. However, its precise measurement in living cells and animals remains a challenge. In this study, a novel bioluminescence H2S probe was designed and synthesized by modifying the 6'-amino group of d-aminoluciferin into a 6'-azido group, which was highly selective against other reactive sulfur, nitrogen, and oxygen species. Our H2S probe azidoluciferin sensitively reacted with H2S to release d-aminoluciferin with a strong bioluminescence signal. On the basis of its high selectivity and sensitivity, the H2S probe was used to detect H2S production in live cancer cells and nude mice. The bioluminescence signal decreased in mice treated with propargylglycine, an inhibitor of H2S, suggesting that our H2S probe can detect endogenous H2S in real time, in vivo. Overall, the excellent sensing properties of the probe combined with its bioimaging capability make it a useful tool to study H2S biological roles.

  1. 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.

  2. 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.

  3. Operational Analysis of Time-Optimal Maneuvering for Imaging Spacecraft

    2013-03-01

    satellites for remote sensing ranges from military applications to tracking global weather patterns, tectonic activity, surface vegetation, ocean...satellite-sensors/pleiades-1.html [7] H. Y. Lee, T. Kim, W. Park, and H. K. Lee, “Extraction of digital elevation models from satellite stereo images

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

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

    2014-06-01

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

  5. Identification of a vacuolar proton channel that triggers the bioluminescent flash in dinoflagellates

    Rodriguez, Juan D.; Haq, Saddef; Bachvaroff, Tsvetan; Nowak, Kristine F.; Nowak, Scott J.; Morgan, Deri; Cherny, Vladimir V.; Sapp, Maredith M.; Bernstein, Steven; Bolt, Andrew; DeCoursey, Thomas E.; Place, Allen R.; Smith, Susan M. E.

    2017-01-01

    In 1972, J. Woodland Hastings and colleagues predicted the existence of a proton selective channel (HV1) that opens in response to depolarizing voltage across the vacuole membrane of bioluminescent dinoflagellates and conducts protons into specialized luminescence compartments (scintillons), thereby causing a pH drop that triggers light emission. HV1 channels were subsequently identified and demonstrated to have important functions in a multitude of eukaryotic cells. Here we report a predicted protein from Lingulodinium polyedrum that displays hallmark properties of bona fide HV1, including time-dependent opening with depolarization, perfect proton selectivity, and characteristic ΔpH dependent gating. Western blotting and fluorescence confocal microscopy of isolated L. polyedrum scintillons immunostained with antibody to LpHV1 confirm LpHV1’s predicted organellar location. Proteomics analysis demonstrates that isolated scintillon preparations contain peptides that map to LpHV1. Finally, Zn2+ inhibits both LpHV1 proton current and the acid-induced flash in isolated scintillons. These results implicate LpHV1 as the voltage gated proton channel that triggers bioluminescence in L. polyedrum, confirming Hastings’ hypothesis. The same channel likely mediates the action potential that communicates the signal along the tonoplast to the scintillon. PMID:28178296

  6. Saccharomyces cerevisiae BLYAS, a New Bioluminescent Bioreporter for Detection of Androgenic Compounds▿

    Eldridge, Melanie L.; Sanseverino, John; Layton, Alice C.; Easter, James P.; Schultz, T. Wayne; Sayler, Gary S.

    2007-01-01

    A Saccharomyces cerevisiae strain, capable of autonomous bioluminescence, was engineered to respond to androgenic chemicals. The strain, S. cerevisiae BLYAS, contains the human androgen receptor in the chromosome and was constructed by inserting a series of androgen response elements between divergent yeast promoters GPD and ADH1 on pUTK401 that constitutively expressed luxA and luxB to create pUTK420. Cotransformation of this plasmid with a second plasmid (pUTK404), containing the genes required for aldehyde synthesis (luxCDE) and FMN reduction (frp), yielded a bioluminescent bioreporter responsive to androgenic chemicals. Using dihydrotestosterone (DHT) as a standard, the response time and the 50% effective concentration values were 3 to 4 h and (9.7 ± 4.6) × 10−9 M, respectively. The lower limit of detection in response to DHT was 2.5 × 10−9 M, and in response to testosterone it was 2.5 × 10−10 M. This strain is suitable for high-throughput screening of chemicals with potential for remote environmental monitoring systems because of the assay speed, sensitivity, and self-containment. PMID:17675419

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

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

    2013-11-01

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

  8. A modified phasor approach for analyzing time-gated fluorescence lifetime images

    Fereidouni, F.; Esposito, A.; Blab, G.; Gerritsen, H.C.

    2011-01-01

    Fluorescence lifetime imaging is a versatile tool that permits mapping the biochemical environment in the cell. Among various fluorescence lifetime imaging techniques, timecorrelated single photon counting and time-gating methods have been demonstrated to be very efficient and robust for the imaging

  9. Avaliação da qualidade microbiológica de bebida láctea e creme de leite UAT por ATP-Bioluminescência Evaluation of microbiological quality of UHT milk drink and UHT milk cream by ATP-Bioluminescence

    A.F. Cunha

    2013-04-01

    Full Text Available Embora métodos tradicionais sejam utilizados na avaliação microbiológica de produtos UAT, metodologias rápidas, baseadas em ATP-Bioluminescência, têm sido desenvolvidas. Os resultados da aplicação dessa técnica em 54 amostras de bebida láctea UAT achocolatada e 12 de creme de leite UAT foram comparados com os resultados de métodos microbiológicos, utilizando-se diferentes meios de cultura e tempos de incubação das referidas amostras. A técnica de ATP-Bioluminescência foi aplicada por meio do sistema MLS, e os resultados foram expressos em unidades relativas de luz (RLU. Em todos os tempos de incubação - 48, 72 e 168 horas - , as amostras apresentaram contagens baixas de microrganismos mesófilos e psicrotróficos aeróbios quando analisadas em meio PCA, BHI, PetrifilmTM AC e por ATP-Bioluminescência (Although traditional methods are used for the microbiological evaluation of UHT products, rapid methodologies based on ATP-Bioluminescence have been developed. The results of applying this technique in 54 samples of chocolate UHT milk drink and 12 of UHT milk cream were compared with the results of microbiological methods, using different culture media and incubation times for the referred samples. The ATP-Bioluminescence technique was applied through the MLS system and the results were expressed as relative light units (RLU. In all incubation times - 48, 72, and 168 hours - , the samples showed lower counts of mesophilic and psychrotrophic aerobic microorganisms when analyzed using PCA, BHI, PetrifilmTM AC and ATP-Bioluminescence (<150RLU, demonstrating the technique's high specificity. Only one sample of UHT milk cream showed a mesophilic aerobic count above the standard established by Brazilian legislation (<100CFU/mL when analyzed in PCA (260 CFU/mL and PetrifilmTM AC (108CFU/mL at 168 hours. This high count of aerobic mesophilic microorganisms was also detected by the ATP-Bioluminescence (416 RLU technique. The results of

  10. 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).

  11. 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.

  12. 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...

  13. An Assessment and Annotated Bibliography of Marine Bioluminescence Research: 1979-1987.

    1993-01-01

    1983). Speculations on the hydrogen peroxide and the photogenic cells are Colours of Marine Bioluminescence. Abstr., 15th associated with a brown...of the taxonomic distribution of Affinity of the Reduced Riboflavin 5’-Phosphate Site. bioluminescence among various groups of organisms Biochemistry...possible biological functions for for reduced riboflavin 5’-phosphate (FMNH,). The bioluminescence are explored. The spectral emission inhibitor was

  14. A study on bioluminescence and photoluminescence in the earthworm Eisenia lucens.

    Pes, O; Midlik, A; Schlaghamersky, J; Zitnan, M; Taborsky, P

    2016-02-01

    Eisenia lucens is an earthworm living in the organic soil layer of decomposing wood. When irritated, the worm expels coelomic fluid through pores in its body wall, exhibiting blue-green bioluminescence. The mechanism of the bioluminescence, which seems to be different from other bioluminescence systems of terrestrial animals, has been studied in this work. Many lines of evidence indicate that riboflavin stored in coelomycetes plays an important role in this glowing reaction.

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

    Seliger, H H

    1976-01-01

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

  16. Model System for Live Imaging of Neuronal Responses to Injury and Repair

    Mathieu Gravel

    2011-11-01

    Full Text Available Although it has been well established that induction of growth-associated protein-43 (GAP-43 during development coincides with axonal outgrowth and early synapse formation, the existence of neuronal plasticity and neurite outgrowth in the adult central nervous system after injuries is more controversial. To visualize the processes of neuronal injury and repair in living animals, we generated reporter mice for bioluminescence and fluorescence imaging bearing the luc (luciferase and gfp (green fluorescent protein reporter genes under the control of the murine GAP-43 promoter. Reporter functionality was first observed during the development of transgenic embryos. Using in vivo bioluminescence and fluorescence imaging, we visualized induction of the GAP-43 signals from live embryos starting at E10.5, as well as neuronal responses to brain and peripheral nerve injuries (the signals peaked at 14 days postinjury. Moreover, three-dimensional analysis of the GAP-43 bioluminescent signal confirmed that it originated from brain structures affected by ischemic injury. The analysis of fluorescence signal at cellular level revealed colocalization between endogenous protein and the GAP-43-driven gfp transgene. Taken together, our results suggest that the GAP-43-luc/gfp reporter mouse represents a valid model system for real-time analysis of neurite outgrowth and the capacity of the adult nervous system to regenerate after injuries.

  17. Off-axis quantitative phase imaging processing using CUDA: toward real-time applications.

    Pham, Hoa; Ding, Huafeng; Sobh, Nahil; Do, Minh; Patel, Sanjay; Popescu, Gabriel

    2011-07-01

    We demonstrate real time off-axis Quantitative Phase Imaging (QPI) using a phase reconstruction algorithm based on NVIDIA's CUDA programming model. The phase unwrapping component is based on Goldstein's algorithm. By mapping the process of extracting phase information and unwrapping to GPU, we are able to speed up the whole procedure by more than 18.8× with respect to CPU processing and ultimately achieve video rate for mega-pixel images. Our CUDA implementation also supports processing of multiple images simultaneously. This enables our imaging system to support high speed, high throughput, and real-time image acquisition and visualization.

  18. All-passive pixel super-resolution of time-stretch imaging

    Chan, Antony C. S.; Ng, Ho-Cheung; Bogaraju, Sharat C. V.; So, Hayden K. H.; Lam, Edmund Y.; Tsia, Kevin K.

    2017-03-01

    Based on image encoding in a serial-temporal format, optical time-stretch imaging entails a stringent requirement of state-of-the-art fast data acquisition unit in order to preserve high image resolution at an ultrahigh frame rate — hampering the widespread utilities of such technology. Here, we propose a pixel super-resolution (pixel-SR) technique tailored for time-stretch imaging that preserves pixel resolution at a relaxed sampling rate. It harnesses the subpixel shifts between image frames inherently introduced by asynchronous digital sampling of the continuous time-stretch imaging process. Precise pixel registration is thus accomplished without any active opto-mechanical subpixel-shift control or other additional hardware. Here, we present the experimental pixel-SR image reconstruction pipeline that restores high-resolution time-stretch images of microparticles and biological cells (phytoplankton) at a relaxed sampling rate (≈2–5 GSa/s)—more than four times lower than the originally required readout rate (20 GSa/s) — is thus effective for high-throughput label-free, morphology-based cellular classification down to single-cell precision. Upon integration with the high-throughput image processing technology, this pixel-SR time-stretch imaging technique represents a cost-effective and practical solution for large scale cell-based phenotypic screening in biomedical diagnosis and machine vision for quality control in manufacturing.

  19. Imaging Rainfall Infiltration Processes with the Time-Lapse Electrical Resistivity Imaging Method

    Zhang, Gang; Zhang, Gui-Bin; Chen, Chien-chih; Chang, Ping-Yu; Wang, Tzu-Pin; Yen, Horng-Yuan; Dong, Jia-Jyun; Ni, Chuen-Fa; Chen, Su-Chin; Chen, Chao-Wei; Jia, Zheng-yuan

    2016-06-01

    Electrical Resistivity Imaging (ERI) was carried out continuously for 10 days to map the subsurface resistivity distribution along a potentially hazardous hillslope at the Jieshou Junior High School in Taoyuan, Taiwan. The reliability of the inverted resistivity structures down to about 25 m depth was examined with synthetic modeling using the same electrode arrangements installed on land surface as in field surveys, together with a DOI (depth-of-investigation) index calculated from the ERI data. The subsurface resistivity distribution is consistent with results from well logging. These ERI recordings were taken daily and provided highly resolved imagery of the resistivity distribution underground and illustrated the dynamical fluid-flow behavior due to heavy rainfall infiltration. Using Archie's law, the resistivity distribution was transformed into a map of relative water saturation (RWS), which is strongly correlated with the rainfall infiltration process. We then found that the averaged RWS is significantly correlated with daily precipitation. Our observations indicate that time-lapse ERI is effective in monitoring subterraneous rainfall infiltration; moreover, the preferential flow paths can be delineated according to the changes in averaged RWS derived from the ERI data.

  20. Tomographic imaging with ultra-wideband noise radar using time-domain data

    Shin, Hee Jung; Narayanan, Ram M.; Rangaswamy, Muralidhar

    2013-05-01

    This paper investigates the feasibility of using a noise waveform in an ultra-wideband (UWB) radar system for two-dimensional tomographic imaging of a stationary object with a multistatic tomographic geometry. Multiple UWB transmitters and receivers are positioned along each side of the imaging area. We perform several numerical simulations in time-domain, and the successful imaging of the target is achieved by visual inspection of the formed images.