WorldWideScience

Sample records for fluorescence molecular tomography

  1. In Vivo Diffuse Optical Tomography and Fluorescence Molecular Tomography

    Directory of Open Access Journals (Sweden)

    Mingze Li

    2010-01-01

    Full Text Available Diffuse optical tomography (DOT and fluorescence molecular tomography (FMT are two attractive imaging techniques for in vivo physiological and psychological research. They have distinct advantages such as non-invasiveness, non-ionizing radiation, high sensitivity and longitudinal monitoring. This paper reviews the key components of DOT and FMT. Light propagation model, mathematical reconstruction algorithm, imaging instrumentation and medical applications are included. Future challenges and perspective on optical tomography are discussed.

  2. Fluorescence molecular tomography in the presence of background fluorescence

    International Nuclear Information System (INIS)

    Soubret, Antoine; Ntziachristos, Vasilis

    2006-01-01

    Fluorescence molecular tomography is an emerging imaging technique that resolves the bio-distribution of engineered fluorescent probes developed for in vivo reporting of specific cellular and sub-cellular targets. The method can detect fluorochromes in picomole amounts or less, imaged through entire animals, but the detection sensitivity and imaging performance drop in the presence of background, non-specific fluorescence. In this study, we carried out a theoretical and an experimental investigation on the effect of background fluorescence on the measured signal and on the tomographic reconstruction. We further examined the performance of three subtraction methods based on physical models of photon propagation, using experimental data on phantoms and small animals. We show that the data pre-processing with subtraction schemes can improve image quality and quantification when non-specific background florescence is present

  3. Fluorescence Molecular Tomography: Principles and Potential for Pharmaceutical Research

    Directory of Open Access Journals (Sweden)

    Florian Stuker

    2011-04-01

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

  4. Laplacian manifold regularization method for fluorescence molecular tomography

    Science.gov (United States)

    He, Xuelei; Wang, Xiaodong; Yi, Huangjian; Chen, Yanrong; Zhang, Xu; Yu, Jingjing; He, Xiaowei

    2017-04-01

    Sparse regularization methods have been widely used in fluorescence molecular tomography (FMT) for stable three-dimensional reconstruction. Generally, ℓ1-regularization-based methods allow for utilizing the sparsity nature of the target distribution. However, in addition to sparsity, the spatial structure information should be exploited as well. A joint ℓ1 and Laplacian manifold regularization model is proposed to improve the reconstruction performance, and two algorithms (with and without Barzilai-Borwein strategy) are presented to solve the regularization model. Numerical studies and in vivo experiment demonstrate that the proposed Gradient projection-resolved Laplacian manifold regularization method for the joint model performed better than the comparative algorithm for ℓ1 minimization method in both spatial aggregation and location accuracy.

  5. Early Detection of Breast Cancer by Fluorescence Molecular Tomography

    National Research Council Canada - National Science Library

    Ntziachristos, Vasilis

    2007-01-01

    .... We have successfully completed all goals and achieved the three major aims of the proposal, i.e. i) the development of appropriate fluorescence imaging methods for highly reliable and quantitative fluorescence imaging ii...

  6. Fast automatic segmentation of anatomical structures in x-ray computed tomography images to improve fluorescence molecular tomography reconstruction.

    Science.gov (United States)

    Freyer, Marcus; Ale, Angelique; Schulz, Ralf B; Zientkowska, Marta; Ntziachristos, Vasilis; Englmeier, Karl-Hans

    2010-01-01

    The recent development of hybrid imaging scanners that integrate fluorescence molecular tomography (FMT) and x-ray computed tomography (XCT) allows the utilization of x-ray information as image priors for improving optical tomography reconstruction. To fully capitalize on this capacity, we consider a framework for the automatic and fast detection of different anatomic structures in murine XCT images. To accurately differentiate between different structures such as bone, lung, and heart, a combination of image processing steps including thresholding, seed growing, and signal detection are found to offer optimal segmentation performance. The algorithm and its utilization in an inverse FMT scheme that uses priors is demonstrated on mouse images.

  7. Improving limited-projection-angle fluorescence molecular tomography using a co-registered x-ray computed tomography scan.

    Science.gov (United States)

    Radrich, Karin; Ale, Angelique; Ermolayev, Vladimir; Ntziachristos, Vasilis

    2012-12-01

    We examine the improvement in imaging performance, such as axial resolution and signal localization, when employing limited-projection-angle fluorescence molecular tomography (FMT) together with x-ray computed tomography (XCT) measurements versus stand-alone FMT. For this purpose, we employed living mice, bearing a spontaneous lung tumor model, and imaged them with FMT and XCT under identical geometrical conditions using fluorescent probes for cancer targeting. The XCT data was employed, herein, as structural prior information to guide the FMT reconstruction. Gold standard images were provided by fluorescence images of mouse cryoslices, providing the ground truth in fluorescence bio-distribution. Upon comparison of FMT images versus images reconstructed using hybrid FMT and XCT data, we demonstrate marked improvements in image accuracy. This work relates to currently disseminated FMT systems, using limited projection scans, and can be employed to enhance their performance.

  8. Fluorescence background subtraction technique for hybrid fluorescence molecular tomography/x-ray computed tomography imaging of a mouse model of early stage lung cancer.

    Science.gov (United States)

    Ale, Angelique; Ermolayev, Vladimir; Deliolanis, Nikolaos C; Ntziachristos, Vasilis

    2013-05-01

    The ability to visualize early stage lung cancer is important in the study of biomarkers and targeting agents that could lead to earlier diagnosis. The recent development of hybrid free-space 360-deg fluorescence molecular tomography (FMT) and x-ray computed tomography (XCT) imaging yields a superior optical imaging modality for three-dimensional small animal fluorescence imaging over stand-alone optical systems. Imaging accuracy was improved by using XCT information in the fluorescence reconstruction method. Despite this progress, the detection sensitivity of targeted fluorescence agents remains limited by nonspecific background accumulation of the fluorochrome employed, which complicates early detection of murine cancers. Therefore we examine whether x-ray CT information and bulk fluorescence detection can be combined to increase detection sensitivity. Correspondingly, we research the performance of a data-driven fluorescence background estimator employed for subtraction of background fluorescence from acquisition data. Using mice containing known fluorochromes ex vivo, we demonstrate the reduction of background signals from reconstructed images and sensitivity improvements. Finally, by applying the method to in vivo data from K-ras transgenic mice developing lung cancer, we find small tumors at an early stage compared with reconstructions performed using raw data. We conclude with the benefits of employing fluorescence subtraction in hybrid FMT-XCT for early detection studies.

  9. Reconstruction for limited-projection fluorescence molecular tomography based on projected restarted conjugate gradient normal residual.

    Science.gov (United States)

    Cao, Xu; Zhang, Bin; Liu, Fei; Wang, Xin; Bai, Jing

    2011-12-01

    Limited-projection fluorescence molecular tomography (FMT) can greatly reduce the acquisition time, which is suitable for resolving fast biology processes in vivo but suffers from severe ill-posedness because of the reconstruction using only limited projections. To overcome the severe ill-posedness, we report a reconstruction method based on the projected restarted conjugate gradient normal residual. The reconstruction results of two phantom experiments demonstrate that the proposed method is feasible for limited-projection FMT. © 2011 Optical Society of America

  10. Compression of Born ratio for fluorescence molecular tomography/x-ray computed tomography hybrid imaging: methodology and in vivo validation.

    Science.gov (United States)

    Mohajerani, Pouyan; Ntziachristos, Vasilis

    2013-07-01

    The 360° rotation geometry of the hybrid fluorescence molecular tomography/x-ray computed tomography modality allows for acquisition of very large datasets, which pose numerical limitations on the reconstruction. We propose a compression method that takes advantage of the correlation of the Born-normalized signal among sources in spatially formed clusters to reduce the size of system model. The proposed method has been validated using an ex vivo study and an in vivo study of a nude mouse with a subcutaneous 4T1 tumor, with and without inclusion of a priori anatomical information. Compression rates of up to two orders of magnitude with minimum distortion of reconstruction have been demonstrated, resulting in large reduction in weight matrix size and reconstruction time.

  11. FMT-XCT: in vivo animal studies with hybrid fluorescence molecular tomography-X-ray computed tomography.

    Science.gov (United States)

    Ale, Angelique; Ermolayev, Vladimir; Herzog, Eva; Cohrs, Christian; de Angelis, Martin Hrabé; Ntziachristos, Vasilis

    2012-06-01

    The development of hybrid optical tomography methods to improve imaging performance has been suggested over a decade ago and has been experimentally demonstrated in animals and humans. Here we examined in vivo performance of a camera-based hybrid fluorescence molecular tomography (FMT) system for 360° imaging combined with X-ray computed tomography (XCT). Offering an accurately co-registered, information-rich hybrid data set, FMT-XCT has new imaging possibilities compared to stand-alone FMT and XCT. We applied FMT-XCT to a subcutaneous 4T1 tumor mouse model, an Aga2 osteogenesis imperfecta model and a Kras lung cancer mouse model, using XCT information during FMT inversion. We validated in vivo imaging results against post-mortem planar fluorescence images of cryoslices and histology data. Besides offering concurrent anatomical and functional information, FMT-XCT resulted in the most accurate FMT performance to date. These findings indicate that addition of FMT optics into the XCT gantry may be a potent upgrade for small-animal XCT systems.

  12. Imaging performance of a hybrid x-ray computed tomography-fluorescence molecular tomography system using priors.

    Science.gov (United States)

    Ale, Angelique; Schulz, Ralf B; Sarantopoulos, Athanasios; Ntziachristos, Vasilis

    2010-05-01

    The performance is studied of two newly introduced and previously suggested methods that incorporate priors into inversion schemes associated with data from a recently developed hybrid x-ray computed tomography and fluorescence molecular tomography system, the latter based on CCD camera photon detection. The unique data set studied attains accurately registered data of high spatially sampled photon fields propagating through tissue along 360 degrees projections. Approaches that incorporate structural prior information were included in the inverse problem by adding a penalty term to the minimization function utilized for image reconstructions. Results were compared as to their performance with simulated and experimental data from a lung inflammation animal model and against the inversions achieved when not using priors. The importance of using priors over stand-alone inversions is also showcased with high spatial sampling simulated and experimental data. The approach of optimal performance in resolving fluorescent biodistribution in small animals is also discussed. Inclusion of prior information from x-ray CT data in the reconstruction of the fluorescence biodistribution leads to improved agreement between the reconstruction and validation images for both simulated and experimental data.

  13. Full-direct method for imaging pharmacokinetic parameters in dynamic fluorescence molecular tomography

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Guanglei, E-mail: guangleizhang@bjtu.edu.cn [Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084 (China); Department of Biomedical Engineering, School of Computer and Information Technology, Beijing Jiaotong University, Beijing 100044 (China); Pu, Huangsheng; Liu, Fei; Bai, Jing [Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084 (China); He, Wei [China Institute of Sport Science, Beijing 100061 (China); Luo, Jianwen, E-mail: luo-jianwen@tsinghua.edu.cn [Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084 (China); Center for Biomedical Imaging Research, School of Medicine, Tsinghua University, Beijing 100084 (China)

    2015-02-23

    Images of pharmacokinetic parameters (also known as parametric images) in dynamic fluorescence molecular tomography (FMT) can provide three-dimensional metabolic information for biological studies and drug development. However, the ill-posed nature of FMT and the high temporal variation of fluorophore concentration together make it difficult to obtain accurate parametric images in small animals in vivo. In this letter, we present a method to directly reconstruct the parametric images from the boundary measurements based on hybrid FMT/X-ray computed tomography (XCT) system. This method can not only utilize structural priors obtained from the XCT system to mitigate the ill-posedness of FMT but also make full use of the temporal correlations of boundary measurements to model the high temporal variation of fluorophore concentration. The results of numerical simulation and mouse experiment demonstrate that the proposed method leads to significant improvements in the reconstruction quality of parametric images.

  14. A three-step reconstruction method for fluorescence molecular tomography based on compressive sensing

    DEFF Research Database (Denmark)

    Zhu, Yansong; Jha, Abhinav K.; Dreyer, Jakob K.

    2017-01-01

    Fluorescence molecular tomography (FMT) is a promising tool for real time in vivo quantification of neurotransmission (NT) as we pursue in our BRAIN initiative effort. However, the acquired image data are noisy and the reconstruction problem is ill-posed. Further, while spatial sparsity of the NT...... matrix coherence. The resultant image data are input to a homotopy-based reconstruction strategy that exploits sparsity via ℓ1 regularization. The reconstructed image is then input to a maximum-likelihood expectation maximization (MLEM) algorithm that retains the sparseness of the input estimate...... and improves upon the quantitation by accurate Poisson noise modeling. The proposed reconstruction method was evaluated in a three-dimensional simulated setup with fluorescent sources in a cuboidal scattering medium with optical properties simulating human brain cortex (reduced scattering coefficient: 9.2 cm-1...

  15. Efficient L1 regularization-based reconstruction for fluorescent molecular tomography using restarted nonlinear conjugate gradient.

    Science.gov (United States)

    Shi, Junwei; Zhang, Bin; Liu, Fei; Luo, Jianwen; Bai, Jing

    2013-09-15

    For the ill-posed fluorescent molecular tomography (FMT) inverse problem, the L1 regularization can protect the high-frequency information like edges while effectively reduce the image noise. However, the state-of-the-art L1 regularization-based algorithms for FMT reconstruction are expensive in memory, especially for large-scale problems. An efficient L1 regularization-based reconstruction algorithm based on nonlinear conjugate gradient with restarted strategy is proposed to increase the computational speed with low memory consumption. The reconstruction results from phantom experiments demonstrate that the proposed algorithm can obtain high spatial resolution and high signal-to-noise ratio, as well as high localization accuracy for fluorescence targets.

  16. Non-stationary reconstruction for dynamic fluorescence molecular tomography with extended kalman filter.

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    Liu, Xin; Wang, Hongkai; Yan, Zhuangzhi

    2016-11-01

    Dynamic fluorescence molecular tomography (FMT) plays an important role in drug delivery research. However, the majority of current reconstruction methods focus on solving the stationary FMT problems. If the stationary reconstruction methods are applied to the time-varying fluorescence measurements, the reconstructed results may suffer from a high level of artifacts. In addition, based on the stationary methods, only one tomographic image can be obtained after scanning one circle projection data. As a result, the movement of fluorophore in imaged object may not be detected due to the relative long data acquisition time (typically >1 min). In this paper, we apply extended kalman filter (EKF) technique to solve the non-stationary fluorescence tomography problem. Especially, to improve the EKF reconstruction performance, the generalized inverse of kalman gain is calculated by a second-order iterative method. The numerical simulation, phantom, and in vivo experiments are performed to evaluate the performance of the method. The experimental results indicate that by using the proposed EKF-based second-order iterative (EKF-SOI) method, we cannot only clearly resolve the time-varying distributions of fluorophore within imaged object, but also greatly improve the reconstruction time resolution (~2.5 sec/frame) which makes it possible to detect the movement of fluorophore during the imaging processes.

  17. Quantitative performance characterization of three-dimensional noncontact fluorescence molecular tomography

    Science.gov (United States)

    Favicchio, Rosy; Psycharakis, Stylianos; Schönig, Kai; Bartsch, Dusan; Mamalaki, Clio; Papamatheakis, Joseph; Ripoll, Jorge; Zacharakis, Giannis

    2016-02-01

    Fluorescent proteins and dyes are routine tools for biological research to describe the behavior of genes, proteins, and cells, as well as more complex physiological dynamics such as vessel permeability and pharmacokinetics. The use of these probes in whole body in vivo imaging would allow extending the range and scope of current biomedical applications and would be of great interest. In order to comply with a wide variety of application demands, in vivo imaging platform requirements span from wide spectral coverage to precise quantification capabilities. Fluorescence molecular tomography (FMT) detects and reconstructs in three dimensions the distribution of a fluorophore in vivo. Noncontact FMT allows fast scanning of an excitation source and noninvasive measurement of emitted fluorescent light using a virtual array detector operating in free space. Here, a rigorous process is defined that fully characterizes the performance of a custom-built horizontal noncontact FMT setup. Dynamic range, sensitivity, and quantitative accuracy across the visible spectrum were evaluated using fluorophores with emissions between 520 and 660 nm. These results demonstrate that high-performance quantitative three-dimensional visible light FMT allowed the detection of challenging mesenteric lymph nodes in vivo and the comparison of spectrally distinct fluorescent reporters in cell culture.

  18. Molecular imaging needles: dual-modality optical coherence tomography and fluorescence imaging of labeled antibodies deep in tissue

    Science.gov (United States)

    Scolaro, Loretta; Lorenser, Dirk; Madore, Wendy-Julie; Kirk, Rodney W.; Kramer, Anne S.; Yeoh, George C.; Godbout, Nicolas; Sampson, David D.; Boudoux, Caroline; McLaughlin, Robert A.

    2015-01-01

    Molecular imaging using optical techniques provides insight into disease at the cellular level. In this paper, we report on a novel dual-modality probe capable of performing molecular imaging by combining simultaneous three-dimensional optical coherence tomography (OCT) and two-dimensional fluorescence imaging in a hypodermic needle. The probe, referred to as a molecular imaging (MI) needle, may be inserted tens of millimeters into tissue. The MI needle utilizes double-clad fiber to carry both imaging modalities, and is interfaced to a 1310-nm OCT system and a fluorescence imaging subsystem using an asymmetrical double-clad fiber coupler customized to achieve high fluorescence collection efficiency. We present, to the best of our knowledge, the first dual-modality OCT and fluorescence needle probe with sufficient sensitivity to image fluorescently labeled antibodies. Such probes enable high-resolution molecular imaging deep within tissue. PMID:26137379

  19. Optimization and performance evaluation of a conical mirror based fluorescence molecular tomography imaging system

    Science.gov (United States)

    Zhao, Yue; Zhang, Wei; Zhu, Dianwen; Li, Changqing

    2016-03-01

    We performed numerical simulations and phantom experiments with a conical mirror based fluorescence molecular tomography (FMT) imaging system to optimize its performance. With phantom experiments, we have compared three measurement modes in FMT: the whole surface measurement mode, the transmission mode, and the reflection mode. Our results indicated that the whole surface measurement mode performed the best. Then, we applied two different neutral density (ND) filters to improve the measurement's dynamic range. The benefits from ND filters are not as much as predicted. Finally, with numerical simulations, we have compared two laser excitation patterns: line and point. With the same excitation position number, we found that the line laser excitation had slightly better FMT reconstruction results than the point laser excitation. In the future, we will implement Monte Carlo ray tracing simulations to calculate multiple reflection photons, and create a look-up table accordingly for calibration.

  20. A penalized linear and nonlinear combined conjugate gradient method for the reconstruction of fluorescence molecular tomography.

    Science.gov (United States)

    Shang, Shang; Bai, Jing; Song, Xiaolei; Wang, Hongkai; Lau, Jaclyn

    2007-01-01

    Conjugate gradient method is verified to be efficient for nonlinear optimization problems of large-dimension data. In this paper, a penalized linear and nonlinear combined conjugate gradient method for the reconstruction of fluorescence molecular tomography (FMT) is presented. The algorithm combines the linear conjugate gradient method and the nonlinear conjugate gradient method together based on a restart strategy, in order to take advantage of the two kinds of conjugate gradient methods and compensate for the disadvantages. A quadratic penalty method is adopted to gain a nonnegative constraint and reduce the illposedness of the problem. Simulation studies show that the presented algorithm is accurate, stable, and fast. It has a better performance than the conventional conjugate gradient-based reconstruction algorithms. It offers an effective approach to reconstruct fluorochrome information for FMT.

  1. A study of MRI-guided diffuse fluorescence molecular tomography for monitoring PDT effects in pancreas cancer

    Science.gov (United States)

    Samkoe, Kimberley S.; Davis, Scott C.; Srinivasan, Subhadra; O'Hara, Julia A.; Hasan, Tayyaba; Pogue, Brian W.

    2009-06-01

    Over the last several decades little progress has been made in the therapy and treatment monitoring of pancreas adenocarcinoma, a devastating and aggressive form of cancer that has a 5-year patient survival rate of 3%. Currently, investigations for the use of interstitial Verteporfin photodynamic therapy (PDT) are being undertaken in both orthotopic xenograft mouse models and in human clinical trials. In the mouse models, magnetic resonance (MR) imaging has been used as a measure of surrogate response to Verteporfin PDT; however, MR imaging alone lacks the molecular information required to assess the metabolic function and growth rates of the tumor immediately after treatment. We propose the implementation of MR-guided fluorescence tomography in conjunction with a fluorescently labeled (IR-Dye 800 CW, LI-COR) epidermal growth factor (EGF) as a molecular measure of surrogate response. To demonstrate the effectiveness of MR-guided diffuse fluorescence tomography for molecular imaging, we have used the AsPC-1 (+EGFR) human pancreatic adenocarcinoma in an orthotopic mouse model. EGF IRDye 800CW was injected 48 hours prior to imaging. MR image sequences were collected simultaneously with the fluorescence data using a MR-coupled diffuse optical tomography system. Image reconstruction was performed multiple times with varying abdominal organ segmentation in order to obtain a optimal tomographic image. It is shown that diffuse fluorescence tomography of the orthotopic pancreas model is feasible, with consideration of confounding fluorescence signals from the multiple organs and tissues surrounding the pancreas. MR-guided diffuse fluorescence tomography will be used to monitor EGF response after photodynamic therapy. Additionally, it provide the opportunity to individualize subsequent therapies based on response to PDT as well as to evaluate the success of combination therapies, such as PDT with chemotherapy, antibody therapy or even radiation.

  2. Enhanced spatial resolution in fluorescence molecular tomography using restarted L1-regularized nonlinear conjugate gradient algorithm.

    Science.gov (United States)

    Shi, Junwei; Liu, Fei; Zhang, Guanglei; Luo, Jianwen; Bai, Jing

    2014-04-01

    Owing to the high degree of scattering of light through tissues, the ill-posedness of fluorescence molecular tomography (FMT) inverse problem causes relatively low spatial resolution in the reconstruction results. Unlike L2 regularization, L1 regularization can preserve the details and reduce the noise effectively. Reconstruction is obtained through a restarted L1 regularization-based nonlinear conjugate gradient (re-L1-NCG) algorithm, which has been proven to be able to increase the computational speed with low memory consumption. The algorithm consists of inner and outer iterations. In the inner iteration, L1-NCG is used to obtain the L1-regularized results. In the outer iteration, the restarted strategy is used to increase the convergence speed of L1-NCG. To demonstrate the performance of re-L1-NCG in terms of spatial resolution, simulation and physical phantom studies with fluorescent targets located with different edge-to-edge distances were carried out. The reconstruction results show that the re-L1-NCG algorithm has the ability to resolve targets with an edge-to-edge distance of 0.1 cm at a depth of 1.5 cm, which is a significant improvement for FMT.

  3. Self-prior strategy for organ reconstruction in fluorescence molecular tomography.

    Science.gov (United States)

    Zhou, Yuan; Chen, Maomao; Su, Han; Luo, Jianwen

    2017-10-01

    The purpose of this study is to propose a strategy for organ reconstruction in fluorescence molecular tomography (FMT) without prior information from other imaging modalities, and to overcome the high cost and ionizing radiation caused by the traditional structural prior strategy. The proposed strategy is designed as an iterative architecture to solve the inverse problem of FMT. In each iteration, a short time Fourier transform (STFT) based algorithm is used to extract the self-prior information in the space-frequency energy spectrum with the assumption that the regions with higher fluorescence concentration have larger energy intensity, then the cost function of the inverse problem is modified by the self-prior information, and lastly an iterative Laplacian regularization algorithm is conducted to solve the updated inverse problem and obtains the reconstruction results. Simulations and in vivo experiments on liver reconstruction are carried out to test the performance of the self-prior strategy on organ reconstruction. The organ reconstruction results obtained by the proposed self-prior strategy are closer to the ground truth than those obtained by the iterative Tikhonov regularization (ITKR) method (traditional non-prior strategy). Significant improvements are shown in the evaluation indexes of relative locational error (RLE), relative error (RE) and contrast-to-noise ratio (CNR). The self-prior strategy improves the organ reconstruction results compared with the non-prior strategy and also overcomes the shortcomings of the traditional structural prior strategy. Various applications such as metabolic imaging and pharmacokinetic study can be aided by this strategy.

  4. Image reconstruction of fluorescent molecular tomography based on the tree structured Schur complement decomposition

    Directory of Open Access Journals (Sweden)

    Wang Jiajun

    2010-05-01

    Full Text Available Abstract Background The inverse problem of fluorescent molecular tomography (FMT often involves complex large-scale matrix operations, which may lead to unacceptable computational errors and complexity. In this research, a tree structured Schur complement decomposition strategy is proposed to accelerate the reconstruction process and reduce the computational complexity. Additionally, an adaptive regularization scheme is developed to improve the ill-posedness of the inverse problem. Methods The global system is decomposed level by level with the Schur complement system along two paths in the tree structure. The resultant subsystems are solved in combination with the biconjugate gradient method. The mesh for the inverse problem is generated incorporating the prior information. During the reconstruction, the regularization parameters are adaptive not only to the spatial variations but also to the variations of the objective function to tackle the ill-posed nature of the inverse problem. Results Simulation results demonstrate that the strategy of the tree structured Schur complement decomposition obviously outperforms the previous methods, such as the conventional Conjugate-Gradient (CG and the Schur CG methods, in both reconstruction accuracy and speed. As compared with the Tikhonov regularization method, the adaptive regularization scheme can significantly improve ill-posedness of the inverse problem. Conclusions The methods proposed in this paper can significantly improve the reconstructed image quality of FMT and accelerate the reconstruction process.

  5. Anatomical image-guided fluorescence molecular tomography reconstruction using kernel method

    Science.gov (United States)

    Baikejiang, Reheman; Zhao, Yue; Fite, Brett Z.; Ferrara, Katherine W.; Li, Changqing

    2017-01-01

    Abstract. Fluorescence molecular tomography (FMT) is an important in vivo imaging modality to visualize physiological and pathological processes in small animals. However, FMT reconstruction is ill-posed and ill-conditioned due to strong optical scattering in deep tissues, which results in poor spatial resolution. It is well known that FMT image quality can be improved substantially by applying the structural guidance in the FMT reconstruction. An approach to introducing anatomical information into the FMT reconstruction is presented using the kernel method. In contrast to conventional methods that incorporate anatomical information with a Laplacian-type regularization matrix, the proposed method introduces the anatomical guidance into the projection model of FMT. The primary advantage of the proposed method is that it does not require segmentation of targets in the anatomical images. Numerical simulations and phantom experiments have been performed to demonstrate the proposed approach’s feasibility. Numerical simulation results indicate that the proposed kernel method can separate two FMT targets with an edge-to-edge distance of 1 mm and is robust to false-positive guidance and inhomogeneity in the anatomical image. For the phantom experiments with two FMT targets, the kernel method has reconstructed both targets successfully, which further validates the proposed kernel method. PMID:28464120

  6. Direct reconstruction of pharmacokinetic parameters in dynamic fluorescence molecular tomography by the augmented Lagrangian method

    Science.gov (United States)

    Zhu, Dianwen; Zhang, Wei; Zhao, Yue; Li, Changqing

    2016-03-01

    Dynamic fluorescence molecular tomography (FMT) has the potential to quantify physiological or biochemical information, known as pharmacokinetic parameters, which are important for cancer detection, drug development and delivery etc. To image those parameters, there are indirect methods, which are easier to implement but tend to provide images with low signal-to-noise ratio, and direct methods, which model all the measurement noises together and are statistically more efficient. The direct reconstruction methods in dynamic FMT have attracted a lot of attention recently. However, the coupling of tomographic image reconstruction and nonlinearity of kinetic parameter estimation due to the compartment modeling has imposed a huge computational burden to the direct reconstruction of the kinetic parameters. In this paper, we propose to take advantage of both the direct and indirect reconstruction ideas through a variable splitting strategy under the augmented Lagrangian framework. Each iteration of the direct reconstruction is split into two steps: the dynamic FMT image reconstruction and the node-wise nonlinear least squares fitting of the pharmacokinetic parameter images. Through numerical simulation studies, we have found that the proposed algorithm can achieve good reconstruction results within a small amount of time. This will be the first step for a combined dynamic PET and FMT imaging in the future.

  7. A generic, geometric cocalibration method for a combined system of fluorescence molecular tomography and microcomputed tomography with arbitrarily shaped objects

    International Nuclear Information System (INIS)

    Fu Jianwei; Yang Xiaoquan; Wang Kan; Luo Qingming; Gong Hui

    2011-01-01

    Purpose: A combined system of fluorescence molecular tomography and microcomputed tomography (FMT and mCT) can provide molecular and anatomical information of small animals in a single study with intrinsically coregistered images. The anatomical information provided by the mCT subsystem is commonly used as a reference to locate the fluorophore distribution or as a priori structural information to improve the performance of FMT. Therefore, the transformation between the coordinate systems of the subsystem needs to be determined in advanced. Methods: A cocalibration method for the combined system of FMT and mCT is proposed. First, linear models are adopted to describe the galvano mirrors and the charge-coupled device (CCD) camera in the FMT subsystem. Second, the position and orientation of the galvano mirrors are determined with the input voltages of the galvano mirrors and the markers, whose positions are predetermined. The position, orientation and normalized pixel size of the CCD camera are obtained by analysing the projections of a point-like marker at different positions. Finally, the orientation and position of sources and the corresponding relationship between the detectors and their projections on the image plane are predicted. Because the positions of the markers are acquired with mCT, the registration of the FMT and mCT could be realized by direct image fusion. Results: The accuracy and consistency of this method in the presence of noise is evaluated by computer simulation. Next, a practical implementation for an experimental FMT and mCT system is carried out and validated. The maximum prediction error of the source positions on the surface of a cylindrical phantom is within 0.375 mm and that of the projections of a point-like marker is within 0.629 pixel. Finally, imaging experiments of the fluorophore distribution in a cylindrical phantom and a phantom with a complex shape demonstrate the feasibility of the proposed method. Conclusions: This method is

  8. Performance dependence of hybrid x-ray computed tomography/fluorescence molecular tomography on the optical forward problem.

    Science.gov (United States)

    Hyde, Damon; Schulz, Ralf; Brooks, Dana; Miller, Eric; Ntziachristos, Vasilis

    2009-04-01

    Hybrid imaging systems combining x-ray computed tomography (CT) and fluorescence tomography can improve fluorescence imaging performance by incorporating anatomical x-ray CT information into the optical inversion problem. While the use of image priors has been investigated in the past, little is known about the optimal use of forward photon propagation models in hybrid optical systems. In this paper, we explore the impact on reconstruction accuracy of the use of propagation models of varying complexity, specifically in the context of these hybrid imaging systems where significant structural information is known a priori. Our results demonstrate that the use of generically known parameters provides near optimal performance, even when parameter mismatch remains.

  9. Imaging of pharmacokinetic rates of indocyanine green in mouse liver with a hybrid fluorescence molecular tomography/x-ray computed tomography system.

    Science.gov (United States)

    Zhang, Guanglei; Liu, Fei; Zhang, Bin; He, Yun; Luo, Jianwen; Bai, Jing

    2013-04-01

    Pharmacokinetic rates have the potential to provide quantitative physiological and pathological information for biological studies and drug development. Fluorescence molecular tomography (FMT) is an attractive imaging tool for three-dimensionally resolving fluorophore distribution in small animals. In this letter, pharmacokinetic rates of indocyanine green (ICG) in mouse liver are imaged with a hybrid FMT and x-ray computed tomography (XCT) system. A recently developed FMT method using structural priors from an XCT system is adopted to improve the quality of FMT reconstruction. In the in vivo experiments, images of uptake and excretion rates of ICG in mouse liver are obtained, which can be used to quantitatively evaluate liver function. The accuracy of the results is validated by a fiber-based fluorescence measurement system.

  10. Three-dimensional fluorescence lifetime tomography

    International Nuclear Information System (INIS)

    Godavarty, Anuradha; Sevick-Muraca, Eva M.; Eppstein, Margaret J.

    2005-01-01

    Near-infrared fluorescence tomography using molecularly targeted lifetime-sensitive, fluorescent contrast agents have applications for early-stage cancer diagnostics. Yet, although the measurement of fluorescent lifetime imaging microscopy (FLIM) is extensively used in microscopy and spectroscopy applications, demonstration of fluorescence lifetime tomography for medical imaging is limited to two-dimensional studies. Herein, the feasibility of three-dimensional fluorescence-lifetime tomography on clinically relevant phantom volumes is established, using (i) a gain-modulated intensified charge coupled device (CCD) and modulated laser diode imaging system, (ii) two fluorescent contrast agents, e.g., Indocyanine green and 3-3'-Diethylthiatricarbocyanine iodide differing in their fluorescence lifetime by 0.62 ns, and (iii) a two stage approximate extended Kalman filter reconstruction algorithm. Fluorescence measurements of phase and amplitude were acquired on the phantom surface under different target to background fluorescence absorption (70:1, 100:1) and fluorescence lifetime (1:1, 2.1:1) contrasts at target depths of 1.4-2 cm. The Bayesian tomography algorithm was employed to obtain three-dimensional images of lifetime and absorption owing to the fluorophores

  11. Integrin-Targeted Hybrid Fluorescence Molecular Tomography/X-ray Computed Tomography for Imaging Tumor Progression and Early Response in Non-Small Cell Lung Cancer

    Directory of Open Access Journals (Sweden)

    Xiaopeng Ma

    2017-01-01

    Full Text Available Integrins play an important role in tumor progression, invasion and metastasis. Therefore we aimed to evaluate a preclinical imaging approach applying ανβ3 integrin targeted hybrid Fluorescence Molecular Tomography/X-ray Computed Tomography (FMT-XCT for monitoring tumor progression as well as early therapy response in a syngeneic murine Non-Small Cell Lung Cancer (NSCLC model. Lewis Lung Carcinomas were grown orthotopically in C57BL/6 J mice and imaged in-vivo using a ανβ3 targeted near-infrared fluorescence (NIRF probe. ανβ3-targeted FMT-XCT was able to track tumor progression. Cilengitide was able to substantially block the binding of the NIRF probe and suppress the imaging signal. Additionally mice were treated with an established chemotherapy regimen of Cisplatin and Bevacizumab or with a novel MEK inhibitor (Refametinib for 2 weeks. While μCT revealed only a moderate slowdown of tumor growth, ανβ3 dependent signal decreased significantly compared to non-treated mice already at one week post treatment. ανβ3 targeted imaging might therefore become a promising tool for assessment of early therapy response in the future.

  12. Ultrasound guided fluorescence molecular tomography with improved quantification by an attenuation compensated born-normalization and in vivo preclinical study of cancer

    International Nuclear Information System (INIS)

    Li, Baoqiang; Berti, Romain; Abran, Maxime; Lesage, Frédéric

    2014-01-01

    Ultrasound imaging, having the advantages of low-cost and non-invasiveness over MRI and X-ray CT, was reported by several studies as an adequate complement to fluorescence molecular tomography with the perspective of improving localization and quantification of fluorescent molecular targets in vivo. Based on the previous work, an improved dual-modality Fluorescence-Ultrasound imaging system was developed and then validated in imaging study with preclinical tumor model. Ultrasound imaging and a profilometer were used to obtain the anatomical prior information and 3D surface, separately, to precisely extract the tissue boundary on both sides of sample in order to achieve improved fluorescence reconstruction. Furthermore, a pattern-based fluorescence reconstruction on the detection side was incorporated to enable dimensional reduction of the dataset while keeping the useful information for reconstruction. Due to its putative role in the current imaging geometry and the chosen reconstruction technique, we developed an attenuation compensated Born-normalization method to reduce the attenuation effects and cancel off experimental factors when collecting quantitative fluorescence datasets over large area. Results of both simulation and phantom study demonstrated that fluorescent targets could be recovered accurately and quantitatively using this reconstruction mechanism. Finally, in vivo experiment confirms that the imaging system associated with the proposed image reconstruction approach was able to extract both functional and anatomical information, thereby improving quantification and localization of molecular targets

  13. The Value of 5-Aminolevulinic Acid in Low-grade Gliomas and High-grade Gliomas Lacking Glioblastoma Imaging Features: An Analysis Based on Fluorescence, Magnetic Resonance Imaging, 18F-Fluoroethyl Tyrosine Positron Emission Tomography, and Tumor Molecular Factors.

    Science.gov (United States)

    Jaber, Mohammed; Wölfer, Johannes; Ewelt, Christian; Holling, Markus; Hasselblatt, Martin; Niederstadt, Thomas; Zoubi, Tarek; Weckesser, Matthias; Stummer, Walter

    2016-03-01

    Approximately 20% of grade II and most grade III gliomas fluoresce after 5-aminolevulinic acid (5-ALA) application. Conversely, approximately 30% of nonenhancing gliomas are actually high grade. The aim of this study was to identify preoperative factors (ie, age, enhancement, 18F-fluoroethyl tyrosine positron emission tomography [F-FET PET] uptake ratios) for predicting fluorescence in gliomas without typical glioblastomas imaging features and to determine whether fluorescence will allow prediction of tumor grade or molecular characteristics. Patients harboring gliomas without typical glioblastoma imaging features were given 5-ALA. Fluorescence was recorded intraoperatively, and biopsy specimens collected from fluorescing tissue. World Health Organization (WHO) grade, Ki-67/MIB-1 index, IDH1 (R132H) mutation status, O-methylguanine DNA methyltransferase (MGMT) promoter methylation status, and 1p/19q co-deletion status were assessed. Predictive factors for fluorescence were derived from preoperative magnetic resonance imaging and F-FET PET. Classification and regression tree analysis and receiver-operating-characteristic curves were generated for defining predictors. Of 166 tumors, 82 were diagnosed as WHO grade II, 76 as grade III, and 8 as glioblastomas grade IV. Contrast enhancement, tumor volume, and F-FET PET uptake ratio >1.85 predicted fluorescence. Fluorescence correlated with WHO grade (P fluorescing grade III gliomas was higher than in nonfluorescing tumors, whereas in fluorescing and nonfluorescing grade II tumors, no differences were noted. Age, tumor volume, and F-FET PET uptake are factors predicting 5-ALA-induced fluorescence in gliomas without typical glioblastoma imaging features. Fluorescence was associated with an increased Ki-67/MIB-1 index and high-grade pathology. Whether fluorescence in grade II gliomas identifies a subtype with worse prognosis remains to be determined.

  14. Fluorescence diffuse tomography of small animals with DsRed2 fluorescent protein

    Science.gov (United States)

    Turchin, I. V.; Plehanov, V. I.; Orlova, A. G.; Kamenskiy, V. A.; Kleshnin, M. S.; Shirmanova, M. V.; Shakhova, N. M.; Balalaeva, I. V.; Savitskiy, A. P.

    2006-05-01

    Fluorescent compounds are used as markers to diagnose oncological diseases, to study molecular processes typical for carcinogenesis, and to investigate metastasis formation and tumor regress under the influence of therapeutics. Different types of tomography, such as continuous wave (CW), frequency-domain (FD), and time-domain (TD) tomography, allow fluorescence imaging of tumors located deep in human or animal tissue. In this work, preliminary results of the frequency domain fluorescent diffuse tomography (FDT) method in application to DsRed2 protein as a fluorescent agent are presented. For the first step of our experiments, we utilized low-frequency amplitude modulation (1 kHz) of second harmonic of Nd: YAG (532 nm). The transilluminative configuration was used in the setup. The results of post mortem experiments with capsules containing DsRed2 inserted inside the esophagus of a 3-day-old hairless rat to simulate tumor are shown. An algorithm of processing fluorescent images based on calculating the zero of maximum curvature has been applied to detect fluorescent inclusion boundaries in the image. This work demonstrates the potential capability of the FDT method for imaging deep fluorescent tumors in human tissue or animal models of human cancer. Improvement of the setup can be accomplished by using high-frequency modulation (using a 110-MHz acoustooptical modulator).

  15. X-ray microtome by fluorescence tomography

    CERN Document Server

    Simionovici, A S; Guenzler, F; Schrör, C; Snigirev, A; Snigireva, I; Tümmler, J; Weitkamp, T

    2001-01-01

    The X-ray fluorescence microtomography method is presented, which is capable of virtually slicing samples to obtain cross-sections of their inner structure. High precision experimental results of fluo-tomography in 'pencil-beam' geometry with up to 1.2 mu m resolution are described. Image reconstructions are based on either a simplified algebraic reconstruction method (ART) or the filtered back-projection method (FBP). Phantoms of inhomogeneous test objects as well as biological samples are successfully analyzed.

  16. Possible Radiation-Induced Damage to the Molecular Structure of Wooden Artifacts Due to Micro-Computed Tomography, Handheld X-Ray Fluorescence, and X-Ray Photoelectron Spectroscopic Techniques

    Directory of Open Access Journals (Sweden)

    Madalena Kozachuk

    2016-05-01

    Full Text Available This study was undertaken to ascertain whether radiation produced by X-ray photoelectron spectroscopy (XPS, micro-computed tomography (μCT and/or portable handheld X-ray fluorescence (XRF equipment might damage wood artifacts during analysis. Changes at the molecular level were monitored by Fourier transform infrared (FTIR analysis. No significant changes in FTIR spectra were observed as a result of μCT or handheld XRF analysis. No substantial changes in the collected FTIR spectra were observed when XPS analytical times on the order of minutes were used. However, XPS analysis collected over tens of hours did produce significant changes in the FTIR spectra.

  17. Radiative transport-based frequency-domain fluorescence tomography

    International Nuclear Information System (INIS)

    Joshi, Amit; Rasmussen, John C; Sevick-Muraca, Eva M; Wareing, Todd A; McGhee, John

    2008-01-01

    We report the development of radiative transport model-based fluorescence optical tomography from frequency-domain boundary measurements. The coupled radiative transport model for describing NIR fluorescence propagation in tissue is solved by a novel software based on the established Attila(TM) particle transport simulation platform. The proposed scheme enables the prediction of fluorescence measurements with non-contact sources and detectors at a minimal computational cost. An adjoint transport solution-based fluorescence tomography algorithm is implemented on dual grids to efficiently assemble the measurement sensitivity Jacobian matrix. Finally, we demonstrate fluorescence tomography on a realistic computational mouse model to locate nM to μM fluorophore concentration distributions in simulated mouse organs

  18. Deep-tissue reporter-gene imaging with fluorescence and optoacoustic tomography: a performance overview.

    Science.gov (United States)

    Deliolanis, Nikolaos C; Ale, Angelique; Morscher, Stefan; Burton, Neal C; Schaefer, Karin; Radrich, Karin; Razansky, Daniel; Ntziachristos, Vasilis

    2014-10-01

    A primary enabling feature of near-infrared fluorescent proteins (FPs) and fluorescent probes is the ability to visualize deeper in tissues than in the visible. The purpose of this work is to find which is the optimal visualization method that can exploit the advantages of this novel class of FPs in full-scale pre-clinical molecular imaging studies. Nude mice were stereotactically implanted with near-infrared FP expressing glioma cells to from brain tumors. The feasibility and performance metrics of FPs were compared between planar epi-illumination and trans-illumination fluorescence imaging, as well as to hybrid Fluorescence Molecular Tomography (FMT) system combined with X-ray CT and Multispectral Optoacoustic (or Photoacoustic) Tomography (MSOT). It is shown that deep-seated glioma brain tumors are possible to visualize both with fluorescence and optoacoustic imaging. Fluorescence imaging is straightforward and has good sensitivity; however, it lacks resolution. FMT-XCT can provide an improved rough resolution of ∼1 mm in deep tissue, while MSOT achieves 0.1 mm resolution in deep tissue and has comparable sensitivity. We show imaging capacity that can shift the visualization paradigm in biological discovery. The results are relevant not only to reporter gene imaging, but stand as cross-platform comparison for all methods imaging near infrared fluorescent contrast agents.

  19. Diffuse fluorescence tomography of exo- and endogenously labeled tumors

    Science.gov (United States)

    Balalaeva, Irina V.; Turchin, Ilya V.; Orlova, Anna G.; Plekhanov, Vladimir I.; Shirmanova, Marina V.; Kleshnin, Michail S.; Fiks, Ilya I.; Zagainova, Elena V.; Kamensky, Vladislav A.

    2007-06-01

    Strong light scattering and absorption limit observation of the internal structure of biological tissue. Only special tools for turbid media imaging, such as optical diffuse tomography, enable noninvasive investigation of the internal biological tissues, including visualization and intravital monitoring of deep tumors. In this work the preliminary results of diffuse fluorescence tomography (DFT) of small animals are presented. Usage of exogenous fluorophores, targeted specifically at tumor cells, and fluorescent proteins expressed endogenously can significantly increase the contrast of obtained images. Fluorescent compounds of different nature, such as sulphonated aluminium phthalocyanine (Photosens), red fluorescing proteins and CdTe/CdSe-core/shell nanocrystals (quantum dots) were applied. We tested diffuse fluorescence tomography method at model media, in post mortem and in vivo experiments. The animal was scanned in transilluminative configuration by low-frequency modulated light (1 kHz) from Nd:YAG laser with second harmonic generation at wavelength of 532 nm or semiconductor laser at wavelength of 655 nm. Quantum dots or protein DsRed2 in glass capsules (inner diameter 2-3 mm) were placed post mortem inside the esophagus of 7-day-old hairless rats to simulate marked tumors. Photosens was injected intravenously to linear mice with metastazing Lewis lung carcinoma. The reconstruction algorithm, based on Algebraic Reconstruction Technique, was created and tested numerically in model experiments. High contrast images of tumor simulating capsules with DsRed2 concentrations about 10 -6 M and quantum dots about 5x10 -11 M have been obtained. Organ distribution of Photosens and its accumulation in tumors and surrounding tissues of animals has been examined. We have conducted the monitoring of tumors, exogenously labeled by photosensitizer. This work demonstrates potential capabilities of DFT method for intravital detection and monitoring of deep fluorescent

  20. Bessel beam fluorescence lifetime tomography of live embryos (Conference Presentation)

    Science.gov (United States)

    Xu, Dongli; Peng, Leilei

    2016-03-01

    Optical tomography allows isotropic 3D imaging of embryos. Scanning-laser optical tomography (SLOT) has superior light collecting efficiency than wide-field optical tomography, making it ideal for fluorescence imaging of live embryos. We previously reported an imaging system that combines SLOT with a novel Fourier-multiplexed fluorescence lifetime imaging (FmFLIM) technique named FmFLIM-SLOT. FmFLIM-SLOT performs multiplexed FLIM-FRET readout of multiple FRET sensors in live embryos. Here we report a recent effort on improving the spatial resolution of the FmFLIM-SLOT system in order to image complex biochemical processes in live embryos at the cellular level. Optical tomography has to compromise between resolution and the depth of view. In SLOT, the commonly-used focused Gaussian beam diverges quickly from the focal plane, making it impossible to achieve high resolution imaging in a large volume specimen. We thus introduce Bessel beam laser-scanning tomography, which illuminates the sample with a spatial-light-modulator-generated Bessel beam that has an extended focal depth. The Bessel beam is scanned across the whole specimen. Fluorescence projection images are acquired at equal angular intervals as the sample rotates. Reconstruction artifacts due to annular-rings of the Bessel beam are removed by a modified 3D filtered back projection algorithm. Furthermore, in combination of Fourier-multiplexing fluorescence lifetime imaging (FmFLIM) method, the Bessel FmFLIM-SLOT system is capable of perform 3D lifetime imaging of live embryos at cellular resolution. The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove that Bessel FmFLIM-SLOT is a promising imaging method in development biology research.

  1. Depth-resolved imaging of colon tumor using optical coherence tomography and fluorescence laminar optical tomography (Conference Presentation)

    Science.gov (United States)

    Tang, Qinggong; Frank, Aaron; Wang, Jianting; Chen, Chao-wei; Jin, Lily; Lin, Jon; Chan, Joanne M.; Chen, Yu

    2016-03-01

    Early detection of neoplastic changes remains a critical challenge in clinical cancer diagnosis and treatment. Many cancers arise from epithelial layers such as those of the gastrointestinal (GI) tract. Current standard endoscopic technology is unable to detect those subsurface lesions. Since cancer development is associated with both morphological and molecular alterations, imaging technologies that can quantitative image tissue's morphological and molecular biomarkers and assess the depth extent of a lesion in real time, without the need for tissue excision, would be a major advance in GI cancer diagnostics and therapy. In this research, we investigated the feasibility of multi-modal optical imaging including high-resolution optical coherence tomography (OCT) and depth-resolved high-sensitivity fluorescence laminar optical tomography (FLOT) for structural and molecular imaging. APC (adenomatous polyposis coli) mice model were imaged using OCT and FLOT and the correlated histopathological diagnosis was obtained. Quantitative structural (the scattering coefficient) and molecular imaging parameters (fluorescence intensity) from OCT and FLOT images were developed for multi-parametric analysis. This multi-modal imaging method has demonstrated the feasibility for more accurate diagnosis with 87.4% (87.3%) for sensitivity (specificity) which gives the most optimal diagnosis (the largest area under receiver operating characteristic (ROC) curve). This project results in a new non-invasive multi-modal imaging platform for improved GI cancer detection, which is expected to have a major impact on detection, diagnosis, and characterization of GI cancers, as well as a wide range of epithelial cancers.

  2. Quantum process tomography by 2D fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Pachón, Leonardo A. [Grupo de Física Atómica y Molecular, Instituto de Física, Facultad de Ciencias Exactas y Naturales, Universidad de Antioquia UdeA, Calle 70 No. 52-21, Medellín (Colombia); Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138 (United States); Marcus, Andrew H. [Department of Chemistry and Biochemistry, Oregon Center for Optics, Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403 (United States); Aspuru-Guzik, Alán [Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138 (United States)

    2015-06-07

    Reconstruction of the dynamics (quantum process tomography) of the single-exciton manifold in energy transfer systems is proposed here on the basis of two-dimensional fluorescence spectroscopy (2D-FS) with phase-modulation. The quantum-process-tomography protocol introduced here benefits from, e.g., the sensitivity enhancement ascribed to 2D-FS. Although the isotropically averaged spectroscopic signals depend on the quantum yield parameter Γ of the doubly excited-exciton manifold, it is shown that the reconstruction of the dynamics is insensitive to this parameter. Applications to foundational and applied problems, as well as further extensions, are discussed.

  3. Quantum process tomography by 2D fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Pachón, Leonardo A.; Marcus, Andrew H.; Aspuru-Guzik, Alán

    2015-01-01

    Reconstruction of the dynamics (quantum process tomography) of the single-exciton manifold in energy transfer systems is proposed here on the basis of two-dimensional fluorescence spectroscopy (2D-FS) with phase-modulation. The quantum-process-tomography protocol introduced here benefits from, e.g., the sensitivity enhancement ascribed to 2D-FS. Although the isotropically averaged spectroscopic signals depend on the quantum yield parameter Γ of the doubly excited-exciton manifold, it is shown that the reconstruction of the dynamics is insensitive to this parameter. Applications to foundational and applied problems, as well as further extensions, are discussed

  4. Small-Animal Imaging Using Diffuse Fluorescence Tomography.

    Science.gov (United States)

    Davis, Scott C; Tichauer, Kenneth M

    2016-01-01

    Diffuse fluorescence tomography (DFT) has been developed to image the spatial distribution of fluorescence-tagged tracers in living tissue. This capability facilitates the recovery of any number of functional parameters, including enzymatic activity, receptor density, blood flow, and gene expression. However, deploying DFT effectively is complex and often requires years of know-how, especially for newer mutlimodal systems that combine DFT with conventional imaging systems. In this chapter, we step through the process of using MRI-DFT imaging of a receptor-targeted tracer in small animals.

  5. Fluorescent scanning x-ray tomography with synchrotron radiation

    Science.gov (United States)

    Takeda, Tohoru; Maeda, Toshikazu; Yuasa, Tetsuya; Akatsuka, Takao; Ito, Tatsuo; Kishi, Kenichi; Wu, Jin; Kazama, Masahiro; Hyodo, Kazuyuki; Itai, Yuji

    1995-02-01

    Fluorescent scanning (FS) x-ray tomography was developed to detect nonradioactive tracer materials (iodine and gadolinium) in a living object. FS x-ray tomography consists of a silicon (111) channel cut monochromator, an x-ray shutter, an x-ray slit system and a collimator for detection, a scanning table for the target organ, and an x-ray detector with pure germanium. The minimal detectable dose of iodine in this experiment was 100 ng in a volume of 2 mm3 and a linear relationship was shown between the photon counts of a fluorescent x ray and the concentration of iodine contrast material. A FS x-ray tomographic image was clearly obtained with a phantom.

  6. Fluorescence based molecular in vivo imaging

    International Nuclear Information System (INIS)

    Ebert, Bernd

    2008-01-01

    Molecular imaging represents a modern research area that allows the in vivo study of molecular biological process kinetics using appropriate probes and visualization methods. This methodology may be defined- apart from the contrast media injection - as non-abrasive. In order to reach an in vivo molecular process imaging as accurate as possible the effects of the used probes on the biological should not be too large. The contrast media as important part of the molecular imaging can significantly contribute to the understanding of molecular processes and to the development of tailored diagnostics and therapy. Since more than 15 years PTB is developing optic imaging systems that may be used for fluorescence based visualization of tissue phantoms, small animal models and the localization of tumors and their predecessors, and for the early recognition of inflammatory processes in clinical trials. Cellular changes occur during many diseases, thus the molecular imaging might be of importance for the early diagnosis of chronic inflammatory diseases. Fluorescent dyes can be used as unspecific or also as specific contrast media, which allow enhanced detection sensitivity

  7. A parallel adaptive finite element simplified spherical harmonics approximation solver for frequency domain fluorescence molecular imaging

    International Nuclear Information System (INIS)

    Lu Yujie; Zhu Banghe; Rasmussen, John C; Sevick-Muraca, Eva M; Shen Haiou; Wang Ge

    2010-01-01

    Fluorescence molecular imaging/tomography may play an important future role in preclinical research and clinical diagnostics. Time- and frequency-domain fluorescence imaging can acquire more measurement information than the continuous wave (CW) counterpart, improving the image quality of fluorescence molecular tomography. Although diffusion approximation (DA) theory has been extensively applied in optical molecular imaging, high-order photon migration models need to be further investigated to match quantitation provided by nuclear imaging. In this paper, a frequency-domain parallel adaptive finite element solver is developed with simplified spherical harmonics (SP N ) approximations. To fully evaluate the performance of the SP N approximations, a fast time-resolved tetrahedron-based Monte Carlo fluorescence simulator suitable for complex heterogeneous geometries is developed using a convolution strategy to realize the simulation of the fluorescence excitation and emission. The validation results show that high-order SP N can effectively correct the modeling errors of the diffusion equation, especially when the tissues have high absorption characteristics or when high modulation frequency measurements are used. Furthermore, the parallel adaptive mesh evolution strategy improves the modeling precision and the simulation speed significantly on a realistic digital mouse phantom. This solver is a promising platform for fluorescence molecular tomography using high-order approximations to the radiative transfer equation.

  8. Laser induced fluorescence of trapped molecular ions

    International Nuclear Information System (INIS)

    Grieman, F.J.

    1979-10-01

    An experimental apparatus for obtaining the optical spectra of molecular ions is described. The experimental technique includes the use of three dimensional ion trapping, laser induced fluorescence, and gated photon counting methods. The ions, which are produced by electron impact, are confined in a radio-frequency quadrupole ion trap of cylindrical design. Because the quadrupole ion trap allows mass selection of the molecular ion desired for study, the analysis of the spectra obtained is greatly simplified. The ion trap also confines the ions to a region easily probed by a laser beam. 18 references

  9. Laser induced fluorescence of trapped molecular ions

    Energy Technology Data Exchange (ETDEWEB)

    Grieman, F.J.

    1979-10-01

    An experimental apparatus for obtaining the optical spectra of molecular ions is described. The experimental technique includes the use of three dimensional ion trapping, laser induced fluorescence, and gated photon counting methods. The ions, which are produced by electron impact, are confined in a radio-frequency quadrupole ion trap of cylindrical design. Because the quadrupole ion trap allows mass selection of the molecular ion desired for study, the analysis of the spectra obtained is greatly simplified. The ion trap also confines the ions to a region easily probed by a laser beam. 18 references.

  10. Frequency domain fluorescence diffuse tomography of small animals

    Science.gov (United States)

    Orlova, Anna G.; Turchin, Ilya V.; Kamensky, Vladislav A.; Plehanov, Vladimir I.; Balalaeva, Irina V.; Sergeeva, Ekaterina A.; Shirmanova, Marina V.; Kleshnin, Michail S.

    2007-05-01

    Fluorescent compounds for selective cancer cell marking are used for development of novel medical diagnostic methods, investigation of the influence of external factors on tumor growth, regress and metastasis. Only special tools for turbid media imaging, such as optical diffusion tomography permit noninvasive monitoring of fluorescent-labeled tumor alterations deep in animal tissue. In this work, the results of preliminary experiments utilizing frequency-domain fluorescent diffusion tomography (FD FDT) experimental setup in small animal are presented. Low-frequency modulated light (1 kHz) from Nd:YAG laser with second harmonic generation at the wavelength of 532 nm was used in the setup. The transilluminative planar configuration was used in the setup. A series of model experiments has been conducted and show good agreement between theoretical and experimental fluorescence intensity. Models of deep tumors were created by two methods: (1) glass capsules containing fluorophore solution were inserted into esophagus of small animals to simulate marked tumors; (2) a suspension of transfected HEΚ293-Turbo-RFP cells was subcutaneously injected to small animal. The conducted experiments have shown that FD FDT allows one to detect the presence of labeled tumor cells in small animals, to determine the volume of an experimental tumor, to perform 3D tumor reconstruction, as well as to conduct monitoring investigations. The obtained results demonstrate the potential capability of the FD FDT method for noninvasive whole-body imaging in cancer studies, diagnostics and therapy.

  11. Fluorescence diffuse tomography for tumor detection and monitoring

    Science.gov (United States)

    Balalaeva, Irina V.; Orlova, Anna G.; Shirmanova, Marina V.; Kibraeva, Elena A.; Zagainova, Elena V.; Turchin, Ilya V.

    2007-05-01

    Strong light scattering and absorption limit visualization of the internal structure of biological tissue. Only special tools for turbid media imaging, such as optical diffuse tomography, enable noninvasive investigation of the internal biological tissues, including visualization and intravital monitoring of deep tumors. In this work the preliminary results of fluorescence diffuse tomography (FDT) of small animals are presented. Using of exogenous fluorophores, targeted specifically at tumor cells, and fluorescent proteins expressed endogenously can significantly increase the contrast of obtained images. Fluorescent compounds of different nature, such as sulphonated aluminium phthalocyanine (Photosens), red fluorescing proteins and CdTe/CdSe-core/shell nanocrystals (quantum dots) were applied. The animal was scanned in the transilluminative configuration by low-frequency modulated light (1 kHz) from Nd:YAG laser with second harmonic generation at the wavelength of 532 nm or semiconductor laser at the wavelength of 655 nm. Photosens was injected intravenously into linear mice with metastazing Lewis lung carcinoma in dose 4 mg/kg. Quantum dots (5x10 -11 M) or protein DsRed2 (1-5x10 -6 M) in glass capsules (inner diameter 2-3 mm) were placed inside the esophagus of 7-day-old hairless rats (18-20 g) to simulate marked tumors. Cells of HEK-293 Phoenix line, transitory transfected with Turbo-RFP protein gene, were injected hypodermically to immunodeficient mice. This work demonstrates potential capabilities of FDT method for detection and monitoring of deep fluorescent-labeled tumors in animal models. Strong advantages of fluorescent proteins and quantum dots over the traditional photosensitizer for FDT imaging are shown.

  12. In vivo quantification of fluorescent molecular markers in real-time by ratio Imaging for diagnostic screening and image-guided surgery

    NARCIS (Netherlands)

    Bogaards, A.; Sterenborg, H. J. C. M.; Trachtenberg, J.; Wilson, B. C.; Lilge, L.

    2007-01-01

    Future applications of "molecular diagnostic screening" and "molecular image-guided surgery" will demand images of molecular markers with high resolution and high throughput (similar to >= 30 frames/second). MRI, SPECT, PET, optical fluorescence tomography, hyper-spectral fluorescence imaging, and

  13. Toward robust high resolution fluorescence tomography: a hybrid row-action edge preserving regularization

    Science.gov (United States)

    Behrooz, Ali; Zhou, Hao-Min; Eftekhar, Ali A.; Adibi, Ali

    2011-02-01

    Depth-resolved localization and quantification of fluorescence distribution in tissue, called Fluorescence Molecular Tomography (FMT), is highly ill-conditioned as depth information should be extracted from limited number of surface measurements. Inverse solvers resort to regularization algorithms that penalize Euclidean norm of the solution to overcome ill-posedness. While these regularization algorithms offer good accuracy, their smoothing effects result in continuous distributions which lack high-frequency edge-type features of the actual fluorescence distribution and hence limit the resolution offered by FMT. We propose an algorithm that penalizes the total variation (TV) norm of the solution to preserve sharp transitions and high-frequency components in the reconstructed fluorescence map while overcoming ill-posedness. The hybrid algorithm is composed of two levels: 1) An Algebraic Reconstruction Technique (ART), performed on FMT data for fast recovery of a smooth solution that serves as an initial guess for the iterative TV regularization, 2) A time marching TV regularization algorithm, inspired by the Rudin-Osher-Fatemi TV image restoration, performed on the initial guess to further enhance the resolution and accuracy of the reconstruction. The performance of the proposed method in resolving fluorescent tubes inserted in a liquid tissue phantom imaged by a non-contact CW trans-illumination FMT system is studied and compared to conventional regularization schemes. It is observed that the proposed method performs better in resolving fluorescence inclusions at higher depths.

  14. Fully integrated high-speed intravascular optical coherence tomography/near-infrared fluorescence structural/molecular imaging in vivo using a clinically available near-infrared fluorescence-emitting indocyanine green to detect inflamed lipid-rich atheromata in coronary-sized vessels.

    Science.gov (United States)

    Lee, Sunki; Lee, Min Woo; Cho, Han Saem; Song, Joon Woo; Nam, Hyeong Soo; Oh, Dong Joo; Park, Kyeongsoon; Oh, Wang-Yuhl; Yoo, Hongki; Kim, Jin Won

    2014-08-01

    Lipid-rich inflamed coronary plaques are prone to rupture. The purpose of this study was to assess lipid-rich inflamed plaques in vivo using fully integrated high-speed optical coherence tomography (OCT)/near-infrared fluorescence (NIRF) molecular imaging with a Food and Drug Administration-approved indocyanine green (ICG). An integrated high-speed intravascular OCT/NIRF imaging catheter and a dual-modal OCT/NIRF system were constructed based on a clinical OCT platform. For imaging lipid-rich inflamed plaques, the Food and Drug Administration-approved NIRF-emitting ICG (2.25 mg/kg) or saline was injected intravenously into rabbit models with experimental atheromata induced by balloon injury and 12- to 14-week high-cholesterol diets. Twenty minutes after injection, in vivo OCT/NIRF imaging of the infrarenal aorta and iliac arteries was acquired only under contrast flushing through catheter (pullback speed up to ≤20 mm/s). NIRF signals were strongly detected in the OCT-visualized atheromata of the ICG-injected rabbits. The in vivo NIRF target-to-background ratio was significantly larger in the ICG-injected rabbits than in the saline-injected controls (Pfluorescence reflectance imaging, which correlated well with the in vivo target-to-background ratios (Pfluorescence microscopy, and histopathology also corroborated the in vivo imaging findings. Integrated OCT/NIRF structural/molecular imaging with a Food and Drug Administration -approved ICG accurately identified lipid-rich inflamed atheromata in coronary-sized vessels. This highly translatable dual-modal imaging approach could enhance our capabilities to detect high-risk coronary plaques. © 2014 American Heart Association, Inc.

  15. Laser induced fluorescence of trapped molecular ions

    International Nuclear Information System (INIS)

    Winn, J.S.

    1980-10-01

    Laser induced fluoresence (LIF) spectra (laser excitation spectra) are conceptually among the most simple spectra to obtain. One need only confine a gaseous sample in a suitable container, direct a laser along one axis of the container, and monitor the sample's fluorescence at a right angle to the laser beam. As the laser wavelength is changed, the changes in fluorescence intensity map the absorption spectrum of the sample. (More precisely, only absorption to states which have a significant radiative decay component are monitored.) For ion spectroscopy, one could benefit in many ways by such an experiment. Most optical ion spectra have been observed by emission techniques, and, aside from the problems of spectral analysis, discharge emission methods often produce the spectra of many species, some of which may be unknown or uncertain. Implicit in the description of LIF given above is certainty as to the chemical identity of the carrier of the spectrum. This article describes a method by which the simplifying aspects of LIF can be extended to molecular ions

  16. The feasibility study on 3-dimensional fluorescent x-ray computed tomography using the pinhole effect for biomedical applications.

    Science.gov (United States)

    Sunaguchi, Naoki; Yuasa, Tetsuya; Hyodo, Kazuyuki; Zeniya, Tsutomu

    2013-01-01

    We propose a 3-dimensional fluorescent x-ray computed tomography (CT) pinhole collimator, aimed at providing molecular imaging with quantifiable measures and sub-millimeter spatial resolution. In this study, we demonstrate the feasibility of this concept and investigate imaging properties such as spatial resolution, contrast resolution and quantifiable measures, by imaging physical phantoms using a preliminary imaging system developed with monochromatic synchrotron x rays constructed at the BLNE-7A experimental line at KEK, Japan.

  17. Patch-based anisotropic diffusion scheme for fluorescence diffuse optical tomography--part 2: image reconstruction.

    Science.gov (United States)

    Correia, Teresa; Koch, Maximilian; Ale, Angelique; Ntziachristos, Vasilis; Arridge, Simon

    2016-02-21

    Fluorescence diffuse optical tomography (fDOT) provides 3D images of fluorescence distributions in biological tissue, which represent molecular and cellular processes. The image reconstruction problem is highly ill-posed and requires regularisation techniques to stabilise and find meaningful solutions. Quadratic regularisation tends to either oversmooth or generate very noisy reconstructions, depending on the regularisation strength. Edge preserving methods, such as anisotropic diffusion regularisation (AD), can preserve important features in the fluorescence image and smooth out noise. However, AD has limited ability to distinguish an edge from noise. We propose a patch-based anisotropic diffusion regularisation (PAD), where regularisation strength is determined by a weighted average according to the similarity between patches around voxels within a search window, instead of a simple local neighbourhood strategy. However, this method has higher computational complexity and, hence, we wavelet compress the patches (PAD-WT) to speed it up, while simultaneously taking advantage of the denoising properties of wavelet thresholding. Furthermore, structural information can be incorporated into the image reconstruction with PAD-WT to improve image quality and resolution. In this case, the weights used to average voxels in the image are calculated using the structural image, instead of the fluorescence image. The regularisation strength depends on both structural and fluorescence images, which guarantees that the method can preserve fluorescence information even when it is not structurally visible in the anatomical images. In part 1, we tested the method using a denoising problem. Here, we use simulated and in vivo mouse fDOT data to assess the algorithm performance. Our results show that the proposed PAD-WT method provides high quality and noise free images, superior to those obtained using AD.

  18. Fluorescence-Guided Probes of Aptamer-Targeted Gold Nanoparticles with Computed Tomography Imaging Accesses for in Vivo Tumor Resection.

    Science.gov (United States)

    Li, Cheng-Hung; Kuo, Tsung-Rong; Su, Hsin-Jan; Lai, Wei-Yun; Yang, Pan-Chyr; Chen, Jinn-Shiun; Wang, Di-Yan; Wu, Yi-Chun; Chen, Chia-Chun

    2015-10-28

    Recent development of molecular imaging probes for fluorescence-guided surgery has shown great progresses for determining tumor margin to execute the tissue resection. Here we synthesize the fluorescent gold nanoparticles conjugated with diatrizoic acid and nucleolin-targeted AS1411 aptamer. The nanoparticle conjugates exhibit high water-solubility, good biocompatibility, visible fluorescence and strong X-ray attenuation for computed tomography (CT) contrast enhancement. The fluorescent nanoparticle conjugates are applied as a molecular contrast agent to reveal the tumor location in CL1-5 tumor-bearing mice by CT imaging. Furthermore, the orange-red fluorescence emitting from the conjugates in the CL1-5 tumor can be easily visualized by the naked eyes. After the resection, the IVIS measurements show that the fluorescence signal of the nanoparticle conjugates in the tumor is greatly enhanced in comparison to that in the controlled experiment. Our work has shown potential application of functionalized nanoparticles as a dual-function imaging agent in clinical fluorescence-guided surgery.

  19. Polarization of fluorescence: a probe of molecular autoionization

    International Nuclear Information System (INIS)

    Leroi, G.E.; Dehmer, J.L.; Parr, A.C.; Poliakoff, E.D.

    1983-01-01

    The polarization of fluorescence from excited-state molecular photoions provides a direct probe of the photoionization dynamics and the symmetry signatures of autoionizing resonances. Measurements on CO 2 and CS 2 are presented as examples

  20. Three-dimensional simultaneous optical coherence tomography and confocal fluorescence microscopy for investigation of lung tissue.

    Science.gov (United States)

    Gaertner, Maria; Cimalla, Peter; Meissner, Sven; Kuebler, Wolfgang M; Koch, Edmund

    2012-07-01

    Although several strategies exist for a minimal-invasive treatment of patients with lung failure, the mortality rate of acute respiratory distress syndrome still reaches 30% at minimum. This striking number indicates the necessity of understanding lung dynamics on an alveolar level. To investigate the dynamical behavior on a microscale, we used three-dimensional geometrical and functional imaging to observe tissue parameters including alveolar size and length of embedded elastic fibers during ventilation. We established a combined optical coherence tomography (OCT) and confocal fluorescence microscopy system that is able to monitor the distension of alveolar tissue and elastin fibers simultaneously within three dimensions. The OCT system can laterally resolve a 4.9 μm line pair feature and has an approximately 11 μm full-width-half-maximum axial resolution in air. confocal fluorescence microscopy visualizes molecular properties of the tissue with a resolution of 0.75 μm (laterally), and 5.9 μm (axially) via fluorescence detection of the dye sulforhodamine B specifically binding to elastin. For system evaluation, we used a mouse model in situ to perform lung distension by application of different constant pressure values within the physiological regime. Our method enables the investigation of alveolar dynamics by helping to reveal basic processes emerging during artificial ventilation and breathing.

  1. Validating Intravascular Imaging with Serial Optical Coherence Tomography and Confocal Fluorescence Microscopy.

    Science.gov (United States)

    Tardif, Pier-Luc; Bertrand, Marie-Jeanne; Abran, Maxime; Castonguay, Alexandre; Lefebvre, Joël; Stähli, Barbara E; Merlet, Nolwenn; Mihalache-Avram, Teodora; Geoffroy, Pascale; Mecteau, Mélanie; Busseuil, David; Ni, Feng; Abulrob, Abedelnasser; Rhéaume, Éric; L'Allier, Philippe; Tardif, Jean-Claude; Lesage, Frédéric

    2016-12-15

    Atherosclerotic cardiovascular diseases are characterized by the formation of a plaque in the arterial wall. Intravascular ultrasound (IVUS) provides high-resolution images allowing delineation of atherosclerotic plaques. When combined with near infrared fluorescence (NIRF), the plaque can also be studied at a molecular level with a large variety of biomarkers. In this work, we present a system enabling automated volumetric histology imaging of excised aortas that can spatially correlate results with combined IVUS/NIRF imaging of lipid-rich atheroma in cholesterol-fed rabbits. Pullbacks in the rabbit aortas were performed with a dual modality IVUS/NIRF catheter developed by our group. Ex vivo three-dimensional (3D) histology was performed combining optical coherence tomography (OCT) and confocal fluorescence microscopy, providing high-resolution anatomical and molecular information, respectively, to validate in vivo findings. The microscope was combined with a serial slicer allowing for the imaging of the whole vessel automatically. Colocalization of in vivo and ex vivo results is demonstrated. Slices can then be recovered to be tested in conventional histology.

  2. Three-dimensional in vivo fluorescence diffuse optical tomography of breast cancer in humans

    Science.gov (United States)

    Corlu, Alper; Choe, Regine; Durduran, Turgut; Rosen, Mark A.; Schweiger, Martin; Arridge, Simon R.; Schnall, Mitchell D.; Yodh, Arjun G.

    2007-05-01

    We present three-dimensional (3D) in vivo images of human breast cancer based on fluorescence diffuse optical tomography (FDOT). To our knowledge, this work represents the first reported 3D fluorescence tomography of human breast cancer in vivo. In our protocol, the fluorophore Indocyanine Green (ICG) is injected intravenously. Fluorescence excitation and detection are accomplished in the soft-compression, parallel-plane, transmission geometry using laser sources at 786 nm and spectrally filtered CCD detection. Phantom and in vivo studies confirm the signals are due to ICG fluorescence, rather than tissue autofluorescence and excitation light leakage. Fluorescence images of breast tumors were in good agreement with those of MRI, and with DOT based on endogenous contrast. Tumorto- normal tissue contrast based on ICG fluorescence was two-to-four-fold higher than contrast based on hemoglobin and scattering parameters. In total the measurements demonstrate that FDOT of breast cancer is feasible and promising.

  3. Multiple X-ray tomography using transmitted, scattered and fluorescent radiation

    International Nuclear Information System (INIS)

    Cesareo, R.; Brunetti, A.; Golosio, B.; Lopes, R.T.; Barroso, R.C.; Donativi, M.; Castellano, A.; Quarta, S.

    2003-01-01

    A multiple CT-scanner is described, which contemporaneously uses transmitted, scattered and fluorescent X-rays for Imaging. The scanner is characterized by a small size X-ray tube and by four detectors: a ''pencil'' X-ray NaI(Tl) for transmitted tomography, a larger size NaI(Tl) for 90 C o Compton tomography, a thermoelectrically cooled Si-PIN or CdZnTe for fluorescent imaging and a CdZnTe for Rayleigh (or diffraction) tomography. Examples of applications are shown

  4. Molecular Iodine Fluorescence Using a Green Helium-Neon Laser

    Science.gov (United States)

    Williamson, J. Charles

    2011-01-01

    Excitation of molecular iodine vapor with a green (543.4 nm) helium-neon laser produces a fluorescence spectrum that is well suited for the upper-level undergraduate physical chemistry laboratory. Application of standard evaluation techniques to the spectrum yields ground electronic-state molecular parameters in good agreement with literature…

  5. In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue.

    Science.gov (United States)

    Kantelhardt, Sven R; Kalasauskas, Darius; König, Karsten; Kim, Ella; Weinigel, Martin; Uchugonova, Aisada; Giese, Alf

    2016-05-01

    High resolution multiphoton tomography and fluorescence lifetime imaging differentiates glioma from adjacent brain in native tissue samples ex vivo. Presently, multiphoton tomography is applied in clinical dermatology and experimentally. We here present the first application of multiphoton and fluorescence lifetime imaging for in vivo imaging on humans during a neurosurgical procedure. We used a MPTflex™ Multiphoton Laser Tomograph (JenLab, Germany). We examined cultured glioma cells in an orthotopic mouse tumor model and native human tissue samples. Finally the multiphoton tomograph was applied to provide optical biopsies during resection of a clinical case of glioblastoma. All tissues imaged by multiphoton tomography were sampled and processed for conventional histopathology. The multiphoton tomograph allowed fluorescence intensity- and fluorescence lifetime imaging with submicron spatial resolution and 200 picosecond temporal resolution. Morphological fluorescence intensity imaging and fluorescence lifetime imaging of tumor-bearing mouse brains and native human tissue samples clearly differentiated tumor and adjacent brain tissue. Intraoperative imaging was found to be technically feasible. Intraoperative image quality was comparable to ex vivo examinations. To our knowledge we here present the first intraoperative application of high resolution multiphoton tomography and fluorescence lifetime imaging of human brain tumors in situ. It allowed in vivo identification and determination of cell density of tumor tissue on a cellular and subcellular level within seconds. The technology shows the potential of rapid intraoperative identification of native glioma tissue without need for tissue processing or staining.

  6. Fluorescent and Colorimetric Molecular Recognition Probe for Hydrogen Bond Acceptors

    OpenAIRE

    Pike, Sarah Jane; Hunter, Christopher Alexander

    2018-01-01

    The association constants for formation of 1 : 1 complexes between a H-bond donor, 1-naphthol, and a diverse range of charged and neutral H-bond acceptors have been measured using UV/vis absorption and fluorescence emission titrations. The performance of 1-naphthol as a dual colorimetric and fluorescent molecular recognition probe for determining the H-bond acceptor (HBA) parameters of charged and neutral solutes has been investigated in three solvents. The data were employed to establish sel...

  7. Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging

    International Nuclear Information System (INIS)

    Ntziachristos, Vasilis; Bremer, Christoph; Weissleder, Ralph

    2003-01-01

    A recent development in biomedical imaging is the non-invasive mapping of molecular events in intact tissues using fluorescence. Underpinning to this development is the discovery of bio-compatible, specific fluorescent probes and proteins and the development of highly sensitive imaging technologies for in vivo fluorescent detection. Of particular interest are fluorochromes that emit in the near infrared (NIR), a spectral window, whereas hemoglobin and water absorb minimally so as to allow photons to penetrate for several centimetres in tissue. In this review article we concentrate on optical imaging technologies used for non-invasive imaging of the distribution of such probes. We illuminate the advantages and limitations of simple photographic methods and turn our attention to fluorescence-mediated molecular tomography (FMT), a technique that can three-dimensionally image gene expression by resolving fluorescence activation in deep tissues. We describe theoretical specifics, and we provide insight into its in vivo capacity and the sensitivity achieved. Finally, we discuss its clinical feasibility. (orig.)

  8. Highly Selective Fluorescent Sensing of Proteins Based on a Fluorescent Molecularly Imprinted Nanosensor

    Directory of Open Access Journals (Sweden)

    Shuo Wang

    2013-09-01

    Full Text Available A fluorescent molecularly imprinted nanosensor was obtained by grafting imprinted polymer onto the surface of multi-wall carbon nanotubes and post-imprinting treatment with fluorescein isothiocyanate (FITC. The fluorescence of lysozyme-imprinted polymer (Lys-MIP was quenched more strongly by Lys than that of nonimprinted polymer (NIP, which indicated that the Lys-MIP could recognize Lys. The resulted imprinted material has the ability to selectively sense a target protein, and an imprinting factor of 3.34 was achieved. The Lys-MIP also showed selective detection for Lys among other proteins such as cytochrome C (Cyt C, hemoglobin (HB and bovine serum albumin (BSA due to the imprinted sites in the Lys-MIP. This approach combines the high selectivity of surface molecular imprinting technology and fluorescence, and converts binding events into detectable signals by monitoring fluorescence spectra. Therefore, it will have further applications for Lys sensing.

  9. Polarized X-ray excitation for scatter reduction in X-ray fluorescence computed tomography.

    Science.gov (United States)

    Vernekohl, Don; Tzoumas, Stratis; Zhao, Wei; Xing, Lei

    2018-05-25

    X-ray fluorescence computer tomography (XFCT) is a new molecular imaging modality which uses X-ray excitation to stimulate the emission of fluorescent photons in high atomic number contrast agents. Scatter contamination is one of the main challenges in XFCT imaging which limits the molecular sensitivity. When polarized X-rays are used, it is possible to reduce the scatter contamination significantly by placing detectors perpendicular to the polarization direction. This study quantifies scatter contamination for polarized and unpolarized X-ray excitation and determines the advantages of scatter reduction. The amount of scatter in preclinical XFCT is quantified in Monte Carlo simulations. The fluorescent X-rays are emitted isotropically, while scattered X-rays propagate in polarization direction. The magnitude of scatter contamination is studied in XFCT simulations of a mouse phantom. In this study, the contrast agent gold is examined as an example but a scatter reduction from polarized excitation is also expected for other elements. The scatter reduction capability is examined for different polarization intensities with a monoenergetic X-ray excitation energy of 82 keV. The study evaluates two different geometrical shapes of CZT detectors which are modeled with an energy resolution of 1 keV FWHM at an X-ray energy of 80 keV. Benefits of a detector placement perpendicular to the polarization direction are shown in iterative and analytic image reconstruction including scatter correction. The contrast to noise ratio (CNR) and the normalized mean square error (NMSE) are analyzed and compared for the reconstructed images. A substantial scatter reduction for common detector sizes was achieved for 100% and 80% linear polarization while lower polarization intensities provide a decreased scatter reduction. By placing the detector perpendicular to the polarization direction, a scatter reduction by factor up to 5.5 can be achieved for common detector sizes. The image

  10. Frequency domain fluorescent diffuse tomography of small animals with DsRed2-expressed tumors

    Science.gov (United States)

    Turchin, Ilya V.; Savitsky, Alexander P.; Kamensky, Vladislav A.; Plehanov, Vladimir I.; Orlova, Anna G.; Sergeeva, Ekaterina A.; Kleshnin, Mikhail S.; Shirmanova, Marina V.

    2006-02-01

    The main applications of fluorescent proteins (FPs) are monitoring tumor growth, angiogenesis, metastases formation and effects of new classes of drugs. Different types of tomography allow fluorescence imaging of tumors located deep in human or animal tissue. These techniques were used for investigation of the distribution of near-infrared fluorescent probes, but only a few works are devoted to fluorescence tomography in visible light. In this work, preliminary results of the frequency domain fluorescent diffuse tomography (FD FDT) method in application to DsRed2 protein as a fluorescent agent are presented. For the first step of our experiments we utilized second harmonic generation of Nd:YAG laser (532 nm) modulated by low frequency (1 kHz) in the experimental setup. The transilluminative planar configuration was used in the setup. A series of model experiments has been conducted and show good agreement between theoretical and experimental fluorescence intensity. Post mortem experiments with capsules containing DsRed2 and scattering solution introduced into esophagus of rats to simulate tumor formation have been conducted. The results of these experiments show that sensitivity of the setup is sufficient to detect DsRed2 in concentrations similar to those in FP-expressed tumor, but the contrast is not enough high to separate fluorescence of DsRed2 and surrounding tissues. The setup can be significantly improved by utilizing high-frequency modulation (110 MHz using acousto-optical modulator) of the excitation light and precise phase measurements due to difference in fluorescence life-time of FPs and surrounding tissues. An algorithm of processing a fluorescent image based on calculating zero of maximum curvature was employed for detection of fluorescent inclusions boundaries in the image.

  11. Protein recognition by a pattern-generating fluorescent molecular probe

    Science.gov (United States)

    Pode, Zohar; Peri-Naor, Ronny; Georgeson, Joseph M.; Ilani, Tal; Kiss, Vladimir; Unger, Tamar; Markus, Barak; Barr, Haim M.; Motiei, Leila; Margulies, David

    2017-12-01

    Fluorescent molecular probes have become valuable tools in protein research; however, the current methods for using these probes are less suitable for analysing specific populations of proteins in their native environment. In this study, we address this gap by developing a unimolecular fluorescent probe that combines the properties of small-molecule-based probes and cross-reactive sensor arrays (the so-called chemical 'noses/tongues'). On the one hand, the probe can detect different proteins by generating unique identification (ID) patterns, akin to cross-reactive arrays. On the other hand, its unimolecular scaffold and selective binding enable this ID-generating probe to identify combinations of specific protein families within complex mixtures and to discriminate among isoforms in living cells, where macroscopic arrays cannot access. The ability to recycle the molecular device and use it to track several binding interactions simultaneously further demonstrates how this approach could expand the fluorescent toolbox currently used to detect and image proteins.

  12. Preclinical, fluorescence and diffuse optical tomography: non-contact instrumentation, modeling and time-resolved 3D reconstruction

    International Nuclear Information System (INIS)

    Nouizi, F.

    2011-09-01

    Time-Resolved Diffuse Optical Tomography (TR-DOT) is a new non-invasive imaging technique increasingly used in the clinical and preclinical fields. It yields optical absorption and scattering maps of the explored organs, and related physiological parameters. Time-Resolved Fluorescence Diffuse Optical Tomography (TR-FDOT) is based on the detection of fluorescence photons. It provides spatio-temporal maps of fluorescent probe concentrations and life times, and allows access to metabolic and molecular imaging which is important for diagnosis and therapeutic monitoring, particularly in oncology. The main goal of this thesis was to reconstruct 3D TR-DOT/TR-FDOT images of small animals using time-resolved optical technology. Data were acquired using optical fibers fixed around the animal without contact with its surface. The work was achieved in four steps: 1)- Setting up an imaging device to record the 3D coordinates of an animal's surface; 2)- Modeling the no-contact approach to solve the forward problem; 3)- Processing of the measured signals taking into account the impulse response of the device; 4)- Implementation of a new image reconstruction method based on a selection of carefully chosen points. As a result, good-quality 3D optical images were obtained owing to reduced cross-talk between absorption and scattering. Moreover, the computation time was cut down, compared to full-time methods using whole temporal profiles. (author)

  13. Multimodality Imaging Probe for Positron Emission Tomography and Fluorescence Imaging Studies

    Directory of Open Access Journals (Sweden)

    Suresh K. Pandey

    2014-05-01

    Full Text Available Our goal is to develop multimodality imaging agents for use in cell tracking studies by positron emission tomography (PET and optical imaging (OI. For this purpose, bovine serum albumin (BSA was complexed with biotin (histologic studies, 5(6- carboxyfluorescein, succinimidyl ester (FAM SE (OI studies, and diethylenetriamine pentaacetic acid (DTPA for chelating gallium 68 (PET studies. For synthesis of BSA-biotin-FAM-DTPA, BSA was coupled to (+-biotin N-hydroxysuccinimide ester (biotin-NHSI. BSA- biotin was treated with DTPA-anhydride and biotin-BSA-DTPA was reacted with FAM. The biotin-BSA-DTPA-FAM was reacted with gallium chloride 3 to 5 mCi eluted from the generator using 0.1 N HCl and was passed through basic resin (AG 11 A8 and 150 mCi (100 μL, pH 7–8 was incubated with 0.1 mg of FAM conjugate (100 μL at room temperature for 15 minutes to give 66Ga-BSA-biotin-DTPA-FAM. A shaved C57 black mouse was injected with FAM conjugate (50 μL at one flank and FAM-68Ga (50 μL, 30 mCi at the other. Immediately after injection, the mouse was placed in a fluorescence imaging system (Kodak In-Vivo F, Bruker Biospin Co., Woodbridge, CT and imaged (Λex: 465 nm, Λem: 535 nm, time: 8 seconds, Xenon Light Source, Kodak. The same mouse was then placed under an Inveon microPET scanner (Siemens Medical Solutions, Knoxville, TN injected (intravenously with 25 μCi of 18F and after a half-hour (to allow sufficient bone uptake was imaged for 30 minutes. Molecular weight determined using matrix-associated laser desorption ionization (MALDI for the BSA sample was 66,485 Da and for biotin-BSA was 67,116 Da, indicating two biotin moieties per BSA molecule; for biotin-BSA-DTPA was 81,584 Da, indicating an average of 30 DTPA moieties per BSA molecule; and for FAM conjugate was 82,383 Da, indicating an average of 1.7 fluorescent moieties per BSA molecule. Fluorescence imaging clearly showed localization of FAM conjugate and FAM-68Ga at respective flanks of the mouse

  14. Review of X-ray Tomography and X-ray Fluorescence Spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Shear, Trevor A. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-03-16

    This literature review will focus on both laboratory and synchrotron based X-ray tomography of materials and highlight the inner workings of these instruments. X-ray fluorescence spectroscopy will also be reviewed and applications of the tandem use of these techniques will be explored. The real world application of these techniques during the internship will also be discussed.

  15. Enhancing early bladder cancer detection with fluorescence-guided endoscopic optical coherence tomography

    Science.gov (United States)

    Pan, Y. T.; Xie, T. Q.; Du, C. W.; Bastacky, S.; Meyers, S.; Zeidel, M. L.

    2003-12-01

    We report an experimental study of the possibility of enhancing early bladder cancer diagnosis with fluorescence-image-guided endoscopic optical coherence tomography (OCT). After the intravesical instillation of a 10% solution of 5-aminolevulinic acid, simultaneous fluorescence imaging (excitation of 380-420 nm, emission of 620-700 nm) and OCT are performed on rat bladders to identify the photochemical and morphological changes associated with uroepithelial tumorigenesis. The preliminary results of our ex vivo study reveal that both fluorescence and OCT can identify early uroepithelial cancers, and OCT can detect precancerous lesions (e.g., hyperplasia) that fluorescence may miss. This suggests that a cystoscope combining 5-aminolevulinic acid fluorescence and OCT imaging has the potential to enhance the efficiency and sensitivity of early bladder cancer diagnosis.

  16. Environment-sensitive behavior of fluorescent molecular rotors

    Directory of Open Access Journals (Sweden)

    Theodorakis Emmanuel A

    2010-09-01

    Full Text Available Abstract Molecular rotors are a group of fluorescent molecules that form twisted intramolecular charge transfer (TICT states upon photoexcitation. When intramolecular twisting occurs, the molecular rotor returns to the ground state either by emission of a red-shifted emission band or by nonradiative relaxation. The emission properties are strongly solvent-dependent, and the solvent viscosity is the primary determinant of the fluorescent quantum yield from the planar (non-twisted conformation. This viscosity-sensitive behavior gives rise to applications in, for example, fluid mechanics, polymer chemistry, cell physiology, and the food sciences. However, the relationship between bulk viscosity and the molecular-scale interaction of a molecular rotor with its environment are not fully understood. This review presents the pertinent theories of the rotor-solvent interaction on the molecular level and how this interaction leads to the viscosity-sensitive behavior. Furthermore, current applications of molecular rotors as microviscosity sensors are reviewed, and engineering aspects are presented on how measurement accuracy and precision can be improved.

  17. Time reversal optical tomography locates fluorescent targets in a turbid medium

    Science.gov (United States)

    Wu, Binlin; Cai, W.; Gayen, S. K.

    2013-03-01

    A fluorescence optical tomography approach that extends time reversal optical tomography (TROT) to locate fluorescent targets embedded in a turbid medium is introduced. It uses a multi-source illumination and multi-detector signal acquisition scheme, along with TR matrix formalism, and multiple signal classification (MUSIC) to construct pseudo-image of the targets. The samples consisted of a single or two small tubes filled with water solution of Indocyanine Green (ICG) dye as targets embedded in a 250 mm × 250 mm × 60 mm rectangular cell filled with Intralipid-20% suspension as the scattering medium. The ICG concentration was 1μM, and the Intralipid-20% concentration was adjusted to provide ~ 1-mm transport length for both excitation wavelength of 790 nm and fluorescence wavelength around 825 nm. The data matrix was constructed using the diffusely transmitted fluorescence signals for all scan positions, and the TR matrix was constructed by multiplying data matrix with its transpose. A pseudo spectrum was calculated using the signal subspace of the TR matrix. Tomographic images were generated using the pseudo spectrum. The peaks in the pseudo images provided locations of the target(s) with sub-millimeter accuracy. Concurrent transmission TROT measurements corroborated fluorescence-TROT findings. The results demonstrate that TROT is a fast approach that can be used to obtain accurate three-dimensional position information of fluorescence targets embedded deep inside a highly scattering medium, such as, a contrast-enhanced tumor in a human breast.

  18. Molecular engineering and fluorescence for the detection of toxic cations

    International Nuclear Information System (INIS)

    Souchon, V.

    2007-11-01

    This work is a part of the 'Toxicologie Nucleaire Environnementale' program which aims at studying the effects on the living of heavy metals or radionuclides involved in nuclear industry. Most particularly, it deals with the design of new fluorescent sensors for the selective detection of Pb 2+ , Cd 2+ and Cs + in biological media. Several fluorescent calixarenes possessing nitrogen atoms were synthesized and their properties as potential lead sensors were investigated. One of them could be used in experimental conditions close to biological media and new target compounds with amide functional groups were proposed. Many approaches were considered for the design of selective fluorescent sensors for cadmium. On the basis of literature results, many chelating compounds incorporating sulfur atoms were synthesized but showed no significant affinity towards cadmium. On the opposite, compounds functionalized with several pyridine-2'-yl-1,2,3-triazol fluorescent moieties linked to a β-cyclodextrin or a calix[4]arene showed good affinity for cadmium in methanol, but the selectivity was found to be insufficient. In contrast, very satisfying results in terms of both selectivity and sensitivity could be obtained with the commercial calcium sensor Rhod-5N in an aqueous medium at neutral pH. Lastly, micromolar detection limits for the selective detection of caesium were reached in an aqueous medium at neutral pH thanks to a new sulfonated fluorescent calixarene with two appended crown-ethers. An original complexation mechanism was proposed and validated by molecular modelling (DFT). (author)

  19. Fluorescence excitation studies of molecular photoionization in external electric fields

    International Nuclear Information System (INIS)

    Poliakoff, E.D.; Dehmer, J.L.; Parr, A.C.; Leroi, G.E.

    1985-01-01

    Using molecular nitrogen as an example, we show that fluorescence excitation spectroscopy can be used to measure partial photoionization cross sections of free molecules in external electric fields. The production of the N 2 + (B 2 Σ/sub u/ + ) state was studied and the threshold for this process was found to shift linearly with the square root of the applied field. This behavior is compared with the hydrogenic case and with previously studied systems

  20. Multimodal optical coherence tomography and fluorescence lifetime imaging with interleaved excitation sources for simultaneous endogenous and exogenous fluorescence.

    Science.gov (United States)

    Shrestha, Sebina; Serafino, Michael J; Rico-Jimenez, Jesus; Park, Jesung; Chen, Xi; Zhaorigetu, Siqin; Walton, Brian L; Jo, Javier A; Applegate, Brian E

    2016-09-01

    Multimodal imaging probes a variety of tissue properties in a single image acquisition by merging complimentary imaging technologies. Exploiting synergies amongst the data, algorithms can be developed that lead to better tissue characterization than could be accomplished by the constituent imaging modalities taken alone. The combination of optical coherence tomography (OCT) with fluorescence lifetime imaging microscopy (FLIM) provides access to detailed tissue morphology and local biochemistry. The optical system described here merges 1310 nm swept-source OCT with time-domain FLIM having excitation at 355 and 532 nm. The pulses from 355 and 532 nm lasers have been interleaved to enable simultaneous acquisition of endogenous and exogenous fluorescence signals, respectively. The multimodal imaging system was validated using tissue phantoms. Nonspecific tagging with Alexa Flour 532 in a Watanbe rabbit aorta and active tagging of the LOX-1 receptor in human coronary artery, demonstrate the capacity of the system for simultaneous acquisition of OCT, endogenous FLIM, and exogenous FLIM in tissues.

  1. Photoacoustic tomography of human hepatic malignancies using intraoperative indocyanine green fluorescence imaging.

    Science.gov (United States)

    Miyata, Akinori; Ishizawa, Takeaki; Kamiya, Mako; Shimizu, Atsushi; Kaneko, Junichi; Ijichi, Hideaki; Shibahara, Junji; Fukayama, Masashi; Midorikawa, Yutaka; Urano, Yasuteru; Kokudo, Norihiro

    2014-01-01

    Recently, fluorescence imaging following the preoperative intravenous injection of indocyanine green has been used in clinical settings to identify hepatic malignancies during surgery. The aim of this study was to evaluate the ability of photoacoustic tomography using indocyanine green as a contrast agent to produce representative fluorescence images of hepatic tumors by visualizing the spatial distribution of indocyanine green on ultrasonographic images. Indocyanine green (0.5 mg/kg, intravenous) was preoperatively administered to 9 patients undergoing hepatectomy. Intraoperatively, photoacoustic tomography was performed on the surface of the resected hepatic specimens (n = 10) under excitation with an 800 nm pulse laser. In 4 hepatocellular carcinoma nodules, photoacoustic imaging identified indocyanine green accumulation in the cancerous tissue. In contrast, in one hepatocellular carcinoma nodule and five adenocarcinoma foci (one intrahepatic cholangiocarcinoma and 4 colorectal liver metastases), photoacoustic imaging delineated indocyanine green accumulation not in the cancerous tissue but rather in the peri-cancerous hepatic parenchyma. Although photoacoustic tomography enabled to visualize spatial distribution of ICG on ultrasonographic images, which was consistent with fluorescence images on cut surfaces of the resected specimens, photoacoustic signals of ICG-containing tissues decreased approximately by 40% even at 4 mm depth from liver surfaces. Photoacoustic tomography using indocyanine green also failed to identify any hepatocellular carcinoma nodules from the body surface of model mice with non-alcoholic steatohepatitis. In conclusion, photoacoustic tomography has a potential to enhance cancer detectability and differential diagnosis by ultrasonographic examinations and intraoperative fluorescence imaging through visualization of stasis of bile-excreting imaging agents in and/or around hepatic tumors. However, further technical advances are needed

  2. Photoacoustic tomography of human hepatic malignancies using intraoperative indocyanine green fluorescence imaging.

    Directory of Open Access Journals (Sweden)

    Akinori Miyata

    Full Text Available Recently, fluorescence imaging following the preoperative intravenous injection of indocyanine green has been used in clinical settings to identify hepatic malignancies during surgery. The aim of this study was to evaluate the ability of photoacoustic tomography using indocyanine green as a contrast agent to produce representative fluorescence images of hepatic tumors by visualizing the spatial distribution of indocyanine green on ultrasonographic images. Indocyanine green (0.5 mg/kg, intravenous was preoperatively administered to 9 patients undergoing hepatectomy. Intraoperatively, photoacoustic tomography was performed on the surface of the resected hepatic specimens (n = 10 under excitation with an 800 nm pulse laser. In 4 hepatocellular carcinoma nodules, photoacoustic imaging identified indocyanine green accumulation in the cancerous tissue. In contrast, in one hepatocellular carcinoma nodule and five adenocarcinoma foci (one intrahepatic cholangiocarcinoma and 4 colorectal liver metastases, photoacoustic imaging delineated indocyanine green accumulation not in the cancerous tissue but rather in the peri-cancerous hepatic parenchyma. Although photoacoustic tomography enabled to visualize spatial distribution of ICG on ultrasonographic images, which was consistent with fluorescence images on cut surfaces of the resected specimens, photoacoustic signals of ICG-containing tissues decreased approximately by 40% even at 4 mm depth from liver surfaces. Photoacoustic tomography using indocyanine green also failed to identify any hepatocellular carcinoma nodules from the body surface of model mice with non-alcoholic steatohepatitis. In conclusion, photoacoustic tomography has a potential to enhance cancer detectability and differential diagnosis by ultrasonographic examinations and intraoperative fluorescence imaging through visualization of stasis of bile-excreting imaging agents in and/or around hepatic tumors. However, further technical

  3. Human thyroid specimen imaging by fluorescent x-ray computed tomography with synchrotron radiation

    Science.gov (United States)

    Takeda, Tohoru; Yu, Quanwen; Yashiro, Toru; Yuasa, Tetsuya; Hasegawa, Yasuo; Itai, Yuji; Akatsuka, Takao

    1999-09-01

    Fluorescent x-ray computed tomography (FXCT) is being developed to detect non-radioactive contrast materials in living specimens. The FXCT system consists of a silicon (111) channel cut monochromator, an x-ray slit and a collimator for fluorescent x ray detection, a scanning table for the target organ and an x-ray detector for fluorescent x-ray and transmission x-ray. To reduce Compton scattering overlapped on the fluorescent K(alpha) line, incident monochromatic x-ray was set at 37 keV. The FXCT clearly imaged a human thyroid gland and iodine content was estimated quantitatively. In a case of hyperthyroidism, the two-dimensional distribution of iodine content was not uniform, and thyroid cancer had a small amount of iodine. FXCT can be used to detect iodine within thyroid gland quantitatively and to delineate its distribution.

  4. Development of Ultrasonic Modulation Probe for Fluorescence Tomography Based on Acousto-Optic Effect

    Directory of Open Access Journals (Sweden)

    Trinh Quang Duc

    2011-01-01

    Full Text Available We have developed an ultrasonic probe for fluorescence modulation to image fluorescence within biological tissues. The probe consists of a focused ultrasonic transducer mounted on actuators for mechanical fan scanning, which can be used in contact with the measuring object aiming for clinical application. The mechanical fan scanning employed in the probe has a beneficial feature of portability. As a result, fluorescent beads, which were localized with the diameter of 2 mm at 20 mm depth in a pork meat tissue, were detected with resolution of 3 mm. The system performance denotes the feasibility of development towards the final goal of ultrasonic fluorescence modulation tomography for clinical applications.

  5. A tumor-targeted polymer theranostics platform for positron emission tomography and fluorescence imaging

    Czech Academy of Sciences Publication Activity Database

    Koziolová, Eva; Goel, S.; Chytil, Petr; Janoušková, Olga; Barnhart, T. E.; Cai, W.; Etrych, Tomáš

    2017-01-01

    Roč. 9, č. 30 (2017), s. 10906-10918 ISSN 2040-3364 R&D Projects: GA ČR(CZ) GA15-02986S; GA MZd(CZ) NV16-28594A; GA MŠk(CZ) LO1507 Institutional support: RVO:61389013 Keywords : N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers * positron emission tomography ( PET ) * fluorescence imaging Subject RIV: CD - Macromolecular Chemistry OBOR OECD: Polymer science Impact factor: 7.367, year: 2016

  6. Fundus auto fluorescence and spectral domain ocular coherence tomography in the early detection of chloroquine retinopathy

    OpenAIRE

    Megan B. Goodman; Ari Ziskind

    2015-01-01

    Purpose: To determine the sensitivity of spectral domain ocular coherence tomography (SD-OCT) and fundus auto fluorescence (FAF) images as a screening test to detect early changes in the retina prior to the onset of chloroquine retinopathy. Method: The study was conducted using patients taking chloroquine (CQ), referred by the Rheumatology Department to the Ophthalmology Department at Tygerberg Academic Hospital. Group A consisted of 59 patients on CQ for less than 5 years, and Group B co...

  7. Use of multiphoton tomography and fluorescence lifetime imaging to investigate skin pigmentation in vivo

    Science.gov (United States)

    Dancik, Yuri; Favre, Amandine; Loy, Chong Jin; Zvyagin, Andrei V.; Roberts, Michael S.

    2013-02-01

    There is a growing body of literature showing the usefulness of multiphoton tomography (MPT) and fluorescence lifetime imaging for in situ characterization of skin constituents and the ensuing development of noninvasive diagnostic tools against skin diseases. Melanin and pigmentation-associated skin cancers constitute some of the major applications. We show that MPT and fluorescence lifetime imaging can be used to measure changes in cutaneous melanin concentration and that these can be related to the visible skin color. Melanin in the skin of African, Indian, Caucasian, and Asian volunteers is detected on the basis of its emission wavelength and fluorescence lifetimes in solution and in a melanocyte-keratinocyte cell culture. Fluorescence intensity is used to characterize the melanin content and distribution as a function of skin type and depth into the skin (stratum granulosum and stratum basale). The measured fluorescence intensities in given skin types agree with melanin amounts reported by others using biopsies. Our results suggest that spatial distribution of melanin in skin can be studied using MPT and fluorescence lifetime imaging, but further studies are needed to ascertain that the method can resolve melanin amount in smaller depth intervals.

  8. Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing

    International Nuclear Information System (INIS)

    Sakhalkar, H S; Dewhirst, M; Oliver, T; Cao, Y; Oldham, M

    2007-01-01

    Optical emission computed tomography (optical-ECT) is a technique for imaging the three-dimensional (3D) distribution of fluorescent probes in biological tissue specimens with high contrast and spatial resolution. In optical-ECT, functional information can be imaged by (i) systemic application of functional labels (e.g. fluorophore labelled proteins) and/or (ii) endogenous expression of fluorescent reporter proteins (e.g. red fluorescent protein (RFP), green fluorescent protein (GFP)) in vivo. An essential prerequisite for optical-ECT is optical clearing, a procedure where tissue specimens are made transparent to light by sequential perfusion with fixing, dehydrating and clearing agents. In this study, we investigate clearing protocols involving a selection of common fixing (4% buffered paraformaldehyde (PFA), methanol and ethanol), dehydrating (methanol and ethanol) and clearing agents (methyl salicylate and benzyl-alcohol-benzyl-benzoate (BABB)) in order to determine a 'fluorescence friendly' clearing procedure. Cell culture experiments were employed to optimize the sequence of chemical treatments that best preserve fluorescence. Texas red (TxRed), fluorescein isothiocyanate (FITC), RFP and GFP were tested as fluorophores and fluorescent reporter proteins of interest. Fluorescent and control cells were imaged on a microscope using a DSred2 and FITC filter set. The most promising clearing protocols of cell culture experiments were applied to whole xenograft tumour specimens, to test their effectiveness in large unsectioned samples. Fluorescence of TxRed/FITC fluorophores was not found to be significantly affected by any of the test clearing protocols. RFP and GFP fluorescence, however, was found to be significantly greater when cell fixation was in ethanol. Fixation in either PFA or methanol resulted in diminished fluorescence. After ethanol fixation, the RFP and GFP fluorescence proved remarkably robust to subsequent exposure to either methyl salicylate or BABB

  9. Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing

    Energy Technology Data Exchange (ETDEWEB)

    Sakhalkar, H S [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Dewhirst, M [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Oliver, T [Department of Cell Biology, Duke University Medical Center, Durham, NC 27710 (United States); Cao, Y [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Oldham, M [Department of Radiation Oncology Physics, and Biomedical Engineering, Duke University Medical Center, Durham, NC 27710 (United States)

    2007-04-21

    Optical emission computed tomography (optical-ECT) is a technique for imaging the three-dimensional (3D) distribution of fluorescent probes in biological tissue specimens with high contrast and spatial resolution. In optical-ECT, functional information can be imaged by (i) systemic application of functional labels (e.g. fluorophore labelled proteins) and/or (ii) endogenous expression of fluorescent reporter proteins (e.g. red fluorescent protein (RFP), green fluorescent protein (GFP)) in vivo. An essential prerequisite for optical-ECT is optical clearing, a procedure where tissue specimens are made transparent to light by sequential perfusion with fixing, dehydrating and clearing agents. In this study, we investigate clearing protocols involving a selection of common fixing (4% buffered paraformaldehyde (PFA), methanol and ethanol), dehydrating (methanol and ethanol) and clearing agents (methyl salicylate and benzyl-alcohol-benzyl-benzoate (BABB)) in order to determine a 'fluorescence friendly' clearing procedure. Cell culture experiments were employed to optimize the sequence of chemical treatments that best preserve fluorescence. Texas red (TxRed), fluorescein isothiocyanate (FITC), RFP and GFP were tested as fluorophores and fluorescent reporter proteins of interest. Fluorescent and control cells were imaged on a microscope using a DSred2 and FITC filter set. The most promising clearing protocols of cell culture experiments were applied to whole xenograft tumour specimens, to test their effectiveness in large unsectioned samples. Fluorescence of TxRed/FITC fluorophores was not found to be significantly affected by any of the test clearing protocols. RFP and GFP fluorescence, however, was found to be significantly greater when cell fixation was in ethanol. Fixation in either PFA or methanol resulted in diminished fluorescence. After ethanol fixation, the RFP and GFP fluorescence proved remarkably robust to subsequent exposure to either methyl salicylate

  10. A 3D imaging system integrating photoacoustic and fluorescence orthogonal projections for anatomical, functional and molecular assessment of rodent models

    Science.gov (United States)

    Brecht, Hans P.; Ivanov, Vassili; Dumani, Diego S.; Emelianov, Stanislav Y.; Anastasio, Mark A.; Ermilov, Sergey A.

    2018-03-01

    We have developed a preclinical 3D imaging instrument integrating photoacoustic tomography and fluorescence (PAFT) addressing known deficiencies in sensitivity and spatial resolution of the individual imaging components. PAFT is designed for simultaneous acquisition of photoacoustic and fluorescence orthogonal projections at each rotational position of a biological object, enabling direct registration of the two imaging modalities. Orthogonal photoacoustic projections are utilized to reconstruct large (21 cm3 ) volumes showing vascularized anatomical structures and regions of induced optical contrast with spatial resolution exceeding 100 µm. The major advantage of orthogonal fluorescence projections is significant reduction of background noise associated with transmitted or backscattered photons. The fluorescence imaging component of PAFT is used to boost detection sensitivity by providing low-resolution spatial constraint for the fluorescent biomarkers. PAFT performance characteristics were assessed by imaging optical and fluorescent contrast agents in tissue mimicking phantoms and in vivo. The proposed PAFT technology will enable functional and molecular volumetric imaging using fluorescent biomarkers, nanoparticles, and other photosensitive constructs mapped with high fidelity over robust anatomical structures, such as skin, central and peripheral vasculature, and internal organs.

  11. Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Breunig, Hans Georg; König, Karsten; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Christopher

    2011-07-01

    Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential.

  12. Counting constituents in molecular complexes by fluorescence photon antibunching

    Energy Technology Data Exchange (ETDEWEB)

    Fore, S; Laurence, T; Hollars, C; Huser, T

    2007-04-17

    Modern single molecule fluorescence microscopy offers new, highly quantitative ways of studying the systems biology of cells while keeping the cells healthy and alive in their natural environment. In this context, a quantum optical technique, photon antibunching, has found a small niche in the continuously growing applications of single molecule techniques to small molecular complexes. Here, we review some of the most recent applications of photon antibunching in biophotonics, and we provide a guide for how to conduct photon antibunching experiments at the single molecule level by applying techniques borrowed from time-correlated single photon counting. We provide a number of new examples for applications of photon antibunching to the study of multichromophoric molecules and small molecular complexes.

  13. Chemical point detection using differential fluorescence from molecularly imprinted polymers

    Science.gov (United States)

    Pestov, Dmitry; Anderson, John E.; Nelson, Jean; Tepper, Gary C.

    2004-12-01

    Fluorescence represents one of the most attractive approaches for chemical sensing due to the abundant light produced by most fluorophores, resulting in excellent detection sensitivity. However, the broad and overlapping emission spectra of target and background species have made it difficult to perform species identification in a field instrument because of the need to perform spectral decomposition and analysis. This paper describes a new chemical sensing strategy based on differential fluorescence measurements from molecularly imprinted polymers, which eliminates the need to perform any spectral analysis. Species identification is accomplished by measuring the differential light output from a pair of polymers-one imprinted to a target species and the other identical, but not imprinted. The imprinted polymer selectively concentrates the target molecule and controls the energy (wavelength) of the emitted fluorescence signal and the differential output eliminates common mode signals associated with non-specific background interference. Because no spectral analysis is required, the sensors can be made extremely small and require very little power. Preliminary performance parameters from a prototype sensor are presented and discussed.

  14. Fluorescence quantum yield measurements of fluorescent proteins: a laboratory experiment for a biochemistry or molecular biophysics laboratory course.

    Science.gov (United States)

    Wall, Kathryn P; Dillon, Rebecca; Knowles, Michelle K

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts absorbed photons into emitted photons and it is necessary to know for assessing what fluorescent protein is the most appropriate for a particular application. In this work, we have designed an upper-level, biochemistry laboratory experiment where students measure the fluorescence quantum yields of fluorescent proteins relative to a standard organic dye. Four fluorescent protein variants, enhanced cyan fluorescent protein (ECFP), enhanced green fluorescent protein (EGFP), mCitrine, and mCherry, were used, however the methods described are useful for the characterization of any fluorescent protein or could be expanded to fluorescent quantum yield measurements of organic dye molecules. The laboratory is designed as a guided inquiry project and takes two, 4 hr laboratory periods. During the first day students design the experiment by selecting the excitation wavelength, choosing the standard, and determining the concentration needed for the quantum yield experiment that takes place in the second laboratory period. Overall, this laboratory provides students with a guided inquiry learning experience and introduces concepts of fluorescence biophysics into a biochemistry laboratory curriculum. © 2014 The International Union of Biochemistry and Molecular Biology.

  15. Fluorescent and colorimetric molecular recognition probe for hydrogen bond acceptors.

    Science.gov (United States)

    Pike, Sarah J; Hunter, Christopher A

    2017-11-22

    The association constants for formation of 1 : 1 complexes between a H-bond donor, 1-naphthol, and a diverse range of charged and neutral H-bond acceptors have been measured using UV/vis absorption and fluorescence emission titrations. The performance of 1-naphthol as a dual colorimetric and fluorescent molecular recognition probe for determining the H-bond acceptor (HBA) parameters of charged and neutral solutes has been investigated in three solvents. The data were employed to establish self-consistent H-bond acceptor parameters (β) for benzoate, azide, chloride, thiocyanate anions, a series of phosphine oxides, phosphate ester, sulfoxide and a tertiary amide. The results demonstrate both the transferability of H-bond parameters between different solvents and the utility of the naphthol-based dual molecular recognition probe to exploit orthogonal spectroscopic techniques to determine the HBA properties of neutral and charged solutes. The benzoate anion is the strongest HBA studied with a β parameter of 15.4, and the neutral tertiary amide is the weakest H-bond acceptor investigated with a β parameter of 8.5. The H-bond acceptor strength of the azide anion is higher than that of chloride (12.8 and 12.2 respectively), and the thiocyanate anion has a β value of 10.8 and thus is a significantly weaker H-bond acceptor than both the azide and chloride anions.

  16. In vivo fluorescence enhanced optical tomography reconstruction of lung cancer of non immersed small animals

    Science.gov (United States)

    Hervé, L.; Koenig, A.; Da Silva, A.; Berger, M.; Boutet, J.; Dinten, J. M.; Peltié, P.; Rizo, P.

    2007-02-01

    Fluorescence enhanced diffuse optical tomography (fDOT) is envisioned to be useful to collect functional information from small animal models. For oncology applications, cancer-targeted fluorescent markers can be used as a surrogate of the cancer activity. We are developing a continuous wave fDOT bench intended to be integrated in systems dedicated to whole body small animal fluorescence analyses. The focus is currently put on the reconstruction of non immersed small animals imaged by a CCD camera. The reconstruction stage already corrects the tissue heterogeneity artifacts through the computation of an optical heterogeneity map. We will show how this formalism coupled with the determination of the animal boundaries performed by a laser scanner, can be used to manage non contact acquisitions. The time of reconstruction for a 10 × 9 laser source positions, 45 × 40 detector elements and 14 × 11 × 14 mesh voxels is typically 10 minutes on a 3GHz PCs corresponding to the acquisition time allowing the two tasks to be performed in parallel. The system is validated on an in vivo experiment performed on three healthy nude mice and a mouse bearing a lung tumor at 10, 12 and 14 days after implantation allowing the follow up of the disease. The 3D fluorescence reconstructions of this mouse are presented and the total fluorescence amounts are compared.

  17. Combined X-ray fluorescence and absorption computed tomography using a synchrotron beam

    International Nuclear Information System (INIS)

    Hall, C

    2013-01-01

    X-ray computed tomography (CT) and fluorescence X-ray computed tomography (FXCT) using synchrotron sources are both useful tools in biomedical imaging research. Synchrotron CT (SRCT) in its various forms is considered an important technique for biomedical imaging since the phase coherence of SR beams can be exploited to obtain images with high contrast resolution. Using a synchrotron as the source for FXCT ensures a fluorescence signal that is optimally detectable by exploiting the beam monochromaticity and polarisation. The ability to combine these techniques so that SRCT and FXCT images are collected simultaneously, would bring distinct benefits to certain biomedical experiments. Simultaneous image acquisition would alleviate some of the registration difficulties which comes from collecting separate data, and it would provide increased information about the sample: functional X-ray images from the FXCT, with the morphological information from the SRCT. A method is presented for generating simultaneous SRCT and FXCT images. Proof of principle modelling has been used to show that it is possible to recover a fluorescence image of a point-like source from an SRCT apparatus by suitably modulating the illuminating planar X-ray beam. The projection image can be successfully used for reconstruction by removing the static modulation from the sinogram in the normal flat and dark field processing. Detection of the modulated fluorescence signal using an energy resolving detector allows the position of a fluorescent marker to be obtained using inverse reconstruction techniques. A discussion is made of particular reconstruction methods which might be applied by utilising both the CT and FXCT data.

  18. Correlative cryo-fluorescence light microscopy and cryo-electron tomography of Streptomyces.

    Science.gov (United States)

    Koning, Roman I; Celler, Katherine; Willemse, Joost; Bos, Erik; van Wezel, Gilles P; Koster, Abraham J

    2014-01-01

    Light microscopy and electron microscopy are complementary techniques that in a correlative approach enable identification and targeting of fluorescently labeled structures in situ for three-dimensional imaging at nanometer resolution. Correlative imaging allows electron microscopic images to be positioned in a broader temporal and spatial context. We employed cryo-correlative light and electron microscopy (cryo-CLEM), combining cryo-fluorescence light microscopy and cryo-electron tomography, on vitrified Streptomyces bacteria to study cell division. Streptomycetes are mycelial bacteria that grow as long hyphae and reproduce via sporulation. On solid media, Streptomyces subsequently form distinct aerial mycelia where cell division leads to the formation of unigenomic spores which separate and disperse to form new colonies. In liquid media, only vegetative hyphae are present divided by noncell separating crosswalls. Their multicellular life style makes them exciting model systems for the study of bacterial development and cell division. Complex intracellular structures have been visualized with transmission electron microscopy. Here, we describe the methods for cryo-CLEM that we applied for studying Streptomyces. These methods include cell growth, fluorescent labeling, cryo-fixation by vitrification, cryo-light microscopy using a Linkam cryo-stage, image overlay and relocation, cryo-electron tomography using a Titan Krios, and tomographic reconstruction. Additionally, methods for segmentation, volume rendering, and visualization of the correlative data are described. © 2014 Elsevier Inc. All rights reserved.

  19. Occlusal overload investigations by noninvasive technology: fluorescence microscopy and en-face optical coherence tomography

    Science.gov (United States)

    Marcauteanu, Corina; Negrutiu, Meda; Sinescu, Cosmin; Demjan, Enikö; Hughes, Michael; Bradu, Adrian; Dobre, George; Podoleanu, Adrian G.

    2009-07-01

    The aim of this study is the early detection and monitoring of occlusal overload in bruxing patients. En-Face Optical coherence tomography (eF-OCT) and fluorescence microscopy (FM) were used for the imaging of several anterior teeth extracted from patients with light active bruxism. We found a characteristic pattern of enamel cracks, that reached the tooth surface. We concluded that the combination of the en-Face OCT and FM is a promising non-invasive alternative technique for reliable monitoring of occlusal overload.

  20. Evaluation of dental enamel caries assessment using Quantitative Light Induced Fluorescence and Optical Coherence Tomography.

    Science.gov (United States)

    Maia, Ana Marly Araújo; de Freitas, Anderson Zanardi; de L Campello, Sergio; Gomes, Anderson Stevens Leônidas; Karlsson, Lena

    2016-06-01

    An in vitro study of morphological alterations between sound dental structure and artificially induced white spot lesions in human teeth, was performed through the loss of fluorescence by Quantitative Light-Induced Fluorescence (QLF) and the alterations of the light attenuation coefficient by Optical Coherence Tomography (OCT). To analyze the OCT images using a commercially available system, a special algorithm was applied, whereas the QLF images were analyzed using the software available in the commercial system employed. When analyzing the sound region against white spot lesions region by QLF, a reduction in the fluorescence intensity was observed, whilst an increase of light attenuation by the OCT system occurred. Comparison of the percentage of alteration between optical properties of sound and artificial enamel caries regions showed that OCT processed images through the attenuation of light enhanced the tooth optical alterations more than fluorescence detected by QLF System. QLF versus OCT imaging of enamel caries: a photonics assessment. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. An eco-friendly molecularly imprinted fluorescence composite material based on carbon dots for fluorescent detection of 4-nitrophenol

    International Nuclear Information System (INIS)

    Hao, Tongfan; Wei, Xiao; Nie, Yijing; Zhou, Zhiping; Xu, Yeqing; Yan, Yongsheng

    2016-01-01

    We on report an eco-friendly molecularly imprinted material based on carbon dots (C-dots) via a facile and efficient sol–gel polymerization for selective fluorescence detection of 4-nitrophenol (4-NP). The amino-modified C-dots were firstly synthesized by a hydrothermal process using citric acid as the carbon source and poly(ethyleneimine) as the surface modifier, and then after a sol–gel molecular imprinting process, the molecularly imprinted fluorescence material was obtained. The material (MIP-C-dots) showed strong fluorescence from C-dots and high selectivity due to the presence of a molecular imprint. After the detection conditions were optimized, the relative fluorescence intensity (F_0/F) of MIP-C-dots presented a good linearity with 4-NP concentrations in the linear range of 0.2 − 50 μmol L"-"1 with a detection limit (3σ/k) of 0.06 μmol L"-"1. In addition, the correlation coefficient was 0.9978 and the imprinting factor was 2.76. The method was applicable to the determination of trace 4-NP in Yangtze River water samples and good recoveries from 92.6–107.3 % were obtained. The present study provides a general strategy to fabricate materials based on C-dots with good fluorescence property for selective fluorescence detection of organic pollutants. (author)

  2. Singular value decomposition metrics show limitations of detector design in diffuse fluorescence tomography.

    Science.gov (United States)

    Leblond, Frederic; Tichauer, Kenneth M; Pogue, Brian W

    2010-11-29

    The spatial resolution and recovered contrast of images reconstructed from diffuse fluorescence tomography data are limited by the high scattering properties of light propagation in biological tissue. As a result, the image reconstruction process can be exceedingly vulnerable to inaccurate prior knowledge of tissue optical properties and stochastic noise. In light of these limitations, the optimal source-detector geometry for a fluorescence tomography system is non-trivial, requiring analytical methods to guide design. Analysis of the singular value decomposition of the matrix to be inverted for image reconstruction is one potential approach, providing key quantitative metrics, such as singular image mode spatial resolution and singular data mode frequency as a function of singular mode. In the present study, these metrics are used to analyze the effects of different sources of noise and model errors as related to image quality in the form of spatial resolution and contrast recovery. The image quality is demonstrated to be inherently noise-limited even when detection geometries were increased in complexity to allow maximal tissue sampling, suggesting that detection noise characteristics outweigh detection geometry for achieving optimal reconstructions.

  3. Fluorescent x-ray computed tomography with synchrotron radiation using fan collimator

    Science.gov (United States)

    Takeda, Tohoru; Akiba, Masahiro; Yuasa, Tetsuya; Kazama, Masahiro; Hoshino, Atsunori; Watanabe, Yuuki; Hyodo, Kazuyuki; Dilmanian, F. Avraham; Akatsuka, Takao; Itai, Yuji

    1996-04-01

    We describe a new system of fluorescent x-ray computed tomography applied to image nonradioactive contrast materials in vivo. The system operates on the basis of computed tomography (CT) of the first generation. The experiment was also simulated using the Monte Carlo method. The research was carried out at the BLNE-5A bending-magnet beam line of the Tristan Accumulation Ring in Kek, Japan. An acrylic cylindrical phantom containing five paraxial channels of 5 and 4 mm diameters was imaged. The channels were filled with a diluted iodine-based contrast material, with iodine concentrations of 2 mg/ml and 500 (mu) g/ml. Spectra obtained with the system's high purity germanium (HPGe) detector separated clearly the K(alpha ) and K(beta 1) x-ray fluorescent lines, and the Compton scattering. CT images were reconstructed from projections generated by integrating the counts in these spectral lines. The method had adequate sensitivity and detection power, as shown by the experiment and predicted by the simulations, to show the iodine content of the phantom channels, which corresponded to 1 and 4 (mu) g iodine content per pixel in the reconstructed images.

  4. Fluorescent X-ray computed tomography using synchrotron radiation for imaging nonradioactive tracer materials

    Energy Technology Data Exchange (ETDEWEB)

    Akiba, Masahiro; Yuasa, Tetsuya; Uchida, Akira; Akatsuka, Takao [Yamagata Univ., Yonezawa (Japan). Electrical and Information of Engineering; Takeda, Tohoru; Hyodo, Kazuyuki; Itai, Yuji

    1997-09-01

    We describe a system of fluorescent X-ray computed tomography using synchrotron radiation (SR-FXCT) to image nonradioactive contrast materials. The system operates on the basis of computed tomography (CT) scanned by the pencil beam. In the previous experiment, we have imaged an acrylic cylindrical phantom with cross-shaped channel, filled with a diluted iodine-based tracer material of 200 {mu}g/ml. This research is aimed to improve image quality, to select the optimum energy of the incident X-ray, to confirm quantitative evaluation of the image, and to demonstrate FXCT image for living body. First, we simulated output energy profile by the Monte Carlo simulation and confirmed to predetermine the incident X-ray energy at 37 keV, in order to separate the fluorescent photons from background scattering components. Next, the imaging experiment was performed by using conventional CT algorithm under the optimum parameter at the Tristan Accumulation Ring, KEK, Japan. An acrylic phantom containing five paraxial channels of 5 and 4 mm in diameter, could be imaged; where each channel was respectively filled with diluted iodine-based contrast materials of 50, 100, 200 and 500 {mu}g/ml. From the reconstructed image, we confirmed quantitativity in the FXCT image. Finally, a rat`s brain was imaged in vitro by FXCT and monochromatic transmission CT. The comparison between these results showed that the iodine-rich region in the FXCT image corresponded with that in the monochromatic transmission CT image. (author)

  5. Study of continuous-wave domain fluorescence diffuse optical tomography for quality control on agricultural produce

    Energy Technology Data Exchange (ETDEWEB)

    Nadhira, Vebi, E-mail: vebi@tf.itb.ac.id; Kurniadi, Deddy, E-mail: vebi@tf.itb.ac.id; Juliastuti, E., E-mail: vebi@tf.itb.ac.id; Sutiswan, Adeline, E-mail: vebi@tf.itb.ac.id [Instrumentation and Control Research Group, Faculty of Industrial Technology, Institute Technology of Bandung, Ganesha 10 40132 Bandung (Indonesia)

    2014-03-24

    The importance of monitoring the quality of vegetables and fruits is prosperity by giving a competitive advantage for producer and providing a more healthy food for consumer. Diffuse Optical Tomography (DOT) is offering the possibility to detect the internal defects of the agricultural produce quality. Fluorescence diffuse optical tomography (FDOT) is the development of DOT, offering the possibilities to improve spatial resolution and to contrast image. The purpose of this research is to compare FDOT and DOT in forward analysis with continuous wave approach. The scattering and absorbing parameters of potatoes are used to represent the real condition. The object was illuminated by the NIR source from some positions on the boundary of object. A set of NIR detector are placed on the peripheral position of the object to measure the intensity of propagated or emitted light. In the simulation, we varied a condition of object then we analyzed the sensitivity of forward problem. The result of this study shows that FDOT has a better sensitivity than DOT and a better potential to monitor internal defects of agricultural produce because of the contrast value between optical and fluorescence properties of agricultural produce normal tissue and defects.

  6. Imaging retinal degeneration in mice by combining Fourier domain optical coherence tomography and fluorescent scanning laser ophthalmoscopy

    Science.gov (United States)

    Hossein-Javaheri, Nima; Molday, Laurie L.; Xu, Jing; Molday, Robert S.; Sarunic, Marinko V.

    2009-02-01

    Visualization of the internal structures of the retina is critical for clinical diagnosis and monitoring of pathology as well as for medical research investigating the root causes of retinal degeneration. Optical Coherence Tomography (OCT) is emerging as the preferred technique for non-contact sub-surface depth-resolved imaging of the retina. The high resolution cross sectional images acquired in vivo by OCT can be compared to histology to visually delineate the retinal layers. The recent demonstration of the significant sensitivity increase obtained through use of Fourier domain (FD) detection with OCT has been used to facilitate high speed scanning for volumetric reconstruction of the retina in software. The images acquired by OCT are purely structural, relying on refractive index differences in the tissue for contrast, and do not provide information on the molecular content of the sample. We have constructed a FDOCT prototype and combined it with a fluorescent Scanning Laser Ophthalmoscope (fSLO) to permit real time alignment of the field of view on the retina. The alignment of the FDOCT system to the specimen is crucial for the registration of measurements taken throughout longitudinal studies. In addition, fluorescence detection has been integrated with the SLO to enable the en face localization of a molecular contrast signal, which is important for retinal angiography, and also for detection of autofluorescence associated with some forms of retinal degeneration, for example autofluorescence lipofuscin accumulations are associated with Stargardt's Macular Dystrophy. The integrated FD OCT/fSLO system was investigated for imaging the retina of the mice in vivo.

  7. Fluorescence diffuse optical tomography: benefits of using the time-resolved modality

    International Nuclear Information System (INIS)

    Ducros, Nicolas

    2009-01-01

    Fluorescence diffuse optical tomography enables the three-dimensional reconstruction of fluorescence markers injected within a biological tissue, with light in the near infrared range. The simple continuous modality uses steady excitation light and operates from the measurements at different positions of the attenuation of the incident beam. This technique is low-cost, non-ionizing, and easy to handle, but subject to low resolution for thick tissues due to diffusion. Hopefully, the time-resolved modality, which provides the time of flight of any detected photon, could overcome this limitation and pave the way to clinical applications. This thesis aims at determining the best way to exploit the time resolved information and at quantifying the advantages of this modality over the standard continuous wave one. Model deviations must be carefully limited when ill-posed problems as fluorescence diffuse optical tomography are considered. As a result, we have first addressed the modelling part of the problem. We have shown that the photons density models to good approximation the measurable quantity that is the quantity measured by an actual acquisition set-up. Then, the moment-based reconstruction scheme has been thoroughly evaluated by means of a theoretical analysis of the moments properties. It was found that the moment-based approach requires high photon counts to be profitable compared to the continuous wave modality. Last, a novel wavelet-based approach, which enables an improved reconstruction quality, has been introduced. This approach has shown good ability to exploit the temporal information at lower photon counts. (author) [fr

  8. Fluorescent x-ray computed tomography to visualize specific material distribution

    Science.gov (United States)

    Takeda, Tohoru; Yuasa, Tetsuya; Hoshino, Atsunori; Akiba, Masahiro; Uchida, Akira; Kazama, Masahiro; Hyodo, Kazuyuki; Dilmanian, F. Avraham; Akatsuka, Takao; Itai, Yuji

    1997-10-01

    Fluorescent x-ray computed tomography (FXCT) is being developed to detect non-radioactive contrast materials in living specimens. The FXCT systems consists of a silicon channel cut monochromator, an x-ray slit and a collimator for detection, a scanning table for the target organ and an x-ray detector for fluorescent x-ray and transmission x-ray. To reduce Compton scattering overlapped on the K(alpha) line, incident monochromatic x-ray was set at 37 keV. At 37 keV Monte Carlo simulation showed almost complete separation between Compton scattering and the K(alpha) line. Actual experiments revealed small contamination of Compton scattering on the K(alpha) line. A clear FXCT image of a phantom was obtained. Using this system the minimal detectable dose of iodine was 30 ng in a volume of 1 mm3, and a linear relationship was demonstrated between photon counts of fluorescent x-rays and the concentration of iodine contrast material. The use of high incident x-ray energy allows an increase in the signal to noise ratio by reducing the Compton scattering on the K(alpha) line.

  9. Multi-spectral and fluorescence diffuse optical tomography of breast cancer

    Science.gov (United States)

    Corlu, Alper

    Multi-spectral and fluorescence diffuse optical tomography (DOT) techniques are explored and applied to image human breast cancer in vivo. Image reconstruction algorithms that utilize first and second order gradient information are described in detail. Breast DOT requires large computational memory and long run times. To this end, parallel computation techniques were developed appropriate to each reconstruction algorithm. A parallel plate DOT instrument developed for breast cancer imaging is described. The system relies heavily on continuous-wave (CW) transmission measurements and utilizes frequency domain (FD) measurements on the reemission side. However, traditional DOT image reconstruction methods based on CW measurements fail to separate tissue absorption and scattering uniquely. In this manuscript, multi-spectral DOT is shown to be capable of minimizing cross-talk and retrieving spectral parameters almost uniquely when the measurement wavelengths are optimized. A theoretical framework to select optimum wavelengths is provided, and tested with computer simulations. Results from phantom spectroscopy experiments and in vivo patient measurements support the notion that multi-spectral methods are superior to traditional DOT image reconstruction schemes. The same breast DOT instrument is improved and utilized to obtain the first in vivo images of human breast cancer based on fluorescence DOT (FDOT). To this end the fluorophore Indocyanine Green (ICG) is injected intravenously and fluorescence excitation and detection are accomplished in the soft-compression, parallel-plane, transmission geometry using laser sources at 786 nm and spectrally filtered CCD detection. Careful phantom and in vivo measurements are carried on to assure that the signals are due to ICG fluorescence, rather than tissue autofluorescence and excitation light leakage. An in vivo measurement protocol is designed to maximize the ICG contrast by acquiring full fluorescence tomographic scan during

  10. Molecular Viscosity Sensors with Two Rotators for Optimizing the Fluorescence Intensity-Contrast Trade-Off.

    Science.gov (United States)

    Lee, Seung-Chul; Lee, Chang-Lyoul; Heo, Jeongyun; Jeong, Chan-Uk; Lee, Gyeong-Hui; Kim, Sehoon; Yoon, Woojin; Yun, Hoseop; Park, Sung O; Kwak, Sang Kyu; Park, Sung-Ha; Kwon, O-Pil

    2018-02-26

    A series of fluorescent molecular rotors obtained by introducing two rotational groups ("rotators"), which exhibit different rotational and electron-donating abilities, are discussed. Whereas the control molecular rotor, PH, includes a single rotator (the widely used phenyl group), the PO molecular rotors consist of two rotators (a phenyl group and an alkoxy group), which exhibit simultaneous strongly electron-donating and easy rotational abilities. Compared with the control rotor PH, PO molecular rotors exhibited one order of magnitude higher quantum yield (fluorescence intensity) and simultaneously exhibited significantly higher fluorescence contrast. These properties are directly related to the strong electron-donating ability and low energy barrier of rotation of the alkoxy group, as confirmed by dynamic fluorescence experiments and quantum chemical calculations. The PO molecular rotors exhibited two fluorescence relaxation pathways, whereas the PH molecular rotor exhibited a single fluorescence relaxation pathway. Cellular fluorescence imaging with PO molecular rotors for mapping cellular viscosity was successfully demonstrated. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. The lymphatic mechanisms of brain cleaning: application of optical coherence tomography and fluorescence microscopy

    Science.gov (United States)

    Glushkovskaya-Semyachkina, O.; Abdurashitov, A.; Fedosov, I.; Namykin, A.; Pavlov, A.; Shirokov, A.; Shushunova, N.; Sindeeva, O.; Khorovodov, A.; Ulanova, M.; Sagatova, V.; Agranovich, I.; Bodrova, A.; Kurths, J.

    2018-04-01

    Here we studied the role of cerebral lymphatic system in the brain clearing using intraparenchymal injection of Evans Blue and gold nanorods assessed by optical coherent tomography and fluorescence microscopy. Our data clearly show that the cerebral lymphatic system plays an important role in the brain cleaning via meningeal lymphatic vessels but not cerebral veins. Meningeal lymphatic vessels transport fluid from the brain into the deep cervical node, which is the first anatomical "station" for lymph outflow from the brain. The lymphatic processes underlying brain clearing are more slowly vs. peripheral lymphatics. These results shed light on the lymphatic mechanisms responsible for brain clearing as well as interaction between the intra- and extracranial lymphatic compartment.

  12. Dynamic fluorescence imaging with molecular agents for cancer detection

    Science.gov (United States)

    Kwon, Sun Kuk

    Non-invasive dynamic optical imaging of small animals requires the development of a novel fluorescence imaging modality. Herein, fluorescence imaging is demonstrated with sub-second camera integration times using agents specifically targeted to disease markers, enabling rapid detection of cancerous regions. The continuous-wave fluorescence imaging acquires data with an intensified or an electron-multiplying charge-coupled device. The work presented in this dissertation (i) assessed dose-dependent uptake using dynamic fluorescence imaging and pharmacokinetic (PK) models, (ii) evaluated disease marker availability in two different xenograft tumors, (iii) compared the impact of autofluorescence in fluorescence imaging of near-infrared (NIR) vs. red light excitable fluorescent contrast agents, (iv) demonstrated dual-wavelength fluorescence imaging of angiogenic vessels and lymphatics associated with a xenograft tumor model, and (v) examined dynamic multi-wavelength, whole-body fluorescence imaging with two different fluorescent contrast agents. PK analysis showed that the uptake of Cy5.5-c(KRGDf) in xenograft tumor regions linearly increased with doses of Cy5.5-c(KRGDf) up to 1.5 nmol/mouse. Above 1.5 nmol/mouse, the uptake did not increase with doses, suggesting receptor saturation. Target to background ratio (TBR) and PK analysis for two different tumor cell lines showed that while Kaposi's sarcoma (KS1767) exhibited early and rapid uptake of Cy5.5-c(KRGDf), human melanoma tumors (M21) had non-significant TBR differences and early uptake rates similar to the contralateral normal tissue regions. The differences may be due to different compartment location of the target. A comparison of fluorescence imaging with NIR vs. red light excitable fluorescent dyes demonstrates that NIR dyes are associated with less background signal, enabling rapid tumor detection. In contrast, animals injected with red light excitable fluorescent dyes showed high autofluorescence. Dual

  13. Adaptation and focusing of optode configurations for fluorescence optical tomography by experimental design methods.

    Science.gov (United States)

    Freiberger, Manuel; Clason, Christian; Scharfetter, Hermann

    2010-01-01

    Fluorescence tomography excites a fluorophore inside a sample by light sources on the surface. From boundary measurements of the fluorescent light, the distribution of the fluorophore is reconstructed. The optode placement determines the quality of the reconstructions in terms of, e.g., resolution and contrast-to-noise ratio. We address the adaptation of the measurement setup. The redundancy of the measurements is chosen as a quality criterion for the optodes and is computed from the Jacobian of the mathematical formulation of light propagation. The algorithm finds a subset with minimum redundancy in the measurements from a feasible pool of optodes. This allows biasing the design in order to favor reconstruction results inside a given region. Two different variations of the algorithm, based on geometric and arithmetic averaging, are compared. Both deliver similar optode configurations. The arithmetic averaging is slightly more stable, whereas the geometric averaging approach shows a better conditioning of the sensitivity matrix and mathematically corresponds more closely with entropy optimization. Adapted illumination and detector patterns are presented for an initial set of 96 optodes placed on a cylinder with focusing on different regions. Examples for the attenuation of fluorophore signals from regions outside the focus are given.

  14. A time-domain fluorescence diffusion optical tomography system for breast tumor diagnosis

    Science.gov (United States)

    Zhang, Wei; Gao, Feng; Wu, LinHui; Ma, Wenjuan; Yang, Fang; Zhou, Zhongxing; Zhang, Limin; Zhao, Huijuan

    2011-02-01

    A prototype time-domain fluorescence diffusion optical tomography (FDOT) system using near-infrared light is presented. The system employs two pulsed light sources, 32 source fibers and 32 detection channels, working separately for acquiring the temporal distribution of the photon flux on the tissue surface. The light sources are provided by low power picosecond pulsed diode lasers at wavelengths of 780 nm and 830 nm, and a 1×32-fiber-optic-switch sequentially directs light sources to the object surface through 32 source fibers. The light signals re-emitted from the object are collected by 32 detection fibers connected to four 8×1 fiber-optic-switch and then routed to four time-resolved measuring channels, each of which consists of a collimator, a filter wheel, a photomultiplier tube (PMT) photon-counting head and a time-correlated single photon counting (TCSPC) channel. The performance and efficacy of the designed multi-channel PMT-TCSPC system are assessed by reconstructing the fluorescent yield and lifetime images of a solid phantom.

  15. Intravascular atherosclerotic imaging with combined fluorescence and optical coherence tomography probe based on a double-clad fiber combiner

    Science.gov (United States)

    Liang, Shanshan; Saidi, Arya; Jing, Joe; Liu, Gangjun; Li, Jiawen; Zhang, Jun; Sun, Changsen; Narula, Jagat; Chen, Zhongping

    2012-07-01

    We developed a multimodality fluorescence and optical coherence tomography probe based on a double-clad fiber (DCF) combiner. The probe is composed of a DCF combiner, grin lens, and micromotor in the distal end. An integrated swept-source optical coherence tomography and fluorescence intensity imaging system was developed based on the combined probe for the early diagnoses of atherosclerosis. This system is capable of real-time data acquisition and processing as well as image display. For fluorescence imaging, the inflammation of atherosclerosis and necrotic core formed with the annexin V-conjugated Cy5.5 were imaged. Ex vivo imaging of New Zealand white rabbit arteries demonstrated the capability of the combined system.

  16. A molecular-sized optical logic circuit for digital modulation of a fluorescence signal

    Science.gov (United States)

    Nishimura, Takahiro; Tsuchida, Karin; Ogura, Yusuke; Tanida, Jun

    2018-03-01

    Fluorescence measurement allows simultaneous detection of multiple molecular species by using spectrally distinct fluorescence probes. However, due to the broad spectra of fluorescence emission, the multiplicity of fluorescence measurement is generally limited. To overcome this limitation, we propose a method to digitally modulate fluorescence output signals with a molecular-sized optical logic circuit by using optical control of fluorescence resonance energy transfer (FRET). The circuit receives a set of optical inputs represented with different light wavelengths, and then it switches high and low fluorescence intensity from a reporting molecule according to the result of the logic operation. By using combinational optical inputs in readout of fluorescence signals, the number of biomolecular species that can be identified is increased. To implement the FRET-based circuits, we designed two types of basic elements, YES and NOT switches. An YES switch produces a high-level output intensity when receiving a designated light wavelength input and a low-level intensity without the light irradiation. A NOT switch operates inversely to the YES switch. In experiments, we investigated the operation of the YES and NOT switches that receive a 532-nm light input and modulate the fluorescence intensity of Alexa Fluor 488. The experimental result demonstrates that the switches can modulate fluorescence signals according to the optical input.

  17. Surface plasmon-enhanced molecular fluorescence induced by gold nanostructures

    International Nuclear Information System (INIS)

    Teng, Y.; Ueno, K.; Shi, X.; Aoyo, D.; Misawa, H.; Qiu, J.

    2012-01-01

    The authors report on surface plasmon-enhanced fluorescence of Eosin Y molecules induced by gold nanostructures. Al 2 O 3 films deposited by atomic layer deposition with sub-nanometer resolution were used as the spacer layer to control the distance between molecules and the gold surface. As the thickness of the Al 2 O 3 film increased, the fluorescence intensity first increased and then decreased. The highest enhancement factor is achieved with a 1 nm Al 2 O 3 film. However, the trend for the fluorescence lifetime is the opposite. It first decreased and then increased. The changes in the fluorescence quantum yield were also calculated. The yield shows a similar trend to the fluorescence intensity. The competition between the surface plasmon-induced increase in the radiative decay rate and the gold-induced fluorescence quenching is responsible for the observed phenomenon. In addition, this competition strongly depends on the thickness of the spacer layer between Eosin Y molecules and the gold surface. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  18. Analysis of fluorescence quenching of pyronin B and pyronin Y by molecular oxygen in aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Celebi, Neslihan [Faculty of Arts and Sciences, Department of Chemistry, Atatuerk University, 25240 Erzurum (Turkey); Arik, Mustafa [Faculty of Arts and Sciences, Department of Chemistry, Atatuerk University, 25240 Erzurum (Turkey); Onganer, Yavuz [Faculty of Arts and Sciences, Department of Chemistry, Atatuerk University, 25240 Erzurum (Turkey)]. E-mail: yonganer@atauni.edu.tr

    2007-09-15

    The fluorescence quenching of pyronin B and pyronin Y molecules by molecular oxygen in aqueous solution was studied by using steady-state and time-resolved fluorescence and UV-Vis absorption spectroscopy techniques. In order to understand the quenching mechanism, fluorescence decays, absorption and fluorescence spectra of the probes were recorded as a function of the oxygen concentration and temperature. The quenching was found to be appreciable and shows positive deviation in the Stern-Volmer representation obtained from the fluorescence intensity ratio. Fluorescence quenching constants (k {sub q}) were calculated from the {tau} {sub o}/{tau} vs. [Q] plots having linear correlation and compared with calculated diffusion-controlled rate constants (k {sub diff}) values. Experimental results were in good agreement with the simultaneous dynamic and static quenching model.

  19. Analysis of fluorescence quenching of pyronin B and pyronin Y by molecular oxygen in aqueous solution

    International Nuclear Information System (INIS)

    Celebi, Neslihan; Arik, Mustafa; Onganer, Yavuz

    2007-01-01

    The fluorescence quenching of pyronin B and pyronin Y molecules by molecular oxygen in aqueous solution was studied by using steady-state and time-resolved fluorescence and UV-Vis absorption spectroscopy techniques. In order to understand the quenching mechanism, fluorescence decays, absorption and fluorescence spectra of the probes were recorded as a function of the oxygen concentration and temperature. The quenching was found to be appreciable and shows positive deviation in the Stern-Volmer representation obtained from the fluorescence intensity ratio. Fluorescence quenching constants (k q ) were calculated from the τ o /τ vs. [Q] plots having linear correlation and compared with calculated diffusion-controlled rate constants (k diff ) values. Experimental results were in good agreement with the simultaneous dynamic and static quenching model

  20. Molecular quantification of genes encoding for green-fluorescent proteins

    DEFF Research Database (Denmark)

    Felske, A; Vandieken, V; Pauling, B V

    2003-01-01

    A quantitative PCR approach is presented to analyze the amount of recombinant green fluorescent protein (gfp) genes in environmental DNA samples. The quantification assay is a combination of specific PCR amplification and temperature gradient gel electrophoresis (TGGE). Gene quantification...... PCR strategy is a highly specific and sensitive way to monitor recombinant DNA in environments like the efflux of a biotechnological plant....

  1. Fundus auto fluorescence and spectral domain ocular coherence tomography in the early detection of chloroquine retinopathy

    Directory of Open Access Journals (Sweden)

    Megan B. Goodman

    2015-08-01

    Full Text Available Purpose: To determine the sensitivity of spectral domain ocular coherence tomography (SD-OCT and fundus auto fluorescence (FAF images as a screening test to detect early changes in the retina prior to the onset of chloroquine retinopathy. Method: The study was conducted using patients taking chloroquine (CQ, referred by the Rheumatology Department to the Ophthalmology Department at Tygerberg Academic Hospital. Group A consisted of 59 patients on CQ for less than 5 years, and Group B consisted of 53 patients on CQ for more than 5 years. A 200 × 200 macula thickness map, 5-line raster SD-OCT on a Carl Zeiss Meditec Cirrus HD-OCT and FAF images on a Carl Zeiss Meditec Visucam 500 were recorded for 223 eyes. Images were reviewed independently, and then those of Groups A and B compared. Results: There were no statistically significant differences between Groups A and B. The criteria included the internal limiting membrane and the retinal pigment epithelium (ILM-RPE thickness, interdigitation zone integrity (p = 0.891, df = 1, χ² = 0.1876, ellipsoid zone integrity (p = 0.095, df = 2, χ² = 4.699 and FAF image irregularities (p = 0.479, df = 1, χ²= 4995978. Conclusion: The inclusion of SD-OCT and FAF as objective tests into the prescribed screening guidelines does not appear to simplify the detection of subclinical injury in patients on chloroquine treatment.

  2. Laboratory Scale X-ray Fluorescence Tomography: Instrument Characterization and Application in Earth and Environmental Science.

    Science.gov (United States)

    Laforce, Brecht; Vermeulen, Bram; Garrevoet, Jan; Vekemans, Bart; Van Hoorebeke, Luc; Janssen, Colin; Vincze, Laszlo

    2016-03-15

    A new laboratory scale X-ray fluorescence (XRF) imaging instrument, based on an X-ray microfocus tube equipped with a monocapillary optic, has been developed to perform XRF computed tomography experiments with both higher spatial resolution (20 μm) and a better energy resolution (130 eV @Mn-K(α)) than has been achieved up-to-now. This instrument opens a new range of possible applications for XRF-CT. Next to the analytical characterization of the setup by using well-defined model/reference samples, demonstrating its capabilities for tomographic imaging, the XRF-CT microprobe has been used to image the interior of an ecotoxicological model organism, Americamysis bahia. This had been exposed to elevated metal (Cu and Ni) concentrations. The technique allowed the visualization of the accumulation sites of copper, clearly indicating the affected organs, i.e. either the gastric system or the hepatopancreas. As another illustrative application, the scanner has been employed to investigate goethite spherules from the Cretaceous-Paleogene boundary, revealing the internal elemental distribution of these valuable distal ejecta layer particles.

  3. Performance Enhancement of Pharmacokinetic Diffuse Fluorescence Tomography by Use of Adaptive Extended Kalman Filtering.

    Science.gov (United States)

    Wang, Xin; Wu, Linhui; Yi, Xi; Zhang, Yanqi; Zhang, Limin; Zhao, Huijuan; Gao, Feng

    2015-01-01

    Due to both the physiological and morphological differences in the vascularization between healthy and diseased tissues, pharmacokinetic diffuse fluorescence tomography (DFT) can provide contrast-enhanced and comprehensive information for tumor diagnosis and staging. In this regime, the extended Kalman filtering (EKF) based method shows numerous advantages including accurate modeling, online estimation of multiparameters, and universal applicability to any optical fluorophore. Nevertheless the performance of the conventional EKF highly hinges on the exact and inaccessible prior knowledge about the initial values. To address the above issues, an adaptive-EKF scheme is proposed based on a two-compartmental model for the enhancement, which utilizes a variable forgetting-factor to compensate the inaccuracy of the initial states and emphasize the effect of the current data. It is demonstrated using two-dimensional simulative investigations on a circular domain that the proposed adaptive-EKF can obtain preferable estimation of the pharmacokinetic-rates to the conventional-EKF and the enhanced-EKF in terms of quantitativeness, noise robustness, and initialization independence. Further three-dimensional numerical experiments on a digital mouse model validate the efficacy of the method as applied in realistic biological systems.

  4. Molecularly imprinted fluorescent probe based on FRET for selective and sensitive detection of doxorubicin

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Zhifeng, E-mail: 897061147@qq.com [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Deng, Peihong; Li, Junhua [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Xu, Li [Department of Applied Chemistry, College of Materials and Energy, South China Agricultural University, Guangzhou 510642 (China); Tang, Siping [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China)

    2017-04-15

    Highlights: • FRET-based molecularly imprinted probe for detection of doxorubicin was prepared. • The detection limit of the probe was 13.8 nM for doxorubicin. • The FRET-based probe had a higher selectivity for the template than ordinary MIMs. - Abstract: In this work, a new type of fluorescent probe for detection of doxorubicin has been constructed by the combined use of fluorescence resonance energy transfer (FRET) technology and molecular imprinting technique (MIT). Using doxorubicin as the template, the molecularly imprinted polymer thin layer was fabricated on the surfaces of carbon dot (CD) modified silica by sol-gel polymerization. The excitation energy of the fluorescent donor (CDs) could be transferred to the fluorescent acceptor (doxorubicin). The FRET based fluorescent probe demonstrated high sensitivity and selectivity for doxorubicin. The detection limit was 13.8 nM. The fluorescent probe was successfully applied for detecting doxorubicin in doxorubicin-spiked plasmas with a recovery of 96.8–103.8%, a relative standard deviation (RSD) of 1.3–2.8%. The strategy for construction of FRET-based molecularly imprinted materials developed in this work is very promising for analytical applications.

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

    Science.gov (United States)

    Qin, Zhengtao

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

  6. X-ray nanoprobes and diffraction-limited storage rings: opportunities and challenges of fluorescence tomography of biological specimens

    Energy Technology Data Exchange (ETDEWEB)

    Jonge, Martin D. de, E-mail: martin.dejonge@synchrotron.org.au [Australian Synchrotron, 800 Blackburn Road, Clayton, Victoria 3168 (Australia); Ryan, Christopher G. [CSIRO Earth Science and Research Engineering, Clayton, Victoria 3168 (Australia); Jacobsen, Chris J. [Advanced Photon Source, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439 (United States); Department of Physics, Chemistry of Life Processes Institute, Northwestern University, 2170 Campus Drive, Evanston, IL 60208 (United States); Chemistry of Life Processes Institute, Northwestern University, 2170 Campus Drive, Evanston, IL 60208 (United States)

    2014-08-27

    Nanoscale X-ray scanning microscopes, or X-ray nanoprobes, will benefit greatly from diffraction-limited storage rings. Here the requirements for nanoscale fluorescence tomography are explored to gain insight into the scientific opportunities and technical challenges that such sources offer. X-ray nanoprobes require coherent illumination to achieve optic-limited resolution, and so will benefit directly from diffraction-limited storage rings. Here, the example of high-resolution X-ray fluorescence tomography is focused on as one of the most voracious demanders of coherent photons, since the detected signal is only a small fraction of the incident flux. Alternative schemes are considered for beam delivery, sample scanning and detectors. One must consider as well the steps before and after the X-ray experiment: sample preparation and examination conditions, and analysis complexity due to minimum dose requirements and self-absorption. By understanding the requirements and opportunities for nanoscale fluorescence tomography, one gains insight into the R&D challenges in optics and instrumentation needed to fully exploit the source advances that diffraction-limited storage rings offer.

  7. Interaction of Chelerythrine with Keyhole Limpet Hemocyanin: a Fluorescence Spectroscopy and Molecular Docking Study

    Science.gov (United States)

    Zhong, M.; Long, R. Q.; Wang, Y. H.; Chen, C. L.

    2018-05-01

    The quenching mechanism between chelerythrine (CHE) and keyhole limpet hemocyanin (KLH) was investigated using fluorescence spectroscopy and molecular docking. The experiments were conducted at three different temperatures (293, 298, and 303 K). The results revealed that the intrinsic fluorescence of KLH was strongly quenched by CHE through a static quenching mechanism. The thermodynamic parameters (ΔG, ΔH, and ΔS) of the interaction were calculated, indicating that the interaction between CHE and KLH was spontaneous and that van der Waals forces and hydrogen bond formation played major roles in the binding process. The intrinsic fluorescence of the tyrosine and tryptophan residues in KLH was studied by synchronous fluorescence, which suggested that CHE changed the conformation of KLH. Finally, molecular docking was used to obtain detailed information on the binding sites and binding affinities between CHE and KLH.

  8. CMOS Time-Resolved, Contact, and Multispectral Fluorescence Imaging for DNA Molecular Diagnostics

    Directory of Open Access Journals (Sweden)

    Nan Guo

    2014-10-01

    Full Text Available Instrumental limitations such as bulkiness and high cost prevent the fluorescence technique from becoming ubiquitous for point-of-care deoxyribonucleic acid (DNA detection and other in-field molecular diagnostics applications. The complimentary metal-oxide-semiconductor (CMOS technology, as benefited from process scaling, provides several advanced capabilities such as high integration density, high-resolution signal processing, and low power consumption, enabling sensitive, integrated, and low-cost fluorescence analytical platforms. In this paper, CMOS time-resolved, contact, and multispectral imaging are reviewed. Recently reported CMOS fluorescence analysis microsystem prototypes are surveyed to highlight the present state of the art.

  9. Strategies of molecular imprinting-based fluorescence sensors for chemical and biological analysis.

    Science.gov (United States)

    Yang, Qian; Li, Jinhua; Wang, Xiaoyan; Peng, Hailong; Xiong, Hua; Chen, Lingxin

    2018-07-30

    One pressing concern today is to construct sensors that can withstand various disturbances for highly selective and sensitive detecting trace analytes in complicated samples. Molecularly imprinted polymers (MIPs) with tailor-made binding sites are preferred to be recognition elements in sensors for effective targets detection, and fluorescence measurement assists in highly sensitive detection and user-friendly control. Accordingly, molecular imprinting-based fluorescence sensors (MI-FL sensors) have attracted great research interest in many fields such as chemical and biological analysis. Herein, we comprehensively review the recent advances in MI-FL sensors construction and applications, giving insights on sensing principles and signal transduction mechanisms, focusing on general construction strategies for intrinsically fluorescent or nonfluorescent analytes and improvement strategies in sensing performance, particularly in sensitivity. Construction strategies are well overviewed, mainly including the traditional indirect methods of competitive binding against pre-bound fluorescent indicators, employment of fluorescent functional monomers and embedding of fluorescence substances, and novel rational designs of hierarchical architecture (core-shell/hollow and mesoporous structures), post-imprinting modification, and ratiometric fluorescence detection. Furthermore, MI-FL sensor based microdevices are discussed, involving micromotors, test strips and microfluidics, which are more portable for rapid point-of-care detection and in-field diagnosing. Finally, the current challenges and future perspectives of MI-FL sensors are proposed. Copyright © 2018 Elsevier B.V. All rights reserved.

  10. Molecular Imaging of Transporters with Positron Emission Tomography

    Science.gov (United States)

    Antoni, Gunnar; Sörensen, Jens; Hall, Håkan

    Positron emission tomography (PET) visualization of brain components in vivo is a rapidly growing field. Molecular imaging with PET is also increasingly used in drug development, especially for the determination of drug receptor interaction for CNS-active drugs. This gives the opportunity to relate clinical efficacy to per cent receptor occupancy of a drug on a certain targeted receptor and to relate drug pharmacokinetics in plasma to interaction with target protein. In the present review we will focus on the study of transporters, such as the monoamine transporters, the P-glycoprotein (Pgp) transporter, the vesicular monoamine transporter type 2, and the glucose transporter using PET radioligands. Neurotransmitter transporters are presynaptically located and in vivo imaging using PET can therefore be used for the determination of the density of afferent neurons. Several promising PET ligands for the noradrenaline transporter (NET) have been labeled and evaluated in vivo including in man, but a really useful PET ligand for NET still remains to be identified. The most promising tracer to date is (S,S)-[18F]FMeNER-D2. The in vivo visualization of the dopamine transporter (DAT) may give clues in the evaluation of conditions related to dopamine, such as Parkinson's disease and drug abuse. The first PET radioligands based on cocaine were not selective, but more recently several selective tracers such as [11C]PE2I have been characterized and shown to be suitable as PET radioligands. Although there are a large number of serotonin transporter inhibitors used today as SSRIs, it was not until very recently, when [11C]McN5652 was synthesized, that this transporter was studied using PET. New candidates as PET radioligands for the SERT have subsequently been developed and [11C]DASB and [11C]MADAM and their analogues are today the most promising ligands. The existing radioligands for Pgp transporters seem to be suitable tools for the study of both peripheral and central drug

  11. L-shell x-ray fluorescence computed tomography (XFCT) imaging of Cisplatin

    International Nuclear Information System (INIS)

    Bazalova, Magdalena; Ahmad, Moiz; Pratx, Guillem; Xing, Lei

    2014-01-01

    X-ray fluorescence computed tomography (XFCT) imaging has been focused on the detection of K-shell x-rays. The potential utility of L-shell x-ray XFCT is, however, not well studied. Here we report the first Monte Carlo (MC) simulation of preclinical L-shell XFCT imaging of Cisplatin. We built MC models for both L- and K-shell XFCT with different excitation energies (15 and 30 keV for L-shell and 80 keV for K-shell XFCT). Two small-animal sized imaging phantoms of 2 and 4 cm diameter containing a series of objects of 0.6 to 2.7 mm in diameter at 0.7 to 16 mm depths with 10 to 250 µg mL −1  concentrations of Pt are used in the study. Transmitted and scattered x-rays were collected with photon-integrating transmission detector and photon-counting detector arc, respectively. Collected data were rearranged into XFCT and transmission CT sinograms for image reconstruction. XFCT images were reconstructed with filtered back-projection and with iterative maximum-likelihood expectation maximization without and with attenuation correction. While K-shell XFCT was capable of providing an accurate measurement of Cisplatin concentration, its sensitivity was 4.4 and 3.0 times lower than that of L-shell XFCT with 15 keV excitation beam for the 2 cm and 4 cm diameter phantom, respectively. With the inclusion of excitation and fluorescence beam attenuation correction, we found that L-shell XFCT was capable of providing fairly accurate information of Cisplatin concentration distribution. With a dose of 29 and 58 mGy, clinically relevant Cisplatin Pt concentrations of 10 µg mg −1  could be imaged with L-shell XFCT inside a 2 cm and 4 cm diameter object, respectively. (paper)

  12. Molecular engineering and fluorescence for the detection of toxic cations; Ingenierie moleculaire et fluorescence pour la reconnaissance de cations toxiques

    Energy Technology Data Exchange (ETDEWEB)

    Souchon, V

    2007-11-15

    This work is a part of the 'Toxicologie Nucleaire Environnementale' program which aims at studying the effects on the living of heavy metals or radionuclides involved in nuclear industry. Most particularly, it deals with the design of new fluorescent sensors for the selective detection of Pb{sup 2+}, Cd{sup 2+} and Cs{sup +} in biological media. Several fluorescent calixarenes possessing nitrogen atoms were synthesized and their properties as potential lead sensors were investigated. One of them could be used in experimental conditions close to biological media and new target compounds with amide functional groups were proposed. Many approaches were considered for the design of selective fluorescent sensors for cadmium. On the basis of literature results, many chelating compounds incorporating sulfur atoms were synthesized but showed no significant affinity towards cadmium. On the opposite, compounds functionalized with several pyridine-2'-yl-1,2,3-triazol fluorescent moieties linked to a {beta}-cyclodextrin or a calix[4]arene showed good affinity for cadmium in methanol, but the selectivity was found to be insufficient. In contrast, very satisfying results in terms of both selectivity and sensitivity could be obtained with the commercial calcium sensor Rhod-5N in an aqueous medium at neutral pH. Lastly, micromolar detection limits for the selective detection of caesium were reached in an aqueous medium at neutral pH thanks to a new sulfonated fluorescent calixarene with two appended crown-ethers. An original complexation mechanism was proposed and validated by molecular modelling (DFT). (author)

  13. Enhanced fluorescence of a molecular dipole near metal nanoparticle

    International Nuclear Information System (INIS)

    Pustovit, Vitaliy N.

    2010-01-01

    We study theoretically radiative and nonradiative decay of a single molecule near small gold nanoparticle. The local field enhancement leads to an increased radiative decay rate while the energy transfer from molecule to optically inactive electronic states in nanoparticle results in a decrease in the fluorescence quantum efficiency for small molecule-nanoparticle distances. We performed a DFT-TDLDA calculation of both the enhancement and the quenching for small nanometersized gold nanoparticles. We found that in close proximity to the surface, the nonradiative decay rate is dominated by generation of electron-hole pairs out of the Fermi sea resulting in a significantly lower quantum efficiency as compared to that obtained from electromagnetic calculations. For large distances, the efficiency is maximal for molecule polarized normal to the surface, whereas for small distances it is maximal for parallel orientation.

  14. Enhanced fluorescence of a molecular dipole near metal nanoparticle

    Energy Technology Data Exchange (ETDEWEB)

    Pustovit, Vitaliy N., E-mail: pustovit@ccmsi.u [Chuiko Institute of Surface Chemistry, National Academy of Sciences of Ukraine, General Naumov Street 17, 03164 Kyiv-164 (Ukraine)

    2010-01-15

    We study theoretically radiative and nonradiative decay of a single molecule near small gold nanoparticle. The local field enhancement leads to an increased radiative decay rate while the energy transfer from molecule to optically inactive electronic states in nanoparticle results in a decrease in the fluorescence quantum efficiency for small molecule-nanoparticle distances. We performed a DFT-TDLDA calculation of both the enhancement and the quenching for small nanometersized gold nanoparticles. We found that in close proximity to the surface, the nonradiative decay rate is dominated by generation of electron-hole pairs out of the Fermi sea resulting in a significantly lower quantum efficiency as compared to that obtained from electromagnetic calculations. For large distances, the efficiency is maximal for molecule polarized normal to the surface, whereas for small distances it is maximal for parallel orientation.

  15. Facilitating in vivo tumor localization by principal component analysis based on dynamic fluorescence molecular imaging

    Science.gov (United States)

    Gao, Yang; Chen, Maomao; Wu, Junyu; Zhou, Yuan; Cai, Chuangjian; Wang, Daliang; Luo, Jianwen

    2017-09-01

    Fluorescence molecular imaging has been used to target tumors in mice with xenograft tumors. However, tumor imaging is largely distorted by the aggregation of fluorescent probes in the liver. A principal component analysis (PCA)-based strategy was applied on the in vivo dynamic fluorescence imaging results of three mice with xenograft tumors to facilitate tumor imaging, with the help of a tumor-specific fluorescent probe. Tumor-relevant features were extracted from the original images by PCA and represented by the principal component (PC) maps. The second principal component (PC2) map represented the tumor-related features, and the first principal component (PC1) map retained the original pharmacokinetic profiles, especially of the liver. The distribution patterns of the PC2 map of the tumor-bearing mice were in good agreement with the actual tumor location. The tumor-to-liver ratio and contrast-to-noise ratio were significantly higher on the PC2 map than on the original images, thus distinguishing the tumor from its nearby fluorescence noise of liver. The results suggest that the PC2 map could serve as a bioimaging marker to facilitate in vivo tumor localization, and dynamic fluorescence molecular imaging with PCA could be a valuable tool for future studies of in vivo tumor metabolism and progression.

  16. Fluorescence-Raman Dual Modal Endoscopic System for Multiplexed Molecular Diagnostics

    Science.gov (United States)

    Jeong, Sinyoung; Kim, Yong-Il; Kang, Homan; Kim, Gunsung; Cha, Myeong Geun; Chang, Hyejin; Jung, Kyung Oh; Kim, Young-Hwa; Jun, Bong-Hyun; Hwang, Do Won; Lee, Yun-Sang; Youn, Hyewon; Lee, Yoon-Sik; Kang, Keon Wook; Lee, Dong Soo; Jeong, Dae Hong

    2015-03-01

    Optical endoscopic imaging, which was recently equipped with bioluminescence, fluorescence, and Raman scattering, allows minimally invasive real-time detection of pathologies on the surface of hollow organs. To characterize pathologic lesions in a multiplexed way, we developed a dual modal fluorescence-Raman endomicroscopic system (FRES), which used fluorescence and surface-enhanced Raman scattering nanoprobes (F-SERS dots). Real-time, in vivo, and multiple target detection of a specific cancer was successful, based on the fast imaging capability of fluorescence signals and the multiplex capability of simultaneously detected SERS signals using an optical fiber bundle for intraoperative endoscopic system. Human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR) on the breast cancer xenografts in a mouse orthotopic model were successfully detected in a multiplexed way, illustrating the potential of FRES as a molecular diagnostic instrument that enables real-time tumor characterization of receptors during routine endoscopic procedures.

  17. A small molecular pH-dependent fluorescent probe for cancer cell imaging in living cell.

    Science.gov (United States)

    Ma, Junbao; Li, Wenqi; Li, Juanjuan; Shi, Rongguang; Yin, Gui; Wang, Ruiyong

    2018-05-15

    A novel pH-dependent two-photon fluorescent molecular probe ABMP has been prepared based on the fluorophore of 2, 4, 6-trisubstituted pyridine. The probe has an absorption wavelength at 354 nm and corresponding emission wavelength at 475 nm with the working pH range from 2.20 to 7.00, especially owning a good liner response from pH = 2.40 to pH = 4.00. ABMP also has excellent reversibility, photostability and selectivity which promotes its ability in analytical application. The probe can be excited with a two-photon fluorescence microscopy and the fluorescence cell imaging indicated that the probe can distinguish Hela cancer cells out of normal cells with a two-photon fluorescence microscopy which suggested its potential application in tumor cell detection. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Selective detection of heavy metal ions by calixarene-based fluorescent molecular sensors

    Science.gov (United States)

    Zhang, Haitao; Faye, Djibril; Zhang, Han; Lefevre, Jean-Pierre; Delaire, J. A.; Leray, Isabelle

    2012-06-01

    The synthesis, spectroscopic characterization and complexing properties of calixarene-based fluorescent sensors are reported. The calixarene bearing four dansyl fluorophores (Calix-DANS4) exhibits a very high affinity for the detection of lead. A fluorimetric micro-device based on the use of a Y-shape microchannel was developed and allows lead detection with a 5 ppb detection limit. For mercury detection, a fluorescent molecular sensor containing a calixarene anchored with four 8-quinolinoloxy groups (Calix-Q) has been synthesized. The absorption and fluorescence spectra of this sensor are sensitive to the presence of metal cations. An efficient fluorescence quenching is observed upon mercury complexation because of a photoinduced electron transfer from the fluorophore to the bound mercury. Calix-Q shows a high selectivity towards Hg2+ over interfering cations (Na+, K+, Ca2+, Cu2+, Zn2+, Cd2+ and Pb2+) and a 70 ppb sensitivity.

  19. Iodinated oil-loaded, fluorescent mesoporous silica-coated iron oxide nanoparticles for magnetic resonance imaging/computed tomography/fluorescence trimodal imaging

    Directory of Open Access Journals (Sweden)

    Xue S

    2014-05-01

    Full Text Available Sihan Xue,1 Yao Wang,1 Mengxing Wang,2 Lu Zhang,1 Xiaoxia Du,2 Hongchen Gu,1 Chunfu Zhang1,31School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, 2Shanghai Key Laboratory of Magnetic Resonance, Department of Physics, East China Normal University, 3State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, People’s Republic of ChinaAbstract: In this study, a novel magnetic resonance imaging (MRI/computed tomography (CT/fluorescence trifunctional probe was prepared by loading iodinated oil into fluorescent mesoporous silica-coated superparamagnetic iron oxide nanoparticles (i-fmSiO4@SPIONs. Fluorescent mesoporous silica-coated superparamagnetic iron oxide nanoparticles (fmSiO4@SPIONs were prepared by growing fluorescent dye-doped silica onto superparamagnetic iron oxide nanoparticles (SPIONs directed by a cetyltrimethylammonium bromide template. As prepared, fmSiO4@SPIONs had a uniform size, a large surface area, and a large pore volume, which demonstrated high efficiency for iodinated oil loading. Iodinated oil loading did not change the sizes of fmSiO4@SPIONs, but they reduced the MRI T2 relaxivity (r2 markedly. I-fmSiO4@SPIONs were stable in their physical condition and did not demonstrate cytotoxic effects under the conditions investigated. In vitro studies indicated that the contrast enhancement of MRI and CT, and the fluorescence signal intensity of i-fmSiO4@SPION aqueous suspensions and macrophages, were intensified with increased i-fmSiO4@SPION concentrations in suspension and cell culture media. Moreover, for the in vivo study, the accumulation of i-fmSiO4@SPIONs in the liver could also be detected by MRI, CT, and fluorescence imaging. Our study demonstrated that i-fmSiO4@SPIONs had great potential for MRI/C/fluorescence trimodal imaging.Keywords: multifunctional probe, SPIONs, mesoporous silica

  20. A distance-dependent metal-enhanced fluorescence sensing platform based on molecular beacon design.

    Science.gov (United States)

    Zhou, Zhenpeng; Huang, Hongduan; Chen, Yang; Liu, Feng; Huang, Cheng Zhi; Li, Na

    2014-02-15

    A new metal-enhanced fluorescence (MEF) based platform was developed on the basis of distance-dependent fluorescence quenching-enhancement effect, which combined the easiness of Ag-thiol chemistry with the MEF property of noble-metal structures as well as the molecular beacon design. For the given sized AgNPs, the fluorescence enhancement factor was found to increase with a d(6) dependency in agreement with fluorescence resonance energy transfer mechanism at shorter distance and decrease with a d(-3) dependency in agreement with plasmonic enhancement mechanism at longer distance between the fluorophore and the AgNP surface. As a proof of concept, the platform was demonstrated by a sensitive detection of mercuric ions, using thymine-containing molecular beacon to tune silver nanoparticle (AgNP)-enhanced fluorescence. Mercuric ions were detected via formation of a thymine-mercuric-thymine structure to open the hairpin, facilitating fluorescence recovery and AgNP enhancement to yield a limit of detection of 1 nM, which is well below the U.S. Environmental Protection Agency regulation of the Maximum Contaminant Level Goal (10nM) in drinking water. Since the AgNP functioned as not only a quencher to reduce the reagent blank signal but also an enhancement substrate to increase fluorescence of the open hairpin when target mercuric ions were present, the quenching-enhancement strategy can greatly improve the detection sensitivity and can in principle be a universal approach for various targets when combined with molecular beacon design. © 2013 Elsevier B.V. All rights reserved.

  1. Confirmation of molecular formulas of metallic complexes through X-ray fluorescence quantitative analysis

    International Nuclear Information System (INIS)

    Filgueiras, C.A.L.; Marques, E.V.; Machado, R.M.

    1984-01-01

    X-ray fluorescence spectrophotometry was employed to determined the metal content in a series of five transition element complexes (Mn, Ti, Zn, V). The results confirmed the molecular formulas of these complexes, already proposed on the basis of elemental microanalysis, solution condutimetry and other analytical methods. (C.L.B.) [pt

  2. Optimized measurements of separations and angles between intra-molecular fluorescent markers

    DEFF Research Database (Denmark)

    Mortensen, Kim; Sung, Jongmin; Flyvbjerg, Henrik

    2015-01-01

    We demonstrate a novel, yet simple tool for the study of structure and function of biomolecules by extending two-colour co-localization microscopy to fluorescent molecules with fixed orientations and in intra-molecular proximity. From each colour-separated microscope image in a time-lapse movie...

  3. Exploring Fluorescence Antibunching in Solution To Determine the Stoichiometry of Molecular Complexes

    Czech Academy of Sciences Publication Activity Database

    Sýkora, Jan; Kaiser, K.; Gregor, I.; Bönigk, W.; Schmalzing, G.; Enderlain, J.

    2007-01-01

    Roč. 79, - (2007), s. 4040-4049 ISSN 0003-2700 R&D Projects: GA MŠk(CZ) LC06063 Institutional research plan: CEZ:AV0Z40400503 Keywords : exploring fluorescence antibunching * molecular complex es * biophysical methods Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 5.287, year: 2007

  4. Spectral triangulation molecular contrast optical coherence tomography with indocyanine green as the contrast agent

    OpenAIRE

    Yang, Changhuei; McGuckin, Laura E. L.; Simon, John D.; Choma, Michael A.; Applegate, Brian E.; Izatt, Joseph A.

    2004-01-01

    We report a new molecular contrast optical coherence tomography (MCOCT) implementation that profiles the contrast agent distribution in a sample by measuring the agent's spectral differential absorption. The method, spectra triangulation MCOCT, can effectively suppress contributions from spectrally dependent scatterings from the sample without a priori knowledge of the scattering properties. We demonstrate molecular imaging with this new MCOCT modality by mapping the distribution of indocyani...

  5. Reconstruction method for fluorescent X-ray computed tomography by least-squares method using singular value decomposition

    Science.gov (United States)

    Yuasa, T.; Akiba, M.; Takeda, T.; Kazama, M.; Hoshino, A.; Watanabe, Y.; Hyodo, K.; Dilmanian, F. A.; Akatsuka, T.; Itai, Y.

    1997-02-01

    We describe a new attenuation correction method for fluorescent X-ray computed tomography (FXCT) applied to image nonradioactive contrast materials in vivo. The principle of the FXCT imaging is that of computed tomography of the first generation. Using monochromatized synchrotron radiation from the BLNE-5A bending-magnet beam line of Tristan Accumulation Ring in KEK, Japan, we studied phantoms with the FXCT method, and we succeeded in delineating a 4-mm-diameter channel filled with a 500 /spl mu/g I/ml iodine solution in a 20-mm-diameter acrylic cylindrical phantom. However, to detect smaller iodine concentrations, attenuation correction is needed. We present a correction method based on the equation representing the measurement process. The discretized equation system is solved by the least-squares method using the singular value decomposition. The attenuation correction method is applied to the projections by the Monte Carlo simulation and the experiment to confirm its effectiveness.

  6. A Dual Reporter Iodinated Labeling Reagent for Cancer Positron Emission Tomography Imaging and Fluorescence-Guided Surgery

    Science.gov (United States)

    2018-01-01

    The combination of early diagnosis and complete surgical resection offers the greatest prospect of curative cancer treatment. An iodine-124/fluorescein-based dual-modality labeling reagent, 124I-Green, constitutes a generic tool for one-step installation of a positron emission tomography (PET) and a fluorescent reporter to any cancer-specific antibody. The resulting antibody conjugate would allow both cancer PET imaging and intraoperative fluorescence-guided surgery. 124I-Green was synthesized in excellent radiochemical yields of 92 ± 5% (n = 4) determined by HPLC with an improved one-pot three-component radioiodination reaction. The A5B7 carcinoembryonic antigen (CEA)-specific antibody was conjugated to 124I-Green. High tumor uptake of the dual-labeled A5B7 of 20.21 ± 2.70, 13.31 ± 0.73, and 10.64 ± 1.86%ID/g was observed in CEA-expressing SW1222 xenograft mouse model (n = 3) at 24, 48, and 72 h post intravenous injection, respectively. The xenografts were clearly visualized by both PET/CT and ex vivo fluorescence imaging. These encouraging results warrant the further translational development of 124I-Green for cancer PET imaging and fluorescence-guided surgery. PMID:29388770

  7. Theoretical analysis of fluorescence signals in filamentation of femtosecond laser pulses in nitrogen molecular gas

    International Nuclear Information System (INIS)

    Arevalo, E.; Becker, A.

    2005-01-01

    We study numerically and analytically the role of the combined effect of self-focusing, geometrical focusing, and the plasma defocusing in the formation of the fluorescence signal during the filamentation of a Ti:sapphire laser pulse in nitrogen molecular gas. Results of numerical simulations are used to estimate the number of excited ions in the focal volume, which is proportional to the fluorescence signal. We find good agreement between the theoretical results and the experimental data, showing that such data can be used to get further insight into the effective focal volume during filamentation of femtosecond laser pulses in transparent media

  8. TU-A-9A-05: First Experimental Demonstration of the Anisotropic Detection Principle in X-Ray Fluorescence Computed Tomography

    International Nuclear Information System (INIS)

    Ahmad, M; Bazalova, M; Fahrig, R; Xing, L

    2014-01-01

    Purpose: To improve the sensitivity of X-ray fluorescence computed tomography (XFCT) for in vivo molecular imaging. Is the maximum sensitivity achieved with an isotropic (4π) detector configuration? We prove that this is not necessarily true, and that a greater sensitivity is possible with anisotropic detector configuration. Methods: An XFCT imaging system was constructed consisting of 1) a collimated pencil beam x-ray source using a fluoroscopy grade x-ray tube; 2) a CdTe x-ray photon counting detector to detect fluorescent x-rays; and 3) a rotation/translation stage for tomographic imaging. We created a 6.5-cm diameter water phantom with 2-cm inserts of low gold concentration (0.25%–1%) to simulate tumors targeted by gold nano-particles. The placement of x-ray fluorescence detector were chosen to minimize scatter x-rays. XFCT imaging was performed at three different detector positions (60°, 90°, 145°) to determine the impact of forward-scatter, side-scatter, and back-scatter on imaging performance. The three data sets were also combined to estimate the imaging performance with an isotropic detector. Results: The highest imaging performance was achieved when the XF detector was in the backscatter 145° configuration. The signal-to-noise ratio (SNR) was 5.5 for the 0.25% gold concentration compared to SNRs of 1.4, 0, and 2.4 for 60°, 90°, and combined (60°+90°+145°) datasets. Only the 145° detector arrangement alone could detect the 0.25% concentration. The imaging dose was 14 mGy for each detector arrangement experiment. Conclusion: This study experimentally proves, for the fist time, the Anisotropic Detection Principle in XF imaging, which holds that optimized anisotropic x-ray fluorescence detection provides greater sensitivity than isotropic detection. The optimized detection arrangement was used to improve the sensitivity of the XFCT experiment. The achieved XFCT sensitivity is the highest ever for a phantom at least this large using a benchtop x

  9. Inflammation Modulates Murine Venous Thrombosis Resolution In Vivo: Assessment by Multimodal Fluorescence Molecular Imaging

    Science.gov (United States)

    Ripplinger, Crystal M.; Kessinger, Chase W.; Li, Chunqiang; Kim, Jin Won; McCarthy, Jason R.; Weissleder, Ralph; Henke, Peter K.; Lin, Charles P.; Jaffer, Farouc A.

    2012-01-01

    Objective Assessment of thrombus inflammation in vivo could provide new insights into deep vein thrombosis (DVT) resolution. Here we develop and evaluate two integrated fluorescence molecular-structural imaging strategies to quantify DVT-related inflammation and architecture, and to assess the effect of thrombus inflammation on subsequent DVT resolution in vivo. Methods and Results Murine DVT were created with topical 5% FeCl3 application to thigh or jugular veins (n=35). On day 3, mice received macrophage and matrix metalloproteinase (MMP) activity fluorescence imaging agents. On day 4, integrated assessment of DVT inflammation and architecture was performed using confocal fluorescence intravital microscopy (IVM). Day 4 analyses showed robust relationships among in vivo thrombus macrophages, MMP activity, and FITC-dextran deposition (r>0.70, pthrombus inflammation at day 4 predicted the magnitude of DVT resolution at day 6 (pthrombus resolution. PMID:22995524

  10. Innovative molecular-based fluorescent nanoparticles for multicolor single particle tracking in cells

    International Nuclear Information System (INIS)

    Daniel, Jonathan; Blanchard-Desce, Mireille; Godin, Antoine G; Palayret, Matthieu; Lounis, Brahim; Cognet, Laurent

    2016-01-01

    Based on an original molecular-based design, we present bright and photostable fluorescent organic nanoparticles (FONs) showing excellent colloidal stability in various aqueous environments. Complementary near-infrared emitting and green emitting FONs were prepared using a simple, fast and robust protocol. Both types of FONs could be simultaneously imaged at the single-particle level in solution as well as in biological environments using a monochromatic excitation and a dual-color fluorescence microscope. No evidence of acute cytotoxicity was found upon incubation of live cells with mixed solutions of FONs, and both types of nanoparticles were found internalized in the cells where their motion could be simultaneously tracked at video-rate up to minutes. These fluorescent organic nanoparticles open a novel non-toxic alternative to existing nanoparticles for imaging biological structures, compatible with live-cell experiments and specially fitted for multicolor single particle tracking. (paper)

  11. Cucurbiturils: molecular nanocapsules for time-resolved fluorescence-based assays.

    Science.gov (United States)

    Marquez, Cesar; Huang, Fang; Nau, Werner M

    2004-03-01

    A new fluorescent host-guest system based on the inclusion of the fluorophore 2,3-diazabicyclo[2.2.2]oct-2-ene (DBO) into the cavity of the molecular container compound cucurbit[7]uril (CB7) has been designed which possesses an exceedingly long-lived emission (690 ns in aerated water). The large binding constant of (4 +/- 1) x 10(5) M(-1) along with the resistance of the CB7.DBO complex toward external fluorescence quenchers allow the use of CB7 as an enhancer in time-resolved fluorescence-based assays, e.g., to screen enzyme activity or inhibition by using DBO-labeled peptides as substrates. The response of CB7.DBO to different environmental conditions and possible quenchers are described.

  12. Bladder cancer diagnosis with fluorescence-image-guided optical coherence tomography

    Science.gov (United States)

    Wang, Z. G.; Durand, D. B.; Adler, H.; Pan, Y. T.

    2006-02-01

    A fluorescence-image-guided OCT (FIG-OCT) system is described, and its ability to enhance the sensitivity and specificity is examined in an animal bladder cancer model. Total 97 specimens were examined by fluorescence imaging, OCT and histological microscopy. The sensitivity and specificity of FIG-OCT is 100% and 93% respectively, compared to 79% and 53% for fluorescence imaging, while the OCT examination time has been dramatically decreased by 3~4 times. In combination of endoscopic OCT, FIG-OCT is a promising technique for effective early bladder cancer diagnosis.

  13. Chemodosimeter-based fluorescent detection of L-cysteine after extracted by molecularly imprinted polymers.

    Science.gov (United States)

    Cai, Xiaoqiang; Li, Jinhua; Zhang, Zhong; Wang, Gang; Song, Xingliang; You, Jinmao; Chen, Lingxin

    2014-03-01

    A chemodosimeter-based fluorescent detection method coupled with molecularly imprinted polymers (MIPs) extraction was developed for determination of L-cysteine (L-Cys) by combining molecular imprinting technique with fluorescent chemodosimeter. The MIPs prepared by precipitation polymerization with L-Cys as template, possessed high specific surface area of 145 m(2)/g and good thermal stability without decomposition lower than 300 °C, and were successfully applied as an adsorbent with excellent selectivity for L-Cys over other amino acids, and enantioselectivity was also demonstrated. A novel chemodosimeter, rhodamine B1, was synthesized for discriminating L-Cys from its structurally similar homocysteine and glutathione as well as various possibly co-existing biospecies in aqueous solutions with notable fluorescence enhancement when adding L-Cys. As L-Cys was added with increasing concentrations, an emission band peaked at 580 nm occurred and significantly increased in fluorescence intensity, by which the L-Cys could be sensed optically. High detectability up to 12.5 nM was obtained. An excellent linearity was found within the wide range of 0.05-50 μM (r=0.9996), and reasonable relative standard deviations ranging from 0.3% to 3.5% were attained. Such typical features as high selectivity, high sensitivity, easy operation and low cost enabled this MIPs-fluorometry to be potentially applicable for routine detection of trace L-Cys. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Molecular engineering of two-photon fluorescent probes for bioimaging applications

    Science.gov (United States)

    Liu, Hong-Wen; Liu, Yongchao; Wang, Peng; Zhang, Xiao-Bing

    2017-03-01

    During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems. In this review, we will give a general introduction of molecular engineering of two-photon fluorescent probes based on different fluorescence response mechanisms for bioimaging applications during the past decade. Inspired by the desired advantages of small-molecule two-photon fluorescent probes in biological imaging applications, we expect that more attention will be devoted to the development of new two-photon fluorophores and applications of TPM in areas of bioanalysis and disease diagnosis.

  15. Tomography

    International Nuclear Information System (INIS)

    1985-01-01

    Already widely accepted in medicine, tomography can also be useful in industry. The theory behind tomography and a demonstration of the technique to inspect a motorcycle carburetor is presented. To demonstrate the potential of computer assisted tomography (CAT) to accurately locate defects in three dimensions, a sectioned 5 cm gate valve with a shrink cavity made visible by the sectioning was tomographically imaged using a Co-60 source. The tomographic images revealed a larger cavity below the sectioned surface. The position of this cavity was located with an in-plane and axial precision of approximately +-1 mm. The volume of the cavity was estimated to be approximately 40 mm 3

  16. Automated microaxial tomography of cell nuclei after specific labelling by fluorescence in situ hybridisation

    Czech Academy of Sciences Publication Activity Database

    Kozubek, Michal; Skalníková, M.; Matula, Pe.; Bártová, Eva; Rauch, J.; Neuhaus, F.; Eipel, H.; Hasmann, M.

    2002-01-01

    Roč. 33, 7-8 (2002), s. 655-665 ISSN 0968-4328 Institutional research plan: CEZ:AV0Z5004920 Keywords : microaxial tomography * automated microscopy * high-resolution cytometry Subject RIV: BO - Biophysics Impact factor: 1.537, year: 2002

  17. High-resolution and high sensitivity mesoscopic fluorescence tomography based on de-scanning EMCCD: System design and thick tissue imaging applications

    Science.gov (United States)

    Ozturk, Mehmet Saadeddin

    Optical microscopy has been one of the essential tools for biological studies for decades, however, its application areas was limited to superficial investigation due to strong scattering in live tissues. Even though advanced techniques such as confocal or multiphoton methods have been recently developed to penetrate beyond a few hundreds of microns deep in tissues, they still cannot perform in the mesoscopic regime (millimeter scale) without using destructive sample preparation protocols such as clearing techniques. They provide rich cellular information; however, they cannot be readily employed to investigate the biological processes at larger scales. Herein, we will present our effort to establish a novel imaging approach that can quantify molecular expression in intact tissues, well beyond the current microscopy depth limits. Mesoscopic Fluorescence Molecular Tomography (MFMT) is an emerging imaging modality that offers unique potential for the non-invasive molecular assessment of thick in-vitro and in-vivo live tissues. This novel imaging modality is based on an optical inverse problem that allows for retrieval of the quantitative spatial distribution of fluorescent tagged bio-markers at millimeter depth. MFMT is well-suited for in-vivo subsurface tissue imaging and thick bio-printed specimens due to its high sensitivity and fast acquisition times, as well as relatively large fields of view. Herein, we will first demonstrate the potential of this technique using our first generation MFMT system applied to multiplexed reporter gene imaging (in-vitro) and determination of Photodynamic Therapy (PDT) agent bio-distribution in a mouse model (in-vivo). Second, we will present the design rationale, in silico benchmarking, and experimental validation of a second generation MFMT (2GMFMT) system. We will demonstrate the gain in resolution and sensitivity achieved due to the de-scanned dense detector configuration implemented. The potential of this novel platform will be

  18. Fluorescence Quantum Yield Measurements of Fluorescent Proteins: A Laboratory Experiment for a Biochemistry or Molecular Biophysics Laboratory Course

    Science.gov (United States)

    Wall, Kathryn P.; Dillon, Rebecca; Knowles, Michelle K.

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts…

  19. Infrared images of reflection nebulae and Orion's bar: Fluorescent molecular hydrogen and the 3.3 micron feature

    International Nuclear Information System (INIS)

    Burton, M.G.; Moorhouse, A.; Brand, P.W.J.L.; Roche, P.F.; Geballe, T.R.

    1989-01-01

    Images were obtained of the (fluorescent) molecular hydrogen 1-0 S(1) line, and of the 3.3 micron emission feature, in Orion's Bar and three reflection nebulae. The emission from these species appears to come from the same spatial locations in all sources observed. This suggests that the 3.3 micron feature is excited by the same energetic UV-photons which cause the molecular hydrogen to fluoresce

  20. 3D imaging of vapour and liquid inclusions from the Mole Granite, Australia, using helical fluorescence tomography

    Energy Technology Data Exchange (ETDEWEB)

    Cauzid, J. [European Synchrotron Radiation Facility, 6 rue Jules Horowitz, BP 220, 38043 Grenoble Cedex (France)], E-mail: jean.cauzid@esrf.fr; Philippot, P. [Geobiosphere Actuelle et Primitive, Institut de Physique du Globe de Paris, CNRS and Universite Denis Diderot, Case 89, 4 place Jussieu, 75252 Paris Cedex 05 (France); Bleuet, P. [European Synchrotron Radiation Facility, 6 rue Jules Horowitz, BP 220, 38043 Grenoble Cedex (France); Simionovici, A. [Laboratoire de Geophysique Interne et Tectonophysique, BP 53, 38041 Grenoble Cedex 9 (France); Somogyi, A. [Synchrotron Soleil, DiffAbs beamline, Saint-Aubin, BP 48, 91192 Gif-sur-Yvette Cedex (France); Golosio, B. [Instituto di Matematica e Fisica, Universita di Sassari, 2 via Vienna, 07100 Sassari (Italy)

    2007-08-15

    World class Cu resources are concentrated in porphyry and epithermal ore deposits. Their formation remains partially understood, however, due to a lack of constraints on the partitioning properties of trace elements in general, and Cu in particular, between vapour and liquid phases evolved from boiling fluids at depth in the Earth's crust. Immiscible liquid and vapour fluid inclusions coexisting in a single quartz grain have been imaged in three dimensions by X-ray Fluorescence Computed Tomography (XFCT). Elemental spatial distributions confirm that Cu, and to a lesser extent As, partition into the vapour phase, whereas Mn, Fe, Zn, Br, Rb, Sr and Pb concentrate in the liquid inclusion. High resolution mapping of the vapour inclusions revealed that Cu is heterogeneously distributed at the scale of a single inclusion and is mostly concentrated as tiny daughter crystals.

  1. Optical design of an optical coherence tomography and multispectral fluorescence imaging endoscope to detect early stage ovarian cancer

    Science.gov (United States)

    Tate, Tyler; Keenan, Molly; Swan, Elizabeth; Black, John; Utzinger, Urs; Barton, Jennifer

    2014-12-01

    The five year survival rate for ovarian cancer is over 90% if early detection occurs, yet no effective early screening method exists. We have designed and are constructing a dual modality Optical Coherence Tomography (OCT) and Multispectral Fluorescence Imaging (MFI) endoscope to optically screen the Fallopian tube and ovary for early stage cancer. The endoscope reaches the ovary via the natural pathway of the vagina, cervix, uterus and Fallopian tube. In order to navigate the Fallopian tube the endoscope must have an outer diameter of 600 μm, be highly flexible, steerable, tracking and nonperforating. The imaging systems consists of six optical subsystems, two from OCT and four from MFI. The optical subsystems have independent and interrelated design criteria. The endoscope will be tested on realistic tissue models and ex vivo tissue to prove feasibility of future human trials. Ultimately the project aims to provide women the first effective ovarian cancer screening technique.

  2. 3D imaging of vapour and liquid inclusions from the Mole Granite, Australia, using helical fluorescence tomography

    Science.gov (United States)

    Cauzid, J.; Philippot, P.; Bleuet, P.; Simionovici, A.; Somogyi, A.; Golosio, B.

    2007-08-01

    World class Cu resources are concentrated in porphyry and epithermal ore deposits. Their formation remains partially understood, however, due to a lack of constraints on the partitioning properties of trace elements in general, and Cu in particular, between vapour and liquid phases evolved from boiling fluids at depth in the Earth's crust. Immiscible liquid and vapour fluid inclusions coexisting in a single quartz grain have been imaged in three dimensions by X-ray Fluorescence Computed Tomography (XFCT). Elemental spatial distributions confirm that Cu, and to a lesser extent As, partition into the vapour phase, whereas Mn, Fe, Zn, Br, Rb, Sr and Pb concentrate in the liquid inclusion. High resolution mapping of the vapour inclusions revealed that Cu is heterogeneously distributed at the scale of a single inclusion and is mostly concentrated as tiny daughter crystals.

  3. Extensively Reversible Thermal Transformations of a Bistable, Fluorescence-Switchable Molecular Solid: Entry into Functional Molecular Phase-Change Materials.

    Science.gov (United States)

    Srujana, P; Radhakrishnan, T P

    2015-06-15

    Functional phase-change materials (PCMs) are conspicuously absent among molecular materials in which the various attributes of inorganic solids have been realized. While organic PCMs are primarily limited to thermal storage systems, the amorphous-crystalline transformation of materials like Ge-Sb-Te find use in advanced applications such as information storage. Reversible amorphous-crystalline transformations in molecular solids require a subtle balance between robust supramolecular assembly and flexible structural elements. We report novel diaminodicyanoquinodimethanes that achieve this transformation by interlinked helical assemblies coupled with conformationally flexible alkoxyalkyl chains. They exhibit highly reversible thermal transformations between bistable (crystalline/amorphous) forms, along with a prominent switching of the fluorescence emission energy and intensity. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Caffeine and sulfadiazine interact differently with human serum albumin: A combined fluorescence and molecular docking study

    Science.gov (United States)

    Islam, Mullah Muhaiminul; Sonu, Vikash K.; Gashnga, Pynsakhiat Miki; Moyon, N. Shaemningwar; Mitra, Sivaprasad

    2016-01-01

    The interaction and binding behavior of the well-known drug sulfadiazine (SDZ) and psychoactive stimulant caffeine (CAF) with human serum albumin (HSA) was monitored by in vitro fluorescence titration and molecular docking calculations under physiological condition. The quenching of protein fluorescence on addition of CAF is due to the formation of protein-drug complex in the ground state; whereas in case of SDZ, the experimental results were explained on the basis of sphere of action model. Although both these compounds bind preferentially in Sudlow's site 1 of the protein, the association constant is approximately two fold higher in case of SDZ (∼4.0 × 104 M-1) in comparison with CAF (∼9.3 × 102 M-1) and correlates well with physico-chemical properties like pKa and lipophilicity of the drugs. Temperature dependent fluorescence study reveals that both SDZ and CAF bind spontaneously with HSA. However, the binding of SDZ with the protein is mainly governed by the hydrophobic forces in contrast with that of CAF; where, the interaction is best explained in terms of electrostatic mechanism. Molecular docking calculation predicts the binding of these drugs in different location of sub-domain IIA in the protein structure.

  5. Fluorescent molecularly imprinted polymer based on Navicula sp. frustules for optical detection of lysozyme.

    Science.gov (United States)

    Lim, Guat Wei; Lim, Jit Kang; Ahmad, Abdul Latif; Chan, Derek Juinn Chieh

    2016-03-01

    The direct correlation between disease and lysozyme (LYZ) levels in human body fluids makes the sensitive and convenient detection of LYZ the focus of scientific research. Fluorescent molecularly imprinted polymer has emerged as a new alternative for LYZ detection in order to resolve the limitation of immunoassays, which are expensive, unstable, require complex preparation, and are time consuming. In this study, a novel fluorescence molecularly imprinted polymer based on Navicula sp. frustules (FITC-MIP) has been synthesized via post-imprinting treatment for LYZ detection. Navicula sp. frustules were used as supported material because of their unique properties of moderate surface area, reproducibility, and biocompatibility, to address the drawbacks of nanoparticle core material with low adsorption capacity. The FITC acts as recognition signal and optical readout, whereas MIP provides LYZ selectivity. The synthesized FITC-MIP showed a response time as short as 5 min depending on the concentration of LYZ. It is found that the LYZ template can significantly quench the fluorescence intensity of FITC-MIP linearly within a concentration range of 0 to 0.025 mg mL(-1), which is well described by Stern-Volmer equation. The FITC-MIP can selectively and sensitively detect down to 0.0015 mg mL(-1) of LYZ concentration. The excellent sensing performance of FITC-MIP suggests that FITC-MIP is a potential biosensor in clinical diagnosis applications.

  6. Cyclopenta[b]naphthalene cyanoacrylate dyes: synthesis and evaluation as fluorescent molecular rotors.

    Science.gov (United States)

    Kocsis, Laura S; Elbel, Kristyna M; Hardigree, Billie A; Brummond, Kay M; Haidekker, Mark A; Theodorakis, Emmanuel A

    2015-03-14

    We describe the design, synthesis and fluorescent profile of a family of environment-sensitive dyes in which a dimethylamino (donor) group is conjugated to a cyanoacrylate (acceptor) unit via a cyclopenta[b]naphthalene ring system. This assembly satisfies the typical D-π-A motif of a fluorescent molecular rotor and exhibits solvatochromic and viscosity-sensitive fluorescence emission. The central naphthalene ring system of these dyes was synthesized via a novel intramolecular dehydrogenative dehydro-Diels-Alder (IDDDA) reaction that permits incorporation of the donor and acceptor groups in variable positions around the aromatic core. A bathochromic shift of excitation and emission peaks was observed with increasing solvent polarity but the dyes exhibited a complex emission pattern with a second red emission band when dissolved in nonpolar solvents. Consistent with other known molecular rotors, the emission intensity increased with increasing viscosity. Interestingly, closer spatial proximity between the donor and the acceptor groups led to decreased viscosity sensitivity combined with an increased quantum yield. This observation indicates that structural hindrance of intramolecular rotation dominates when the donor and acceptor groups are in close proximity. The examined compounds give insight into how excited state intramolecular rotation can be influenced by both the solvent and the chemical structure.

  7. Multimodal fluorescence molecular imaging for in vivo characterization of skin cancer using endogenous and exogenous fluorophores

    Science.gov (United States)

    Miller, Jessica P.; Habimana-Griffin, LeMoyne; Edwards, Tracy S.; Achilefu, Samuel

    2017-06-01

    Similarity of skin cancer with many benign skin pathologies requires reliable methods to detect and differentiate the different types of these lesions. Previous studies have explored the use of disparate optical techniques to identify and estimate the invasive nature of melanoma and basal cell carcinoma with varying outcomes. Here, we used a concerted approach that provides complementary information for rapid screening and characterization of tumors, focusing on squamous cell carcinoma (SCC) of the skin. Assessment of in vivo autofluorescence lifetime (FLT) imaging of endogenous fluorophores that are excitable at longer wavelengths (480 nm) than conventional NADH and FAD revealed a decrease in the short FLT component for SCC compared to normal skin, with mean values of 0.57±0.026 ns and 0.61±0.021 ns, respectively (p=0.004). Subsequent systemic administration of a near-infrared fluorescent molecular probe in SCC bearing mice, followed by the implementation of image processing methods on data acquired from two-dimensional and three-dimensional fluorescence molecular imaging, allowed us to estimate the tumor volume and depth, as well as quantify the fluorescent probe in the tumor. The result suggests the involvement of lipofuscin-like lipopigments and riboflavin in SCC metabolism and serves as a model for staging SCC.

  8. Co-registration of fluorescence diffuse optical tomography (fDOT) with positron emission tomography (PET) and development of multi-angle fDOT

    International Nuclear Information System (INIS)

    Tong, X.

    2012-01-01

    This thesis concerns the image processing of fluorescence diffuse optical tomography (fDOT), following two axes: fDOT image co-registration with PET (positron emission tomography) image and improvement of fDOT image reconstructions using mirrors to collect additional projections. It is presented in two parts:In the first part, an automatic method to co-register the fDOT images with PET images has been developed to correlate all the information from each modality. This co-registration method is based on automatic detection of fiducial markers (FM) present in both modalities. The particularity of this method is the use of optical surface image obtained in fDOT imaging system, which serves to identify the Z position of FM in optical images. We tested this method on a model of mice bearing tumor xenografts of MEN2A cancer cells that mimic a human medullary thyroid carcinoma, after a double injection of radiotracer [ 18 F] 2-fluoro-2-Deoxy-D-glucose (FDG) for PET imaging and optical fluorescent infrared tracer Sentidye. With the accuracy of our method, we can demonstrate that the signal of Sentidye is present both in the tumor and surrounding vessels.The fDOT reconstruction image quality is degraded along the Z axis due to a limited number of projections for reconstruction. In the second part, the work is oriented towards a new method of fDOT image reconstruction with a new multi-angle data acquisition system in placing two mirrors on each side of the animal. This work was conducted in collaboration with the CS Department of University College London (UCL), a partner of the European project FMT-XCT. TOAST software developed by this team was used as source code for the reconstruction algorithm, and was modified to adapt to the concerned problem. After several tests on the adjustment of program parameters, we applied this method on a phantom that simulating the biological tissue and on mice. The results showed an improvement in the reconstructed image of a semi

  9. Three-dimensional organization of pKi-67: a comparative fluorescence and electron tomography study using FluoroNanogold.

    Science.gov (United States)

    Cheutin, Thierry; O'Donohue, Marie-Françoise; Beorchia, Adrien; Klein, Christophe; Kaplan, Hervé; Ploton, Dominique

    2003-11-01

    The monoclonal antibody (MAb) Ki-67 is routinely used in clinical studies to estimate the growth fraction of tumors. However, the role of pKi-67, the protein detected by the Ki-67 MAb, remains elusive, although some biochemical data strongly suggest that it might organize chromatin. To better understand the functional organization of pKi-67, we studied its three-dimensional distribution in interphase cells by confocal microscopy and electron tomography. FluoroNanogold, a single probe combining a dense marker with a fluorescent dye, was used to investigate pKi-67 organization at the optical and ultrastructural levels. Observation by confocal microscopy followed by 3D reconstruction showed that pKi-67 forms a shell around the nucleoli. Double labeling experiments revealed that pKi-67 co-localizes with perinucleolar heterochromatin. Electron microscopy studies confirmed this close association and demonstrated that pKi-67 is located neither in the fibrillar nor in the granular components of the nucleolus. Finally, spatial analyses by electron tomography showed that pKi-67 forms cords 250-300 nm in diameter, which are themselves composed of 30-50-nm-thick fibers. These detailed comparative in situ analyses strongly suggest the involvement of pKi-67 in the higher-order organization of perinucleolar chromatin.

  10. The one-sample PARAFAC approach reveals molecular size distributions of fluorescent components in dissolved organic matter

    DEFF Research Database (Denmark)

    Wünsch, Urban; Murphy, Kathleen R.; Stedmon, Colin

    2017-01-01

    Molecular size plays an important role in dissolved organic matter (DOM) biogeochemistry, but its relationship with the fluorescent fraction of DOM (FDOM) remains poorly resolved. Here high-performance size exclusion chromatography (HPSEC) was coupled to fluorescence emission-excitation (EEM...... but not their spectral properties. Thus, in contrast to absorption measurements, bulk fluorescence is unlikely to reliably indicate the average molecular size of DOM. The one-sample approach enables robust and independent cross-site comparisons without large-scale sampling efforts and introduces new analytical...... opportunities for elucidating the origins and biogeochemical properties of FDOM...

  11. Tomography

    International Nuclear Information System (INIS)

    Barrett, H.H.; Gordon, S.; Swindell, W.

    1980-01-01

    Apparatus is described for generating a two-dimensional back-projected image of a slice of an object in tomography. The apparatus uses optical techniques to perform the functions of filtering and back projection. Central to the technique is a cylindrical drum which rotates at a fast rate and whose rotational axis tilts at a slower rate. The novel method overcomes the problem of image blurring due to motion which occurs in many tomographic techniques. It also has the advantages of being less expensive and simpler compared to tomography using digital processing techniques which require fast computers. (UK)

  12. Concept of a selective tumour therapy and its evaluation by near-infrared fluorescence imaging and flat-panel volume computed tomography in mice.

    Science.gov (United States)

    Alves, Frauke; Dullin, Christian; Napp, Joanna; Missbach-Guentner, Jeannine; Jannasch, Katharina; Mathejczyk, Julia; Pardo, Luis A; Stühmer, Walter; Tietze, Lutz-F

    2009-05-01

    Conventional chemotherapy of cancer has its limitations, especially in advanced and disseminated disease and suffers from lack of specificity. This results in a poor therapeutic index and considerable toxicity to normal organs. Therefore, many efforts are made to develop novel therapeutic tools against cancer with the aim of selectively targeting the drug to the tumour site. Drug delivery strategies fundamentally rely on the identification of good-quality biomarkers, allowing unequivocal discrimination between cancer and healthy tissue. At present, antibodies or antibody fragments have clearly proven their value as carrier molecules specific for a tumour-associated molecular marker. This present review draws attention to the use of near-infrared fluorescence (NIRF) imaging to investigate binding specificity and kinetics of carrier molecules such as monoclonal antibodies. In addition, flat-panel volume computed tomography (fpVCT) will be presented to monitor anatomical structures in tumour mouse models over time in a non-invasive manner. Each imaging device sheds light on a different aspect; functional imaging is applied to optimise the dose schedule and the concept of selective tumour therapies, whereas anatomical imaging assesses preclinically the efficacy of novel tumour therapies. Both imaging techniques in combination allow the visualisation of functional information obtained by NIRF imaging within an adequate anatomic framework.

  13. Optical coherent tomography and fluorescent microscopy for the study of meningeal lymphatic systems

    Science.gov (United States)

    Semyachkina-Glushkovskaya, O.; Abdurashitov, A.; Namykin, A.; Fedosov, I.; Pavlov, A.; Karavaev, A.; Sindeeva, O.; Shirokov, A.; Ulanova, M.; Shushunova, N.; Khorovodov, A.; Agranovich, I.; Bodrova, A.; Sagatova, M.; Shareef, Ali Esmat; Saranceva, E.; Dvoryatkina, M.; Tuchin, V.

    2018-04-01

    The development of novel technologies for the imaging of meningeal lymphatic vessels is one of the amazing trends of biophotonics thanks to discovery of brain lymphatics over several years ago. However, there is the limited technologies exist for the study of lymphatics in vivo because lymphatic vessels are transparent with a low speed flow of lymph. Here we demonstrate the successful application of fluorescent microscopy for the imaging of lymphatic system in the mouse brain in vivo.

  14. Recent Advances in Cardiac Computed Tomography: Dual Energy, Spectral and Molecular CT Imaging

    Science.gov (United States)

    Danad, Ibrahim; Fayad, Zahi A.; Willemink, Martin J.; Min, James K.

    2015-01-01

    Computed tomography (CT) evolved into a powerful diagnostic tool and it is impossible to imagine current clinical practice without CT imaging. Due to its widespread availability, ease of clinical application, superb sensitivity for detection of CAD, and non-invasive nature, CT has become a valuable tool within the armamentarium of the cardiologist. In the last few years, numerous technological advances in CT have occurred—including dual energy CT (DECT), spectral CT and CT-based molecular imaging. By harnessing the advances in technology, cardiac CT has advanced beyond the mere evaluation of coronary stenosis to an imaging modality tool that permits accurate plaque characterization, assessment of myocardial perfusion and even probing of molecular processes that are involved in coronary atherosclerosis. Novel innovations in CT contrast agents and pre-clinical spectral CT devices have paved the way for CT-based molecular imaging. PMID:26068288

  15. Quantitative study of luminescence optical tomography. Application to sources localisation in molecular imaging

    International Nuclear Information System (INIS)

    Boffety, Matthieu

    2010-01-01

    Molecular imaging is a major modality in the field of preclinical research. Among the existing methods, techniques based on optical detection of visible or near infrared radiation are the most recent and are mainly represented by luminescence optical tomography techniques. These methods allow for 3D characterization of a biological medium by reconstructing maps of concentration or localisation of luminescent beacons sensitive to biological and chemical processes at the molecular or cellular scale. Luminescence optical tomography is based on a model of light propagation in tissues, a protocol for acquiring surface signal and a numerical inversion procedure used to reconstruct the parameters of interest. This thesis is structured around these three axes and provides an answer to each problem. The main objective of this study is to introduce and present the tools to evaluate the theoretical performances of optical tomography methods. One of its major outcomes is the realisation of experimental tomographic reconstructions from images acquired by an optical imager designed for 2D planar imaging and developed by the company Quidd. In a first step we develop the theory of transport in scattering medium to establish the concept on which our work will rely. We present two different propagation models as well as resolution methods and theoretical difficulties associated with them. In a second part we introduce the statistical tools used to characterise tomographic systems. We define and apply a procedure to simple situations in luminescence optical tomography. The last part of this work presents the development of an inversion procedure. After introducing the theoretical framework we validate the procedure from numerical data before successfully applying it to experimental measurements. (author) [fr

  16. Tomography

    International Nuclear Information System (INIS)

    Allan, C.J.; Keller, N.A.; Lupton, L.R.; Taylor, T.; Tonner, P.D.

    1984-10-01

    Tomography is a non-intrusive imaging technique being developed at CRNL as an industrial tool for generating quantitative cross-sectional density maps of objects. Of most interest is tomography's ability to: distinguish features within complex geometries where other NDT techniques fail because of the complexity of the geometry; detect/locate small density changes/defects within objects, e.g. void fraction measurements within thick-walled vessels, shrink cavities in castings, etc.; provide quantitative data that can be used in analyses, e.g. of complex processes, or fracture mechanics; and provide objective quantitative data that can be used for (computer-based) quality assurance decisions, thereby reducing and in some cases eliminating the present subjectivity often encountered in NDT. The CRNL program is reviewed and examples are presented to illustrate the potential and the limitations of the technology

  17. Temperature dependence of the anisotropy of fluorescence in ring molecular systems

    International Nuclear Information System (INIS)

    Herman, Pavel; Barvik, Ivan

    2007-01-01

    The time dependence of the anisotropy of fluorescence after an impulsive excitation in the molecular ring (resembling the B850 ring of the purple bacterium Rhodopseudomonas acidophila) is calculated. Fast fluctuations of the environment are simulated by dynamic disorder and slow fluctuations by uncorrelated static disorder. Without dynamic disorder modest degrees of static disorder are sufficient to cause the experimentally found initial drop of the anisotropy on a sub-100 fs time scale. In the present investigation we are comparing results for the time-dependent optical anisotropy of the molecular ring for four models of the uncorrelated static disorder: Gaussian disorder in the local energies (model A), Gaussian disorder in the transfer integrals (model B), Gaussian disorder in radial positions of molecules (model C) and Gaussian disorder in angular positions of molecules (model D). Both types of disorder-static and dynamic-are taken into account simultaneously

  18. Comparative Phenotypical and Molecular Analyses of Arabidopsis Grown under Fluorescent and LED Light

    Directory of Open Access Journals (Sweden)

    Franka Seiler

    2017-06-01

    Full Text Available Comparative analyses of phenotypic and molecular traits of Arabidopsis thaliana grown under standardised conditions is still a challenge using climatic devices supplied with common light sources. These are in most cases fluorescent lights, which have several disadvantages such as heat production at higher light intensities, an invariable spectral output, and relatively rapid “ageing”. This results in non-desired variations of growth conditions and lowers the comparability of data acquired over extended time periods. In this study, we investigated the growth behaviour of Arabidopsis Col0 under different light conditions, applying fluorescent compared to LED lamps, and we conducted physiological as well as gene expression analyses. By changing the spectral composition and/or light intensity of LEDs we can clearly influence the growth behaviour of Arabidopsis and thereby study phenotypic attributes under very specific light conditions that are stable and reproducible, which is not necessarily given for fluorescent lamps. By using LED lights, we can also roughly mimic the sun light emission spectrum, enabling us to study plant growth in a more natural-like light set-up. We observed distinct growth behaviour under the different light regimes which was reflected by physiological properties of the plants. In conclusion, LEDs provide variable emission spectra for studying plant growth under defined, stable light conditions.

  19. Fluorescent molecular probes based on excited state prototropism in lipid bilayer membrane

    Science.gov (United States)

    Mohapatra, Monalisa; Mishra, Ashok K.

    2012-03-01

    Excited state prototropism (ESPT) is observed in molecules having one or more ionizable protons, whose proton transfer efficiency is different in ground and excited states. The interaction of various ESPT molecules like naphthols and intramolecular ESPT (ESIPT) molecules like hydroxyflavones etc. with different microheterogeneous media have been studied in detail and excited state prototropism as a probe concept has been gaining ground. The fluorescence of different prototropic forms of such molecules, on partitioning to an organized medium like lipid bilayer membrane, often show sensitive response to the local environment with respect to the local structure, physical properties and dynamics. Our recent work using 1-naphthol as an ESPT fluorescent molecular probe has shown that the incorporation of monomeric bile salt molecules into lipid bilayer membranes composed from dipalmitoylphosphatidylcholine (DPPC, a lung surfactant) and dimyristoylphosphatidylcholine (DMPC), in solid gel and liquid crystalline phases, induce appreciable wetting of the bilayer up to the hydrocarbon core region, even at very low (fisetin, an ESIPT molecule having antioxidant properties, in lipid bilayer membrane has been sensitively monitored from its intrinsic fluorescence behaviour.

  20. Fluorescence in situ hybridization and molecular studies in infertile men with dysplasia of the fibrous sheath.

    Science.gov (United States)

    Baccetti, Baccio; Collodel, Giulia; Gambera, Laura; Moretti, Elena; Serafini, Francesca; Piomboni, Paola

    2005-07-01

    To perform fluorescence in situ hybridization (FISH) and molecular analysis in patients with the genetic sperm defect "dysplasia of the fibrous sheath" (DFS). Retrospective study. Regional Referral Center for Male Infertility, Siena, Italy. Twelve infertile patients with DFS sperm defects. Family history, lymphocytic karyotype, physical and hormonal assays, semen analysis. The DFS sperm phenotype was defined by light, fluorescent, and electron microscopy. Sperm chromosomal constitution was examined by FISH. Gene deletions were tested by polymerase chain reaction. The genetic sperm defect DFS was determined by transmission and scanning electron microscopy. Immunofluorescence staining of A-kinase anchoring protein 4 (AKAP4) showed a moderate and diffuse signal, revealing a disorganized and incompletely assembled fibrous sheath. In 11 of 12 DFS patients, polymerase chain reaction for detecting the presence of partial sequence of AKAP4/AKAP3 binding regions gave positive results. Fluorescence in situ hybridization was performed in decondensed sperm nuclei with probes for chromosomes 18, X, and Y. The mean disomy frequency of chromosome 18 was in the normal range, whereas the mean disomy frequencies of sex chromosomes and diploidies were twice those of controls. These results should be considered when DFS sperm are used in assisted reproductive technology, owing to the high risk of transmission of chromosomal unbalance and of DFS sperm defects to male offspring.

  1. The combination design for open and endoscopic surgery using fluorescence molecular imaging technology

    Science.gov (United States)

    Mao, Yamin; Jiang, Shixin; Ye, Jinzuo; An, Yu; Yang, Xin; Chi, Chongwei; Tian, Jie

    2015-03-01

    For clinical surgery, it is still a challenge to objectively determine tumor margins during surgery. With the development of medical imaging technology, fluorescence molecular imaging (FMI) method can provide real-time intraoperative tumor margin information. Furthermore, surgical navigation system based on FMI technology plays an important role for the aid of surgeons' precise tumor margin decision. However, detection depth is the most limitation exists in the FMI technique and the method convenient for either macro superficial detection or micro deep tissue detection is needed. In this study, we combined advantages of both open surgery and endoscopic imaging systems with FMI technology. Indocyanine green (ICG) experiments were performed to confirm the feasibility of fluorescence detection in our system. Then, the ICG signal was photographed in the detection area with our system. When the system connected with endoscope lens, the minimum quantity of ICG detected by our system was 0.195 ug. For aspect of C mount lens, the sensitivity of ICG detection with our system was 0.195ug. Our experiments results proved that it was feasible to detect fluorescence images with this combination method. Our system shows great potential in the clinical applications of precise dissection of various tumors

  2. 3D ion velocity distribution function measurement in an electric thruster using laser induced fluorescence tomography

    Science.gov (United States)

    Elias, P. Q.; Jarrige, J.; Cucchetti, E.; Cannat, F.; Packan, D.

    2017-09-01

    Measuring the full ion velocity distribution function (IVDF) by non-intrusive techniques can improve our understanding of the ionization processes and beam dynamics at work in electric thrusters. In this paper, a Laser-Induced Fluorescence (LIF) tomographic reconstruction technique is applied to the measurement of the IVDF in the plume of a miniature Hall effect thruster. A setup is developed to move the laser axis along two rotation axes around the measurement volume. The fluorescence spectra taken from different viewing angles are combined using a tomographic reconstruction algorithm to build the complete 3D (in phase space) time-averaged distribution function. For the first time, this technique is used in the plume of a miniature Hall effect thruster to measure the full distribution function of the xenon ions. Two examples of reconstructions are provided, in front of the thruster nose-cone and in front of the anode channel. The reconstruction reveals the features of the ion beam, in particular on the thruster axis where a toroidal distribution function is observed. These findings are consistent with the thruster shape and operation. This technique, which can be used with other LIF schemes, could be helpful in revealing the details of the ion production regions and the beam dynamics. Using a more powerful laser source, the current implementation of the technique could be improved to reduce the measurement time and also to reconstruct the temporal evolution of the distribution function.

  3. Improving x-ray fluorescence signal for benchtop polychromatic cone-beam x-ray fluorescence computed tomography by incident x-ray spectrum optimization: a Monte Carlo study.

    Science.gov (United States)

    Manohar, Nivedh; Jones, Bernard L; Cho, Sang Hyun

    2014-10-01

    To develop an accurate and comprehensive Monte Carlo (MC) model of an experimental benchtop polychromatic cone-beam x-ray fluorescence computed tomography (XFCT) setup and apply this MC model to optimize incident x-ray spectrum for improving production/detection of x-ray fluorescence photons from gold nanoparticles (GNPs). A detailed MC model, based on an experimental XFCT system, was created using the Monte Carlo N-Particle (MCNP) transport code. The model was validated by comparing MC results including x-ray fluorescence (XRF) and scatter photon spectra with measured data obtained under identical conditions using 105 kVp cone-beam x-rays filtered by either 1 mm of lead (Pb) or 0.9 mm of tin (Sn). After validation, the model was used to investigate the effects of additional filtration of the incident beam with Pb and Sn. Supplementary incident x-ray spectra, representing heavier filtration (Pb: 2 and 3 mm; Sn: 1, 2, and 3 mm) were computationally generated and used with the model to obtain XRF/scatter spectra. Quasimonochromatic incident x-ray spectra (81, 85, 90, 95, and 100 keV with 10 keV full width at half maximum) were also investigated to determine the ideal energy for distinguishing gold XRF signal from the scatter background. Fluorescence signal-to-dose ratio (FSDR) and fluorescence-normalized scan time (FNST) were used as metrics to assess results. Calculated XRF/scatter spectra for 1-mm Pb and 0.9-mm Sn filters matched (r ≥ 0.996) experimental measurements. Calculated spectra representing additional filtration for both filter materials showed that the spectral hardening improved the FSDR at the expense of requiring a much longer FNST. In general, using Sn instead of Pb, at a given filter thickness, allowed an increase of up to 20% in FSDR, more prominent gold XRF peaks, and up to an order of magnitude decrease in FNST. Simulations using quasimonochromatic spectra suggested that increasing source x-ray energy, in the investigated range of 81-100 ke

  4. SU-E-T-20: A Novel Hybrid CBCT, Bioluminescence and Fluorescence Tomography System for Preclinical Radiation Research

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, B; Eslami, S; Iordachita, I [Johns Hopkins University, Baltimore, Maryland (United States); Yang, Y [University of Miami School of Medicine, Miami, FL (United States); Patterson, M [Hamilton Regional Cancer Ctr., Hamilton, ON (Canada); Wong, J [Johns Hopkins University, Baltimore, MD (United States); Wang, K [Johns Hopkins Hospital, Baltimore, MD (United States)

    2014-06-01

    Purpose: A novel standalone bioluminescence and fluorescence tomography (BLT and FT) system equipped with high resolution CBCT has been built in our group. In this work, we present the system calibration method and validate our system in both phantom and in vivo environment. Methods: The CBCT is acquired by rotating the animal stage while keeping the x-ray source and detector panel static. The optical signal is reflected by the 3-mirror system to a multispectral filter set and then delivered to the CCD camera with f/1.4 lens mounted. Nine fibers passing through the stage and in contact with the mouse skin serve as the light sources for diffuse optical tomography (DOT) and FT. The anatomical information and optical properties acquired from the CBCT and DOT, respectively, are used as the priori information to improve the BLT/FT reconstruction accuracy. Flat field correction for the optical system was acquired at multiple wavelengths. A home-built phantom is used to register the optical and CBCT coordinates. An absolute calibration relating the CCD photon counts rate to the light fluence rate emitted at animal surface was developed to quantify the bioluminescence power or fluorophore concentration. Results: An optical inhomogeneous phantom with 2 light sources (3mm separation) imbedded is used to test the system. The optical signal is mapped onto the mesh generated from CBCT for optical reconstruction. Our preliminary results show that the center of mass can be reconstructed within 2.8mm accuracy. A live mouse with the light source imbedded is also used to validate our system. Liver or lung metastatic luminescence tumor model will be used for further testing. Conclusion: This hybrid system transforms preclinical research to a level that even sub-palpable volume of cells can be imaged rapidly and non-invasively, which largely extends the scope of radiobiological research. The research is supported by the NCI grant R01CA158100-01.

  5. Image-guided intraocular injection using multimodality optical coherence tomography and fluorescence confocal scanning laser ophthalmoscopy in rodent ophthalmological models

    Science.gov (United States)

    Terrones, Benjamin D.; Benavides, Oscar R.; Leeburg, Kelsey C.; Mehanathan, Sankarathi B.; Levine, Edward M.; Tao, Yuankai K.

    2018-02-01

    Intraocular injections are routinely performed for delivery of anti-VEGF and anti-inflammatory therapies in humans. While these injections are also performed in mice to develop novel models of ophthalmic diseases and screen novel therapeutics, the injection location and volume are not well-controlled and reproducible. We overcome limitations of conventional injections methods by developing a multimodality, long working distance, non-contact optical coherence tomography (OCT) and fluorescence confocal scanning laser ophthalmoscopy (cSLO) system for retinal imaging before and after injections. Our OCT+cSLO system combines a custom-built spectraldomain OCT engine (875+/-85 nm) with 125 kHz line-rate with a modified commercial cSLO with a maximum frame-rate of 30 fps (512 x 512 pix.). The system was designed for an overlapping OCT+cSLO field-of-view of 1.1 mm with a 7.76 mm working distance to the pupil. cSLO excitation light sources and filters were optimized for simultaneous GFP and tdTomato imaging. Lateral resolution was 3.02 µm for OCT and 2.74 μm for cSLO. Intravitreal injections of 5%, 10%, and 20% intralipid with Alex Fluor 488 were manually injected intraocularly in C57BL/6 mice. Post-injection imaging showed structural changes associated with retinal puncture, including the injection track, a retinal elevation, and detachment of the posterior hyaloid. OCT enables quantitative analysis of injection location and volumes whereas complementary cSLO improves specificity for identifying fluorescently labeled injected compounds and transgenic cells. The long working distance of our non-contact OCT+cSLO system is uniquely-suited for concurrent imaging with intraocular injections and may be applied for imaging of ophthalmic surgical dynamics and real-time image-guided injections.

  6. Enhancement of molecular sensitivity in positron emission tomography with quantum correlation of γ-ray photons

    Science.gov (United States)

    Sato, K.; Kobayashi, Y.

    2015-05-01

    Enhancement of molecular sensitivity in positron emission tomography (PET) has long been discussed with respect to imaging instrumentation and algorithms for data treatment. Here, the molecular sensitivity in PET is discussed on the basis of 2-dimensional coincident measurements of 511 keV γ ray photons resultant from two-photon annihilation. Introduction of an additional selection window based on the energy sum and difference of the coincidently measured γ ray photons, without any significant instrumental and algorithmic changes, showed an improvement in the signal-to-noise ratio (SNR) by an order of magnitude. Improvement of performance characteristics in the PET imaging system was demonstrated by an increase in the noise equivalent count rate (NECR) which takes both the SNR and the detection efficiency into consideration. A further improvement of both the SNR and the NECR is expected for the present system in real clinical and in-vivo environments, where much stronger positron sources are employed.

  7. Enhancement of molecular sensitivity in positron emission tomography with quantum correlation of γ-ray photons

    International Nuclear Information System (INIS)

    Sato, K.; Kobayashi, Y.

    2015-01-01

    Enhancement of molecular sensitivity in positron emission tomography (PET) has long been discussed with respect to imaging instrumentation and algorithms for data treatment. Here, the molecular sensitivity in PET is discussed on the basis of 2-dimensional coincident measurements of 511 keV γ ray photons resultant from two-photon annihilation. Introduction of an additional selection window based on the energy sum and difference of the coincidently measured γ ray photons, without any significant instrumental and algorithmic changes, showed an improvement in the signal-to-noise ratio (SNR) by an order of magnitude. Improvement of performance characteristics in the PET imaging system was demonstrated by an increase in the noise equivalent count rate (NECR) which takes both the SNR and the detection efficiency into consideration. A further improvement of both the SNR and the NECR is expected for the present system in real clinical and in-vivo environments, where much stronger positron sources are employed

  8. Fluorescence and NMR spectroscopy together with molecular simulations reveal amphiphilic characteristics of a Burkholderia biofilm exopolysaccharide.

    Science.gov (United States)

    Kuttel, Michelle M; Cescutti, Paola; Distefano, Marco; Rizzo, Roberto

    2017-06-30

    Biofilms are a collective mode of bacterial life in which a self-produced matrix confines cells in close proximity to each other. Biofilms confer many advantages, including protection from chemicals (including antibiotics), entrapment of useful extracellular enzymes and nutrients, as well as opportunities for efficient recycling of molecules from dead cells. Biofilm matrices are aqueous gel-like structures composed of polysaccharides, proteins, and DNA stabilized by intermolecular interactions that may include non-polar connections. Recently, polysaccharides extracted from biofilms produced by species of the Burkholderia cepacia complex were shown to possess clusters of rhamnose, a 6-deoxy sugar with non-polar characteristics. Molecular dynamics simulations are well suited to characterizing the structure and dynamics of polysaccharides, but only relatively few such studies exist of their interaction with non-polar molecules. Here we report an investigation into the hydrophobic properties of the exopolysaccharide produced by Burkholderia multivorans strain C1576. Fluorescence experiments with two hydrophobic fluorescent probes established that this polysaccharide complexes hydrophobic species, and NMR experiments confirmed these interactions. Molecular simulations to model the hydrodynamics of the polysaccharide and the interaction with guest species revealed a very flexible, amphiphilic carbohydrate chain that has frequent dynamic interactions with apolar molecules; both hexane and a long-chain fatty acid belonging to the quorum-sensing system of B. multivorans were tested. A possible role of the non-polar domains of the exopolysaccharide in facilitating the diffusion of aliphatic species toward specific targets within the biofilm aqueous matrix is proposed. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. Comparison of Cherenkov excited fluorescence and phosphorescence molecular sensing from tissue with external beam irradiation.

    Science.gov (United States)

    Lin, Huiyun; Zhang, Rongxiao; Gunn, Jason R; Esipova, Tatiana V; Vinogradov, Sergei; Gladstone, David J; Jarvis, Lesley A; Pogue, Brian W

    2016-05-21

    Ionizing radiation delivered by a medical linear accelerator (LINAC) generates Cherenkov emission within the treated tissue. A fraction of this light, in the 600-900 nm wavelength region, propagates through centimeters of tissue and can be used to excite optical probes in vivo, enabling molecular sensing of tissue analytes. The success of isolating the emission signal from this Cherenkov excitation background is dependent on key factors such as: (i) the Stokes shift of the probe spectra; (ii) the excited state lifetime; (iii) the probe concentration; (iv) the depth below the tissue surface; and (v) the radiation dose used. Previous studies have exclusively focused on imaging phosphorescent dyes, rather than fluorescent dyes. However there are only a few biologically important phosphorescent dyes and yet in comparison there are thousands of biologically relevant fluorescent dyes. So in this study the focus was a study of efficacy of Cherenkov-excited luminescence using fluorescent commercial near-infrared probes, IRDye 680RD, IRDye 700DX, and IRDye 800CW, and comparing them to the well characterized phosphorescent probe Oxyphor PtG4, an oxygen sensitive dye. Each probe was excited by Cherenkov light from a 6 MV external radiation beam, and measured in continuous wave or time-gated modes. The detection was performed by spectrally resolving the luminescence signals, and measuring them with spectrometer-based separation on an ICCD detector. The results demonstrate that IRDye 700DX and PtG4 allowed for the maximal signal to noise ratio. In the case of the phosphorescent probe, PtG4, with emission decays on the microsecond (μs) time scale, time-gated acquisition was possible, and it allowed for higher efficacy in terms of the probe concentration and detection depth. Phantoms containing the probe at 5 mm depth could be detected at concentrations down to the nanoMolar range, and at depths into the tissue simulating phantom near 3 cm. In vivo studies showed that 5

  10. Assessing pharmacokinetics of indocyanine green-loaded nanoparticle in tumor with a dynamic diffuse fluorescence tomography system

    Science.gov (United States)

    Zhang, Yanqi; Yin, Guoyan; Zhao, Huijuan; Ma, Wenjuan; Gao, Feng; Zhang, Limin

    2018-02-01

    Real-time and continuous monitoring of drug release in vivo is an important task in pharmaceutical development. Here, we devoted to explore a real-time continuous study of the pharmacokinetics of free indocyanine green (ICG) and ICG loaded in the shell-sheddable nanoparticles in tumor based on a dynamic diffuse fluorescence tomography (DFT) system: A highly-sensitive dynamic DFT system of CT-scanning mode generates informative and instantaneous sampling datasets; An analysis procedure extracts the pharmacokinetic parameters from the reconstructed time curves of the mean ICG concentration in tumor, using the Gauss-Newton scheme based on two-compartment model. Compared with the pharmacokinetic parameters of free ICG in tumor, the ICG loaded in the shell-sheddable nanoparticles shows efficient accumulation in tumor. The results demonstrate our proposed dynamic-DFT can provide an integrated and continuous view of the drug delivery of the injected agents in different formulations, which is helpful for the development of diagnosis and therapy for tumors.

  11. Suppression of Kasha's rule as a mechanism for fluorescent molecular rotors and aggregation-induced emission

    Science.gov (United States)

    Qian, Hai; Cousins, Morgan E.; Horak, Erik H.; Wakefield, Audrey; Liptak, Matthew D.; Aprahamian, Ivan

    2017-01-01

    Although there are some proposed explanations for aggregation-induced emission, a phenomenon with applications that range from biosensors to organic light-emitting diodes, current understanding of the quantum-mechanical origin of this photophysical behaviour is limited. To address this issue, we assessed the emission properties of a series of BF2-hydrazone-based dyes as a function of solvent viscosity. These molecules turned out to be highly efficient fluorescent molecular rotors. This property, in addition to them being aggregation-induced emission luminogens, enabled us to probe deeper into their emission mechanism. Time-dependent density functional theory calculations and experimental results showed that the emission is not from the S1 state, as predicted from Kasha's rule, but from a higher energy (>S1) state. Furthermore, we found that suppression of internal conversion to the dark S1 state by restricting the rotor rotation enhances fluorescence, which leads to the proposal that suppression of Kasha's rule is the photophysical mechanism responsible for emission in both viscous solution and the solid state.

  12. A Far-ultraviolet Fluorescent Molecular Hydrogen Emission Map of the Milky Way Galaxy

    Energy Technology Data Exchange (ETDEWEB)

    Jo, Young-Soo; Min, Kyoung-Wook [Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701 (Korea, Republic of); Seon, Kwang-Il; Han, Wonyong [Korea Astronomy and Space Science Institute (KASI), 776 Daedeokdae-ro, Yuseong-gu, Daejeon, 305-348 (Korea, Republic of); Edelstein, Jerry, E-mail: stspeak@gmail.com [Space Sciences Laboratory, University of California, Berkeley, CA, 94720 (United States)

    2017-08-01

    We present the far-ultraviolet (FUV) fluorescent molecular hydrogen (H{sub 2}) emission map of the Milky Way Galaxy obtained with FIMS/SPEAR covering ∼76% of the sky. The extinction-corrected intensity of the fluorescent H{sub 2} emission has a strong linear correlation with the well-known tracers of the cold interstellar medium (ISM), including color excess E(B–V) , neutral hydrogen column density N (H i), and H α emission. The all-sky H{sub 2} column density map was also obtained using a simple photodissociation region model and interstellar radiation fields derived from UV star catalogs. We estimated the fraction of H{sub 2} ( f {sub H2}) and the gas-to-dust ratio (GDR) of the diffuse ISM. The f {sub H2} gradually increases from <1% at optically thin regions where E(B–V) < 0.1 to ∼50% for E(B–V)  = 3. The estimated GDR is ∼5.1 × 10{sup 21} atoms cm{sup −2} mag{sup −1}, in agreement with the standard value of 5.8 × 10{sup 21} atoms cm{sup −2} mag{sup −1}.

  13. Nitrogen-doped graphene quantum dots-based fluorescence molecularly imprinted sensor for thiacloprid detection.

    Science.gov (United States)

    Liu, Yang; Cao, Nan; Gui, Wenying; Ma, Qiang

    2018-06-01

    In this paper, a test strip-based sensor was developed for thiacloprid quantitative detection based on PDA molecularly imprinted polymer (MIP) and nitrogen-doped graphene quantum dots (N-GQDs). Thiacloprid is a new type of nicotine insecticide, which can block the normal neurotransmitter delivery process in insects. In the sensing system, N-GQDs were immersed into filter paper at first. Then, dopamine (DA) with thiacloprid can be self-polymerized on test strip surface to form the uniform PDA film. After removed thiacloprid template, the established poly dopamine (PDA) MIP can selectively recognize thiacloprid. As a result, captured thiacloprid can enhance the fluorescence intensity of N-GQDs into the test strip. As a result, the fluorescence intensity of N-GQDs can be linearly related within a certain range of thiacloprid concentration. Under the optimum conditions, the proposed sensor for thiacloprid detection exhibited a linear ranging from 0.1 mg/L to 10 mg/L with a low detection limit of 0.03 mg/L. The N-GQDs based test strip-based sensor for thiaclopridis reported for the first time. The sensing system has high selectivity to thiacloprid and provides new opportunities in the pesticide detection. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Fluorescent Molecular Rotor-in-Paraffin Waxes for Thermometry and Biometric Identification.

    Science.gov (United States)

    Jin, Young-Jae; Dogra, Rubal; Cheong, In Woo; Kwak, Giseop

    2015-07-08

    Novel thermoresponsive sensor systems consisting of a molecular rotor (MR) and paraffin wax (PW) were developed for various thermometric and biometric identification applications. Polydiphenylacetylenes (PDPAs) coupled with long alkyl chains were used as MRs, and PWs of hydrocarbons having 16-20 carbons were utilized as phase-change materials. The PDPAs were successfully dissolved in the molten PWs and did not act as an impurity that prevents phase transition of the PWs. These PDPA-in-PW hybrids had almost the same enthalpies and phase-transition temperatures as the corresponding pure PWs. The hybrids exhibited highly reversible fluorescence (FL) changes at the critical temperatures during phase transition of the PWs. These hybrids were impregnated into common filter paper in the molten state by absorption or were encapsulated into urea resin to enhance their mechanical integrity and cyclic stability during repeated use. The wax papers could be utilized in highly advanced applications including FL image writing/erasing, an array-type thermo-indicator, and fingerprint/palmprint identification. The present findings should facilitate the development of novel fluorescent sensor systems for biometric identification and are potentially applicable for biological and biomedical thermometry.

  15. Simple fluorescence-based detection of protein kinase A activity using a molecular beacon probe.

    Science.gov (United States)

    Ma, Changbei; Lv, Xiaoyuan; Wang, Kemin; Jin, Shunxin; Liu, Haisheng; Wu, Kefeng; Zeng, Weimin

    2017-11-02

    Protein kinase A was detected by quantifying the amount of ATP used after a protein kinase reaction. The ATP assay was performed using the T4 DNA ligase and a molecular beacon (MB). In the presence of ATP, DNA ligase catalyzed the ligation of short DNA. The ligation product then hybridized to MB, resulting in a fluorescence enhancement of the MB. This assay was capable of determining protein kinase A in the range of 12.5∼150 nM, with a detection limit of 1.25 nM. Furthermore, this assay could also be used to investigate the effect of genistein on protein kinase A. It was a universal, non-radioisotopic, and homogeneous method for assaying protein kinase A.

  16. Tomography

    International Nuclear Information System (INIS)

    Brown, B.H.; Barber, D.C.; Freeston, I.L.

    1983-01-01

    Tomography images of a body are constructed by placing a plurality of surface electrodes at spaced intervals on the body, causing currents to flow in the body (e.g. by applying a potential between each pair of electrodes in turn, or by induction), and measuring the potential between pairs of electrodes, calculating the potential expected in each case on the assumption that the body consists of a medium of uniform impedance, plotting the isopotentials corresponding to the calculated results to create a uniform image of the body, obtaining the ratio between the measured potential and the calculated potential in each case, and modifying the image in accordance with the respective ratios by increasing the assumed impedance along an isopotential in proportion to a ratio greater than unity or decreasing the assumed impedance in proportion to a ratio less than unity. The modified impedances along the isopotentials for each pair of electrodes are superimposed. The calculations are carried out using a computer and the plotting is carried out by a visual display unit and/or a print-out unit. (author)

  17. Positron emission tomography in amyotrophic lateral sclerosis: Towards targeting of molecular pathological hallmarks

    International Nuclear Information System (INIS)

    Willekens, Stefanie M.A.; Weehaeghe, Donatienne van; Damme, Philip van; Laere, Koen van

    2017-01-01

    During the past decades, extensive efforts have been made to expand the knowledge of amyotrophic lateral sclerosis (ALS). However, clinical translation of this research, in terms of earlier diagnosis and improved therapy, remains challenging. Since more than 30% of motor neurons are lost when symptoms become clinically apparent, techniques allowing non-invasive, in vivo detection of motor neuron degeneration are needed in the early, pre-symptomatic disease stage. Furthermore, it has become apparent that non-motor signs play an important role in the disease and there is an overlap with cognitive disorders, such as frontotemporal dementia (FTD). Radionuclide imaging, such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT), form an attractive approach to quantitatively monitor the ongoing neurodegenerative processes. Although ["1"8F]-FDG has been recently proposed as a potential biomarker for ALS, active targeting of the underlying pathologic molecular processes is likely to unravel further valuable disease information and may help to decipher the pathogenesis of ALS. In this review, we provide an overview of radiotracers that have already been applied in ALS and discuss possible novel targets for in vivo imaging of various pathogenic processes underlying ALS onset and progression. (orig.)

  18. Positron emission tomography in amyotrophic lateral sclerosis: Towards targeting of molecular pathological hallmarks

    Energy Technology Data Exchange (ETDEWEB)

    Willekens, Stefanie M.A.; Weehaeghe, Donatienne van [University Hospitals Leuven and KU Leuven, Division of Nuclear Medicine, Department of Imaging and Pathology, Leuven (Belgium); Damme, Philip van [University Hospitals Leuven, Department of Neurology, Leuven (Belgium); KU Leuven, Department of Neurosciences, Experimental Neurology, Leuven (Belgium); Leuven Research Institute for Neuroscience and Disease (LIND), Leuven (Belgium); VIB, Vesalius Research Center, Laboratory of Neurobiology, Leuven (Belgium); Laere, Koen van [University Hospitals Leuven and KU Leuven, Division of Nuclear Medicine, Department of Imaging and Pathology, Leuven (Belgium); Leuven Research Institute for Neuroscience and Disease (LIND), Leuven (Belgium)

    2017-03-15

    During the past decades, extensive efforts have been made to expand the knowledge of amyotrophic lateral sclerosis (ALS). However, clinical translation of this research, in terms of earlier diagnosis and improved therapy, remains challenging. Since more than 30% of motor neurons are lost when symptoms become clinically apparent, techniques allowing non-invasive, in vivo detection of motor neuron degeneration are needed in the early, pre-symptomatic disease stage. Furthermore, it has become apparent that non-motor signs play an important role in the disease and there is an overlap with cognitive disorders, such as frontotemporal dementia (FTD). Radionuclide imaging, such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT), form an attractive approach to quantitatively monitor the ongoing neurodegenerative processes. Although [{sup 18}F]-FDG has been recently proposed as a potential biomarker for ALS, active targeting of the underlying pathologic molecular processes is likely to unravel further valuable disease information and may help to decipher the pathogenesis of ALS. In this review, we provide an overview of radiotracers that have already been applied in ALS and discuss possible novel targets for in vivo imaging of various pathogenic processes underlying ALS onset and progression. (orig.)

  19. First molecular identification of the transgene red fluorescent protein (RFP in transgenic ornamental zebrafish (Danio rerio introduced in Peru

    Directory of Open Access Journals (Sweden)

    Carlos Scotto

    2013-09-01

    Full Text Available In this paper the transgenic fluorescent red, orange and pink zebra fish (Danio rerio, found in local aquariums in Peru, were identified using the PCR technique to amplify the transgene RFP sea anemone belonging to Discosoma spp. The gene expression of the red fluorescent protein (RFP transgene was found to determine different gradients-of-bioluminescence (shades in color in each GMO fish analyzed. We performed sequence analysis of the two variants of the RFP along with six variants of the existing fluorescent protein GFP from the Genbank, this could help identify quickly if they are new genes or variants thereof as these novel fluorescent proteins may be introduced in aquatic GMO in the future. Thus, developing and improving biosecurity measures through its timely detection at the molecular genetic level.

  20. Relating Trp-Glu dipeptide fluorescence to molecular conformation: the role of the discrete Chi 1 and Chi 2 angles.

    Science.gov (United States)

    Eisenberg, Azaria Solomon; Juszczak, Laura J

    2013-07-05

    Molecular dynamics (MD), coupled with fluorescence data for charged dipeptides of tryptophanyl glutamic acid (Trp-Glu), reveal a detailed picture of how specific conformation affects fluorescence. Fluorescence emission spectra and time-resolved emission measurements have been collected for all four charged species. MD simulations 20 to 30 ns in length have also been carried out for the Trp-Glu species, as simulation provides aqueous phase conformational data that can be correlated with the fluorescence data. The calculations show that each dipeptide species is characterized by a similar set of six, discrete Chi 1, Chi 2 dihedral angle pairs. The preferred Chi 1 angles--60°, 180°, and 300°--play the significant role in positioning the terminal amine relative to the indole ring. A Chi 1 angle of 60° results in the arching of the backbone over the indole ring and no interaction of the ring with the terminal amine. Chi 1 values of 180° and 300° result in an extension of the backbone away from the indole ring and a NH3 cation-π interaction with indole. This interaction is believed responsible for charge transfer quenching. Two fluorescence lifetimes and their corresponding amplitudes correlate with the Chi 1 angle probability distribution for all four charged Trp-Glu dipeptides. Fluorescence emission band maxima are also consistent with the proposed pattern of terminal amine cation quenching of fluorescence. Copyright © 2013 Wiley Periodicals, Inc.

  1. In vivo molecular photoacoustic tomography of melanomas targeted by bioconjugated gold nanocages.

    Science.gov (United States)

    Kim, Chulhong; Cho, Eun Chul; Chen, Jingyi; Song, Kwang Hyun; Au, Leslie; Favazza, Christopher; Zhang, Qiang; Cobley, Claire M; Gao, Feng; Xia, Younan; Wang, Lihong V

    2010-08-24

    Early diagnosis, accurate staging, and image-guided resection of melanomas remain crucial clinical objectives for improving patient survival and treatment outcomes. Conventional techniques cannot meet this demand because of the low sensitivity, low specificity, poor spatial resolution, shallow penetration, and/or ionizing radiation. Here we overcome such limitations by combining high-resolution photoacoustic tomography (PAT) with extraordinarily optical absorbing gold nanocages (AuNCs). When bioconjugated with [Nle(4),D-Phe(7)]-alpha-melanocyte-stimulating hormone, the AuNCs can serve as a novel contrast agent for in vivo molecular PAT of melanomas with both exquisite sensitivity and high specificity. The bioconjugated AuNCs enhanced contrast approximately 300% more than the control, PEGylated AuNCs. The in vivo PAT quantification of the amount of AuNCs accumulated in melanomas was further validated with inductively coupled plasma mass spectrometry (ICP-MS).

  2. In vivo molecular photoacoustic tomography of melanomas targeted by bio-conjugated gold nanocages

    Science.gov (United States)

    Kim, Chulhong; Cho, Eun Chul; Chen, Jingyi; Song, Kwang Hyun; Au, Leslie; Favazza, Christopher; Zhang, Qiang; Cobley, Claire M.; Gao, Feng; Xia, Younan; Wang, Lihong V.

    2010-01-01

    Early diagnosis, accurate staging, and image-guided resection of melanomas remain crucial clinical objectives for improving patient survival and treatment outcomes. Conventional techniques cannot meet this demand because of the low sensitivity, low specificity, poor spatial resolution, shallow penetration, and/or ionizing radiation. Here we overcome such limitations by combining high-resolution photoacoustic tomography (PAT) with extraordinarily optical absorbing gold nanocages (AuNCs). When bio-conjugated with [Nle4,D-Phe7]-α-melanocyte-stimulating hormone, the AuNCs can serve as a novel contrast agent for in vivo molecular PAT of melanomas with both exquisite sensitivity and high specificity. The bio-conjugated AuNCs enhanced contrast ~300% more than the control, PEGylated AuNCs. The in vivo PAT quantification of the amount of AuNCs accumulated in melanomas was further validated with inductively coupled plasma mass spectrometry (ICP-MS). PMID:20731439

  3. A quencher-free molecular beacon design based on pyrene excimer fluorescence using pyrene-labeled UNA (unlocked nucleic acid)

    DEFF Research Database (Denmark)

    Karlsen, Kasper Kannegård; Okholm, Anders Hauge; Kjems, Jørgen

    2013-01-01

    A quencher-free molecular beacon capable of generating pyrene excimer fluorescence has been constructed using strategically positioned pyrene-UNA monomers. Hybridization of a fully complementary RNA target was accompanied by a pyrene excimer emission increase of more than 900%, and detection of RNA...

  4. Fluorescence- and capillary electrophoresis (CE)-based SSR DNA fingerprinting and a molecular identity database for the Louisiana sugarcane industry

    Science.gov (United States)

    A database of Louisiana sugarcane molecular identity has been constructed and is being updated annually using FAM or HEX or NED fluorescence- and capillary electrophoresis (CE)-based microsatellite (SSR) fingerprinting information. The fingerprints are PCR-amplified from leaf DNA samples of current ...

  5. Noninvasive imaging of multiple myeloma using near infrared fluorescent molecular probe

    Science.gov (United States)

    Hathi, Deep; Zhou, Haiying; Bollerman-Nowlis, Alex; Shokeen, Monica; Akers, Walter J.

    2016-03-01

    Multiple myeloma is a plasma cell malignancy characterized by monoclonal gammopathy and osteolytic bone lesions. Multiple myeloma is most commonly diagnosed in late disease stages, presenting with pathologic fracture. Early diagnosis and monitoring of disease status may improve quality of life and long-term survival for multiple myeloma patients from what is now a devastating and fatal disease. We have developed a near-infrared targeted fluorescent molecular probe with high affinity to the α4β1 integrin receptor (VLA-4)overexpressed by a majority of multiple myeloma cells as a non-radioactive analog to PET/CT tracer currently being developed for human diagnostics. A near-infrared dye that emits about 700 nm was conjugated to a high affinity peptidomimmetic. Binding affinity and specificity for multiple myeloma cells was investigated in vitro by tissue staining and flow cytometry. After demonstration of sensitivity and specificity, preclinical optical imaging studies were performed to evaluate tumor specificity in murine subcutaneous and metastatic multiple myeloma models. The VLA-4-targeted molecular probe showed high affinity for subcutaneous MM tumor xenografts. Importantly, tumor cells specific accumulation in the bone marrow of metastatic multiple myeloma correlated with GFP signal from transfected cells. Ex vivo flow cytometry of tumor tissue and bone marrow further corroborated in vivo imaging data, demonstrating the specificity of the novel agent and potential for quantitative imaging of multiple myeloma burden in these models.

  6. Biochemical activity of a fluorescent dye rhodamine 6G: Molecular modeling, electrochemical, spectroscopic and thermodynamic studies.

    Science.gov (United States)

    Al Masum, Abdulla; Chakraborty, Maharudra; Ghosh, Soumen; Laha, Dipranjan; Karmakar, Parimal; Islam, Md Maidul; Mukhopadhyay, Subrata

    2016-11-01

    Interaction of CT DNA with Rhodamine 6G (R6G) has been studied using molecular docking, electrochemical, spectroscopic and thermodynamic methods. From the study, it was illustrated that Rhodamine 6G binds to the minor groove of CT DNA. The binding was cooperative in nature. Circular voltametric study showed significant change in peak current and peak potential due to complexation. All the studies showed that the binding constant was in the order of 10 6 M -1 . Circular dichroic spectra showed significant conformational change on binding and DNA unwind during binding. Thermodynamic study showed that binding was favored by negative enthalpy and positive entropy change. From thermodynamic study it was also observed that several positive and negative free energies played significant role during binding and the unfavorable conformational free energy change was overcame by highly negative hydrophobic and salt dependent free energy changes. The experimental results were further validated using molecular docking study and the effect of structure on binding has been studied theoretically. From docking study it was found that the hydrophobic interaction and hydrogen bonds played a significant role during binding. The dye was absorbed by cell and this phenomenon was studied using fluorescent microscope. Cell survivability test showed that the dye active against Human Breast Cancer cells MDA-MB 468. ROS study showed that the activity is due to the production of reactive oxygen. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Graphene oxide-sensitized molecularly imprinted opto-polymers for charge-transfer fluorescent sensing of cyanoguanidine.

    Science.gov (United States)

    Liu, Huilin; Zhou, Kaiwen; Chen, Xiaomo; Wang, Jing; Wang, Shuo; Sun, Baoguo

    2017-11-15

    The hierarchical structuring of materials offers exciting opportunities to construct functional sensors. Multiple processes were combined to create complex materials for the selective detection of cyanoguanidine (CYA) using graphene oxide-sensitized molecularly imprinted opto-polymers (MIOP). Molecular imprinting was used to construct molecular-scale analyte-selective cavities, graphene oxide was introduced to provide a platform for the polymerization, and increase the stability and binding kinetic properties, and 3-methacryloxy propyl trimethoxy silane-modified quantum dots were combined with a functional monomer to increase the fluorescence quantum yield. Polymer cross-linking and fluorescence intensity were optimized for molecular recognition and opto-sensing detection. Selective and sensitive, fluorescence sensing of CYA was possible at concentrations as low as to 1.6μM. It could be applied to the rapid and cost-effective monitoring of CYA in infant formula. The approach is generic and applicable to many molecules and conventional opto-sensors, based on molecularly imprinted polymer formulations, individually or in multiplexed arrays. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Molecular recognition of DNA-protein complexes: A straightforward method combining scanning force and fluorescence microscopy

    NARCIS (Netherlands)

    H. Sanchez (Humberto); R. Kanaar (Roland); C. Wyman (Claire)

    2010-01-01

    textabstractCombining scanning force and fluorescent microscopy allows simultaneous identification of labeled biomolecules and analysis of their nanometer level architectural arrangement. Fluorescent polystyrene nano-spheres were used as reliable objects for alignment of optical and topographic

  9. A sensitive fluorescent nanosensor for chloramphenicol based on molecularly imprinted polymer-capped CdTe quantum dots.

    Science.gov (United States)

    Amjadi, Mohammad; Jalili, Roghayeh; Manzoori, Jamshid L

    2016-05-01

    A novel fluorescent nanosensor using molecularly imprinted silica nanospheres embedded CdTe quantum dots (CdTe@SiO2 @MIP) was developed for detection and quantification of chloramphenicol (CAP). The imprinted sensor was prepared by synthesis of molecularly imprinting polymer (MIP) on the hydrophilic CdTe quantum dots via reverse microemulsion method using small amounts of solvents. The resulting CdTe@SiO2 @MIP nanoparticles were characterized by fluorescence, UV-vis absorption and FT-IR spectroscopy and transmission electron microscopy. They preserved 48% of fluorescence quantum yield of the parent quantum dots. CAP remarkably quenched the fluorescence of prepared CdTe@SiO2 @MIP, probably via electron transfer mechanism. Under the optimal conditions, the relative fluorescence intensity of CdTe@SiO2 @MIP decreased with increasing CAP by a Stern-Volmer type equation in the concentration range of 40-500 µg L(-1). The corresponding detection limit was 5.0 µg L(-1). The intra-day and inter-day values for the precision of the proposed method were all <4%. The developed sensor had a good selectivity and was applied to determine CAP in spiked human and bovine serum and milk samples with satisfactory results. Copyright © 2015 John Wiley & Sons, Ltd.

  10. [Molecular beacon based PNA-FISH method combined with fluorescence scanning for rapid detection of Listeria monocytogenes].

    Science.gov (United States)

    Wu, Shan; Zhang, Xiaofeng; Shuai, Jiangbing; Li, Ke; Yu, Huizhen; Jin, Chenchen

    2016-07-04

    To simplify the PNA-FISH (Peptide nucleic acid-fluorescence in situ hybridization) test, molecular beacon based PNA probe combined with fluorescence scanning detection technology was applied to replace the original microscope observation to detect Listeria monocytogenes The 5′ end and 3′ end of the L. monocytogenes specific PNA probes were labeled with the fluorescent group and the quenching group respectively, to form a molecular beacon based PNA probe. When PNA probe used for fluorescence scanning and N1 treatment as the control, the false positive rate was 11.4%, and the false negative rate was 0; when N2 treatment as the control, the false positive rate decreased to 4.3%, but the false negative rate rose to 18.6%. When beacon based PNA probe used for fluorescence scanning, taken N1 treatment as blank control, the false positive rate was 8.6%, and the false negative rate was 1.4%; taken N2 treatment as blank control, the false positive rate was 5.7%, and the false negative rate was 1.4%. Compared with PNA probe, molecular beacon based PNA probe can effectively reduce false positives and false negatives. The success rates of hybridization of the two PNA probes were 83.3% and 95.2% respectively; and the rates of the two beacon based PNA probes were 91.7% and 90.5% respectively, which indicated that labeling the both ends of the PNA probe dose not decrease the hybridization rate with the target bacteria. The combination of liquid phase PNA-FISH and fluorescence scanning method, can significantly improve the detection efficiency.

  11. Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay.

    Science.gov (United States)

    Hu, Pan; Yang, Bin

    2016-01-15

    Due to its unique features such as high sensitivity, homogeneous format, and independence on fluorescent intensity, fluorescence anisotropy (FA) assay has become a hotspot of study in oligonucleotide-based bioassays. However, until now most FA probes require carefully customized structure designs, and thus are neither generalizable for different sensing systems nor effective to obtain sufficient signal response. To address this issue, a cleavable DNA-protein hybrid molecular beacon was successfully engineered for signal amplified FA bioassay, via combining the unique stable structure of molecular beacon and the large molecular mass of streptavidin. Compared with single DNA strand probe or conventional molecular beacon, the DNA-protein hybrid molecular beacon exhibited a much higher FA value, which was potential to obtain high signal-background ratio in sensing process. As proof-of-principle, this novel DNA-protein hybrid molecular beacon was further applied for FA bioassay using DNAzyme-Pb(2+) as a model sensing system. This FA assay approach could selectively detect as low as 0.5nM Pb(2+) in buffer solution, and also be successful for real samples analysis with good recovery values. Compatible with most of oligonucleotide probes' designs and enzyme-based signal amplification strategies, the molecular beacon can serve as a novel signal translator to expand the application prospect of FA technology in various bioassays. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Problems of fluorescent imaging and its solution using nanofluorophores. Part I: Advantages of fluorescent nanoparticles over conventional organic fluorophores

    International Nuclear Information System (INIS)

    Zhelev, Z.; Hadjidekov, G.; Zlateva, G.; Spasov, L.; Bakalova, R.

    2011-01-01

    The application of fluorescence in deep-tissue imaging is rapidly expanding in fast several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecules in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With development of novel bright fluorophores based on nano-technologies and fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. This review outlines the current status and future trends of fluorescent nanoparticles - quantum dots (QDs), as a new generation of fluorophores in experimental and pre-clinical fluorescent imaging diagnostic. Part 1 focuses on the advantages of quantum dots over conventional organic fluorophores and defines the major requirements to the 'perfect' fluorophore for fluorescent deep-tissue imaging diagnostic. The analysis is based on the limitations of fluorescent imaging in vivo and overcome by using quantum dots

  13. Brilliant molecular nanocrystals emerging from sol-gel thin films: towards a new generation of fluorescent biochips

    International Nuclear Information System (INIS)

    Dubuisson, E; Monnier, V; Sanz-Menez, N; Ibanez, A; Boury, B; Usson, Y; Pansu, R B

    2009-01-01

    To develop highly sensitive biosensors, we made directly available to biological aqueous solutions organic nanocrystals previously grown in the pores of sol-gel films. Through the controlled dissolution of the sol-gel surface, we obtained emerging nanocrystals that remained strongly anchored to the sol-gel coating for good mechanical stability of the final sensing device. We demonstrated that in the presence of a solution of DNA functionalized with a molecular probe, the nanocrystal fluorescence is strongly quenched by Foerster resonance energy transfer thus opening the way towards very sensitive fluorescent biosensors through biomolecules grafted onto fluorescent nanocrystals. Finally, this controlled dissolution, involving weak concentrated NaOH solution, is a generic process that can be used for the thinning of any kind of sol-gel layer.

  14. Anomalous doping of a molecular crystal monitored with confocal fluorescence microscopy: Terrylene in a p-terphenyl crystal

    Science.gov (United States)

    Białkowska, Magda; Deperasińska, Irena; Makarewicz, Artur; Kozankiewicz, Bolesław

    2017-09-01

    Highly terrylene doped single crystals of p-terphenyl, obtained by co-sublimation of both components, showed bright spots in the confocal fluorescence images. Polarization of the fluorescence excitation spectra, blinking and bleaching, and saturation behavior allowed us to attribute them to single molecules of terrylene anomalously embedded between two neighbor layers of the host crystal, in the (a,b) plane. Such an orientation of terrylene molecules results in much more efficient absorption and collection of the fluorescence photons than in the case of previously investigated molecules embedded in the substitution sites. The above conclusion was supported by quantum chemistry calculations. We postulate that the kind of doping considered in this work should be possible in other molecular crystals where the host molecules are organized in a herringbone pattern.

  15. A Conjugate of Pentamethine Cyanine and 18F as a Positron Emission Tomography/Near-Infrared Fluorescence Probe for Multimodality Tumor Imaging

    Directory of Open Access Journals (Sweden)

    Fei-Fei An

    2017-06-01

    Full Text Available The novel synthesis of a dual-modality, pentamethine cyanine (Cy5 fluorescent, 18F positron emission tomography (PET imaging probe is reported. The probe shows a large extinction coefficient and large quantum yield in the biologically transparent, near-infrared window (650–900 nm for in vivo fluorescent imaging. This fluorophore bears the isotope, 18F, giving a 18F-PET/near-infrared fluorescent (NIRF, bi-modal imaging probe, that combines the long-term stability of NIRF and the unlimited penetration depth of PET imaging. The bi-modal probe is labeled with 18F in a quick, one-step reaction, which is important in working with the rapid decay of 18F. The bi-modal probe bears a free carboxyl group, highlighting a PET/NIRF synthon that can be conjugated onto many advanced biomolecules for biomarker-specific in vivo dual-modal PET/NIR tumor imaging, confocal histology, and utility in multi-fluorophore, fluorescence-guided surgery. Its potential in vivo biocompatibility is explored in a quick proof-of-principal in vivo study. The dye is delivered to A549 xenograft flank-tumors to generate PET and NIRF signals at the tumor site. The tumor distribution is confirmed in ex vivo gamma counting and imaging. Pentamethine cyanine (Cy5 has the ability to preferentially accumulate in tumor xenografts. We substitute the PET/NIRF probe for Cy5, and explore this phenomenon.

  16. Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

    Directory of Open Access Journals (Sweden)

    Davidson Michael W

    2008-03-01

    Full Text Available Abstract Background In the 15 years that have passed since the cloning of Aequorea victoria green fluorescent protein (avGFP, the expanding set of fluorescent protein (FP variants has become entrenched as an indispensable toolkit for cell biology research. One of the latest additions to the toolkit is monomeric teal FP (mTFP1, a bright and photostable FP derived from Clavularia cyan FP. To gain insight into the molecular basis for the blue-shifted fluorescence emission we undertook a mutagenesis-based study of residues in the immediate environment of the chromophore. We also employed site-directed and random mutagenesis in combination with library screening to create new hues of mTFP1-derived variants with wavelength-shifted excitation and emission spectra. Results Our results demonstrate that the protein-chromophore interactions responsible for blue-shifting the absorbance and emission maxima of mTFP1 operate independently of the chromophore structure. This conclusion is supported by the observation that the Tyr67Trp and Tyr67His mutants of mTFP1 retain a blue-shifted fluorescence emission relative to their avGFP counterparts (that is, Tyr66Trp and Tyr66His. Based on previous work with close homologs, His197 and His163 are likely to be the residues with the greatest contribution towards blue-shifting the fluorescence emission. Indeed we have identified the substitutions His163Met and Thr73Ala that abolish or disrupt the interactions of these residues with the chromophore. The mTFP1-Thr73Ala/His163Met double mutant has an emission peak that is 23 nm red-shifted from that of mTFP1 itself. Directed evolution of this double mutant resulted in the development of mWasabi, a new green fluorescing protein that offers certain advantages over enhanced avGFP (EGFP. To assess the usefulness of mTFP1 and mWasabi in live cell imaging applications, we constructed and imaged more than 20 different fusion proteins. Conclusion Based on the results of our

  17. Flexibility of cold- and heat-adapted subtilisin-like serine proteinases evaluated with fluorescence quenching and molecular dynamics

    DEFF Research Database (Denmark)

    Sigtryggsdóttir, Asta Rós; Papaleo, Elena; Thorbjarnardóttir, Sigríður H.

    2014-01-01

    activity of cold adapted enzymes when compared to homologues from thermophiles, reflects their higher molecular flexibility. To assess a potential difference in molecular flexibility between the two homologous proteinases, we have measured their Trp fluorescence quenching by acrylamide at different......The subtilisin-like serine proteinases, VPR, from a psychrotrophic Vibrio species and aqualysin I (AQUI) from the thermophile Thermus aquaticus, are structural homologues, but differ significantly with respect to stability and catalytic properties. It has been postulated that the higher catalytic...... to Trp (Y191W). A lower quenching effect of acrylamide on the intrinsic fluorescence of the thermophilic AQUI_Y191W was observed at all temperatures measured (10-55°C), suggesting that it possesses a more rigid structure than VPR. The MD analysis (Cα rmsf profiles) showed that even though VPR and AQUI...

  18. Detection of trace tetracycline in fish via synchronous fluorescence quenching with carbon quantum dots coated with molecularly imprinted silica

    Science.gov (United States)

    Yang, Ji; Lin, Zheng-Zhong; Nur, A.-Zha; Lu, Yan; Wu, Ming-Hui; Zeng, Jun; Chen, Xiao-Mei; Huang, Zhi-Yong

    2018-02-01

    A novel fluorescence-based sensor combining synchronous fluorescence spectroscopy (SFS) with molecularly imprinted polymers (MIPs) was fabricated with reverse microemulsion method. Tetracycline (TC), (3-aminopropyl) triethoxysilane (APTES), tetraethyl orthosilicate (TEOS) and carbon quantum dots (CDs) were used as template, functional monomer, cross-linker and signal sources respectively in the probe preparation. A synchronous fluorescence emission (λem) at 355 nm was observed for the prepared MIP-coated CDs (MIP@CDs) particles when the wavelength interval (Δλ) was set as 70 nm, and the synchronous fluorescence intensity could be rapidly and efficiently quenched by TC based on inner filter effect (IFE). The quenching efficiencies of synchronous fluorescence intensity was linearly fitted with tetracycline (TC) concentrations ranging from 0.1 to 50 μmol L- 1 with a detection limit (DL) of 9 nmol L- 1 (3σ, n = 9). The MIP@CDs was used as a probe to detect TC in fish samples with the recoveries ranging from 98.4% to 103.1% and the relative standard deviation less than 6.0%. The results illustrated that the as-prepared MIP@CDs could be applied to the detection of trace TC in fish samples with rapidity, high sensitivity and accuracy.

  19. Bilayered near-infrared fluorescent nanoparticles based on low molecular weight PEI for tumor-targeted in vivo imaging

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Hao; Li, Ke [Xi’an Jiaotong University, Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology (China); Xu, Liang [The University of Kansas, Department of Molecular Biosciences (United States); Wu, Daocheng, E-mail: wudaocheng@mail.xjtu.edu.cn [Xi’an Jiaotong University, Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology (China)

    2014-12-15

    To improve the tumor fluorescent imaging results in vivo, bilayered nanoparticles encapsulating a lipophilic near-infrared (NIR) fluorescent dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotri-carbocyanine iodide (DiR) were prepared using low molecular weight stearic acid-grafted polyethyleneimine and hyaluronic acid (DiR-PgSHA nanoparticles), which were investigated as a novel NIR fluorescent nano-probe for in vivo tumor-targeted optical imaging. These nanoparticles were characterized by transmission electron microscopy (TEM), infrared (IR) spectra, UV-visual absorption, and fluorescent emission spectra. Their cytotoxicity in vitro and hepatotoxicity in vivo were tested by MTT assay and histological study, respectively. In vivo NIR fluorescence imaging of the DiR-PgSHA nanoparticles was performed using a Carestream imaging system. The DiR-PgSHA nanoparticles were sphere shaped with a diameter of approximately 50 nm according to the TEM images. The DiR-PgSHA nanoparticles had a low cytotoxicity in vitro according to the MTT assay and low hepatotoxicity in vivo as determined in histological studies. The fluorescent emission of DiR-PgSHA nanoparticles was stable in pH values of 5–9 in solution, with only slight blue-shifts of the emission maxima at the basic pH range. The DiR-PgSHA nanoparticles exhibited a substantial tumor-targeting ability in the optical imaging with the use of tumor-bearing mice. These results demonstrated that the DiR-PgSHA nanoparticle is an excellent biocompatible nano-probe for in vivo tumor-targeted NIR fluorescence imaging with a potential for clinical applications.

  20. Bilayered near-infrared fluorescent nanoparticles based on low molecular weight PEI for tumor-targeted in vivo imaging

    International Nuclear Information System (INIS)

    Liu, Hao; Li, Ke; Xu, Liang; Wu, Daocheng

    2014-01-01

    To improve the tumor fluorescent imaging results in vivo, bilayered nanoparticles encapsulating a lipophilic near-infrared (NIR) fluorescent dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotri-carbocyanine iodide (DiR) were prepared using low molecular weight stearic acid-grafted polyethyleneimine and hyaluronic acid (DiR-PgSHA nanoparticles), which were investigated as a novel NIR fluorescent nano-probe for in vivo tumor-targeted optical imaging. These nanoparticles were characterized by transmission electron microscopy (TEM), infrared (IR) spectra, UV-visual absorption, and fluorescent emission spectra. Their cytotoxicity in vitro and hepatotoxicity in vivo were tested by MTT assay and histological study, respectively. In vivo NIR fluorescence imaging of the DiR-PgSHA nanoparticles was performed using a Carestream imaging system. The DiR-PgSHA nanoparticles were sphere shaped with a diameter of approximately 50 nm according to the TEM images. The DiR-PgSHA nanoparticles had a low cytotoxicity in vitro according to the MTT assay and low hepatotoxicity in vivo as determined in histological studies. The fluorescent emission of DiR-PgSHA nanoparticles was stable in pH values of 5–9 in solution, with only slight blue-shifts of the emission maxima at the basic pH range. The DiR-PgSHA nanoparticles exhibited a substantial tumor-targeting ability in the optical imaging with the use of tumor-bearing mice. These results demonstrated that the DiR-PgSHA nanoparticle is an excellent biocompatible nano-probe for in vivo tumor-targeted NIR fluorescence imaging with a potential for clinical applications

  1. Intravascular near-infrared fluorescence molecular imaging of atherosclerosis: toward coronary arterial visualization of biologically high-risk plaques

    Science.gov (United States)

    Calfon, Marcella A.; Vinegoni, Claudio; Ntziachristos, Vasilis; Jaffer, Farouc A.

    2010-01-01

    New imaging methods are urgently needed to identify high-risk atherosclerotic lesions prior to the onset of myocardial infarction, stroke, and ischemic limbs. Molecular imaging offers a new approach to visualize key biological features that characterize high-risk plaques associated with cardiovascular events. While substantial progress has been realized in clinical molecular imaging of plaques in larger arterial vessels (carotid, aorta, iliac), there remains a compelling, unmet need to develop molecular imaging strategies targeted to high-risk plaques in human coronary arteries. We present recent developments in intravascular near-IR fluorescence catheter-based strategies for in vivo detection of plaque inflammation in coronary-sized arteries. In particular, the biological, light transmission, imaging agent, and engineering principles that underlie a new intravascular near-IR fluorescence sensing method are discussed. Intravascular near-IR fluorescence catheters appear highly translatable to the cardiac catheterization laboratory, and thus may offer a new in vivo method to detect high-risk coronary plaques and to assess novel atherosclerosis biologics.

  2. High-resolution emission tomography of small laboratory animals: physics and gamma-astronomy meet molecular biology

    International Nuclear Information System (INIS)

    Beekman, F.J.; Colijn, A.P.; Vastenhouw, B.; Wiegant, V.M.; Gerrits, M.A.F.M.

    2003-01-01

    Molecular imaging can be defined as the characterization and measurement of biological processes in living animals, model systems and humans at the cellular and molecular level using remote imaging detectors. An example concerns the mapping of the distributions of radioactively labeled molecules in laboratory animals which is of crucial importance for life sciences. Tomographic methods like Single Photon Emission Computed Tomography (SPECT) offer a possibility to visualize distributions of radioactively labeled molecules in living animals. Miniature tomography systems, derived from their clinical counterparts, but with a much higher image resolution are under development in several institutes. An example is U-SPECT that will be discussed in the present paper. Such systems are expected to accelerate several biomedical research procedures, the understanding of gene and protein function, as well as pharmaceutical development

  3. Photolysis study of octyl p-methoxycinnamate loaded microemulsion by molecular fluorescence and chemometric approach

    Science.gov (United States)

    Nascimento, Danielle Silva; Insausti, Matías; Band, Beatriz Susana Fernández; Grünhut, Marcos

    2018-02-01

    Octyl p-methoxycinnamate (OMC) is one of the most widely used sunscreen agents. However, the efficiency of OMC as UV filter over time is affected due to the formation of the cis-isomer which presents a markedly lower extinction coefficient (εcis = 12,600 L mol- 1 cm- 1 at 291 nm) than the original trans-isomer (εtrans = 24,000 L mol- 1 cm- 1 at 310 nm). In this work, a novel carrier for OMC based on an oil-in-water microemulsion is proposed in order to improve the photostability of this sunscreen. The formulation was composed of 29.2% (w/w) of a 3:1 mixture of ethanol (co-surfactant) and decaethylene glycol mono-dodecyl ether (surfactant), 1.5% (w/w) of oleic acid (oil phase) and 69.2% (w/w) of water. This microemulsion was prepared in a simple way, under moderate stirring at 25 °C and using acceptable, biocompatible and accessible materials for topical use. OMC was incorporated in the vehicle at a final concentration of 5.0% (w/w), taking into account the maximum permitted levels established by international norms. Then, a photolysis study of the loaded formulation was performed using a continuous flow system. The direct photolysis was monitored over time by molecular fluorescence. The recorded spectra data between 370 y 490 nm were analyzed by multivariate curve resolution-alternating least squares algorithm. The kinetic rate constants corresponding to the photolysis of the trans-OMC were calculated from the concentration profiles, resulting in 0.0049 s- 1 for the trans-OMC loaded microemulsion and 0.0131 s- 1 for the trans-OMC in aqueous media. These results demonstrate a higher photostability of the trans-OMC when loaded in the proposed vehicle with respect to the free trans-OMC in aqueous media.

  4. Multimodality optical coherence tomography and fluorescence confocal scanning laser ophthalmoscopy for image-guided feedback of intraocular injections in mouse models

    Science.gov (United States)

    Benavides, Oscar R.; Terrones, Benjamin D.; Leeburg, Kelsey C.; Mehanathan, Sankarathi B.; Levine, Edward M.; Tao, Yuankai K.

    2018-02-01

    Rodent models are robust tools for understanding human retinal disease and function because of their similarities with human physiology and anatomy and availability of genetic mutants. Optical coherence tomography (OCT) has been well-established for ophthalmic imaging in rodents and enables depth-resolved visualization of structures and image-based surrogate biomarkers of disease. Similarly, fluorescence confocal scanning laser ophthalmoscopy (cSLO) has demonstrated utility for imaging endogenous and exogenous fluorescence and scattering contrast in the mouse retina. Complementary volumetric scattering and en face fluorescence contrast from OCT and cSLO, respectively, enables cellular-resolution longitudinal imaging of changes in ophthalmic structure and function. We present a non-contact multimodal OCT+cSLO small animal imaging system with extended working distance to the pupil, which enables imaging during and after intraocular injection. While injections are routinely performed in mice to develop novel models of ophthalmic diseases and screen novel therapeutics, the location and volume delivered is not precisely controlled and difficult to reproduce. Animals were imaged using a custom-built OCT engine and scan-head combined with a modified commercial cSLO scan-head. Post-injection imaging showed structural changes associated with retinal puncture, including the injection track, a retinal elevation, and detachment of the posterior hyaloid. When combined with imagesegmentation, we believe OCT can be used to precisely identify injection locations and quantify injection volumes. Fluorescence cSLO can provide complementary contrast for either fluorescently labeled compounds or transgenic cells for improved specificity. Our non-contact OCT+cSLO system is uniquely-suited for concurrent imaging with intraocular injections, which may be used for real-time image-guided injections.

  5. Comparison of molecular imprinted particles prepared using precipitation polymerization in water and chloroform for fluorescent detection of nitroaromatics

    Energy Technology Data Exchange (ETDEWEB)

    Stringer, R. Cody, E-mail: rcsm84@mail.mizzou.edu [Department of Biological Engineering, University of Missouri, Columbia, MO (United States); Gangopadhyay, Shubhra, E-mail: gangopadhyays@missouri.edu [Department of Electrical and Computer Engineering, University of Missouri, Columbia, MO (United States); Grant, Sheila A., E-mail: grantsa@missouri.edu [Department of Biological Engineering, University of Missouri, Columbia, MO (United States)

    2011-10-10

    Highlights: {yields} Imprinted polymers prepared using precipitation polymerization. {yields} Comparison of chloroform and water as polymerization solvent. {yields} Imprinted polymer doped with quantum dots for fluorescent sensor. {yields} Fluorescent imprinted polymer used to detect nitroaromatic explosives. {yields} Chloroform is ideal solvent for molecular imprinting of nitroaromatics. - Abstract: A comparative study was conducted to study the effects that two different polymerization solvents would have on the properties of imprinted polymer microparticles prepared using precipitation polymerization. Microparticles prepared in chloroform, which previous results indicated was the optimal solvent for molecular imprinting of nitroaromatic explosive compounds, were compared to water, which was hypothesized to decrease water swelling of the polymer and allow enhanced rebinding of aqueous template. The microparticles were characterized and were integrated into a fluorescence sensing mechanism for detection of nitroaromatic explosive compounds. The performance of the sensing mechanisms was compared to illustrate which polymerization solvent produced optimal imprinted polymer microparticles for detection of nitroaromatic molecules. Results indicated that the structures of microparticles synthesized in chloroform versus water varied greatly. Sensor performance studies showed that the microparticles prepared in chloroform had greater imprinting efficiency and higher template rebinding than those prepared in water. For detection of 2,4,6-trinitrotoluene, the chloroform-based fluorescent microparticles achieved a lower limit of detection of 0.1 {mu}M, as compared to 100 {mu}M for the water-based fluorescent microparticles. Detection limits for 2,4-dinitrotoluene, as well as time response studies, also demonstrated that the chloroform-based particles are more effective for detection of nitroaromatic compounds than water-based particles. These results illustrate that the

  6. Comparison of molecular imprinted particles prepared using precipitation polymerization in water and chloroform for fluorescent detection of nitroaromatics

    International Nuclear Information System (INIS)

    Stringer, R. Cody; Gangopadhyay, Shubhra; Grant, Sheila A.

    2011-01-01

    Highlights: → Imprinted polymers prepared using precipitation polymerization. → Comparison of chloroform and water as polymerization solvent. → Imprinted polymer doped with quantum dots for fluorescent sensor. → Fluorescent imprinted polymer used to detect nitroaromatic explosives. → Chloroform is ideal solvent for molecular imprinting of nitroaromatics. - Abstract: A comparative study was conducted to study the effects that two different polymerization solvents would have on the properties of imprinted polymer microparticles prepared using precipitation polymerization. Microparticles prepared in chloroform, which previous results indicated was the optimal solvent for molecular imprinting of nitroaromatic explosive compounds, were compared to water, which was hypothesized to decrease water swelling of the polymer and allow enhanced rebinding of aqueous template. The microparticles were characterized and were integrated into a fluorescence sensing mechanism for detection of nitroaromatic explosive compounds. The performance of the sensing mechanisms was compared to illustrate which polymerization solvent produced optimal imprinted polymer microparticles for detection of nitroaromatic molecules. Results indicated that the structures of microparticles synthesized in chloroform versus water varied greatly. Sensor performance studies showed that the microparticles prepared in chloroform had greater imprinting efficiency and higher template rebinding than those prepared in water. For detection of 2,4,6-trinitrotoluene, the chloroform-based fluorescent microparticles achieved a lower limit of detection of 0.1 μM, as compared to 100 μM for the water-based fluorescent microparticles. Detection limits for 2,4-dinitrotoluene, as well as time response studies, also demonstrated that the chloroform-based particles are more effective for detection of nitroaromatic compounds than water-based particles. These results illustrate that the enhanced chemical properties of

  7. Molecular Level Understanding of Sodium Dodecyl Sulfate (SDS) Induced Sol-Gel Transition of Pluronic F127 Using Fisetin as a Fluorescent Molecular Probe.

    Science.gov (United States)

    Mishra, Jhili; Swain, Jitendriya; Mishra, Ashok Kumar

    2018-01-11

    The thermoreversible sol-gel transition of pluronic F127 is markedly altered even with addition of submicellar concentration of sodium dodecyl sulfate (SDS) surfactant. Multiple fluorescence parameters like fluorescence intensity, fluorescence anisotropy and fluorescence lifetime of both the prototropic forms (anion (A - *) and phototautomer FT*) of the photoprototropic fluorescent probe fisetin has been efficiently used to understand the molecular level properties like polarity and microviscosity of the PF127-SDS system as a function of temperature. The SDS-induced increase in the interfacial hydrophobicity level is seen to affect the sol-gel phase transition of PF127 (21-18 °C). The E T (30) polarity parameter value of anionic emission of fisetin suggests that there is a considerable decrease in the polarity of the PF127 medium with increase in temperature and with the addition of SDS. The microviscosity progressively increases from ∼5 mPa s (sol state, 10 °C) to ∼22.01 mPa s (gel state 35 °C) in aqueous solution of PF127. The variation in microviscosity with addition of SDS in PF127-SDS mixed system is significant in sol phase whereas in gel phase this variation is significantly less. Temperature dependent fluorescence lifetime of FT* indicates that there is heterogeneity in distribution of fisetin molecules at different domains of PF127. This work also show-cases the sensitivity of fisetin toward change in polarity and change in sol-gel transition temperature of copolymer PF127 with variation in temperature (both forward and reverse directions) and SDS.

  8. Fluorescent angiography and optical coherence tomography with angiography of the ocular fundus in patients with "the wet" form of an age-related macular degeneration.

    Directory of Open Access Journals (Sweden)

    Virsta A.M.

    2017-06-01

    Full Text Available Purpose: to investigate the informative value of fluorescent angiography (FA and optical coherence tomography with fundus angiography (angio-OCT in the diagnosis of "wet" form of age-related macular degeneration (AMD. Material and methods. The present study included 20 patients (20 eyes diagnosed with degeneration of macula and posterior pole of the eye, the "wet" form (late stage age-related macular degeneration, AREDS category 4. The study used machines: optical coherence tomography, Spectralis HRA+OCT (Heidelberg Engeneering, Germany, optical со- herence tomography-angiography CIRRUS HD-OCT MODEL 5000 (Carl Zeiss, Germany. Results. When conducting the FA, in 11 patients found the ill-defined zone of small leakage of dye in 7 patients revealed a clearly defined area of hyperfluorescence in the early phase, and marked leakage of dye in the late phase, 2 patients — uncertain indices. In connection with the received data questionable PHAGE in 11 patients, all were held angio-OCT, to clarify the localization of choroidal neovascularization (CNV. When performing angio-OCT in 11 patients revealed that "wet" form of AMD with occult choroidal neovascularization. In 7 patients there had been discovered classic CNV in 2 patients combined. Conclusion. Angio-OCT gives a clearer picture about the presence of a choroidal neovascular membrane that plays a significant role in determining the course of treatment of patients with wet age-related macular degeneration.

  9. Dual-Modal Colorimetric/Fluorescence Molecular Probe for Ratiometric Sensing of pH and Its Application.

    Science.gov (United States)

    Wu, Luling; Li, Xiaolin; Huang, Chusen; Jia, Nengqin

    2016-08-16

    As traditional pH meters cannot work well for minute regions (such as subcellular organelles) and in harsh media, molecular pH-sensitive devices for monitoring pH changes in diverse local heterogeneous environments are urgently needed. Here, we report a new dual-modal colorimetric/fluorescence merocyanine-based molecular probe (CPH) for ratiometric sensing of pH. Compared with previously reported pH probes, CPH bearing the benzyl group at the nitrogen position of the indolium group and the phenol, which is used as the acceptor for proton, could respond to pH changes immediately through both the ratiometric fluorescence signal readout and naked-eye colorimetric observation. The sensing process was highly stable and reversible. Most importantly, the suitable pKa value (6.44) allows CPH to presumably accumulate in lysosomes and become a lysosome-target fluorescent probe. By using CPH, the intralysosomal pH fluctuation stimulated by antimalaria drug chloroquine was successfully tracked in live cells through the ratiometric fluorescence images. Additionally, CPH could be immobilized on test papers, which exhibited a rapid and reversible colorimetric response to acid/base vapor through the naked-eye colorimetric analysis. This proof-of-concept study presents the potential application of CPH as a molecular tool for monitoring intralysosomal pH fluctuation in live cells, as well as paves the way for developing the economic, reusable, and fast-response optical pH meters for colorimetric sensing acid/base vapor with direct naked-eye observation.

  10. Three dimensional subsurface elemental identification of minerals using confocal micro-X-ray fluorescence and micro-X-ray computed tomography

    International Nuclear Information System (INIS)

    Cordes, Nikolaus L.; Seshadri, Srivatsan; Havrilla, George J.; Yuan, Xiaoli; Feser, Michael; Patterson, Brian M.

    2015-01-01

    Current non-destructive elemental characterization methods, such as scanning electron microscopy-based energy dispersive spectroscopy (SEM–EDS) and micro-X-ray fluorescence spectroscopy (MXRF), are limited to either elemental identification at the surface (SEM–EDS) or suffer from an inability to discriminate between surface or depth information (MXRF). Thus, a non-destructive elemental characterization of individual embedded particles beneath the surface is impossible with either of these techniques. This limitation can be overcome by using laboratory-based 3D confocal micro-X-ray fluorescence spectroscopy (confocal MXRF). This technique utilizes focusing optics on the X-ray source and detector which allows for spatial discrimination in all three dimensions. However, the voxel-by-voxel serial acquisition of a 3D elemental scan can be very time-intensive (~ 1 to 4 weeks) if it is necessary to locate individual embedded particles of interest. As an example, if each point takes a 5 s measurement time, a small volume of 50 × 50 × 50 pixels leads to an acquisition time of approximately 174 h, not including sample stage movement time. Initially screening the samples for particles of interest using micro-X-ray computed tomography (micro-CT) can significantly reduce the time required to spatially locate these particles. Once located, these individual particles can be elementally characterized with confocal MXRF. Herein, we report the elemental identification of high atomic number surface and subsurface particles embedded in a mineralogical matrix by coupling micro-CT and confocal MXRF. Synergistically, these two X-ray based techniques first rapidly locate and then elementally identify individual subsurface particles. - Highlights: • Coupling of confocal X-ray fluorescence spectroscopy and X-ray computed tomography • Qualitative elemental identification of surface and subsurface mineral particles • Non-destructive particle size measurements • Utilization of

  11. Three dimensional subsurface elemental identification of minerals using confocal micro-X-ray fluorescence and micro-X-ray computed tomography

    Energy Technology Data Exchange (ETDEWEB)

    Cordes, Nikolaus L., E-mail: ncordes@lanl.gov [Polymers and Coatings Group, Material Science and Technology Division, Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Seshadri, Srivatsan, E-mail: srivatsan.seshadri@zeiss.com [Carl Zeiss X-ray Microscopy, Inc., Pleasanton, CA 94588 (United States); Havrilla, George J. [Chemical Diagnostics and Engineering, Chemistry Division, Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Yuan, Xiaoli [Julius Kruttschnitt Mineral Research Centre, University of Queensland, Indooroopilly, Brisbane, QLD 4068 (Australia); Feser, Michael [Carl Zeiss X-ray Microscopy, Inc., Pleasanton, CA 94588 (United States); Patterson, Brian M. [Polymers and Coatings Group, Material Science and Technology Division, Los Alamos National Laboratory, Los Alamos, NM 87545 (United States)

    2015-01-01

    Current non-destructive elemental characterization methods, such as scanning electron microscopy-based energy dispersive spectroscopy (SEM–EDS) and micro-X-ray fluorescence spectroscopy (MXRF), are limited to either elemental identification at the surface (SEM–EDS) or suffer from an inability to discriminate between surface or depth information (MXRF). Thus, a non-destructive elemental characterization of individual embedded particles beneath the surface is impossible with either of these techniques. This limitation can be overcome by using laboratory-based 3D confocal micro-X-ray fluorescence spectroscopy (confocal MXRF). This technique utilizes focusing optics on the X-ray source and detector which allows for spatial discrimination in all three dimensions. However, the voxel-by-voxel serial acquisition of a 3D elemental scan can be very time-intensive (~ 1 to 4 weeks) if it is necessary to locate individual embedded particles of interest. As an example, if each point takes a 5 s measurement time, a small volume of 50 × 50 × 50 pixels leads to an acquisition time of approximately 174 h, not including sample stage movement time. Initially screening the samples for particles of interest using micro-X-ray computed tomography (micro-CT) can significantly reduce the time required to spatially locate these particles. Once located, these individual particles can be elementally characterized with confocal MXRF. Herein, we report the elemental identification of high atomic number surface and subsurface particles embedded in a mineralogical matrix by coupling micro-CT and confocal MXRF. Synergistically, these two X-ray based techniques first rapidly locate and then elementally identify individual subsurface particles. - Highlights: • Coupling of confocal X-ray fluorescence spectroscopy and X-ray computed tomography • Qualitative elemental identification of surface and subsurface mineral particles • Non-destructive particle size measurements • Utilization of

  12. Low molecular weight dextran provides similar optical coherence tomography coronary imaging compared to radiographic contrast media.

    Science.gov (United States)

    Frick, Kyle; Michael, Tesfaldet T; Alomar, Mohammed; Mohammed, Atif; Rangan, Bavana V; Abdullah, Shuaib; Grodin, Jerrold; Hastings, Jeffrey L; Banerjee, Subhash; Brilakis, Emmanouil S

    2014-11-01

    Optical coherence tomography (OCT) coronary imaging requires displacement of red blood cells from the vessel lumen. This is usually accomplished using radiographic contrast. Low molecular weight dextran has low cost and is safe in low volumes. In the present study, we compared dextran with contrast for coronary OCT imaging. Fifty-one vessels in 26 patients were sequentially imaged using manual injection of radiographic contrast (iodixanol) and dextran. OCT images were analyzed at 1 mm intervals to determine the image clarity (defined as a visible lumen border > 270°) and to measure the lumen area and lumen diameter. To correct for the refractive index of dextran, the dextran area measurements were multiplied by 1.117 and the dextran length measurements were multiplied by 1.057. A total of 3,418 cross-sections (1,709 with contrast and 1,709 with dextran) were analyzed. There were no complications related to OCT imaging or to contrast or dextran administration. Clear image segments were observed in 97.0% vs. 96.7% of the cross-sections obtained with contrast and dextran, respectively (P = 0.45). The mean lumen areas were also similar: 6.69 ± 1.95 mm(2) with iodixanol vs. 7.06 ± 2.06 mm(2) with dextran (correlation coefficient 0.984). The image quality and measurements during OCT image acquisition are similar for dextran and contrast. Dextran could be used instead of contrast for OCT imaging, especially in patients in whom contrast load minimization is desired. © 2013 Wiley Periodicals, Inc.

  13. Multimodal Imaging of Integrin Receptor-Positive Tumors by Bioluminescence, Fluorescence, Gamma Scintigraphy, and Single-Photon Emission Computed Tomography Using a Cyclic RGD Peptide Labeled with a Near-Infrared Fluorescent Dye and a Radionuclide

    Directory of Open Access Journals (Sweden)

    W. Barry Edwards

    2009-03-01

    Full Text Available Integrins, particularly the αvβ3 heterodimers, play important roles in tumor-induced angiogenesis and invasiveness. To image the expression pattern of the αvβ3 integrin in tumors through a multimodality imaging paradigm, we prepared a cyclic RGDyK peptide analogue (LS308 bearing a tetraazamacrocycle 1,4,7,10-tetraazacyclododecane-N, N′, N″, N‴-tetraacetic acid (DOTA and a lipophilic near-infrared (NIR fluorescent dye cypate. The αvβ3 integrin binding affinity and the internalization properties of LS308 mediated by the αvβ3 integrin in 4t1luc cells were investigated by receptor binding assay and fluorescence microscopy, respectively. The in vivo distribution of 111In-labeled LS308 in a 4t1luc tumor-bearing mouse model was studied by fluorescence, bioluminescence, planar gamma, and single-photon emission computed tomography (SPECT. The results show that LS308 has high affinity for αvβ3 integrin and internalized preferentially via the αvβ3 integrin-mediated endocytosis in 4t1luc cells. We also found that LS308 selectively accumulated in αvβ3-positve tumors in a receptor-specific manner and was visualized by the four imaging methods. Whereas the endogenous bioluminescence imaging identified the ensemble of the tumor tissue, the fluorescence and SPECT methods with the exogenous contrast agent LS308 reported the local expression of αvβ3 integrin. Thus, the multimodal imaging approach could provide important complementary diagnostic information for monitoring the efficacy of new antiangiogenic drugs.

  14. A disposable evanescent wave fiber optic sensor coated with a molecularly imprinted polymer as a selective fluorescence probe.

    Science.gov (United States)

    Ton, Xuan-Anh; Acha, Victor; Bonomi, Paolo; Tse Sum Bui, Bernadette; Haupt, Karsten

    2015-02-15

    We have developed a disposable evanescent wave fiber optic sensor by coating a molecularly imprinted polymer (MIP) containing a fluorescent signaling group on a 4-cm long polystyrene optical waveguide. The MIP is composed of a naphthalimide-based fluorescent monomer, which shows fluorescence enhancement upon binding with carboxyl-containing molecules. The herbicide 2,4-dichlorophenoxyacetic acid and the mycotoxin citrinin were used as model analytes. The coating of the MIP was either performed ex-situ, by dip-coating the fiber with MIP particles synthesized beforehand, or in-situ by evanescent-wave photopolymerization on the fiber. The sensing element was interrogated with a fiber-coupled spectrofluorimeter. The fiber optic sensor detects targets in the low nM range and exhibits specific and selective recognition over structural analogs and non-related carboxyl-containing molecules. This technology can be extended to other carboxyl-containing analytes, and to a broader spectrum of targets using different fluorescent monomers. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Binding analysis for interaction of diacetylcurcumin with β-casein nanoparticles by using fluorescence spectroscopy and molecular docking calculations

    Science.gov (United States)

    Mehranfar, Fahimeh; Bordbar, Abdol-Khalegh; Fani, Najme; Keyhanfar, Mehrnaz

    2013-11-01

    The interaction of diacetylcurcumin (DAC), as a novel synthetic derivative of curcumin, with bovine β-casein (an abundant milk protein that is highly amphiphilic and self assembles into stable micellar nanoparticles in aqueous solution) was investigated using fluorescence quenching experiments, Forster energy transfer measurements and molecular docking calculations. The fluorescence quenching measurements revealed the presence of a single binding site on β-casein for DAC with the binding constant value equals to (4.40 ± 0.03) × 104 M-1. Forster energy transfer measurements suggested that the distance between bound DAC and Trp143 residue is higher than the respective critical distance, hence, the static quenching is more likely responsible for fluorescence quenching other than the mechanism of non-radiative energy transfer. Our results from molecular docking calculations indicated that binding of DAC to β-casein predominantly occurred through hydrophobic contacts in the hydrophobic core of protein. Additionally, in vitro investigation of the cytotoxicity of free DAC and DAC-β-casein complex in human breast cancer cell line MCF7 revealed the higher cytotoxic effect of DAC-β-casein complex.

  16. LED-FISH: Fluorescence microscopy based on light emitting diodes for the molecular analysis of Her-2/neu oncogene amplification

    Directory of Open Access Journals (Sweden)

    Vollmer Ekkehard

    2008-12-01

    experiences emphasize the high potential of this technology to provide a serious alternative to conventional fluorescence microscopy in routine pathology; representing a sustainable technological progress, this low-cost technology will clearly give direction also to the growing field of molecular pathology. * AFTER = Amplified Fluorescence by transmitted Excitation of Radiation

  17. Molecular Detection of Bladder Cancer by Fluorescence Microsatellite Analysis and an Automated Genetic Analyzing System

    Directory of Open Access Journals (Sweden)

    Sarel Halachmi

    2007-01-01

    Full Text Available To investigate the ability of an automated fluorescent analyzing system to detect microsatellite alterations, in patients with bladder cancer. We investigated 11 with pathology proven bladder Transitional Cell Carcinoma (TCC for microsatellite alterations in blood, urine, and tumor biopsies. DNA was prepared by standard methods from blood, urine and resected tumor specimens, and was used for microsatellite analysis. After the primers were fluorescent labeled, amplification of the DNA was performed with PCR. The PCR products were placed into the automated genetic analyser (ABI Prism 310, Perkin Elmer, USA and were subjected to fluorescent scanning with argon ion laser beams. The fluorescent signal intensity measured by the genetic analyzer measured the product size in terms of base pairs. We found loss of heterozygocity (LOH or microsatellite alterations (a loss or gain of nucleotides, which alter the original normal locus size in all the patients by using fluorescent microsatellite analysis and an automated analyzing system. In each case the genetic changes found in urine samples were identical to those found in the resected tumor sample. The studies demonstrated the ability to detect bladder tumor non-invasively by fluorescent microsatellite analysis of urine samples. Our study supports the worldwide trend for the search of non-invasive methods to detect bladder cancer. We have overcome major obstacles that prevented the clinical use of an experimental system. With our new tested system microsatellite analysis can be done cheaper, faster, easier and with higher scientific accuracy.

  18. "Molecular beacon"-hosted thioflavin T: Applications for label-free fluorescent detection of iodide and logic operations.

    Science.gov (United States)

    Li, Yan-Yun; Jiang, Xiao-Qin; Lu, Ling-Fei; Zhang, Min; Shi, Guoyue

    2016-04-01

    In this work, we presented a simple, label-free and rapid-responsive fluorescence assay for iodide (I(-)) detection based on "molecular beacon (MB)"-hosted thioflavin T (ThT), achieving a limit of detection as low as 158 nM. The proposed method exhibited very good selectivity to I(-) ions over other anions interference due to the strong binding force between I(-) ions with Hg(2+). Upon the addition of I(-) ions, it would capture Hg(2+) from a T-Hg(2+)-T complex belonging to the MB-like DNA hairpin structure, which eventually quenched the initial fluorescence as output. In addition, it was successfully applied for operation of an integrated DNA logic gate system and to the determination of I(-) in real samples such as human urine. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Molecular imaging of atherosclerosis in mice with MRI and near-infrared fluorescence imaging

    International Nuclear Information System (INIS)

    Lu Tong; Wen Song; Zhou Guanhui; Ju Shenghong; Teng Gaojun

    2012-01-01

    Objective: To explore the feasibility of detecting atherosclerotic plaques with 7.0 T MRI and near-infrared fluorescence imaging (NIRF) using molecular imaging probes. Methods: Atherosclerotic plaques were established in male atherosclerotic apolipoprotein E knockout (ApoE-/-) mice fed with high-cholesterol diet for 20 weeks. Wild-type C57BL/6 mice were used as negative controls. 7.0 T MRI was performed before and 36 h after intravenously administration of ultrasmall superparamagnetic particle of iron oxide (USPIO). NIR 797 was conjugated with anti-mouse-oxidized modified low density lipoprotein (oxLDL) antibodies to construct an anti-oxLDL-Ab-NIR 797 probe while non-specific IgG-NIR 797 and PBS used as controls. NIRF was performed 24 h after tail vein injection of the probe. Independent sample t-test and one-way analysis of variance were used to analyze the data by SPSS 17.0. Results: In APOE-/-mice, in vivo 36 h post-USPIO T 2 WI images revealed strong focal signal loss in the abdominal aorta than that of pre-USPIO, with relative signal intensity 0.70 ± 0.04 and 1.28 ± 0.06, respectively (t=3.376, P<0.05). The percent of signal reduced was (-56.58 ± 4.25)%. The Prussian blue staining confirmed the depositions of iron particles in the plaque lesions. Significant fluorochrome accumulation in atherosclerotic plaques was demonstrated in aortic root, aortic arch and the starting of descending aorta 24 h after injection of the anti-oxLDL-Ab-NIR 797 probe. Minimal antibody uptake was observed in normal vessels from wild-type mice receiving the anti-oxLDL-Ab-NIR 797 (SNR: 2.29 ± 1.11) and in atherosclerotic vessels from ApoE-/- mice receiving the non-specific IgG-NIR 797 (19.58 ±3.06) or PBS (4.19 ±0.82), which was significantly different from the uptake of anti-oxLDL-Ab-NIR 797 group (42.51 ±5.24, F=25.104, P<0.05). Comparison between oil red O staining and NIRF 24 h after injection of NIR 797 labeled oxLDL-antibody revealed a significant correlation (r=0.738, P

  20. Balanced cross-rate model for saturated molecular fluorescence in flames using a nanosecond pulse length laser

    International Nuclear Information System (INIS)

    Lucht, R.P.; Sweeney, D.W.; Laurendeau, N.M.

    1980-01-01

    The balanced cross-rate model is proposed to analyze laser-induced molecular fluorescence signals when the laser pulse length is of the order of nanoseconds. Nanosecond pulse length lasers. specifically Q-switched Nd:YAG-pumped dye lasers, are attractive for saturated molecular fluorescence spectroscopy because of their high peak power and because of their short pulse length minimizes the risk of laser-induced chemistry. In the balanced cross-rate model, single upper and lower rotational levels are assumed to be directly coupled by the laser radiation. Because the laser-induced processes which couple these levels are so fast at saturation intensities, a steady state is established between the two levels within picoseconds. Provided that the total population of the two laser-coupled rotational levels is constant during the laser pulse, the total molecular population can be calculated from the observed upper rotational level population using a two-level saturation model and Boltzmann statistics. Numerical simulation of the laser excitation dynamics of OH in an atmospheric pressure H 2 /O 2 /N 2 flame indicates that the balanced cross-rate model will give accurate results provided that the rotational relaxation rates in the upper and lower sets of rotational levels are approximately equal

  1. Investigation of the interaction between isomeric derivatives and human serum albumin by fluorescence spectroscopy and molecular modeling

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ruiyong, E-mail: wangry@zzu.edu.cn; Dou, Huanjing; Yin, Yujing; Xie, Yuanzhe; Sun, Li; Liu, Chunmei; Dong, Jingjing; Huang, Gang; Zhu, Yanyan; Song, Chuanjun, E-mail: chjsong@zzu.edu.cn; Chang, Junbiao, E-mail: changjunbiao@zzu.edu.cn

    2014-10-15

    In this paper, we have synthesized 9H-pyrrolo[1,2-a]indol-9-ones and the isomeric indeno[2,1-b]pyrrol-8-ones. The interactions of human serum albumin with series of isomeric derivatives have been studied by spectrophotometric methods. Results show the intrinsic fluorescence is quenched by the derivatives with a static quenching procedure. The thermodynamics parameters indicate that van der Waals forces and hydrogen bonds play a major role in the interactions. The results of synchronous fluorescence spectra demonstrate that the microenvironments of Trp residue of human serum albumin are disturbed by most derivatives. Thermodynamic results showed that the 9H-pyrrolo[1,2-a]indol-9-ones are stronger quenchers and bind to human serum albumin with the higher affinity than isomeric indeno[2,1-b]pyrrol-8-ones. The influence of molecular structure on the binding aspects has been investigated. - Highlights: • The interactions between isomeric derivatives and HSA have been investigated. • Results reveal that 9H-pyrrolo[1,2-a]indol-9-ones are stronger quenchers for HSA. • Hydrogen bonds and van der Waals forces play major role in the binding process. • The influence of molecular structure on the binding aspects has been investigated. • The binding study was also modeled by molecular docking.

  2. Investigation of the interaction between isomeric derivatives and human serum albumin by fluorescence spectroscopy and molecular modeling

    International Nuclear Information System (INIS)

    Wang, Ruiyong; Dou, Huanjing; Yin, Yujing; Xie, Yuanzhe; Sun, Li; Liu, Chunmei; Dong, Jingjing; Huang, Gang; Zhu, Yanyan; Song, Chuanjun; Chang, Junbiao

    2014-01-01

    In this paper, we have synthesized 9H-pyrrolo[1,2-a]indol-9-ones and the isomeric indeno[2,1-b]pyrrol-8-ones. The interactions of human serum albumin with series of isomeric derivatives have been studied by spectrophotometric methods. Results show the intrinsic fluorescence is quenched by the derivatives with a static quenching procedure. The thermodynamics parameters indicate that van der Waals forces and hydrogen bonds play a major role in the interactions. The results of synchronous fluorescence spectra demonstrate that the microenvironments of Trp residue of human serum albumin are disturbed by most derivatives. Thermodynamic results showed that the 9H-pyrrolo[1,2-a]indol-9-ones are stronger quenchers and bind to human serum albumin with the higher affinity than isomeric indeno[2,1-b]pyrrol-8-ones. The influence of molecular structure on the binding aspects has been investigated. - Highlights: • The interactions between isomeric derivatives and HSA have been investigated. • Results reveal that 9H-pyrrolo[1,2-a]indol-9-ones are stronger quenchers for HSA. • Hydrogen bonds and van der Waals forces play major role in the binding process. • The influence of molecular structure on the binding aspects has been investigated. • The binding study was also modeled by molecular docking

  3. New insights into heat induced structural changes of pectin methylesterase on fluorescence spectroscopy and molecular modeling basis

    Science.gov (United States)

    Nistor, Oana Viorela; Stănciuc, Nicoleta; Aprodu, Iuliana; Botez, Elisabeta

    2014-07-01

    Heat-induced structural changes of Aspergillus oryzae pectin methylesterase (PME) were studied by means of fluorescence spectroscopy and molecular modeling, whereas the functional enzyme stability was monitored by inactivation studies. The fluorescence spectroscopy experiments were performed at two pH value (4.5 and 7.0). At both pH values, the phase diagrams were linear, indicating the presence of two molecular species induced by thermal treatment. A red shift of 7 nm was observed at neutral pH by increasing temperature up to 60 °C, followed by a blue shift of 4 nm at 70 °C, suggesting significant conformational rearrangements. The quenching experiments using acrylamide and iodide demonstrate a more flexible conformation of enzyme with increasing temperature, especially at neutral pH. The experimental results were complemented with atomic level observations on PME model behavior after performing molecular dynamics simulations at different temperatures. The inactivation kinetics of PME in buffer solutions was fitted using a first-order kinetics model, resulting in activation energy of 241.4 ± 7.51 kJ mol-1.

  4. Dynamic Assessment of the Endothelialization of Tissue-Engineered Blood Vessels Using an Optical Coherence Tomography Catheter-Based Fluorescence Imaging System.

    Science.gov (United States)

    Gurjarpadhye, Abhijit Achyut; DeWitt, Matthew R; Xu, Yong; Wang, Ge; Rylander, Marissa Nichole; Rylander, Christopher G

    2015-07-01

    Lumen endothelialization of bioengineered vascular scaffolds is essential to maintain small-diameter graft patency and prevent thrombosis postimplantation. Unfortunately, nondestructive imaging methods to visualize this dynamic process are lacking, thus slowing development and clinical translation of these potential tissue-engineering approaches. To meet this need, a fluorescence imaging system utilizing a commercial optical coherence tomography (OCT) catheter was designed to visualize graft endothelialization. C7 DragonFly™ intravascular OCT catheter was used as a channel for delivery and collection of excitation and emission spectra. Poly-dl-lactide (PDLLA) electrospun scaffolds were seeded with endothelial cells (ECs). Seeded cells were exposed to Calcein AM before imaging, causing the living cells to emit green fluorescence in response to blue laser. By positioning the catheter tip precisely over a specimen using high-fidelity electromechanical components, small regions of the specimen were excited selectively. The resulting fluorescence intensities were mapped on a two-dimensional digital grid to generate spatial distribution of fluorophores at single-cell-level resolution. Fluorescence imaging of endothelialization on glass and PDLLA scaffolds was performed using the OCT catheter-based imaging system as well as with a commercial fluorescence microscope. Cell coverage area was calculated for both image sets for quantitative comparison of imaging techniques. Tubular PDLLA scaffolds were maintained in a bioreactor on seeding with ECs, and endothelialization was monitored over 5 days using the OCT catheter-based imaging system. No significant difference was observed in images obtained using our imaging system to those acquired with the fluorescence microscope. Cell area coverage calculated using the images yielded similar values. Nondestructive imaging of endothelialization on tubular scaffolds showed cell proliferation with cell coverage area increasing from

  5. Quantum dots versus organic fluorophores in fluorescent deep-tissue imaging--merits and demerits.

    Science.gov (United States)

    Bakalova, Rumiana; Zhelev, Zhivko; Gadjeva, Veselina

    2008-12-01

    The use of fluorescence in deep-tissue imaging is rapidly expanding in last several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecular targets in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With the development of novel bright fluorophores based on nanotechnologies and 3D fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. The fluorescent imaging has also a potential to give a real map of human anatomy and physiology. The current review outlines the advantages of fluorescent nanoparticles over conventional organic dyes in deep-tissue imaging in vivo and defines the major requirements to the "perfect fluorophore". The analysis proceeds from the basic principles of fluorescence and major characteristics of fluorophores, light-tissue interactions, and major limitations of fluorescent deep-tissue imaging. The article is addressed to a broad readership - from specialists in this field to university students.

  6. A matter of collection and detection for intraoperative and noninvasive near-infrared fluorescence molecular imaging: To see or not to see?

    Science.gov (United States)

    Zhu, Banghe; Rasmussen, John C.; Sevick-Muraca, Eva M.

    2014-01-01

    Purpose: Although fluorescence molecular imaging is rapidly evolving as a new combinational drug/device technology platform for molecularly guided surgery and noninvasive imaging, there remains no performance standards for efficient translation of “first-in-humans” fluorescent imaging agents using these devices. Methods: The authors employed a stable, solid phantom designed to exaggerate the confounding effects of tissue light scattering and to mimic low concentrations (nM–pM) of near-infrared fluorescent dyes expected clinically for molecular imaging in order to evaluate and compare the commonly used charge coupled device (CCD) camera systems employed in preclinical studies and in human investigational studies. Results: The results show that intensified CCD systems offer greater contrast with larger signal-to-noise ratios in comparison to their unintensified CCD systems operated at clinically reasonable, subsecond acquisition times. Conclusions: Camera imaging performance could impact the success of future “first-in-humans” near-infrared fluorescence imaging agent studies. PMID:24506637

  7. Imaging breast adipose and fibroglandular tissue molecular signatures by using hybrid MRI-guided near-infrared spectral tomography

    Science.gov (United States)

    Brooksby, Ben; Pogue, Brian W.; Jiang, Shudong; Dehghani, Hamid; Srinivasan, Subhadra; Kogel, Christine; Tosteson, Tor D.; Weaver, John; Poplack, Steven P.; Paulsen, Keith D.

    2006-06-01

    Magnetic resonance (MR)-guided near-infrared spectral tomography was developed and used to image adipose and fibroglandular breast tissue of 11 normal female subjects, recruited under an institutional review board-approved protocol. Images of hemoglobin, oxygen saturation, water fraction, and subcellular scattering were reconstructed and show that fibroglandular fractions of both blood and water are higher than in adipose tissue. Variation in adipose and fibroglandular tissue composition between individuals was not significantly different across the scattered and dense breast categories. Combined MR and near-infrared tomography provides fundamental molecular information about these tissue types with resolution governed by MR T1 images. hemoglobin | magnetic resonance imaging | water | fat | oxygen saturation

  8. Molecular pathology in vulnerable carotid plaques: correlation with [18]-fluorodeoxyglucose positron emission tomography (FDG-PET)

    DEFF Research Database (Denmark)

    Graebe, M; Pedersen, Sune Folke; Borgwardt, L

    2008-01-01

    OBJECTIVES: Atherosclerosis is recognised as an inflammatory disease, and new diagnostic tools are warranted to evaluate plaque inflammatory activity and risk of cardiovascular events. We investigated [18]-fluorodeoxyglucose (FDG) uptake in vulnerable carotid plaques visualised by positron emission...... tomography (PET). Uptake was correlated to quantitative gene expression of known markers of inflammation and plaque vulnerability. METHODS: Ten patients with recent transient ischaemic attack and carotid artery stenosis (>50%) underwent combined FDG-PET and computed tomography angiography (CTA) the day...

  9. Design and synthesis of a fluorescent molecular imprinted polymer for use in an optical fibre-based cocaine sensor

    Science.gov (United States)

    Wren, Stephen P.; Piletsky, Sergey A.; Karim, Kal; Gascoine, Paul; Lacey, Richard; Sun, Tong; Grattan, Kenneth T. V.

    2014-05-01

    Previously, we have developed chemical sensors using fibre optic-based techniques for the detection of Cocaine, utilising molecularly imprinted polymers (MIPs) containing fluorescein moieties as the signalling groups. Here, we report the computational design of a fluorophore which was incorporated into a MIP for the generation of a novel sensor that offers improved sensitivity for Cocaine with a detection range of 1-100μM. High selectivity for Cocaine over a suite of known Cocaine interferants (25μM) was also demonstrated by measuring changes in the intensity of fluorescence signals received from the sensor.

  10. How to measure separation and angles between inter-molecular fluorescent markers

    DEFF Research Database (Denmark)

    Flyvbjerg, Henrik

    Structure and function of an individual biomolecule can be explored with minimum two fluorescent markers of different colors. Since the light of such markers can be spec- trally separated and imaged simultaneously, the markers can be colocalized. Here, we describe the method used for such two......-color colocalization microscopy. Then we extend it to fluorescent markers with fixed orientations and in intramolecular proximity. Our benchmarking of this extension produced two extra results: (a) we established short double-labeled DNA molecules as probes of 3D orientation of anything to which one can attach them...

  11. Incoherent-scatter computed tomography with monochromatic synchrotron x ray: feasibility of multi-CT imaging system for simultaneous measurement-of fluorescent and incoherent scatter x rays

    Science.gov (United States)

    Yuasa, T.; Akiba, M.; Takeda, T.; Kazama, M.; Hoshino, A.; Watanabe, Y.; Hyodo, K.; Dilmanian, F. A.; Akatsuka, T.; Itai, Y.

    1997-10-01

    We describe a new system of incoherent scatter computed tomography (ISCT) using monochromatic synchrotron X rays, and we discuss its potential to be used in in vivo imaging for medical use. The system operates on the basis of computed tomography (CT) of the first generation. The reconstruction method for ISCT uses the least squares method with singular value decomposition. The research was carried out at the BLNE-5A bending magnet beam line of the Tristan Accumulation Ring in KEK, Japan. An acrylic cylindrical phantom of 20-mm diameter containing a cross-shaped channel was imaged. The channel was filled with a diluted iodine solution with a concentration of 200 /spl mu/gI/ml. Spectra obtained with the system's high purity germanium (HPGe) detector separated the incoherent X-ray line from the other notable peaks, i.e., the iK/sub /spl alpha// and K/sub /spl beta/1/ X-ray fluorescent lines and the coherent scattering peak. CT images were reconstructed from projections generated by integrating the counts In the energy window centering around the incoherent scattering peak and whose width was approximately 2 keV. The reconstruction routine employed an X-ray attenuation correction algorithm. The resulting image showed more homogeneity than one without the attenuation correction.

  12. In situ detection of atomic and molecular iodine using Resonance and Off-Resonance Fluorescence by Lamp Excitation: ROFLEX

    Directory of Open Access Journals (Sweden)

    J. C. Gómez Martín

    2011-01-01

    Full Text Available We demonstrate a new instrument for in situ detection of atmospheric iodine atoms and molecules based on atomic and molecular resonance and off-resonance ultraviolet fluorescence excited by lamp emission. The instrument combines the robustness, light weight, low power consumption and efficient excitation of radio-frequency discharge light sources with the high sensitivity of the photon counting technique. Calibration of I2 fluorescence is achieved via quantitative detection of the molecule by Incoherent Broad Band Cavity-enhanced Absorption Spectroscopy. Atomic iodine fluorescence signal is calibrated by controlled broad band photolysis of known I2 concentrations in the visible spectral range at atmospheric pressure. The instrument has been optimised in laboratory experiments to reach detection limits of 1.2 pptv for I atoms and 13 pptv for I2, for S/N = 1 and 10 min of integration time. The ROFLEX system has been deployed in a field campaign in northern Spain, representing the first concurrent observation of ambient mixing ratios of iodine atoms and molecules in the 1–350 pptv range.

  13. Two-Photon Absorption Properties of Gold Fluorescent Protein: A Combined Molecular Dynamics and Quantum Chemistry Study.

    Science.gov (United States)

    Simsek, Yusuf; Brown, Alex

    2018-05-09

    Molecular dynamic (MD) simulations were carried out to obtain the conformational changes of the chromophore in the gold fluorescent protein (PDB ID: 1OXF). To obtain two-photon absorption (TPA) cross-sections, time dependent density functional theory (TD-DFT) computations were performed for chromophore geometries sampled along the trajectory. The TD-DFT computations used the CAM-B3LYP functional and 6-31+G(d) basis set with the conductor-like polarizable continuum model (PCM) with parameters for water. Results showed that two dihedral angles change remarkably over the simulation time. TPA cross-sections were found to average 20 GM for the excitation to S1 between 430 and 460 nm; however, the maximal and minimal values were 35GM and 5GM, respectively. Besides the effects of the dihedrals on the spectroscopic properties, some bond lengths affected the excitation energies and the TPA cross-sections significantly (up to ±25-30%) while the effects of bond angles were smaller (±5%). Overall the present results provide insight in the effects of conformational exibility on TPA (with gold fluorescent protein as a specific example) and suggest that further experimental measurements of TPA for gold fluorescent protein should be undertaken.

  14. Adaptation of Tri-molecular fluorescence complementation allows assaying of regulatory Csr RNA-protein interactions in bacteria.

    Science.gov (United States)

    Gelderman, Grant; Sivakumar, Anusha; Lipp, Sarah; Contreras, Lydia

    2015-02-01

    sRNAs play a significant role in controlling and regulating cellular metabolism. One of the more interesting aspects of certain sRNAs is their ability to make global changes in the cell by interacting with regulatory proteins. In this work, we demonstrate the use of an in vivo Tri-molecular Fluorescence Complementation assay to detect and visualize the central regulatory sRNA-protein interaction of the Carbon Storage Regulatory system in E. coli. The Carbon Storage Regulator consists primarily of an RNA binding protein, CsrA, that alters the activity of mRNA targets and of an sRNA, CsrB, that modulates the activity of CsrA. We describe the construction of a fluorescence complementation system that detects the interactions between CsrB and CsrA. Additionally, we demonstrate that the intensity of the fluorescence of this system is able to detect changes in the affinity of the CsrB-CsrA interaction, as caused by mutations in the protein sequence of CsrA. While previous methods have adopted this technique to study mRNA or RNA localization, this is the first attempt to use this technique to study the sRNA-protein interaction directly in bacteria. This method presents a potentially powerful tool to study complex bacterial RNA protein interactions in vivo. © 2014 Wiley Periodicals, Inc.

  15. Preliminary study on X-ray fluorescence computed tomography imaging of gold nanoparticles: Acceleration of data acquisition by multiple pinholes scheme

    Science.gov (United States)

    Sasaya, Tenta; Sunaguchi, Naoki; Seo, Seung-Jum; Hyodo, Kazuyuki; Zeniya, Tsutomu; Kim, Jong-Ki; Yuasa, Tetsuya

    2018-04-01

    Gold nanoparticles (GNPs) have recently attracted attention in nanomedicine as novel contrast agents for cancer imaging. A decisive tomographic imaging technique has not yet been established to depict the 3-D distribution of GNPs in an object. An imaging technique known as pinhole-based X-ray fluorescence computed tomography (XFCT) is a promising method that can be used to reconstruct the distribution of GNPs from the X-ray fluorescence emitted by GNPs. We address the acceleration of data acquisition in pinhole-based XFCT for preclinical use using a multiple pinhole scheme. In this scheme, multiple projections are simultaneously acquired through a multi-pinhole collimator with a 2-D detector and full-field volumetric beam to enhance the signal-to-noise ratio of the projections; this enables fast data acquisition. To demonstrate the efficacy of this method, we performed an imaging experiment using a physical phantom with an actual multi-pinhole XFCT system that was constructed using the beamline AR-NE7A at KEK. The preliminary study showed that the multi-pinhole XFCT achieved a data acquisition time of 20 min at a theoretical detection limit of approximately 0.1 Au mg/ml and at a spatial resolution of 0.4 mm.

  16. 3D characterisation of the gaseous and liquid phase using laser-induced exciplex fluorescence (LIEF) tomography; Dreidimensionale Charakterisierung der Gas- und Fluessigphase mittels laserinduzierter Exciplexfluoreszenz (LIEF) Tomografie

    Energy Technology Data Exchange (ETDEWEB)

    Rogler, P.; Grzeszik, R.; Arndt, S. [Robert Bosch GmbH, Stuttgart (Germany); Waidmann, C. [Fachhochschule Aalen (Germany); Aigner, M. [DLR, Stuttgart (Germany). Inst. fuer Verbrennungstechnik

    2007-07-01

    The quality of mixture formation in gasoline engines has a significant influence on combustion, emissions and mileage. The measurement technique used for mixture formation analysis of both liquid and vapor phase is laser induced exciplex fluorescence (LIEF), where the aromatics TMPD and naphthalene are added to a non-fluorescing model fuel free of aromatic compounds. For spatially high-resolved measurements of liquid and vapor phase this technique was extended from planar to volume information via tomography. This new approach requires fundamental studies of tracer behavior and new evaluation algorithms for reliable signal interpretation of fluid dynamics in sprays. Using this measurement technique GDI sprays are investigated at a typical stratified engine operation point at part load. For the different injector types, e.g. multi hole and annular orifice injector, an optimal illumination is crucial. With the collected data the spray volume can be calculated, mixture homogeneity can be evaluated and, using a few assumptions, the air/fuel ratio {lambda} can be computed. (orig.)

  17. Investigation of three flavonoids binding to bovine serum albumin using molecular fluorescence technique

    International Nuclear Information System (INIS)

    Bi Shuyun; Yan Lili; Pang Bo; Wang Yu

    2012-01-01

    The three flavonoids including naringenin, hesperetin and apigenin binding to bovine serum albumin (BSA) at pH 7.4 was studied by fluorescence quenching, synchronous fluorescence and UV-vis absorption spectroscopic techniques. The results obtained revealed that naringenin, hesperetin and apigenin strongly quenched the intrinsic fluorescence of BSA. The Stern-Volmer curves suggested that these quenching processes were all static quenching processes. At 291 K, the value and the order of the binding constant were K A n aringenin) =4.08x10 4 A(hesperetin) =5.40x10 4 ∼K A(apigenin) =5.32x10 4 L mol -1 . The main binding force between the flavonoid and BSA was hydrophobic and electrostatic force. According to the Foerster theory of non-radiation energy transfer, the binding distances (r 0 ) were obtained as 3.36, 3.47 and 3.30 nm for naringenin-BSA, hesperetin-BSA and apigenin-BSA, respectively. The effect of some common ions such as Fe 3+ , Cu 2+ , Mg 2+ , Mn 2+ , Zn 2+ and Ca 2+ on the binding was also studied in detail. The competition binding was also performed. The apparent binding constant (K' A ) obtained suggested that one flavonoid had an obvious effect on the binding of another flavonoid to protein when they coexisted in BSA solution. - Highlights: → Quenchings of BSA fluorescence by the flavonoids was all static quenchings. → Synchronous fluorescence was applied to study the structural change of BSA. → Binding constant, binding site and binding force were determined. → Competition binding experiments were performed. → One flavonoid had an obvious effect on the binding of another one to BSA.

  18. Molecular and structural characterization of fluorescent human parvovirus B19 virus-like particles

    International Nuclear Information System (INIS)

    Gilbert, Leona; Toivola, Jouni; White, Daniel; Ihalainen, Teemu; Smith, Wesley; Lindholm, Laura; Vuento, Matti; Oker-Blom, Christian

    2005-01-01

    Although sharing a T = 1 icosahedral symmetry with other members of the Parvoviridae family, it has been suggested that the fivefold channel of the human parvovirus B19 VP2 capsids is closed at its outside end. To investigate the possibility of placing a relatively large protein moiety at this site of B19, fluorescent virus-like particles (fVLPs) of B19 were developed. The enhanced green fluorescent protein (EGFP) was inserted at the N-terminus of the structural protein VP2 and assembly of fVLPs from this fusion protein was obtained. Electron microscopy revealed that these fluorescent protein complexes were very similar in size when compared to wild-type B19 virus. Further, fluorescence correlation spectroscopy showed that an average of nine EGFP domains were associated with these virus-like structures. Atomic force microscopy and immunoprecipitation studies showed that EGFP was displayed on the surface of these fVLPs. Confocal imaging indicated that these chimeric complexes were targeted to late endosomes when expressed in insect cells. The fVLPs were able to efficiently enter cancer cells and traffic to the nucleus via the microtubulus network. Finally, immunoglobulins present in human parvovirus B19 acute and past-immunity serum samples were able to detect antigenic epitopes present in these fVLPs. In summary, we have developed fluorescent virus-like nanoparticles displaying a large heterologous entity that should be of help to elucidate the mechanisms of infection and pathogenesis of human parvovirus B19. In addition, these B19 nanoparticles serve as a model in the development of targetable vehicles designed for delivery of biomolecules

  19. Is hybridic positron emission tomography/computerized tomography the only option? The future of nuclear medicine and molecular imaging.

    Science.gov (United States)

    Grammaticos, Philip; Zerva, Cherry; Asteriadis, Ioannis; Trontzos, Christos; Hatziioannou, Kostas

    2007-01-01

    sources of radiation" b) nuclear radiation and c) molecular nuclear medicine. The "European Journal of Nuclear Medicine and Molecular Imaging" shall have to erase the three last words of its title and be renamed. As Professor Abass Alavi et al (2007), have mentioned: "Is PET/CT the only option?" In favor of PET/CT are the following: Attenuation correction (AC) and better anatomical localization of lesions visualized with PET. Also PET/CT can be used as a diagnostic CT scanner (dCT). Against using the PET/CT scanners are the following arguments: a) This equipment is not necessary because we can always ask the Radiologists for a dCT scan. Many patients have already done a dCT scan at the time they are referred for a PET scan to the Nuclear Medicine Department. b) The absolute clinical indications for PET/CT with the use of a contrast agent, are under investigation. c) Although there is at present a list of indications suggested for the PET/CT scanner, there are studies disputing some of these indications, as for example in metastatic colon cancer where a high diagnostic accuracy for PET study alone, has been reported. d) The option of AC performed by the PET/CT scanner has also been questioned. Artifacts may be up to 84%. e) The PET/CT is expensive, time consuming, space occupying, and needs additional medical and technical personnel. f) Not to mention the extra radiation dose to the patients. g) Shall we inform those young medical students who wish to become nuclear medicine physicians, to hold their decision till the content of future Nuclear Medicine is clarified? We may suggest that: Our specialty could be renamed as: "Clinical Nuclear Medicine" and include additional "proper certified education" on the PET/CT equipment. The PET/CT scanner should remain in the Nuclear Medicine Department where Radiologists could act as advisors.

  20. Multimodal nanoparticles as alignment and correlation markers in fluorescence/soft X-ray cryo-microscopy/tomography of nucleoplasmic reticulum and apoptosis in mammalian cells

    International Nuclear Information System (INIS)

    Hagen, Christoph; Werner, Stephan; Carregal-Romero, Susana; Malhas, Ashraf N.; Klupp, Barbara G.; Guttmann, Peter; Rehbein, Stefan; Henzler, Katja; Mettenleiter, Thomas C.

    2014-01-01

    Correlative fluorescence and soft X-ray cryo-microscopy/tomography on flat sample holders is perfectly suited to study the uncompromised physiological status of adherent cells at its best possible preservation by imaging after fast cryo-immobilization. To understand the mechanism by which herpesviruses induce nucleoplasmic reticulum, i.e. invaginations of the nuclear envelope, during their egress from the host cell nucleus, morphologically similar structures found in laminopathies and after chemical induction were investigated as a potentially more easily accessible model system. For example, anti-retroviral protease inhibitors like Saquinavir also induce invaginations of the nuclear membranes. With the help of newly designed multimodal nanoparticles as alignment and correlation markers, and by optimizing fluorescence cryo-microscopy data acquisition, an elaborate three-dimensional network of nucleoplasmic reticulum was demonstrated in nuclei of Saquinavir-treated rabbit kidney cells expressing a fluorescently labeled inner nuclear membrane protein. In part of the protease inhibitor-treated samples, nuclei exhibited dramatic ultrastructural changes indicative of programmed cell death/apoptosis. This unexpected observation highlights another unique feature of soft X-ray microscopy, i.e. high absorption contrast information not relying on labeled cellular components, at a 3D resolution of approximately 40 nm (half-pitch) and through a sample thickness of several micrometers. These properties make it a valuable part of the cell biology imaging toolbox to visualize the cellular ultrastructure in its completeness. - Highlights: • Nucleoplasmic reticulum was demonstrated in nuclei of Saquinavir-treated cells. • New polyelectrolyte-Qdot ® 605 coated gold beads were employed as fiducials. • Saquinavir can induce a strong apoptotic phenotype in the nucleus. • CryoXT is an auspicious imaging technique in apoptosis research

  1. The feasibility of polychromatic cone-beam x-ray fluorescence computed tomography (XFCT) imaging of gold nanoparticle-loaded objects: a Monte Carlo study.

    Science.gov (United States)

    Jones, Bernard L; Cho, Sang Hyun

    2011-06-21

    A recent study investigated the feasibility to develop a bench-top x-ray fluorescence computed tomography (XFCT) system capable of determining the spatial distribution and concentration of gold nanoparticles (GNPs) in vivo using a diagnostic energy range polychromatic (i.e. 110 kVp) pencil-beam source. In this follow-up study, we examined the feasibility of a polychromatic cone-beam implementation of XFCT by Monte Carlo (MC) simulations using the MCNP5 code. In the current MC model, cylindrical columns with various sizes (5-10 mm in diameter) containing water loaded with GNPs (0.1-2% gold by weight) were inserted into a 5 cm diameter cylindrical polymethyl methacrylate phantom. The phantom was then irradiated by a lead-filtered 110 kVp x-ray source, and the resulting gold fluorescence and Compton-scattered photons were collected by a series of energy-sensitive tallies after passing through lead parallel-hole collimators. A maximum-likelihood iterative reconstruction algorithm was implemented to reconstruct the image of GNP-loaded objects within the phantom. The effects of attenuation of both the primary beam through the phantom and the gold fluorescence photons en route to the detector were corrected during the image reconstruction. Accurate images of the GNP-containing phantom were successfully reconstructed for three different phantom configurations, with both spatial distribution and relative concentration of GNPs well identified. The pixel intensity of regions containing GNPs was linearly proportional to the gold concentration. The current MC study strongly suggests the possibility of developing a bench-top, polychromatic, cone-beam XFCT system for in vivo imaging.

  2. Molecularly imprinted polymer based on MWCNT-QDs as fluorescent biomimetic sensor for specific recognition of target protein

    Energy Technology Data Exchange (ETDEWEB)

    Ding, Zhaoqiang [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Annie Bligh, S.W. [Department of Life Sciences, Faculty of Science and Technology, University of Westminster, 115 New Cavendish Street, London W1W 6UW (United Kingdom); Tao, Lei; Quan, Jing [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Nie, Huali, E-mail: niehuali@dhu.edu.cn [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Zhu, Limin, E-mail: lzhu@dhu.edu.cn [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Gong, Xiao [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China)

    2015-03-01

    A novel molecularly imprinted optosensing material based on multi-walled carbon nanotube-quantum dots (MWCNT-QDs) has been designed and synthesized for its high selectivity, sensitivity and specificity in the recognition of a target protein bovine serum albumin (BSA). Molecularly imprinted polymer coated MWCNT-QDs using BSA as the template (BMIP-coated MWCNT-QDs) exhibits a fast mass-transfer speed with a response time of 25 min. It is found that the BSA as a target protein can significantly quench the luminescence of BMIP-coated MWCNT-QDs in a concentration-dependent manner that is best described by a Stern–Volmer equation. The K{sub SV} for BSA is much higher than bovine hemoglobin and lysozyme, implying a highly selective recognition of the BMIP-coated MWCNT-QDs to BSA. Under optimal conditions, the relative fluorescence intensity of BMIP-coated MWCNT-QDs decreases linearly with the increasing target protein BSA in the concentration range of 5.0 × 10{sup −7}–35.0 × 10{sup −7} M with a detection limit of 80 nM. - Highlights: • A novel fluorescent biomimetic sensor based on MWCNT-QDs was designed. • The sensor exhibited a fast mass-transfer speed with a response time of 25 min. • The sensor possessed a highly selective recognition to BSA.

  3. Pre-recombination quenching of the radiation induced fluorescence as the approach to study kinetics of ion-molecular reactions

    International Nuclear Information System (INIS)

    Borovkov, V.I.; Ivanishko, I.S.

    2011-01-01

    This study deals with the geminate ion recombination in the presence of bulk scavengers, that is the so-called scavenger problem, as well as with the effect of the scavenging reaction on the radiation-induced recombination fluorescence. have proposed a method to determine the rate constant of the bulk reaction between neutral scavengers and one of the geminate ions if the ion-molecular reaction prevented the formation of electronically excited states upon recombination involving a newly formed ion. If such pre-recombination quenching of the radiation-induced fluorescence took place, it manifested itself as a progressive decrease in the decay of the fluorescence intensity. The relative change in the fluorescence decay as caused by the scavengers was believed to be closely related to the kinetics of the scavenging reaction. The goal of the present study is to support this method, both computationally and experimentally because there are two factors, which cast doubt on the intuitively obvious approach to the scavenger problem: spatial correlations between the particles involved and the drift of the charged reagent in the electric field of its geminate partner. Computer simulation of geminate ions recombination with an explicit modeling of the motion trajectories of scavengers has been performed for media of low dielectric permittivity, i.e. for the maximal Coulomb interaction between the ions. The simulation has shown that upon continuous diffusion of the particles involved, the joint effect of the two above factors can be considered as insignificant with a high accuracy. Besides, it is concluded then that the method of pre-recombination quenching could be applied to study parallel and consecutive reactions where the yields of excited states in the reaction pathways are different with the use of very simple analytical relations of the formal chemical kinetics. The conclusion has been confirmed experimentally by the example of the reactions of electron transfer from

  4. DHA-fluorescent probe is sensitive to membrane order and reveals molecular adaptation of DHA in ordered lipid microdomains☆

    Science.gov (United States)

    Teague, Heather; Ross, Ron; Harris, Mitchel; Mitchell, Drake C.; Shaikh, Saame Raza

    2012-01-01

    Docosahexaenoic acid (DHA) disrupts the size and order of plasma membrane lipid microdomains in vitro and in vivo. However, it is unknown how the highly disordered structure of DHA mechanistically adapts to increase the order of tightly packed lipid microdomains. Therefore, we studied a novel DHA-Bodipy fluorescent probe to address this issue. We first determined if the DHA-Bodipy probe localized to the plasma membrane of primary B and immortal EL4 cells. Image analysis revealed that DHA-Bodipy localized into the plasma membrane of primary B cells more efficiently than EL4 cells. We then determined if the probe detected changes in plasma membrane order. Quantitative analysis of time-lapse movies established that DHA-Bodipy was sensitive to membrane molecular order. This allowed us to investigate how DHA-Bodipy physically adapted to ordered lipid microdomains. To accomplish this, we employed steady-state and time-resolved fluorescence anisotropy measurements in lipid vesicles of varying composition. Similar to cell culture studies, the probe was highly sensitive to membrane order in lipid vesicles. Moreover, these experiments revealed, relative to controls, that upon incorporation into highly ordered microdomains, DHA-Bodipy underwent an increase in its fluorescence lifetime and molecular order. In addition, the probe displayed a significant reduction in its rotational diffusion compared to controls. Altogether, DHA-Bodipy was highly sensitive to membrane order and revealed for the first time that DHA, despite its flexibility, could become ordered with less rotational motion inside ordered lipid microdomains. Mechanistically, this explains how DHA acyl chains can increase order upon formation of lipid microdomains in vivo. PMID:22841541

  5. High sensitive determination of zinc with novel water-soluble small molecular fluorescent sensor

    International Nuclear Information System (INIS)

    Weng Ying; Chen Zilin; Wang Fang; Xue Lin; Jiang Hua

    2009-01-01

    A high sensitive method of quantitative analysis for the determination of zinc in the nutrition supplements has been developed by using a novel water-soluble fluorescent sensor HQ3: (8-pyridylmethyloxy-2-methyl-quinoline). Under the optimized condition of 67 mM phosphate buffer, pH 7.4, and 5% (v/v) DMSO, the zinc concentration showed good linear relationship with fluorescence intensity in the range of 7.5 x 10 -8 to 2.5 x 10 -5 M with the detection limit of 1.5 x 10 -8 M. HQ3 exhibited high selectivity to zinc comparing with other metal ions except for cadmium. The developed analytical method was successfully used for determining the content of zinc in a real sample of zinc gluconate solution of Sanchine.

  6. A fluorescence study of the molecular interactions of harmane with the nucleobases, their nucleosides and mononucleotides.

    Science.gov (United States)

    Balón, M; Muñoz, M A; Carmona, C; Guardado, P; Galán, M

    1999-07-19

    Fluorescence binding studies of harmane to the elemental components of the nucleic acids were undertaken to investigate the origin of the interaction between the drug and DNA. Most of the tested substrates have been found to induce hypochromism in the absorption spectrum of harmane and to quench its fluorescence. The quenching process induced by the nucleobases and their nucleosides is mainly due to the formation of ground state 1:1 complexes. However, in the case of the mononucleotides a dynamic quenching component is also observed. This quenching component is likely due to the excited state interaction of harmane with the phosphate group of the nucleotides. UV-vis spectral changes and quenching measurements have been used to quantify the ground state association constants of the complexes and the quenching rate constants.

  7. A Novel Molecular Fluorescent Technique for Imaging the Somatostatin Receptor 2, Using a DOTATOC Lanthanide Conjugate

    DEFF Research Database (Denmark)

    Andersen, Rune Wiik; Prakash, Vineet; Stensballe, Allan

    -functional DOTATOC component.                       METHOD AND MATERIALS            The chelation of Europium and Samarium to DOTATOC was proven using MALDI-TOF Mass Spectrometry. The rise in quantum yield between unchelated lanthanides and those bound by DOTATOC was examined using fluorescence spectroscopy...

  8. Orbital tomography: Molecular band maps, momentum maps and the imaging of real space orbitals of adsorbed molecules

    Energy Technology Data Exchange (ETDEWEB)

    Offenbacher, Hannes; Lüftner, Daniel; Ules, Thomas; Reinisch, Eva Maria; Koller, Georg, E-mail: georg.koller@uni-graz.at; Puschnig, Peter; Ramsey, Michael G., E-mail: michael.ramsey@uni-graz.at

    2015-10-01

    Highlights: • Orbital tomography within the plane wave final state approximation. • One electron orbital predictions versus angle resolved photoemission experiment. • Geometric and electronic structure of organic thin films elucidated by ARUPS. • Influence of molecular conformation and orientation on ARUPS. • Retrieval of sexiphenyl and pentacene orbitals in real space. - Abstract: The frontier orbitals of molecules are the prime determinants of their chemical, optical and electronic properties. Arguably, the most direct method of addressing the (filled) frontier orbitals is ultra-violet photoemission spectroscopy (UPS). Although UPS is a mature technique from the early 1970s on, the angular distribution of the photoemitted electrons was thought to be too complex to be analysed quantitatively. Recently angle resolved UPS (ARUPS) work on conjugated molecules both, in ordered thick films and chemisorbed monolayers, has shown that the angular (momentum) distribution of the photocurrent from orbital emissions can be simply understood. The approach, based on the assumption of a plane wave final state is becoming known as orbital tomography. Here we will demonstrate, with selected examples of pentacene (5A) and sexiphenyl (6P), the potential of orbital tomography. First it will be shown how the full angular distribution of the photocurrent (momentum map) from a specific orbital is related to the real space orbital by a Fourier transform. Examples of the reconstruction of 5A orbitals will be given and the procedure for recovering the lost phase information will be outlined. We then move to examples of sexiphenyl where we interrogate the original band maps of thick sexiphenyl in the light of our understanding of orbital tomography that has developed since then. With comparison to theoretical simulations of the molecular band maps, the molecular conformation and orientation will be concluded. New results for the sexiphenyl monolayer on Al(1 1 0) will then be

  9. Molecularly Imprinted Core-Shell CdSe@SiO2/CDs as a Ratiometric Fluorescent Probe for 4-Nitrophenol Sensing

    Science.gov (United States)

    Liu, Mingyue; Gao, Zhao; Yu, Yanjun; Su, Rongxin; Huang, Renliang; Qi, Wei; He, Zhimin

    2018-01-01

    4-Nitrophenol (4-NP) is a priority pollutant in water and is both carcinogenic and genotoxic to humans and wildlife even at very low concentrations. Thus, we herein fabricated a novel molecularly imprinted core-shell nanohybrid as a ratiometric fluorescent sensor for the highly sensitive and selective detection of 4-NP. This sensor was functioned by the transfer of fluorescence resonance energy between photoluminescent carbon dots (CDs) and 4-NP. This sensor was synthesized by linking organosilane-functionalized CDs to silica-coated CdSe quantum dots (CdSe@SiO2) via Si-O bonds. The nanohybrids were further modified by anchoring a molecularly imprinted polymer (MIP) layer on the ratiometric fluorescent sensor through a facile sol-gel polymerization method. The morphology, chemical structure, and optical properties of the resulting molecularly imprinted dual-emission fluorescent probe were characterized by transmission electron microscopy and spectroscopic analysis. The probe was then applied in the detection of 4-NP and exhibited good linearity between 0.051 and 13.7 μg/mL, in addition to a low detection limit of 0.026 μg/mL. Furthermore, the simplicity, reliability, high selectivity, and high sensitivity of the developed sensor demonstrate that the combination of MIPs and ratiometric fluorescence allows the preparation of excellent fluorescent sensors for the detection of trace or ultra-trace analytes.

  10. Use of fluorescence spectroscopy to measure molecular autofluorescence in diabetic subjects

    International Nuclear Information System (INIS)

    Gomes, Cinthia Zanini

    2011-01-01

    Diabetes Mellitus (DM) comprises a complex metabolic syndrome, caused by reduced or absent secretion of insulin by pancreatic beta cells, leading to hyperglycemia. Hyperglycemia promotes glycation of proteins and, consequently, the appearance of advanced glycation end products (AGEs). Currently, diabetic patients are monitored by determining levels of glucose and glycated hemoglobin (HbA1c). The complications caused by hyperglycemia may be divided into micro and macrovascular complications, represented by retinopathy, nephropathy, neuropathy and cardiovascular disease. Albumin (HSA) is the most abundant serum protein in the human body and is subject to glycation. The Protoporphyrin IX (PpIX) is the precursor molecule of heme synthesis, structural component of hemoglobin. The in vitro and animals studies have indicated that hyperglycemia promotes a decrease in its concentration in erythrocytes. The fluorescence spectroscopy is a technique widely used in biomedical field. The autofluorescence corresponds to the intrinsic fluorescence present in some molecules, this being associated with the same structure. The aim of this study was to use fluorescence spectroscopy to measure levels of erythrocyte PpIX autofluorescence and AGE-HSA in diabetic and healthy subjects and compare them with levels of blood glucose and HbA1c. This study was conducted with 151 subjects (58 controls and 93 diabetics). Epidemiological data of patients and controls were obtained from medical records. For control subjects, blood glucose levels were obtained from medical records and levels of Hb1Ac obtained by using commercial kits. The determination of the PpIX autofluorescence was performed with excitation at 405 nm and emission at 632 nm. Determination of AGE-HSA was performed with excitation at 370 nm and emission at 455 nm. Approximately 50% of diabetic had micro and macrovascular lesions resulting from hyperglycemia. There were no significant differences in the PpIX emission intensity values

  11. Synthesis of molecularly imprinted dye-silica nanocomposites with high selectivity and sensitivity: Fluorescent imprinted sensor for rapid and efficient detection of τ-fluvalinate in vodka.

    Science.gov (United States)

    Wang, Yunyun; Wang, Jixiang; Cheng, Rujia; Sun, Lin; Dai, Xiaohui; Yan, Yongsheng

    2018-04-01

    An imprinted fluorescent sensor was fabricated based on SiO 2 nanoparticles encapsulated with a molecularly imprinted polymer containing allyl fluorescein. High fluorine cypermethirin as template molecules, methyl methacrylate as functional monomer, and allyl fluorescein as optical materials synthesized a core-shell fluorescent molecular imprinted sensor, which showed a high and rapid sensitivity and selectivity for the detection of τ-fluvalinate. The sensor presented appreciable sensitivity with a limit of 13.251 nM, rapid detection that reached to equilibrium within 3 min, great linear relationship in the relevant concentration range from 0 to 150 nM, and excellent selectivity over structural analogues. In addition, the fluorescent sensor demonstrated desirable regeneration ability (eight cycling operations). The molecularly imprinted polymers ensured specificity, while the fluorescent dyes provided the stabile sensitivity. Finally, an effective application of the sensor was implemented by the detection of τ-fluvalinate in real samples from vodka. The molecularly imprinted fluorescent sensor showed a promising potential in environmental monitoring and food safety. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Fluorescent bioaerosol particle, molecular tracer, and fungal spore concentrations during dry and rainy periods in a semi-arid forest

    Directory of Open Access Journals (Sweden)

    M. I. Gosselin

    2016-12-01

    Full Text Available Bioaerosols pose risks to human health and agriculture and may influence the evolution of mixed-phase clouds and the hydrological cycle on local and regional scales. The availability and reliability of methods and data on the abundance and properties of atmospheric bioaerosols, however, are rather limited. Here we analyze and compare data from different real-time ultraviolet laser/light-induced fluorescence (UV-LIF instruments with results from a culture-based spore sampler and offline molecular tracers for airborne fungal spores in a semi-arid forest in the southern Rocky Mountains of Colorado. Commercial UV-APS (ultraviolet aerodynamic particle sizer and WIBS-3 (wideband integrated bioaerosol sensor, version 3 instruments with different excitation and emission wavelengths were utilized to measure fluorescent aerosol particles (FAPs during both dry weather conditions and periods heavily influenced by rain. Seven molecular tracers of bioaerosols were quantified by analysis of total suspended particle (TSP high-volume filter samples using a high-performance anion-exchange chromatography system with pulsed amperometric detection (HPAEC-PAD. From the same measurement campaign, Huffman et al. (2013 previously reported dramatic increases in total and fluorescent particle concentrations during and immediately after rainfall and also showed a strong relationship between the concentrations of FAPs and ice nuclei (Huffman et al., 2013; Prenni et al., 2013. Here we investigate molecular tracers and show that during rainy periods the atmospheric concentrations of arabitol (35.2 ± 10.5 ng m−3 and mannitol (44.9 ± 13.8 ng m−3 were 3–4 times higher than during dry periods. During and after rain, the correlations between FAP and tracer mass concentrations were also significantly improved. Fungal spore number concentrations on the order of 104 m−3, accounting for 2–5 % of TSP mass during dry periods and 17–23 % during rainy

  13. Pentamethinium fluorescent probes: The impact of molecular structure on photophysical properties and subcellular localization

    Czech Academy of Sciences Publication Activity Database

    Bříza, T.; Rimpelová, S.; Králová, Jarmila; Záruba, K.; Kejík, Z.; Ruml, T.; Martásek, P.; Král, V.

    2014-01-01

    Roč. 107, August 2014 (2014), s. 51-59 ISSN 0143-7208 R&D Projects: GA AV ČR KAN200100801; GA ČR(CZ) GAP303/11/1291; GA MŠk(CZ) CZ.01.05/2.1.00/01.00.30; GA MŠk(CZ) CZ.1.07/2.300/30.0060 Institutional support: RVO:68378050 Keywords : Pentamethinium salts * Fluorescent probes * Mitochondria * Cardiolipin Photostability * Organelle imaging Subject RIV: CE - Biochemistry Impact factor: 3.966, year: 2014

  14. Molecular fluorescence enhancement in plasmonic environments: exploring the role of nonlocal effects

    DEFF Research Database (Denmark)

    Tserkezis, Christos; Stefanou, Nikolaos; Wubs, Martijn

    2016-01-01

    in the nanoparticle vicinity. Here we explore the influence of hitherto disregarded nonclassical effects in the description of emitter-plasmon hybrids, focusing on the roles of metal nonlocal response and especially size-dependent plasmon damping. Through extensive modelling of metallic nanospheres and nanoshells...... a more complete physical description combining electron convection and diffusion, we show that not only are fluorescence rates dramatically reduced as compared to the predictions of the local description and the common hydrodynamic Drude model, but the optimum emitter-nanoparticle distance is also...

  15. Applications of nucleoside-based molecular probes for the in vivo assessment of tumour biochemistry using positron emission tomography (PET

    Directory of Open Access Journals (Sweden)

    Leonard I. Wiebe

    2007-05-01

    Full Text Available Positron emission tomography (PET is a non-invasive nuclear imaging technique. In PET, radiolabelled molecules decay by positron emission. The gamma rays resulting from positron annihilation are detected in coincidence and mapped to produce three dimensional images of radiotracer distribution in the body. Molecular imaging with PET refers to the use of positron-emitting biomolecules that are highly specific substrates for target enzymes, transport proteins or receptor proteins. Molecular imaging with PET produces spatial and temporal maps of the target-related processes. Molecular imaging is an important analytical tool in diagnostic medical imaging, therapy monitoring and the development of new drugs. Molecular imaging has its roots in molecular biology. Originally, molecular biology meant the biology of gene expression, but now molecular biology broadly encompasses the macromolecular biology and biochemistry of proteins, complex carbohydrates and nucleic acids. To date, molecular imaging has focused primarily on proteins, with emphasis on monoclonal antibodies and their derivative forms, small-molecule enzyme substrates and components of cell membranes, including transporters and transmembrane signalling elements. This overview provides an introduction to nucleosides, nucleotides and nucleic acids in the context of molecular imaging.A tomografia por emissão de pósitrons (TEP é uma técnica de imagem não invasiva da medicina nuclear. A TEP utiliza moléculas marcadas com emissores de radiação beta positiva (pósitrons. As radiações gama medidas que resultam do aniquilamento dos pósitrons são detectadas por um sistema de coincidência e mapeadas para produzir uma imagem tridimensional da distribuição do radiotraçador no corpo. A imagem molecular com TEP refere-se ao uso de biomoléculas marcadas com emissor de pósitron que são substratos altamente específicos para alvos como enzimas, proteínas transportadoras ou receptores prot

  16. Quantifying the Assembly of Multicomponent Molecular Machines by Single-Molecule Total Internal Reflection Fluorescence Microscopy.

    Science.gov (United States)

    Boehm, E M; Subramanyam, S; Ghoneim, M; Washington, M Todd; Spies, M

    2016-01-01

    Large, dynamic macromolecular complexes play essential roles in many cellular processes. Knowing how the components of these complexes associate with one another and undergo structural rearrangements is critical to understanding how they function. Single-molecule total internal reflection fluorescence (TIRF) microscopy is a powerful approach for addressing these fundamental issues. In this article, we first discuss single-molecule TIRF microscopes and strategies to immobilize and fluorescently label macromolecules. We then review the use of single-molecule TIRF microscopy to study the formation of binary macromolecular complexes using one-color imaging and inhibitors. We conclude with a discussion of the use of TIRF microscopy to examine the formation of higher-order (i.e., ternary) complexes using multicolor setups. The focus throughout this article is on experimental design, controls, data acquisition, and data analysis. We hope that single-molecule TIRF microscopy, which has largely been the province of specialists, will soon become as common in the tool box of biophysicists and biochemists as structural approaches have become today. © 2016 Elsevier Inc. All rights reserved.

  17. Chiral recognition of proteins having L-histidine residues on the surface with lanthanide ion complex incorporated-molecularly imprinted fluorescent nanoparticles

    International Nuclear Information System (INIS)

    Uzun, Lokman; Uzek, Recep; Şenel, Serap; Say, Ridvan; Denizli, Adil

    2013-01-01

    In this study, lanthanide ion complex incorporated molecularly imprinted fluorescent nanoparticles were synthesized. A combination of three novel approaches was applied for the purpose. First, lanthanide ions [Terbium(III)] were complexed with N-methacryloyl-L-histidine (MAH), polymerizable derivative of L-histidine amino acid, in order to incorporate the complex directly into the polymeric backbone. At the second stage, L-histidine molecules imprinted nanoparticles were utilized instead of whole protein imprinting in order to avoid whole drawbacks such as fragility, complexity, denaturation tendency, and conformation dependency. At the third stage following the first two steps mentioned above, imprinted L-histidine was coordinated with cupric ions [Cu(II)] to conduct the study under mild conditions. Then, molecularly imprinted fluorescent nanoparticles synthesized were used for L-histidine adsorption from aqueous solution to optimize conditions for adsorption and fluorimetric detection. Finally, usability of nanoparticles was investigated for chiral biorecognition using stereoisomer, D-histidine, racemic mixture, D,L-histidine, proteins with surface L-histidine residue, lysozyme, cytochrome C, or without ribonuclease A. The results revealed that the proposed polymerization strategy could make significant contribution to the solution of chronic problems of fluorescent component introduction into polymers. Additionally, the fluorescent nanoparticles reported here could be used for selective separation and fluorescent monitoring purposes. Highlights: • Lanthanide ion complex incorporated molecularly imprinted fluorescent nanoparticles • Direct incorporation of the fluorescent complex into polymeric backbone. • Imprinting by assistance of cupric ion coordination into nanoparticles • Evaluation of the chiral biorecognition ability of nanoparticles • Simultaneous selective separation and fluorescent monitoring

  18. Chiral recognition of proteins having L-histidine residues on the surface with lanthanide ion complex incorporated-molecularly imprinted fluorescent nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Uzun, Lokman, E-mail: lokman@hacettepe.edu.tr [Hacettepe University, Department of Chemistry, 06381, Ankara (Turkey); Uzek, Recep; Şenel, Serap [Hacettepe University, Department of Chemistry, 06381, Ankara (Turkey); Say, Ridvan [Anadolu University, Department of Chemistry, 26470, Eskisehir (Turkey); Denizli, Adil [Hacettepe University, Department of Chemistry, 06381, Ankara (Turkey)

    2013-08-01

    In this study, lanthanide ion complex incorporated molecularly imprinted fluorescent nanoparticles were synthesized. A combination of three novel approaches was applied for the purpose. First, lanthanide ions [Terbium(III)] were complexed with N-methacryloyl-L-histidine (MAH), polymerizable derivative of L-histidine amino acid, in order to incorporate the complex directly into the polymeric backbone. At the second stage, L-histidine molecules imprinted nanoparticles were utilized instead of whole protein imprinting in order to avoid whole drawbacks such as fragility, complexity, denaturation tendency, and conformation dependency. At the third stage following the first two steps mentioned above, imprinted L-histidine was coordinated with cupric ions [Cu(II)] to conduct the study under mild conditions. Then, molecularly imprinted fluorescent nanoparticles synthesized were used for L-histidine adsorption from aqueous solution to optimize conditions for adsorption and fluorimetric detection. Finally, usability of nanoparticles was investigated for chiral biorecognition using stereoisomer, D-histidine, racemic mixture, D,L-histidine, proteins with surface L-histidine residue, lysozyme, cytochrome C, or without ribonuclease A. The results revealed that the proposed polymerization strategy could make significant contribution to the solution of chronic problems of fluorescent component introduction into polymers. Additionally, the fluorescent nanoparticles reported here could be used for selective separation and fluorescent monitoring purposes. Highlights: • Lanthanide ion complex incorporated molecularly imprinted fluorescent nanoparticles • Direct incorporation of the fluorescent complex into polymeric backbone. • Imprinting by assistance of cupric ion coordination into nanoparticles • Evaluation of the chiral biorecognition ability of nanoparticles • Simultaneous selective separation and fluorescent monitoring.

  19. Efficient and Scalable Synthesis of 4-Carboxy-Pennsylvania Green Methyl Ester: A Hydrophobic Building Block for Fluorescent Molecular Probes.

    Science.gov (United States)

    Woydziak, Zachary R; Fu, Liqiang; Peterson, Blake R

    2014-01-01

    Fluorinated fluorophores are valuable tools for studies of biological systems. However, amine-reactive single-isomer derivatives of these compounds are often very expensive. To provide an inexpensive alternative, we report a practical synthesis of 4-carboxy-Pennsylvania Green methyl ester. Derivatives of this hydrophobic fluorinated fluorophore, a hybrid of the dyes Oregon Green and Tokyo Green, are often cell permeable, enabling labeling of intracellular targets and components. Moreover, the low pKa of Pennsylvania Green (4.8) confers bright fluorescence in acidic cellular compartments such as endosomes, enhancing its utility for chemical biology investigations. To improve access to the key intermediate 2,7-difluoro-3,6-dihydroxyxanthen-9-one, we subjected bis-(2,4,5-trifluorophenyl)methanone to iterative nucleophilic aromatic substitution by hydroxide on scales of > 40 g. This intermediate was used to prepare over 15 grams of pure 4-carboxy-Pennsylvania Green methyl ester in 28% overall yield without requiring chromatography. This compound can be converted into the amine reactive N -hydroxysuccinimidyl ester in essentially quantitative yield for the synthesis of a wide variety of fluorescent molecular probes.

  20. Evaluating the binding efficiency of pheromone binding protein with its natural ligand using molecular docking and fluorescence analysis

    Science.gov (United States)

    Ilayaraja, Renganathan; Rajkumar, Ramalingam; Rajesh, Durairaj; Muralidharan, Arumugam Ramachandran; Padmanabhan, Parasuraman; Archunan, Govindaraju

    2014-06-01

    Chemosignals play a crucial role in social and sexual communication among inter- and intra-species. Chemical cues are bound with protein that is present in the pheromones irrespective of sex are commonly called as pheromone binding protein (PBP). In rats, the pheromone compounds are bound with low molecular lipocalin protein α2u-globulin (α2u). We reported farnesol is a natural endogenous ligand (compound) present in rat preputial gland as a bound volatile compound. In the present study, an attempt has been made through computational method to evaluating the binding efficiency of α2u with the natural ligand (farnesol) and standard fluorescent molecule (2-naphthol). The docking analysis revealed that the binding energy of farnesol and 2-naphthol was almost equal and likely to share some binding pocket of protein. Further, to extrapolate the results generated through computational approach, the α2u protein was purified and subjected to fluorescence titration and binding assay. The results showed that the farnesol is replaced by 2-naphthol with high hydrophobicity of TYR120 in binding sites of α2u providing an acceptable dissociation constant indicating the binding efficiency of α2u. The obtained results are in corroboration with the data made through computational approach.

  1. Molecular basis of photochromism of a fluorescent protein revealed by direct 13C detection under laser illumination

    International Nuclear Information System (INIS)

    Mizuno, Hideaki; Mal, Tapas Kumar; Waelchli, Markus; Fukano, Takashi; Ikura, Mitsuhiko; Miyawaki, Atsushi

    2010-01-01

    Dronpa is a green fluorescent protein homologue with a photochromic property. A green laser illumination reversibly converts Dronpa from a green-emissive bright state to a non-emissive dark state, and ultraviolet illumination converts it to the bright state. We have employed solution NMR to understand the underlying molecular mechanism of the photochromism. The detail characterization of Dronpa is hindered as it is metastable in the dark state and spontaneously converts to the bright state. To circumvent this issue, we have designed in magnet laser illumination device. By combining the device with a 150-mW argon laser at 514.5 nm, we have successfully converted and maintained Dronpa in the dark state in the NMR tube by continuous illumination during the NMR experiments. We have employed direct-detection of 13 C nuclei from the carbon skeleton of the chromophore for detailed characterization of chromophore in both states of Dronpa by using the Bruker TCI cryoprobe. The results from NMR data have provided direct evidence of the double bond formation between C α and C β of Y63 in the chromophore, the β-barrel structure in solution, and the ionized and protonated state of Y63 hydroxyl group in the bright and dark states, respectively. These studies have also revealed that a part of β-barrel around the chromophore becomes polymorphic only in the dark state, which may be critical to make the fluorescence dim by increasing the contribution of non-emissive vibrational relaxation pathways.

  2. Semiconductor quantum dots as fluorescent probes for in vitro and in vivo bio-molecular and cellular imaging

    Directory of Open Access Journals (Sweden)

    Sarwat B. Rizvi

    2010-08-01

    Full Text Available Over the years, biological imaging has seen many advances, allowing scientists to unfold many of the mysteries surrounding biological processes. The ideal imaging resolution would be in nanometres, as most biological processes occur at this scale. Nanotechnology has made this possible with functionalised nanoparticles that can bind to specific targets and trace processes at the cellular and molecular level. Quantum dots (QDs or semiconductor nanocrystals are luminescent particles that have the potential to be the next generation fluorophores. This paper is an overview of the basics of QDs and their role as fluorescent probes for various biological imaging applications. Their potential clinical applications and the limitations that need to be overcome have also been discussed.

  3. Hypoxia-Targeting Fluorescent Nanobodies for Optical Molecular Imaging of Pre-Invasive Breast Cancer

    NARCIS (Netherlands)

    van Brussel, Aram S A; Adams, Arthur; Oliveira, Sabrina; Dorresteijn, Bram; El Khattabi, Mohamed; Vermeulen, J. F.; van der Wall, Elsken; Mali, Willem P Th M; Derksen, Patrick W B; van Diest, Paul J; van Bergen En Henegouwen, Paul M P

    PURPOSE: The aim of this work was to develop a CAIX-specific nanobody conjugated to IRDye800CW for molecular imaging of pre-invasive breast cancer. PROCEDURES: CAIX-specific nanobodies were selected using a modified phage display technology, conjugated site-specifically to IRDye800CW and evaluated

  4. Hypoxia-Targeting Fluorescent Nanobodies for Optical Molecular Imaging of Pre-Invasive Breast Cancer

    NARCIS (Netherlands)

    van Brussel, Aram S A; Adams, Arthur; Oliveira, Sabrina; Dorresteijn, Bram; El Khattabi, Mohamed; Vermeulen, Jeroen F.; van der Wall, Elsken; Mali, W.P.T.M.; Derksen, Patrick W B; van Diest, Paul J.; van Bergen En Henegouwen, Paul M P

    Purpose: The aim of this work was to develop a CAIX-specific nanobody conjugated to IRDye800CW for molecular imaging of pre-invasive breast cancer. Procedures: CAIX-specific nanobodies were selected using a modified phage display technology, conjugated site-specifically to IRDye800CW and evaluated

  5. Determination of low molecular weight thiols using monobromobimane fluorescent labeling and high-performance liquid chromatography

    Science.gov (United States)

    Fahey, Robert C.; Newton, Gerald L.

    1988-01-01

    Methods are described for the preparation and high-performance liquid chromatography (HPLC) analysis of monobromobimane derivatives of low molecular weight thiols in extracts of biological samples. Typical problems encountered in the development and application of these methods are discussed. Analysis of mung bean extract is used as an example.

  6. Magnetic resonance and fluorescence studies on pyruvate dehydrogenase complexes and their small molecular weight constituents

    NARCIS (Netherlands)

    Grande, H.J.

    1976-01-01

    The articles presented in this thesis do not describe at first glance one well-defined subject. They are, however, in fact connected by one central theme: the study of large enzyme aggregates by molecular physical methods. Chosen was the pyruvate dehydrogenase complex (PDC) because of its

  7. FIB and MIP: understanding nanoscale porosity in molecularly imprinted polymers via 3D FIB/SEM tomography.

    Science.gov (United States)

    Neusser, G; Eppler, S; Bowen, J; Allender, C J; Walther, P; Mizaikoff, B; Kranz, C

    2017-10-05

    We present combined focused ion beam/scanning electron beam (FIB/SEM) tomography as innovative method for differentiating and visualizing the distribution and connectivity of pores within molecularly imprinted polymers (MIPs) and non-imprinted control polymers (NIPs). FIB/SEM tomography is used in cell biology for elucidating three-dimensional structures such as organelles, but has not yet been extensively applied for visualizing the heterogeneity of nanoscopic pore networks, interconnectivity, and tortuosity in polymers. To our best knowledge, the present study is the first application of this strategy for analyzing the nanoscale porosity of MIPs. MIPs imprinted for propranolol - and the corresponding NIPs - were investigated establishing FIB/SEM tomography as a viable future strategy complementing conventional isotherm studies. For visualizing and understanding the properties of pore networks in detail, polymer particles were stained with osmium tetroxide (OsO 4 ) vapor, and embedded in epoxy resin. Staining with OsO 4 provides excellent contrast during high-resolution SEM imaging. After optimizing the threshold to discriminate between the stained polymer matrix, and pores filled with epoxy resin, a 3D model of the sampled volume may be established for deriving not only the pore volume and pore surface area, but also to visualize the interconnectivity and tortuosity of the pores within the sampled polymer volume. Detailed studies using different types of cross-linkers and the effect of hydrolysis on the resulting polymer properties have been investigated. In comparison of MIP and NIP, it could be unambiguously shown that the interconnectivity of the visualized pores in MIPs is significantly higher vs. the non-imprinted polymer, and that the pore volume and pore area is 34% and approx. 35% higher within the MIP matrix. This confirms that the templating process not only induces selective binding sites, but indeed also affects the physical properties of such

  8. Dimensional metrology of lab-on-a-chip internal structures: a comparison of optical coherence tomography with confocal fluorescence microscopy.

    Science.gov (United States)

    Reyes, D R; Halter, M; Hwang, J

    2015-07-01

    The characterization of internal structures in a polymeric microfluidic device, especially of a final product, will require a different set of optical metrology tools than those traditionally used for microelectronic devices. We demonstrate that optical coherence tomography (OCT) imaging is a promising technique to characterize the internal structures of poly(methyl methacrylate) devices where the subsurface structures often cannot be imaged by conventional wide field optical microscopy. The structural details of channels in the devices were imaged with OCT and analyzed with an in-house written ImageJ macro in an effort to identify the structural details of the channel. The dimensional values obtained with OCT were compared with laser-scanning confocal microscopy images of channels filled with a fluorophore solution. Attempts were also made using confocal reflectance and interferometry microscopy to measure the channel dimensions, but artefacts present in the images precluded quantitative analysis. OCT provided the most accurate estimates for the channel height based on an analysis of optical micrographs obtained after destructively slicing the channel with a microtome. OCT may be a promising technique for the future of three-dimensional metrology of critical internal structures in lab-on-a-chip devices because scans can be performed rapidly and noninvasively prior to their use. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  9. New developments in molecular imaging: positron emission tomography time-of-flight (TOF-PET)

    International Nuclear Information System (INIS)

    Aguilar, P.; Couce, B.; Iglesias, A.; Lois, C.

    2011-01-01

    Positron Emission tomography (PET) in increasingly being used in oncology for the diagnosis and staging of disease, as well as in monitoring response to therapy. One of the last advances in PET is the incorporation of Time-of-Flight (TOF) information, which improves the tomographic reconstruction process and subsequently the quality of the final image. In this work, we explain the principles of PET and the fundamentals of TOF-PET. Clinical images are shown in order to illustrate how TOF-PET improves the detectability of small lesions, particularly in patients with high body mass index. (Author) 20 refs

  10. Molecular Imaging and Precision Medicine: PET/Computed Tomography and Therapy Response Assessment in Oncology.

    Science.gov (United States)

    Sheikhbahaei, Sara; Mena, Esther; Pattanayak, Puskar; Taghipour, Mehdi; Solnes, Lilja B; Subramaniam, Rathan M

    2017-01-01

    A variety of methods have been developed to assess tumor response to therapy. Standardized qualitative criteria based on 18F-fluoro-deoxyglucose PET/computed tomography have been proposed to evaluate the treatment effectiveness in specific cancers and these allow more accurate therapy response assessment and survival prognostication. Multiple studies have addressed the utility of the volumetric PET biomarkers as prognostic indicators but there is no consensus about the preferred segmentation methodology for these metrics. Heterogeneous intratumoral uptake was proposed as a novel PET metric for therapy response assessment. PET imaging techniques will be used to study the biological behavior of cancers during therapy. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Controllable molecular aggregation and fluorescence properties of 1,3,4-oxadiazole derivative

    KAUST Repository

    Li, Min

    2015-10-14

    The molecular self-assembly behaviour of 2,2’-Bis-(4-hexyloxyphenyl)-bi-1,3,4-oxadiazole (BOXD-6) in solution, on surfaces and in bulk crystals, and its photo-physical properties were studied via a combination of experimental techniques and theoretical calculations. It is found that BOXD-6 molecules self-assemble into both H- and J-aggregates at moderate concentration (~10-4 M) and then transit to exclusive J-aggregates at higher concentration (~10-3 M) in tetrahydrofuran. In H-aggregation (α polymorph), BOXD-6 adopts a linear conformation and forms a one- dimensional layered structure; in J-aggregation (β polymorph), it adopts a Z-shaped conformation and form a more ordered two-dimensional layered structure. A π-stacking structure is observed in both cases, and adjacent molecules in the J-aggregation show larger displacement along the molecular long axis direction than that in H-aggregation. Although J-aggregates are almost the only component in concentrated solutions (10-3 M), both H- and J-aggregates can be obtained if concentrated solution is transformed onto substrates through a simple drop-casting method. Such a phase transition during film formation can be easily avoided by adding water as precipitator; a film with pure J-aggregates is then obtained. In order to get more information on molecular self-assembly, intermolecular interaction potential energy surfaces (PES) were evaluated via theoretical calculations at the DFT level (M062x/6-31G**). The PES not only confirm the molecular stacking structures found in crystals but also predict some other likely structures, which will be the target of future experiments.

  12. A novel water-soluble 1,8-naphthalimide as a fluorescent pH-probe and a molecular logic circuit

    International Nuclear Information System (INIS)

    Georgiev, Nikolai I.; Dimitrova, Margarita D.; Krasteva, Paoleta V.; Bojinov, Vladimir B.

    2017-01-01

    A novel highly water-soluble fluorescence sensing 1,8-naphthalimide is synthesized and investigated. The novel compound is designed on the “fluorophore-receptor 1 -spacer-receptor 2 ” model as a molecular fluorescence probe for determination of ions in 100% aqueous media. The novel probe comprising hydrazide and N-methylpiperazine substituents is capable of operating simultaneously via ICT and PET signaling mechanism and of recognizing selectively protons and hydroxyl anions over the representative metal ions and anions. Due to the remarkable fluorescence changes as a function of pH the system is able to act as a three output combinatorial logic circuit with two chemical inputs. Two INHIBIT gates in fluorescence and absorption mode as well as an IMPLICATION logic gate are obtained. Because of the parallel action of both INHIBIT gates a magnitude digital comparator is achieved for the first time in this way.

  13. Synchrotron-based X-ray Fluorescence Microscopy in Conjunction with Nanoindentation to Study Molecular-Scale Interactions of Phenol–Formaldehyde in Wood Cell Walls

    Science.gov (United States)

    Joseph E. Jakes; Christopher G. Hunt; Daniel J. Yelle; Linda Lorenz; Kolby Hirth; Sophie-Charlotte Gleber; Stefan Vogt; Warren Grigsby; Charles R. Frihart

    2015-01-01

    Understanding and controlling molecular-scale interactions between adhesives and wood polymers are critical to accelerate the development of improved adhesives for advanced wood-based materials. The submicrometer resolution of synchrotron-based X-ray fluorescence microscopy (XFM) was found capable of mapping and quantifying infiltration of Br-labeled phenol−...

  14. NIR-Cyanine Dye Linker: a Promising Candidate for Isochronic Fluorescence Imaging in Molecular Cancer Diagnostics and Therapy Monitoring.

    Science.gov (United States)

    Komljenovic, Dorde; Wiessler, Manfred; Waldeck, Waldemar; Ehemann, Volker; Pipkorn, Ruediger; Schrenk, Hans-Hermann; Debus, Jürgen; Braun, Klaus

    2016-01-01

    Personalized anti-cancer medicine is boosted by the recent development of molecular diagnostics and molecularly targeted drugs requiring rapid and efficient ligation routes. Here, we present a novel approach to synthetize a conjugate able to act simultaneously as an imaging and as a chemotherapeutic agent by coupling functional peptides employing solid phase peptide synthesis technologies. Development and the first synthesis of a fluorescent dye with similarity in the polymethine part of the Cy7 molecule whose indolenine-N residues were substituted with a propylene linker are described. Methylating agent temozolomide is functionalized with a tetrazine as a diene component whereas Cy7-cell penetrating peptide conjugate acts as a dienophilic reaction partner for the inverse Diels-Alder click chemistry-mediated ligation route yielding a theranostic conjugate, 3-mercapto-propionic-cyclohexenyl-Cy7-bis-temozolomide-bromide-cell penetrating peptide. Synthesis route described here may facilitate targeted delivery of the therapeutic compound to achieve sufficient local concentrations at the target site or tissue. Its versatility allows a choice of adequate imaging tags applicable in e.g. PET, SPECT, CT, near-infrared imaging, and therapeutic substances including cytotoxic agents. Imaging tags and therapeutics may be simultaneously bound to the conjugate applying click chemistry. Theranostic compound presented here offers a solid basis for a further improvement of cancer management in a precise, patient-specific manner.

  15. Multimodality optical coherence tomography and fluorescence confocal scanning laser ophthalmoscopy in a zebrafish model of retinal vascular occlusion and remodeling

    Science.gov (United States)

    Li, Xiaoyue; Spitz, Kathleen; Bozic, Ivan; Tao, Yuankai K.

    2018-02-01

    Neovascularization in diabetic retinopathy (DR) and age-related macular degeneration (AMD) result in severe vision-loss and are two of the leading causes of blindness. The structural, metabolic, and vascular changes underlying retinal neovascularization are unknown and, thus, there is an unmet need to identify mechanisms of pathogenesis and novel anti-angiogenic therapies. Zebrafish is a robust ophthalmological model because its retina has comparable structure to the human retina and its fecundity and life-cycle enable development of mutant phenotypes of human pathologies. Here, we perform multimodal imaging with OCT and fluorescence confocal scanning laser ophthalmoscopy (cSLO) to identify changes in retinal structure and function in a zebrafish model of vascular leakage. Transgenic zebrafish with EGFP tagged plasma protein were imaged longitudinally at six time points over two weeks to visualize vascular perfusion changes from diethylaminobenzaldehyde (DEAB) treatment. Complementary contrast from OCT-A perfusion maps and cSLO imaging of plasma protein EGFP shows vascular occlusions posttreatment. cSLO images confirm presence of vessels despite loss of OCT-A signal. Plasma protein EGFP contrast also shows significant changes in vessel structure as compared to baseline images. OCT structural volumes show empty vessel cross-sections confirming non-perfusion. In addition, we present algorithms for automated biometric identification of OCT datasets using OCT-A vascular patterns in the presence of significant vascular perfusion changes. These results establish a framework for large-scale in vivo assays to identify novel anti-angiogenic compounds and understand the mechanisms ofneovascularization associated with retinal ocular pathologies.

  16. Illuminating the origins of spectral properties of green fluorescent proteins via proteochemometric and molecular modeling.

    Science.gov (United States)

    Nantasenamat, Chanin; Simeon, Saw; Owasirikul, Wiwat; Songtawee, Napat; Lapins, Maris; Prachayasittikul, Virapong; Wikberg, Jarl E S

    2014-10-15

    Green fluorescent protein (GFP) has immense utility in biomedical imaging owing to its autofluorescent nature. In efforts to broaden the spectral diversity of GFP, there have been several reports of engineered mutants via rational design and random mutagenesis. Understanding the origins of spectral properties of GFP could be achieved by means of investigating its structure-activity relationship. The first quantitative structure-property relationship study for modeling the spectral properties, particularly the excitation and emission maximas, of GFP was previously proposed by us some years ago in which quantum chemical descriptors were used for model development. However, such simplified model does not consider possible effects that neighboring amino acids have on the conjugated π-system of GFP chromophore. This study describes the development of a unified proteochemometric model in which the GFP chromophore and amino acids in its vicinity are both considered in the same model. The predictive performance of the model was verified by internal and external validation as well as Y-scrambling. Our strategy provides a general solution for elucidating the contribution that specific ligand and protein descriptors have on the investigated spectral property, which may be useful in engineering novel GFP variants with desired characteristics. Copyright © 2014 Wiley Periodicals, Inc.

  17. Excitation and deexcitation of N2 molecular levels. Induced fluorescence by electrons and laser

    International Nuclear Information System (INIS)

    Perez Fernandez-Mayoralas, A.

    1989-01-01

    The electron impact excitation followed by fluorescence induced by N 2 -laser absorption was used to study the lifetime of the lowest vibrational level of the B 3 π g electronic state of N 2 . The experimental result of this work is 13 + 1 μs. To measure the lifetime of B 3 π g (v=2,3,5,6,7,8) levels the delayed coincidence method by electron impact was use. The lifetime values were compared with recent experimental and theoretical results. The relative intensi-ties of 3 π g --- A 3 Σ Ω + system bands, in the range (6540-10500 A o ) was measured using a hollow cathode lamp as spectral source. The relative transition moments and its dependence versus the r-centroid was obtained. Total cross sections for electron scattering by N molecules in the range 600 - 5000 eV have been obtained from measurements of the attenuation of a linear electron beam. The results have been compared with available experimental cross sections and with theoretical calculations based on the first Born approximation. (Author)

  18. In vivo photoacoustic tumor tomography using a quinoline-annulated porphyrin as NIR molecular contrast agent.

    Science.gov (United States)

    Luciano, Michael; Erfanzadeh, Mohsen; Zhou, Feifei; Zhu, Hua; Bornhütter, Tobias; Röder, Beate; Zhu, Quing; Brückner, Christian

    2017-01-25

    The synthesis and photophysical properties of a tetra-PEG-modified and freely water-soluble quinoline-annulated porphyrin are described. We previously demonstrated the ability of quinoline-annulated porphyrins to act as an in vitro NIR photoacoustic imaging (PAI) contrast agent. The solubility of the quinoline-annulated porphyrin derivative in serum now allowed the assessment of the efficacy of the PEGylated derivative as an in vivo NIR contrast agent for the PAI of an implanted tumor in a mouse model. A multi-fold contrast enhancement when compared to the benchmark dye ICG could be shown, a finding that could be traced to its photophysical properties (short triplet lifetimes, low fluorescence and singlet oxygen sensitization quantum yields). A NIR excitation wavelength of 790 nm could be used, fully taking advantage of the optical window of tissue. Rapid renal clearance of the dye was observed. Its straight-forward synthesis, optical properties with the possibility for further optical fine-tuning, nontoxicity, favorable elimination rates, and contrast enhancement make this a promising PAI contrast agent. The ability to conjugate the PAI chromophore with a fluorescent tag using a facile and general conjugation strategy was also demonstrated.

  19. Evaluation of optical coherence tomography in the diagnosis of age related macula degeneration compared with fluorescence angiography

    Directory of Open Access Journals (Sweden)

    Fröschl, Barbara

    2007-02-01

    Full Text Available Background: In industrialised nations age-related macular degeneration (AMD is the most common cause of blindness and severe visual impairment. AMD is a disease of the retina characterized by the accumulation of metabolic products in the macula. In early stages drusen and pigment disorders occur, in late stages a dry form is distinguished from the exsudative form with choroidal neovascularisation. AMD causes vision disorders such as blurred vision of the central part of the visual field, leading finally to a dark spot. Several therapies are available for the exsudative form, however an exact diagnosis is partially essential. The gold standard for the diagnosis of AMD is fluorescein angiography (FA, an invasive investigation with intravenous application of a dye. Optical coherence tomography (OCT is a more recent non-invasive procedure. Objectives: The aim of this HTA report is to investigate the efficacy and efficiency of OCT compared to FA. Ethical, societal and legal aspects are also considered. Methods: A systematic literature search was performed in 34 international databases which yielded 2324 articles. Eight publications were included for assessment, according to predefined selection criteria. Results: The number of studies investigating OCT compared to FA in patients with AMD is presently very limited and the quality of the studies is generally low. The number of investigated patients is below 35 in four publications and in only one publication it is above 100. Moreover in most of the articles very selected patient groups are studied. Economic studies concerning the efficiency of OCT compared to FA cannot be identified. DiscussionEven though the patient groups investigated and the objectives of the studies are very heterogenous, all publications uniformly show that OCT cannot replace FA. However, OCT yields additional diagnostic findings and may verify unclear findings of FA. Therefore the application of OCT in addition to FA is useful in

  20. Synaptic molecular imaging in spared and deprived columns of mouse barrel cortex with array tomography.

    Science.gov (United States)

    Weiler, Nicholas C; Collman, Forrest; Vogelstein, Joshua T; Burns, Randal; Smith, Stephen J

    2014-01-01

    A major question in neuroscience is how diverse subsets of synaptic connections in neural circuits are affected by experience dependent plasticity to form the basis for behavioral learning and memory. Differences in protein expression patterns at individual synapses could constitute a key to understanding both synaptic diversity and the effects of plasticity at different synapse populations. Our approach to this question leverages the immunohistochemical multiplexing capability of array tomography (ATomo) and the columnar organization of mouse barrel cortex to create a dataset comprising high resolution volumetric images of spared and deprived cortical whisker barrels stained for over a dozen synaptic molecules each. These dataset has been made available through the Open Connectome Project for interactive online viewing, and may also be downloaded for offline analysis using web, Matlab, and other interfaces.

  1. From fast fluorescence imaging to molecular diffusion law on live cell membranes in a commercial microscope.

    Science.gov (United States)

    Di Rienzo, Carmine; Gratton, Enrico; Beltram, Fabio; Cardarelli, Francesco

    2014-10-09

    It has become increasingly evident that the spatial distribution and the motion of membrane components like lipids and proteins are key factors in the regulation of many cellular functions. However, due to the fast dynamics and the tiny structures involved, a very high spatio-temporal resolution is required to catch the real behavior of molecules. Here we present the experimental protocol for studying the dynamics of fluorescently-labeled plasma-membrane proteins and lipids in live cells with high spatiotemporal resolution. Notably, this approach doesn't need to track each molecule, but it calculates population behavior using all molecules in a given region of the membrane. The starting point is a fast imaging of a given region on the membrane. Afterwards, a complete spatio-temporal autocorrelation function is calculated correlating acquired images at increasing time delays, for example each 2, 3, n repetitions. It is possible to demonstrate that the width of the peak of the spatial autocorrelation function increases at increasing time delay as a function of particle movement due to diffusion. Therefore, fitting of the series of autocorrelation functions enables to extract the actual protein mean square displacement from imaging (iMSD), here presented in the form of apparent diffusivity vs average displacement. This yields a quantitative view of the average dynamics of single molecules with nanometer accuracy. By using a GFP-tagged variant of the Transferrin Receptor (TfR) and an ATTO488 labeled 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine (PPE) it is possible to observe the spatiotemporal regulation of protein and lipid diffusion on µm-sized membrane regions in the micro-to-milli-second time range.

  2. Molecular Imaging of Cancer Using X-ray Computed Tomography with Protease Targeted Iodinated Activity-Based Probes.

    Science.gov (United States)

    Gaikwad, Hanmant K; Tsvirkun, Darya; Ben-Nun, Yael; Merquiol, Emmanuelle; Popovtzer, Rachela; Blum, Galia

    2018-03-14

    X-ray computed tomography (CT) is a robust, precise, fast, and reliable imaging method that enables excellent spatial resolution and quantification of contrast agents throughout the body. However, CT is largely inadequate for molecular imaging applications due mainly to its low contrast sensitivity that forces the use of large concentrations of contrast agents for detection. To overcome this limitation, we generated a new class of iodinated nanoscale activity-based probes (IN-ABPs) that sufficiently accumulates at the target site by covalently binding cysteine cathepsins that are exceptionally highly expressed in cancer. The IN-ABPs are comprised of a short targeting peptide selective to specific cathepsins, an electrophilic moiety that allows activity-dependent covalent binding, and tags containing dendrimers with up to 48 iodine atoms. IN-ABPs selectively bind and inhibit activity of recombinant and intracellular cathepsin B, L, and S. We compared the in vivo kinetics, biodistribution, and tumor accumulation of IN-ABPs bearing 18 and 48 iodine atoms each, and their control counterparts lacking the targeting moiety. Here we show that although both IN-ABPs bind specifically to cathepsins within the tumor and produce detectable CT contrast, the 48-iodine bearing IN-ABP was found to be optimal with signals over 2.1-fold higher than its nontargeted counterpart. In conclusion, this study shows the synthetic feasibility and potential utility of IN-ABPs as potent contrast agents that enable molecular imaging of tumors using CT.

  3. Chiral recognition of proteins having L-histidine residues on the surface with lanthanide ion complex incorporated-molecularly imprinted fluorescent nanoparticles.

    Science.gov (United States)

    Uzun, Lokman; Uzek, Recep; Senel, Serap; Say, Ridvan; Denizli, Adil

    2013-08-01

    In this study, lanthanide ion complex incorporated molecularly imprinted fluorescent nanoparticles were synthesized. A combination of three novel approaches was applied for the purpose. First, lanthanide ions [Terbium(III)] were complexed with N-methacryloyl-L-histidine (MAH), polymerizable derivative of L-histidine amino acid, in order to incorporate the complex directly into the polymeric backbone. At the second stage, L-histidine molecules imprinted nanoparticles were utilized instead of whole protein imprinting in order to avoid whole drawbacks such as fragility, complexity, denaturation tendency, and conformation dependency. At the third stage following the first two steps mentioned above, imprinted L-histidine was coordinated with cupric ions [Cu(II)] to conduct the study under mild conditions. Then, molecularly imprinted fluorescent nanoparticles synthesized were used for L-histidine adsorption from aqueous solution to optimize conditions for adsorption and fluorimetric detection. Finally, usability of nanoparticles was investigated for chiral biorecognition using stereoisomer, D-histidine, racemic mixture, D,L-histidine, proteins with surface L-histidine residue, lysozyme, cytochrome C, or without ribonuclease A. The results revealed that the proposed polymerization strategy could make significant contribution to the solution of chronic problems of fluorescent component introduction into polymers. Additionally, the fluorescent nanoparticles reported here could be used for selective separation and fluorescent monitoring purposes. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Design considerations for highly effective fluorescence excitation and detection optical systems for molecular diagnostics

    Science.gov (United States)

    Kasper, Axel; Van Hille, Herbert; Kuk, Sola

    2018-02-01

    Modern instruments for molecular diagnostics are continuously optimized for diagnostic accuracy, versatility and throughput. The latest progress in LED technology together with tailored optics solutions allows developing highly efficient photonics engines perfectly adapted to the sample under test. Super-bright chip-on-board LED light sources are a key component for such instruments providing maximum luminous intensities in a multitude of narrow spectral bands. In particular the combination of white LEDs with other narrow band LEDs allows achieving optimum efficiency outperforming traditional Xenon light sources in terms of energy consumption, heat dissipation in the system, and switching time between spectral channels. Maximum sensitivity of the diagnostic system can only be achieved with an optimized optics system for the illumination and imaging of the sample. The illumination beam path must be designed for optimum homogeneity across the field while precisely limiting the angular distribution of the excitation light. This is a necessity for avoiding spill-over to the detection beam path and guaranteeing the efficiency of the spectral filtering. The imaging optics must combine high spatial resolution, high light collection efficiency and optimized suppression of excitation light for good signal-to-noise ratio. In order to achieve minimum cross-talk between individual wells in the sample, the optics design must also consider the generation of stray light and the formation of ghost images. We discuss what parameters and limitations have to be considered in an integrated system design approach covering the full path from the light source to the detector.

  5. A novel clinically translatable fluorescent nanoparticle for targeted molecular imaging of tumors in living subjects.

    Science.gov (United States)

    Gao, Jinhao; Chen, Kai; Luong, Richard; Bouley, Donna M; Mao, Hua; Qiao, Tiecheng; Gambhir, Sanjiv S; Cheng, Zhen

    2012-01-11

    The use of quantum dots (QDs) in biomedical research has grown tremendously, yet successful examples of clinical applications are absent due to many clinical concerns. Here, we report on a new type of stable and biocompatible dendron-coated InP/ZnS core/shell QD as a clinically translatable nanoprobe for molecular imaging applications. The QDs (QD710-Dendron) were demonstrated to hold several significant features: near-infrared (NIR) emission, high stability in biological media, suitable size with possible renal clearance, and ability of extravasation. More importantly, a pilot mouse toxicity study confirmed that QD710-Dendron lacks significant toxicity at the doses tested. The acute tumor uptake of QD710-Dendron resulted in good contrast from the surrounding nontumorous tissues, indicating the possibility of passive targeting of the QDs. The highly specific targeting of QD710-Dendron-RGD(2) to integrin α(v)β(3)-positive tumor cells resulted in high tumor uptake and long retention of the nanoprobe at tumor sites. In summary, QD710-Dendron and RGD-modified nanoparticles demonstrate small size, high stability, biocompatibility, favorable in vivo pharmacokinetics, and successful tumor imaging properties. These features satisfy the requirements for clinical translation and should promote efforts to further investigate the possibility of using QD710-Dendron-based nanoprobes in the clinical setting in the near future. © 2011 American Chemical Society

  6. Molecular architecture of axonemal microtubule doublets revealedby cryo-electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Sui, Haixin; Downing, Kenneth H.

    2006-05-22

    The axoneme, which forms the core of eukaryotic flagella and cilia, is one of the largest macromolecular machines with a structure that is largely conserved from protists to mammals. Microtubule doublets are structural components of axonemes containing a number of proteins besides tubulin, and are usually found in arrays of nine doublets arranged around two singlet microtubules. Coordinated sliding of adjacent doublets, which involves a host of other proteins in the axoneme, produces periodic beating movements of the axoneme. We have obtained a 3D density map of intact microtubule doublets using cryo-electron tomography and image averaging. Our map, with a resolution of about 3 nm, provides insights into locations of particular proteins within the doublets and the structural features of the doublets that define their mechanical properties. We identify likely candidates for several of these non-tubulin components of the doublets. This work offers novel insight on how tubulin protofilaments and accessory proteins attach together to form the doublets and provides a structural basis for understanding doublet function in axonemes.

  7. Positron emission tomography molecular imaging of dopaminergic system in drug addiction.

    Science.gov (United States)

    Hou, Haifeng; Tian, Mei; Zhang, Hong

    2012-05-01

    Dopamine (DA) is involved in drug reinforcement, but its role in drug addiction remains unclear. Positron emission tomography (PET) is the first technology used for the direct measurement of components of the dopaminergic system in the living human brain. In this article, we reviewed the major findings of PET imaging studies on the involvement of DA in drug addiction, especially in heroin addiction. Furthermore, we summarized PET radiotracers that have been used to study the role of DA in drug addiction. To investigate presynaptic function in drug addiction, PET tracers have been developed to measure DA synthesis and transport. For the investigation of postsynaptic function, several radioligands targeting dopamine one (D1) receptor and dopamine two (D2) receptor are extensively used in PET imaging studies. Moreover, we also summarized the PET imaging findings of heroin addiction studies, including heroin-induced DA increases and the reinforcement, role of DA in the long-term effects of heroin abuse, DA and vulnerability to heroin abuse and the treatment implications. PET imaging studies have corroborated the role of DA in drug addiction and increase our understanding the mechanism of drug addiction. Copyright © 2012 Wiley Periodicals, Inc.

  8. ICT-Isomerization-Induced Turn-On Fluorescence Probe with a Large Emission Shift for Mercury Ion: Application in Combinational Molecular Logic.

    Science.gov (United States)

    Bhatta, Sushil Ranjan; Mondal, Bijan; Vijaykumar, Gonela; Thakur, Arunabha

    2017-10-02

    A unique turn-on fluorescent device based on a ferrocene-aminonaphtholate derivative specific for Hg 2+ cation was developed. Upon binding with Hg 2+ ion, the probe shows a dramatic fluorescence enhancement (the fluorescence quantum yield increases 58-fold) along with a large red shift of 68 nm in the emission spectrum. The fluorescence enhancement with a red shift may be ascribed to the combinational effect of C═N isomerization and an extended intramolecular charge transfer (ICT) mechanism. The response was instantaneous with a detection limit of 2.7 × 10 -9 M. Upon Hg 2+ recognition, the ferrocene/ferrocenium redox peak was anodically shifted by ΔE 1/2 = 72 mV along with a "naked eye" color change from faint yellow to pale orange for this metal cation. Further, upon protonation of the imine nitrogen, the present probe displays a high fluorescence output due to suppression of the C═N isomerization process. Upon deprotonation using strong base, the fluorescence steadily decreases, which indicates that H + and OH - can be used to regulate the off-on-off fluorescence switching of the present probe. Density functional theory studies revealed that the addition of acid leads to protonation of the imine N (according to natural bond orbital analysis), and the resulting iminium proton forms a strong H-bond (2.307 Å) with one of the triazole N atoms to form a five-membered ring, which makes the molecule rigid; hence, enhancement of the ICT process takes place, thereby leading to a fluorescence enhancement with a red shift. The unprecedented combination of H + , OH - , and Hg 2+ ions has been used to generate a molecular system exhibiting the INHIBIT-OR combinational logic operation.

  9. Optical molecular fluorescence determination of ultra-trace beryllium in occupational and environmental samples using highly alkaline conditions.

    Science.gov (United States)

    Adams, Lori; Agrawal, Anoop; Cronin, John P; Ashley, Kevin

    2017-01-01

    Exposures to beryllium (Be), even at extremely low levels, can cause severe health effects in a percentage of those exposed; consequently, occupational exposure limits (OELs) promulgated for this element are the lowest established for any element. This work describes the advantages of using highly alkaline dye solutions for determination of Be in occupational hygiene and environmental samples by means of an optical molecular fluorescence technique after sample extraction in 1-3% (w˖w -1 ) aqueous ammonium bifluoride (NH 4 HF 2 ). Improved attributes include the ability to further enhance the detection limits of Be in extraction solutions of high acidity with minimal dilution, which is particularly beneficial when NH 4 HF 2 solutions of higher concentration are used for extraction of Be from soil samples. Significant improvements in Be method detection limits (MDLs) are obtained at levels many-fold below those reported previously for this methodology. Notably, MDLs for Be of health organizations and regulatory agencies in the USA and internationally. Applications of enhanced Be measurements to air filter samples, surface wipe samples, soils and newly-designed occupational air sampler inserts are illustrated.

  10. Fluorescent silver nanoclusters capped by polyethyleneimine with different molecular weights: Universal synthesis and application as a temperature sensor

    Energy Technology Data Exchange (ETDEWEB)

    Qu, Fei, E-mail: qufei3323@163.com [The Key Laboratory of Life-Organic Analysis, Qufu Normal University, Qufu 273165, Shandong (China); Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, Qufu Normal University, Qufu 273165, Shandong (China); Li, Qingjin [The Key Laboratory of Life-Organic Analysis, Qufu Normal University, Qufu 273165, Shandong (China); Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, Qufu Normal University, Qufu 273165, Shandong (China); You, Jinmao, E-mail: jmyou6304@163.com [The Key Laboratory of Life-Organic Analysis, Qufu Normal University, Qufu 273165, Shandong (China); Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, Qufu Normal University, Qufu 273165, Shandong (China); Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001 (China)

    2016-09-15

    In this paper, we developed a universal, applicable and simple synthetic method of Ag nanoclusters capped by polyethyleneimine (PEI) with different molecular weights (AgNC-PEIs), including Mw 600, 1300, 1800, 2000, 10,000, 25,000, 70,000, and 750,000. Using formaldehyde as the sole reducing agent, silver nanoclusters could be successfully prepared by using these templates. Subsequently, several characterization techniques were employed to investigate the properties of AgNC-PEIs, and the results suggested that these AgNC-PEIs had similar sizes, structures, and optical features. However, besides the common characteristics, different temperature sensitivities were found for these nanoclusters, in which AgNC-PEI 25000 was proper to be applied as a temperature sensor. With increasing temperature, the fluorescence quenched dramatically, and this change could be readily observed by naked eyes under UV light. Injection of these temperature sensitive nanoclusters into a glass tube, a simple thermometer could be fabricated easily, thus AgNC-PEI 25000 would be a promising candidate for temperature sensing as a visible indicator.

  11. Rapid molecular cytogenetic analysis of X-chromosomal microdeletions: Fluorescence in situ hybridization (FISH) for complex glycerol kinase deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Worley, K.C.; Lindsay, E.A.; McCabe, E.R.B. [Baylor College of Medicine, Houston, TX (United States)] [and others

    1995-07-17

    Diagnosis of X-chromosomal microdeletions has relied upon the traditional methods of Southern blotting and DNA amplification, with carrier identification requiring time-consuming and unreliable dosage calculations. In this report, we describe rapid molecular cytogenetic identification of deleted DNA in affected males with the Xp21 contiguous gene syndrome (complex glycerol kinase deficiency, CGKD) and female carriers for this disorder. CGKD deletions involve the genes for glycerol kinase, Duchenne muscular dystrophy, and/or adrenal hypoplasia congenita. We report an improved method for diagnosis of deletions in individuals with CGKD and for identification of female carriers within their families using fluorescence in situ hybridization (FISH) with a cosmid marker (cosmid 35) within the glycerol kinase gene. When used in combination with an Xq control probe, affected males demonstrate a single signal from the control probe, while female carriers demonstrate a normal chromosome with two signals, as well as a deleted chromosome with a single signal from the control probe. FISH analysis for CGKD provides the advantages of speed and accuracy for evaluation of submicroscopic X-chromosome deletions, particularly in identification of female carriers. In addition to improving carrier evaluation, FISH will make prenatal diagnosis of CGKD more readily available. 17 refs., 2 figs.

  12. Carbon-11 and fluorine-18 chemistry devoted to molecular probes for imaging the brain with positron emission tomography.

    Science.gov (United States)

    Dollé, Frédéric

    2013-01-01

    Exploration of the living human brain in real-time and in a noninvasive way was for centuries only a dream, made, however, possible today with the remarkable development during the four last decades of powerful molecular imaging techniques, and especially positron emission tomography (PET). Molecular PET imaging relies, from a chemical point of view, on the use and preparation of a positron-emitting radiolabelled probe or radiotracer, notably compounds incorporating one of two short-lived radionuclides fluorine-18 (T1/2 : 109.8 min) and carbon-11 (T1/2 : 20.38 min). The growing availability and interest for the radiohalogen fluorine-18 in radiopharmaceutical chemistry undoubtedly results from its convenient half-life and the successful use in clinical oncology of 2-[(18) F]fluoro-2-deoxy-d-glucose ([(18) F]FDG). The special interest of carbon-11 is not only that carbon is present in virtually all biomolecules and drugs allowing therefore for isotopic labelling of their chemical structures but also that a given molecule could be radiolabelled at different functions or sites, permitting to explore (or to take advantage of) in vivo metabolic pathways. PET chemistry includes production of these short-lived radioactive isotopes via nuclear transmutation reactions using a cyclotron, and is directed towards the development of rapid synthetic methods, at the trace level, for the introduction of these nuclides into a molecule, as well as the use of fast purification, analysis and formulation techniques. PET chemistry is the driving force in molecular PET imaging, and this special issue of the Journal of Labelled Compounds and Radiopharmaceuticals, which is strongly chemistry and radiochemistry-oriented, aims at illustrating, be it in part only, the state-of-the-art arsenal of reactions currently available and its potential for the research and development of specific molecular probes labelled with the positron emitters carbon-11 and fluorine-18, with optimal imaging

  13. A novel cyanide-selective colorimetric and fluorescent chemosensor: First molecular security keypad lock based on phosphotungstic acid and CN{sup −} inputs

    Energy Technology Data Exchange (ETDEWEB)

    Tavallali, Hossein, E-mail: Tavallali@pnu.ac.ir; Deilamy-Rad, Gohar; Parhami, Abolfath; Hasanli, Nahid

    2014-02-15

    Highlights: • Our probe is commercially available with good photo stability and high quantum yield. • Both color and fluorescence change with long emission wavelength in aqueous media. • Characteristics of an ON–OFF–ON fluorescence switch. • The simple receptor for CN{sup -} detection with low detection limit (≪WHO). • Mimic the function of a security keypad lock on sequential addition of PTA and CN{sup −}. -- Abstract: Rhodamine B (Rh{sub B}) an available dye has been developed as novel and efficient colorimetric and fluorometric chemosensor for cyanide ions in an absolutely aqueous media. The UV–vis absorption and fluorescent emission titrations experiments have been employed to study the sensing process. Rh{sub B} could act as an efficient “ON–OFF” fluorescent response for phosphotungstic acid (H{sub 3}PW{sub 12}O{sub 40} or PTA) based on an ion associate process. Also (Rh{sub B}{sup +}){sub 3}·PTA{sup 3−} could operate as an “OFF–ON” fluorescent sensor for cyanide anions based on a ligand substitution process. It has been identified as highly sensitive probe for CN{sup −} which responds at 0.3 and 0.04 μmol L{sup −1} concentration levels by absorption and fluorescent method respectively. Depending upon the sequence of addition of PTA and CN{sup −} ions into the solution, Rh{sub B} could be as a molecular security keypad lock with PTA and CN{sup −} inputs. The ionic inputs to new fluorophore have been mimicked as a superimposed electronic molecular keypad lock. The results were compared successfully (>96%) with the data of a spectrophotometry approved method (EPA 9014-1) for cyanide ions.

  14. Single particle and molecular assembly analysis of polyribosomes by single- and double-tilt cryo electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Myasnikov, Alexander G. [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Department of Integrative Structural Biology, Centre National de la Recherche Scientifique (CNRS) UMR 7104/ Institut National de la Santé de la Recherche Médicale INSERM U964/ Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch (France); Afonina, Zhanna A. [Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region (Russian Federation); Klaholz, Bruno P., E-mail: klaholz@igbmc.fr [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Department of Integrative Structural Biology, Centre National de la Recherche Scientifique (CNRS) UMR 7104/ Institut National de la Santé de la Recherche Médicale INSERM U964/ Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch (France)

    2013-03-15

    Cryo electron tomography (cryo-ET) can provide cellular and molecular structural information on various biological samples. However, the detailed interpretation of tomograms reconstructed from single-tilt data tends to suffer from low signal-to-noise ratio and artefacts caused by some systematically missing angular views. While these can be overcome by sub-tomogram averaging, they remain limiting for the analysis of unique structures. Double-tilt ET can improve the tomogram quality by acquiring a second tilt series after an in-plane rotation, but its usage is not widespread yet because it is considered technically demanding and it is rarely used under cryo conditions. Here we show that double-tilt cryo-ET improves the quality of 3D reconstructions so significantly that even single particle analysis can be envisaged despite of the intrinsically low image contrast obtained from frozen-hydrated specimens. This is illustrated by the analysis of eukaryotic polyribosomes in which individual ribosomes were reconstructed using single-tilt, partial and full double-tilt geometries. The improved tomograms favour the faster convergence of iterative sub-tomogram averaging and allow a better 3D classification using multivariate statistical analysis. Our study of single particles and molecular assemblies within polysomes illustrates that the dual-axis approach is particularly useful for cryo applications of ET, both for unique objects and for structures that can be classified and averaged. - Highlights: ► Double-tilt cryo-ET improves 3D reconstructions thus making single particle analysis possible. ► Dual-axis cryo-ET data favour a faster convergence of iterative sub-tomogram averaging. ► Individual ribosomes were reconstructed from single-tilt, partial/ full double-tilt geometries. ► Double-tilt cryo-ET facilitates analysis of larger molecular assemblies such as in cell sections. ► Dual-axis cryo-ET is applicable to unique objects and to structures that can be

  15. Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET)

    NARCIS (Netherlands)

    Lidke, D.S.; Nagy, P.; Barisas, B.G.; Heintzmann, R.; Post, Janine Nicole; Lidke, K.A.; Clayton, A.H.A.; Arndt-jovin, D.J.; Jovin, T.M.

    2003-01-01

    We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow

  16. A Fluorescent Molecular Probe for the Detection of Hydrogen Based on Oxidative Addition Reactions with Crabtree-Type Hydrogenation Catalysts.

    Science.gov (United States)

    Kos, Pavlo; Plenio, Herbert

    2015-11-02

    A Crabtree-type Ir(I) complex tagged with a fluorescent dye (bodipy) was synthesized. The oxidative addition of H2 converts the weakly fluorescent Ir(I) complex (Φ=0.038) into a highly fluorescent Ir(III) species (Φ=0.51). This fluorogenic reaction can be utilized for the detection of H2 and to probe the oxidative addition step in the catalytic hydrogenation of olefins. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Synthesis of new molecular probes radiolabelled with fluorine-18 for imaging neuro-inflammation with Positon Emission Tomography

    International Nuclear Information System (INIS)

    Medran-Navarrete, Vincent

    2014-01-01

    The work presented in this manuscript aims to describe the synthesis of new ligands of the translocation protein 18 kDa (TSPO), their in vitro evaluation and, for the most promising candidates, their isotopic radiolabelling with the short-lived positron emitter fluorine-18 (t 1/2 : 109.8 minutes). The ultimate goal of this work consists in developing new molecular probes, or bio-markers, for imaging neuro-inflammation in a non-invasive and atraumatic manor using Positron Emission Tomography (PET). Neuro-inflammatory processes have been identified in Alzheimer and Parkinson diseases, MS and various psychiatric pathologies. The radioligand of choice for imaging TSPO is currently [ 18 F]DPA-714, a pyr-azolo[1,5-a]pyrimidine radiolabelled with fluorine-18 which has been recently prepared in our laboratories. However, [ 18 F]DPA-714 undergoes a rapid in vivo loss of the radioactive fluorine by cleavage of the fluoro-alkoxy chain as demonstrated in metabolic studies. Therefore, my PhD project aimed to design and develop new structurally related analogues of DPA-714 where the linkage between the main backbone and the fluorine-18 would be reinforced. To this extent, nineteen compounds were prepared and their affinity towards the TSPO was evaluated. Two promising candidates, coded DPA-C5yne and CfO-DPA-714, were radiolabelled with fluorine-18 with good radiochemical yields (20-30 %) and high specific radioactivities (50-90 GBq/μmol). These radioligands were also evaluated by PET imaging at the preclinical stage and displayed equivalent or slightly improved results when compared to [ 18 F]DPA- 714. (author) [fr

  18. Fluorescence and room temperature phosphorescence of 6-bromo-2-naphthol in {beta}-cyclodextrin solution and its selective molecular recognition for cyclohexane

    Energy Technology Data Exchange (ETDEWEB)

    Zhai Yanqiang; Zhang Shuzhen; Xie Jianwei; Liu Changsong

    2003-10-08

    The room temperature phosphorescence (RTP) and fluorescence behavior of 6-bromo-2-naphthol (BN) in water and {beta}-cyclodextrin ({beta}-CD) aerated aqueous solution was investigated. The study of fluorescence behavior at different pH values indicated that three kinds of species of BN (protonated, uncharged and anionic species) formed 1:1 inclusion complexes with {beta}-CD, and RTP and fluorescence emission depended on the pH of the solution. The inclusion complex constants were 430{+-}25 l mol{sup -1} (pH 1.80), 840{+-}25 l mol{sup -1} (pH 5.80), 1850{+-}75 l mol{sup -1} (pH 11.50), respectively. Experimental results elucidated that RTP of the BN/{beta}-CD/cyclohexane solution came from the protonated and uncharged species of BN, but not from the anionic species, though the inclusion constant of the anionic species of BN with {beta}-CD was larger than that of the other two species of BN Selective molecular recognition of BN/{beta}-CD as an RTP sensor for 28 small organic molecules was studied, it was shown that BN/{beta}-CD could be develop as a new RTP sensor with high selectivity molecular recognition ability for cyclohexane.

  19. Fluorescence and room temperature phosphorescence of 6-bromo-2-naphthol in β-cyclodextrin solution and its selective molecular recognition for cyclohexane

    International Nuclear Information System (INIS)

    Zhai Yanqiang; Zhang Shuzhen; Xie Jianwei; Liu Changsong

    2003-01-01

    The room temperature phosphorescence (RTP) and fluorescence behavior of 6-bromo-2-naphthol (BN) in water and β-cyclodextrin (β-CD) aerated aqueous solution was investigated. The study of fluorescence behavior at different pH values indicated that three kinds of species of BN (protonated, uncharged and anionic species) formed 1:1 inclusion complexes with β-CD, and RTP and fluorescence emission depended on the pH of the solution. The inclusion complex constants were 430±25 l mol -1 (pH 1.80), 840±25 l mol -1 (pH 5.80), 1850±75 l mol -1 (pH 11.50), respectively. Experimental results elucidated that RTP of the BN/β-CD/cyclohexane solution came from the protonated and uncharged species of BN, but not from the anionic species, though the inclusion constant of the anionic species of BN with β-CD was larger than that of the other two species of BN Selective molecular recognition of BN/β-CD as an RTP sensor for 28 small organic molecules was studied, it was shown that BN/β-CD could be develop as a new RTP sensor with high selectivity molecular recognition ability for cyclohexane

  20. Dual fluorescent molecular substrates selectively report the activation, sustainability and reversibility of cellular PKB/Akt activity.

    Science.gov (United States)

    Shen, Duanwen; Bai, Mingfeng; Tang, Rui; Xu, Baogang; Ju, Xiaoming; Pestell, Richard G; Achilefu, Samuel

    2013-01-01

    Using a newly developed near-infrared (NIR) dye that fluoresces at two different wavelengths (dichromic fluorescence, DCF), we discovered a new fluorescent substrate for Akt, also known as protein kinase B, and a method to quantitatively report this enzyme's activity in real time. Upon insulin activation of cellular Akt, the enzyme multi-phosphorylated a single serine residue of a diserine DCF substrate in a time-dependent manner, culminating in monophospho- to triphospho-serine products. The NIR DCF probe was highly selective for the Akt1 isoform, which was demonstrated using Akt1 knockout cells derived from MMTV-ErbB2 transgenic mice. The DCF mechanism provides unparalleled potential to assess the stimulation, sustainability, and reversibility of Akt activation longitudinally. Importantly, NIR fluorescence provides a pathway to translate findings from cells to living organisms, a condition that could eventually facilitate the use of these probes in humans.

  1. Enhanced fluorescence of tetrasulfonated zinc phthalocyanine by graphene quantum dots and its application in molecular sensing/imaging.

    Science.gov (United States)

    Wang, Jian; Zhang, Yanjun; Ye, Jiqing; Jiang, Zhou

    2017-06-01

    When excited at 435 nm, tetra-sulfonate zinc phthalocyanine (ZnPcS 4 ) emitted dual fluorescence at 495 and 702 nm. The abnormal fluorescence at 495 nm was experimentally studied and analyzed in detail for the first time. The abnormal fluorescence at 495 nm was deduced to originate from triplet-triplet (T-T) energy transfer of excited phthalocyanine ( 3 *ZnPcS 4 ). Furthermore, graphene quantum dots (GQDs) enhanced the 495 nm fluorescence quantum yield (Q) of ZnPcS 4 . The fluorescence properties of ZnPcS 4 -GQDs conjugate were retained in a cellular environment. Based on the fluorescence of ZnPcS 4 -GQDs conjugate, we designed and prepared an Apt29/thrombin/Apt15 sandwich thrombin sensor with high specificity and affinity. This cost-saving, simple operational sensing strategy can be extended to use in sensing/imaging of other biomolecules. Copyright © 2016 John Wiley & Sons, Ltd.

  2. Single photon emission tomography

    International Nuclear Information System (INIS)

    Buvat, Irene

    2011-09-01

    The objective of this lecture is to present the single photon emission computed tomography (SPECT) imaging technique. Content: 1 - Introduction: anatomic, functional and molecular imaging; Principle and role of functional or molecular imaging; 2 - Radiotracers: chemical and physical constraints, main emitters, radioisotopes production, emitters type and imaging techniques; 3 - Single photon emission computed tomography: gamma cameras and their components, gamma camera specifications, planar single photon imaging characteristics, gamma camera and tomography; 4 - Quantification in single photon emission tomography: attenuation, scattering, un-stationary spatial resolution, partial volume effect, movements, others; 5 - Synthesis and conclusion

  3. Vibronic coupling in molecular crystals: A Franck-Condon Herzberg-Teller model of H-aggregate fluorescence based on quantum chemical cluster calculations

    Energy Technology Data Exchange (ETDEWEB)

    Wykes, M., E-mail: mikewykes@gmail.com; Parambil, R.; Gierschner, J. [Madrid Institute for Advanced Studies, IMDEA Nanoscience, Calle Faraday 9, Campus Cantoblanco, 28049 Madrid (Spain); Beljonne, D. [Laboratory for Chemistry of Novel Materials, University of Mons, Place du Parc 20, 7000 Mons (Belgium)

    2015-09-21

    Here, we present a general approach to treating vibronic coupling in molecular crystals based on atomistic simulations of large clusters. Such clusters comprise model aggregates treated at the quantum chemical level embedded within a realistic environment treated at the molecular mechanics level. As we calculate ground and excited state equilibrium geometries and vibrational modes of model aggregates, our approach is able to capture effects arising from coupling to intermolecular degrees of freedom, absent from existing models relying on geometries and normal modes of single molecules. Using the geometries and vibrational modes of clusters, we are able to simulate the fluorescence spectra of aggregates for which the lowest excited state bears negligible oscillator strength (as is the case, e.g., ideal H-aggregates) by including both Franck-Condon (FC) and Herzberg-Teller (HT) vibronic transitions. The latter terms allow the adiabatic excited state of the cluster to couple with vibrations in a perturbative fashion via derivatives of the transition dipole moment along nuclear coordinates. While vibronic coupling simulations employing FC and HT terms are well established for single-molecules, to our knowledge this is the first time they are applied to molecular aggregates. Here, we apply this approach to the simulation of the low-temperature fluorescence spectrum of para-distyrylbenzene single-crystal H-aggregates and draw comparisons with coarse-grained Frenkel-Holstein approaches previously extensively applied to such systems.

  4. Molecular beacon-decorated polymethylmethacrylate core-shell fluorescent nanoparticles for the detection of survivin mRNA in human cancer cells.

    Science.gov (United States)

    Adinolfi, Barbara; Pellegrino, Mario; Giannetti, Ambra; Tombelli, Sara; Trono, Cosimo; Sotgiu, Giovanna; Varchi, Greta; Ballestri, Marco; Posati, Tamara; Carpi, Sara; Nieri, Paola; Baldini, Francesco

    2017-02-15

    One of the main goals of nanomedicine in cancer is the development of effective drug delivery systems, primarily nanoparticles. Survivin, an overexpressed anti-apoptotic protein in cancer, represents a pharmacological target for therapy and a Molecular Beacon (MB) specific for survivin mRNA is available. In this study, the ability of polymethylmethacrylate nanoparticles (PMMA-NPs) to promote survivin MB uptake in human A549 cells was investigated. Fluorescent and positively charged core PMMA-NPs of nearly 60nm, obtained through an emulsion co-polymerization reaction, and the MB alone were evaluated in solution, for their analytical characterization; then, the MB specificity and functionality were verified after adsorption onto the PMMA-NPs. The carrier ability of PMMA-NPs in A549 was examined by confocal microscopy. With the optimized protocol, a hardly detectable fluorescent signal was obtained after incubation of the cells with the MB alone (fluorescent spots per cell of 1.90±0.40 with a mean area of 1.04±0.20µm 2 ), while bright fluorescent spots inside the cells were evident by using the MB loaded onto the PMMA-NPs. (27.50±2.30 fluorescent spots per cell with a mean area of 2.35±0.16µm 2 ). These results demonstrate the ability of the PMMA-NPs to promote the survivin-MB internalization, suggesting that this complex might represent a promising strategy for intracellular sensing and for the reduction of cancer cell proliferation. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Quenching of the OH and nitrogen molecular emission by methane addition in an Ar capacitively coupled plasma to remove spectral interference in lead determination by atomic fluorescence spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Frentiu, T., E-mail: ftibi@chem.ubbcluj.r [Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University, Arany Janos 11, 400028 Cluj-Napoca (Romania); Ponta, M., E-mail: mponta@chem.ubbcluj.r [Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University, Arany Janos 11, 400028 Cluj-Napoca (Romania); Mihaltan, A.I., E-mail: alinblaj2005@yahoo.co [National Institute for Research and Development of Optoelectronics Bucharest - Research Institute for Analytical Instrumentation, Donath 67, 400293 Cluj-Napoca (Romania); Darvasi, E., E-mail: edarvasi@chem.ubbcluj.r [Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University, Arany Janos 11, 400028 Cluj-Napoca (Romania); Frentiu, M., E-mail: frentiu.maria@yahoo.co [National Institute for Research and Development of Optoelectronics Bucharest - Research Institute for Analytical Instrumentation, Donath 67, 400293 Cluj-Napoca (Romania); Cordos, E., E-mail: emilcordos@gmail.co [National Institute for Research and Development of Optoelectronics Bucharest - Research Institute for Analytical Instrumentation, Donath 67, 400293 Cluj-Napoca (Romania)

    2010-07-15

    A new method is proposed to remove the spectral interference on elements in atomic fluorescence spectrometry by quenching of the molecular emission of the OH radical (A{sup 2{Sigma}+} {yields} X{sup 2{Pi}}) and N{sub 2} second positive system (C{sup 3{Pi}}{sub u} {yields} B{sup 3{Sigma}}{sub g}) in the background spectrum of medium power Ar plasmas. The experiments were carried out in a radiofrequency capacitively coupled plasma (275 W, 27.12 MHz) by CH{sub 4} addition. The quenching is the result of the high affinity of OH radical for a hydrogen atom from the CH{sub 4} molecule and the collisions of the second kind between nitrogen excited molecules and CH{sub 4}, respectively. The decrease of the emission of N{sub 2} second positive system in the presence of CH{sub 4} is also the result of the deactivation of the metastable argon atoms that could excite the nitrogen molecules. For flow rates of 0.7 l min{sup -1} Ar with addition of 7.5 ml min{sup -1} CH{sub 4}, the molecular emission of OH and N{sub 2} was completely removed from the plasma jet spectrum at viewing heights above 60 mm. The molecular emission associated to CH and CH{sub 2} species was not observed in the emission spectrum of Ar/CH{sub 4} plasma in the ultraviolet range. The method was experimented for the determination of Pb at 283.31 nm by atomic fluorescence spectrometry with electrodeless discharge lamp and a multichannel microspectrometer. The detection limit was 35 ng ml{sup -1}, 2-3 times better than in atomic emission spectrometry using the same plasma source, and similar to that in hollow cathode lamp microwave plasma torch atomic fluorescence spectrometry.

  6. Molecular Imaging of β-Amyloid Plaques with Near-Infrared Boron Dipyrromethane (BODIPY-Based Fluorescent Probes

    Directory of Open Access Journals (Sweden)

    Hiroyuki Watanabe

    2013-07-01

    Full Text Available The formation of β-amyloid (Aβ plaques is a critical neurodegenerative change in Alzheimer disease (AD. We designed and synthesized novel boron dipyrromethane (BODIPY-based Aβ probes (BAPs and evaluated their utility for near-infrared fluorescence imaging of Aβ plaques in the brain. In binding experiments in vitro, BAPs showed high affinity for synthetic Aβ aggregates (Kd = 18–149 nM. Furthermore, BAPs clearly stained Aβ plaques in sections of Tg2576 mice. In mouse brain tissue, BAPs showed sufficient uptake for optical imaging. In addition, ex vivo fluorescent staining of brain sections from Tg2576 mice after the injection of BAP-2 showed selective binding of Aβ plaques with little nonspecific binding. BAPs may be useful as a near-infrared fluorescent probe for imaging Aβ plaques.

  7. Two dimensional laser induced fluorescence in the gas phase: a spectroscopic tool for studying molecular spectroscopy and dynamics

    Science.gov (United States)

    Gascooke, Jason R.; Lawrance, Warren D.

    2017-11-01

    Two dimensional laser induced fluorescence (2D-LIF) extends the usual laser induced fluorescence technique by adding a second dimension, the wavelength at which excited states emit, thereby significantly enhancing the information that can be extracted. It allows overlapping absorption features, whether they arise from within the same molecule or from different molecules in a mixture, to be associated with their appropriate "parent" state and/or molecule. While the first gas phase version of the technique was published a decade ago, the technique is in its infancy, having been exploited by only a few groups to date. However, its potential in gas phase spectroscopy and dynamics is significant. In this article we provide an overview of the technique and illustrate its potential with examples, with a focus on those utilising high resolution in the dispersed fluorescence dimension.

  8. Intramembranous bone healing process subsequent to tooth extraction in mice: micro-computed tomography, histomorphometric and molecular characterization.

    Science.gov (United States)

    Vieira, Andreia Espindola; Repeke, Carlos Eduardo; Ferreira Junior, Samuel de Barros; Colavite, Priscila Maria; Biguetti, Claudia Cristina; Oliveira, Rodrigo Cardoso; Assis, Gerson Francisco; Taga, Rumio; Trombone, Ana Paula Favaro; Garlet, Gustavo Pompermaier

    2015-01-01

    Bone tissue has a significant potential for healing, which involves a significant the interplay between bone and immune cells. While fracture healing represents a useful model to investigate endochondral bone healing, intramembranous bone healing models are yet to be developed and characterized. In this study, a micro-computed tomography, histomorphometric and molecular (RealTimePCRarray) characterization of post tooth-extraction alveolar bone healing was performed on C57Bl/6 WT mice. After the initial clot dominance (0 h), the development of a provisional immature granulation tissue is evident (7 d), characterized by marked cell proliferation, angiogenesis and inflammatory cells infiltration; associated with peaks of growth factors (BMP-2-4-7,TGFβ1,VEGFa), cytokines (TNFα, IL-10), chemokines & receptors (CXCL12, CCL25, CCR5, CXCR4), matrix (Col1a1-2, ITGA4, VTN, MMP1a) and MSCs (CD105, CD106, OCT4, NANOG, CD34, CD146) markers expression. Granulation tissue is sequentially replaced by more mature connective tissue (14 d), characterized by inflammatory infiltrate reduction along the increased bone formation, marked expression of matrix remodeling enzymes (MMP-2-9), bone formation/maturation (RUNX2, ALP, DMP1, PHEX, SOST) markers, and chemokines & receptors associated with healing (CCL2, CCL17, CCR2). No evidences of cartilage cells or tissue were observed, strengthening the intramembranous nature of bone healing. Bone microarchitecture analysis supports the evolving healing, with total tissue and bone volumes as trabecular number and thickness showing a progressive increase over time. The extraction socket healing process is considered complete (21 d) when the dental socket is filled by trabeculae bone with well-defined medullary canals; it being the expression of mature bone markers prevalent at this period. Our data confirms the intramembranous bone healing nature of the model used, revealing parallels between the gene expression profile and the

  9. Intramembranous bone healing process subsequent to tooth extraction in mice: micro-computed tomography, histomorphometric and molecular characterization.

    Directory of Open Access Journals (Sweden)

    Andreia Espindola Vieira

    Full Text Available Bone tissue has a significant potential for healing, which involves a significant the interplay between bone and immune cells. While fracture healing represents a useful model to investigate endochondral bone healing, intramembranous bone healing models are yet to be developed and characterized. In this study, a micro-computed tomography, histomorphometric and molecular (RealTimePCRarray characterization of post tooth-extraction alveolar bone healing was performed on C57Bl/6 WT mice. After the initial clot dominance (0 h, the development of a provisional immature granulation tissue is evident (7 d, characterized by marked cell proliferation, angiogenesis and inflammatory cells infiltration; associated with peaks of growth factors (BMP-2-4-7,TGFβ1,VEGFa, cytokines (TNFα, IL-10, chemokines & receptors (CXCL12, CCL25, CCR5, CXCR4, matrix (Col1a1-2, ITGA4, VTN, MMP1a and MSCs (CD105, CD106, OCT4, NANOG, CD34, CD146 markers expression. Granulation tissue is sequentially replaced by more mature connective tissue (14 d, characterized by inflammatory infiltrate reduction along the increased bone formation, marked expression of matrix remodeling enzymes (MMP-2-9, bone formation/maturation (RUNX2, ALP, DMP1, PHEX, SOST markers, and chemokines & receptors associated with healing (CCL2, CCL17, CCR2. No evidences of cartilage cells or tissue were observed, strengthening the intramembranous nature of bone healing. Bone microarchitecture analysis supports the evolving healing, with total tissue and bone volumes as trabecular number and thickness showing a progressive increase over time. The extraction socket healing process is considered complete (21 d when the dental socket is filled by trabeculae bone with well-defined medullary canals; it being the expression of mature bone markers prevalent at this period. Our data confirms the intramembranous bone healing nature of the model used, revealing parallels between the gene expression profile and the

  10. A fluorescent molecular sensor for pH windows in traditional and polymeric biocompatible micelles: comicellization of anionic species to shift and reshape the ON window.

    Science.gov (United States)

    Cavallaro, Gennara; Giammona, Gaetano; Pasotti, Luca; Pallavicini, Piersandro

    2011-09-12

    A new approach is presented to obtain fluorescent sensors for pH windows that work in water and under biomimetic conditions. A single molecule that features all-covalently linked components is used, thus making it capable of working as a fluorescent sensor with an OFF/ON/OFF response to pH value. The components are a tertiary amine, a pyridine, and a fluorophore (pyrene). The forms with both protonated bases or both neutral bases quench the pyrene fluorescence, whereas the form with the neutral pyridine and protonated amine groups is fluorescent. The molecular sensor is also equipped with a long alkyl chain to make it highly hydrophobic in all its protonated and unprotonated forms, that is, either when neutral or charged. Accordingly, it can be confined at any pH value either in traditional (i.e., low-molecular-weight) nonionic surfactant micelles or inside polymeric, biocompatible micellar containers. Relevant for future applications in vivo, thanks to its strong hydrophobicity, no leakage of the molecular sensor is observed from the polymeric biocompatible micelles. Due to the proximity of the pyridine and amine functions in the molecular structure and the poor hydration inside the micelles, the observed pK(a) values are low so that the ON window is positioned at very low pH values. However, the window can be shifted to biologically relevant values by comicellization of anionic species. In particular, in the micelles of the nonionic surfactant TritonX-100, a shift of the ON window to pH 4-6 is obtained by addition of the anionic sodium dodecyl sulphate surfactant, whose negative charge promotes the stability of the protonated forms of the pyridine and amine fragments. In the case of the polymeric micelles, we introduce the use of the amphiphilic polystyrene sulfonate anionic polyelectrolyte, the comicellization of which induces a shift and sharpening of the ON window that is centered at pH 4. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Time-Resolved Fluorescence Anisotropy of Bicyclo[1.1.1]pentane/Tolane-Based Molecular Rods Included in Tris(o-phenylenedioxy)cyclotriphosphazene (TPP).

    Science.gov (United States)

    Cipolloni, Marco; Kaleta, Jiří; Mašát, Milan; Dron, Paul I; Shen, Yongqiang; Zhao, Ke; Rogers, Charles T; Shoemaker, Richard K; Michl, Josef

    2015-04-23

    We examine the fluorescence anisotropy of rod-shaped guests held inside the channels of tris( o -phenylenedioxy)cyclotriphosphazene (TPP) host nanocrystals, characterized by powder X-ray diffraction and solid state NMR spectroscopy. We address two issues: (i) are light polarization measurements on an aqueous colloidal solution of TPP nanocrystals meaningful, or is depolarization by scattering excessive? (ii) Can measurements of the rotational mobility of the included guests be performed at low enough loading levels to suppress depolarization by intercrystallite energy transfer? We find that meaningful measurements are possible and demonstrate that the long axis of molecular rods included in TPP channels performs negligible vibrational motion.

  12. Crystallography and Molecular Arrangement of Polymorphic Monolayer J-Aggregates of a Cyanine Dye: Multiangle Polarized Light Fluorescence Optical Microscopy Study.

    Science.gov (United States)

    Prokhorov, Valery V; Pozin, Sergey I; Perelygina, Olga M; Mal'tsev, Eugene I

    2018-04-24

    The molecular orientation in monolayer J-aggregates of 3,3-di(γ-sulfopropyl)-5,5-dichlorotiamonomethinecyanine dye has been precisely estimated using improved linear polarization measurements in the fluorescence microscope in which a multiangle set of polarization data is obtained using sample rotation. The estimated molecular orientation supplemented with the previously established crystallographic constraints based on the analysis of the well-developed two-dimensional J-aggregate shapes unambiguously indicate the staircase type of molecular arrangement for striplike J-aggregates with the staircases oriented along strips. The molecular transition dipoles are inclined at an angle of ∼25° to the strip direction, whereas the characteristic strip vertex angle ∼45° is formed by the [100] and [1-10] directions of the monoclinic unit cell. Measurements of the geometry of partially unwound tubes and their polarization properties support the model of tube formation by close-packed helical winding of flexible monolayer strips. In the tubes, the long molecular axes are oriented at a small angle in the range of 5-15° to the normal to the tube axis providing low bending energy. At a nanoscale, high-resolution atomic force microscopy imaging of J-aggregate monolayers reveals a complex quasi-one-dimensional organization.

  13. Absorption and Fluorescence of PRODAN in Phospholipid Bilayers: A Combined Quantum Mechanics and Classical Molecular Dynamics Study

    Czech Academy of Sciences Publication Activity Database

    Cwiklik, Lukasz; Aquino, A. J. A.; Vazdar, Mario; Jurkiewicz, Piotr; Pittner, Jiří; Hof, Martin; Lischka, H.

    2011-01-01

    Roč. 115, č. 41 (2011), s. 11428-11437 ISSN 1089-5639 R&D Projects: GA AV ČR IAA400400810 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z40550506 Keywords : fluorescence * density functional theory * charge transfer Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.946, year: 2011

  14. A Turn-on Fluorescence Sensor for Heparin Detection Based on a Release of Taiwan Cobra Cardiotoxin from a DNA Aptamer or Adenosine-Based Molecular Beacon.

    Science.gov (United States)

    Shi, Yi-Jun; Wang, Liang-Jun; Lee, Yuan-Chin; Huang, Chia-Hui; Hu, Wan-Ping; Chang, Long-Sen

    2018-02-19

    This study presents two sensitive fluorescent assays for sensing heparin on the basis of the electrostatic interaction between heparin and Naja naja atra cardiotoxin 3 (CTX3). Owing to CTX3-induced folded structure of an adenosine-based molecular beacon (MB) or a DNA aptamer against CTX3, a reduction in the fluorescent signal of the aptamer or MB 5'-end labeled with carboxyfluorescein (FAM) and 3'-end labeled with 4-([4-(dimethylamino)phenyl]azo)-benzoic acid (DABCYL) was observed upon the addition of CTX3. The presence of heparin and formation of the CTX3-heparin complex caused CTX3 detachment from the MB or aptamer, and restoration of FAM fluorescence of the 5'-FAM-and-3'-DABCYL-labeled MB and aptamer was subsequently noted. Moreover, the detection of heparin with these CTX3-aptamer and CTX3-MB sensors showed high sensitivity and selectivity toward heparin over chondroitin sulfate and hyaluronic acid regardless of the presence of plasma. The limit of detection for heparin in plasma was determined to be 16 ng/mL and 15 ng/mL, respectively, at a signal-to-noise ratio of 3. This study validates the practical utility of the CTX3-aptamer and CTX3-MB systems for determining the concentration of heparin in a biological matrix.

  15. PEG-phospholipid-encapsulated bismuth sulfide and CdSe/ZnS quantum dot core–shell nanoparticle and its computed tomography/fluorescence performance

    International Nuclear Information System (INIS)

    Chen, Jun; Yang, Xiao-Quan; Qin, Meng-Yao; Zhang, Xiao-Shuai; Xuan, Yang; Zhao, Yuan-Di

    2015-01-01

    In this paper, polyethylene glycol-phospholipid structure is used to synthesize hybrid cluster of 40–50 nm diameter that contains hydrophobic bismuth sulfide nanoparticles and CdSe/ZnS quantum dots. The composite probe’s toxicity, CT imaging, and fluorescence imaging performance are also studied. Experimental results show that the nanocomposite hybrid cluster has obvious CT contrast enhancement and fluorescence imaging capability in vitro even after cellular uptake. It gives a CT number of 700 (Hounsfield units) at 15 mg/mL, higher than that of the current iobitridol CT contrast agent. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide experiment reveals that it has low cytotoxicity at concentration up to of 3.14 mg/mL of Bi, indicating the composite probe has potential ability for CT and fluorescence bimodal imaging

  16. Enhancing molecular logic through modulation of temporal and spatial constraints with quantum dot-based systems that use fluorescent (Förster) resonance energy transfer

    Science.gov (United States)

    Claussen, Jonathan C.; Algar, W. Russ; Hildebrandt, Niko; Susumu, Kimihiro; Ancona, Mario G.; Medintz, Igor L.

    2013-10-01

    Luminescent semiconductor nanocrystals or quantum dots (QDs) contain favorable photonic properties (e.g., resistance to photobleaching, size-tunable PL, and large effective Stokes shifts) that make them well-suited for fluorescence (Förster) resonance energy transfer (FRET) based applications including monitoring proteolytic activity, elucidating the effects of nanoparticles-mediated drug delivery, and analyzing the spatial and temporal dynamics of cellular biochemical processes. Herein, we demonstrate how unique considerations of temporal and spatial constraints can be used in conjunction with QD-FRET systems to open up new avenues of scientific discovery in information processing and molecular logic circuitry. For example, by conjugating both long lifetime luminescent terbium(III) complexes (Tb) and fluorescent dyes (A647) to a single QD, we can create multiple FRET lanes that change temporally as the QD acts as both an acceptor and donor at distinct time intervals. Such temporal FRET modulation creates multi-step FRET cascades that produce a wealth of unique photoluminescence (PL) spectra that are well-suited for the construction of a photonic alphabet and photonic logic circuits. These research advances in bio-based molecular logic open the door to future applications including multiplexed biosensing and drug delivery for disease diagnostics and treatment.

  17. Bioresponsive and fluorescent hyaluronic acid-iodixanol nanogels for targeted X-ray computed tomography imaging and chemotherapy of breast tumors

    NARCIS (Netherlands)

    Zhu, Yaqin; Wang, Xinhui; Wang, X.; Chen, J.; Meng, Fenghua; Deng, D.; Cheng, R.; Feijen, Jan; Zhong, Zhiyuan

    2016-01-01

    Nanotheranostics is a rapidly growing field combining disease diagnosis and therapy, which ultimately may add in the development of ‘personalized medicine’. Here, we designed and developed bioresponsive and fluorescent hyaluronic acid-iodixanol nanogels (HAI-NGs) for targeted X-ray computed

  18. The effect of nonlocal dielectric response on the surface-enhanced Raman and fluorescence spectra of molecular systems

    Science.gov (United States)

    Wei, Yong; Pei, Huan; Li, Li; Zhu, Yanying

    2018-06-01

    We present a theoretical study on the influence of the nonlocal dielectric response on surface-enhanced resonant Raman scattering (SERRS) and fluorescence (SEF) spectra of a model molecule confined in the center of a Ag nanoparticle (NP) dimer. In the simulations, the nonlocal dielectric response caused by the electron–hole pair generation in Ag NPs was computed with the d-parameter theory, and the scattering spectra of a model molecule representing the commonly used fluorescent dye rhodamine 6G (R6G) were obtained by density-matrix calculations. The influence of the separation between Ag NP dimers on the damping rate and scattering spectra with and without the nonlocal response were systematically analyzed. The results show that the nonlocal dielectric response is very sensitive to the gap distance of the NP dimers, and it undergoes much faster decay with the increase of the separation than the radiative and energy transfer rates. The Raman and fluorescence peaks as simulated with the nonlocal dielectric response are relative weaker than that without the nonlocal effect for smaller NP separations because the extra decay rates of the nonlocal effect could reduce both the population of the excited state and the interband coherence between the ground and excited states. Our result also indicates that the nonlocal effect is more prominent on the SEF process than the SERRS process.

  19. Influence of excitation light rejection on forward model mismatch in optical tomography

    International Nuclear Information System (INIS)

    Hwang, K; Pan, T; Joshi, A; Rasmussen, J C; Bangerth, W; Sevick-Muraca, E M

    2006-01-01

    Fluorescence enhanced tomography for molecular imaging requires low background for detection and accurate image reconstruction. In this contribution, we show that excitation light leakage is responsible for elevated background and can be minimized with the use of gradient index (GRIN) lenses when using fibre optics to collect propagated fluorescence light from tissue or other biological media. We show that the model mismatch between frequency-domain photon migration (FDPM) measurements and the diffusion approximation prediction is decreased when GRIN lenses are placed prior to the interference filters to provide efficient excitation light rejection. Furthermore, model mismatch is correlated to the degree of excitation light leakage. This work demonstrates the importance of proper light filtering when designing fluorescence optical imaging and tomography

  20. Study of molecularly imprinted solid-phase extraction of gonyautoxins 2,3 in the cultured dinoflagellate Alexandrium tamarense by high-performance liquid chromatography with fluorescence detection

    International Nuclear Information System (INIS)

    Lian, Zi-Ru; Wang, Jiang-Tao

    2013-01-01

    A highly selective sample cleanup procedure combined with molecularly imprinted solid-phase extraction (MISPE) was developed for the isolation of gonyautoxins 2,3 (GTX2,3) from Alexandrium tamarense sample. The molecularly imprinted polymer microspheres (MIPMs) were prepared by suspension polymerization using caffeine as the dummy template molecule, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker and polyvinyl alcohol as the dispersive reagent. The polymer microspheres were used as a selective sorbent for the solid-phase extraction of gonyautoxins 2,3. An off-line MISPE method followed by high-performance liquid chromatography (HPLC) with fluorescence detection for the analysis of gonyautoxins 2,3 was established. Finally, the extract samples from Alexandrium tamarense were analyzed. The results showed the imprinted polymer microspheres exhibited high affinity and selectivity for gonyautoxins 2,3. The interference matrix in the extract were obviously cleaned by MISPE and the extraction efficiency of gonyautoxins 2,3 in the sample ranged from 81.74% to 85.86%. -- Graphical abstract: This is the SEM photograph of molecularly imprinted polymer microspheres (MIPMs). MIPMs were prepared by suspension polymerization and used as selective sorbents for the solid-phase extraction of gonyautoxins 2,3. An off-line MISPE method followed by high-performance liquid chromatography with fluorescence detection for the analysis of gonyautoxins 2,3 was established. The extract samples from Alexandrium tamarense were analyzed by molecularly imprinted solid-phase extraction. Highlights: •The molecularly imprinted polymer microspheres (MIPMs) for GTX2,3 were prepared. •The characteristics and regeneration property of MIPMs were studied. •An off-line method using MIPMs as solid-phase extraction (SPE) sorbents was developed. •GTX2,3 from Alexandrium tamarense extract was successfully isolated by MIPMs-SPE. -- MIPMs for GTX2,3 were

  1. Clinical applicability and prognostic significance of molecular response assessed by fluorescent-PCR of immunoglobulin genes in multiple myeloma. Results from a GEM/PETHEMA study.

    Science.gov (United States)

    Martinez-Lopez, Joaquin; Fernández-Redondo, Elena; García-Sánz, Ramón; Montalbán, María Angeles; Martínez-Sánchez, Pilar; Pavia, Bruno; Mateos, María Victoria; Rosiñol, Laura; Martín, Marisa; Ayala, Rosa; Martínez, Rafael; Blanchard, María Jesus; Alegre, Adrian; Besalduch, Joan; Bargay, Joan; Hernandez, Miguel T; Sarasquete, María Eugenia; Sanchez-Godoy, Pedro; Fernández, Manuela; Blade, Joan; San Miguel, Jesús F; Lahuerta, Juan Jose

    2013-12-01

    Minimal residual disease monitoring is becoming increasingly important in multiple myeloma (MM), but multiparameter flow cytometry (MFC) and allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) techniques are not routinely available. This study investigated the prognostic influence of achieving molecular response assessed by fluorescent-PCR (F-PCR) in 130 newly diagnosed MM patients from Grupo Español Multidisciplinar de Melanoma (GEM)2000/GEM05 trials (NCT00560053, NCT00443235, NCT00464217) who achieved almost very good partial response after induction therapy. As a reference, we used the results observed with simultaneous MFC. F-PCR at diagnosis was performed on DNA using three different multiplex PCRs: IGH D-J, IGK V-J and KDE rearrangements. The applicability of F-PCR was 91·5%. After induction therapy, 64 patients achieved molecular response and 66 non-molecular response; median progression-free survival (PFS) was 61 versus 36 months, respectively (P = 0·001). Median overall survival (OS) was not reached (NR) in molecular response patients (5-year survival: 75%) versus 66 months in the non-molecular response group (P = 0·03). The corresponding PFS and OS values for patients with immunophenotypic versus non-immunophenotypic response were 67 versus 42 months (P = 0·005) and NR (5-year survival: 95%) versus 69 months (P = 0·004), respectively. F-PCR analysis is a rapid, affordable, and easily performable technique that, in some circumstances, may be a valid approach for minimal residual disease investigations in MM. © 2013 John Wiley & Sons Ltd.

  2. Synthesis of molecular imprinted polymers for selective extraction of domperidone from human serum using high performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Salehi, Simin; Rasoul-Amini, Sara; Adib, Noushin; Shekarchi, Maryam

    2016-08-01

    In this study a novel method is described for selective quantization of domperidone in biological matrices applying molecular imprinted polymers (MIPs) as a sample clean up procedure using high performance liquid chromatography coupled with a fluorescence detector. MIPs were synthesized with chloroform as the porogen, ethylene glycol dimethacrylate as the crosslinker, methacrylic acid as the monomer, and domperidone as the template molecule. The new imprinted polymer was used as a molecular sorbent for separation of domperidone from serum. Molecular recognition properties, binding capacity and selectivity of MIPs were determined. The results demonstrated exceptional affinity for domperidone in biological fluids. The domperidone analytical method using MIPs was verified according to validation parameters, such as selectivity, linearity (5-80ng/mL, r(2)=0.9977), precision and accuracy (10-40ng/mL, intra-day=1.7-5.1%, inter-day=4.5-5.9%, and accuracy 89.07-98.9%).The limit of detection (LOD) and quantization (LOQ) of domperidone was 0.0279 and 0.092ng/mL, respectively. The simplicity and suitable validation parameters makes this a highly valuable selective bioequivalence method for domperidone analysis in human serum. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Evaluation of the interactions between human serum albumin (HSA and warfarin or diflunisal by using molecular fluorescence using two approaches

    Directory of Open Access Journals (Sweden)

    Susana Amézqueta

    2018-03-01

    Full Text Available Serum albumin is the main drug transporter of the bloodstream and contains two main binding sites:  Sudlow I or acidic drug binding site, and Sudlow II or benzodiazepine binding site. Warfarin, a well-known anticoagulant drug commonly used in the prevention of thrombosis and thromboembolism, binds to Sudlow I site, whereas non-steroidal antiinflammatory drugs (NSAIDs such as diflunisal bind preferentially to Sudlow II site.  Albumin is a fluorophore that modifies its fluorescence (quenching or enhancement effect when it is bound to a drug. The application of the double logarithm Stern-Volmer equation allows the calculation of the stoichiometry and the binding constant of the process. This procedure does not consider the possible interferences coming from the fluorescence of the drug though. Another strategy to evaluate the binding constants is to consider the whole spectrum, taking into account all the possible species in equilibrium; in this case we have used an extended version of the STAR program, which can deal with 300 spectra, each containing up to 300 data points. The aim of this work is to compare both approaches to evaluate the interaction between warfarin (Sudlow I and diflunisal (Sudlow II and HSA: the double logarithm Stern-Volmer equation and the STAR program.

  4. Spectroscopic and TD-DFT studies on the dual mode fluorescent chemosensors based on pyrene thiosemicarbazones, and its application as molecular-scale logic devices

    Energy Technology Data Exchange (ETDEWEB)

    Basheer, Sabeel M. [Department of Chemistry, National Institute of Technology, Tiruchirappalli 620 015 (India); Willis, Anthony C. [Research School of Chemistry, The Australian National University, Canberra, ACT 2601 (Australia); Sreekanth, Anandaram, E-mail: sreekanth@nitt.edu [Department of Chemistry, National Institute of Technology, Tiruchirappalli 620 015 (India)

    2017-03-15

    Two newly synthesised pyrene based molecules are hereby reported as molecular switches. The absorption and emission response for receptors with and without F{sup −}, CN{sup −} and Cu{sup 2+} ions can mimic multiple logic gate such as AND, NOR, XNOR, OR, XOR, INHIBITION and TRANSFER gates. The fluorescence reversibility was checked with the alternative addition of fluoride and calcium ions, which can be explained by the “Read-Erase-Read-Write” logic loop. The calculated binding constant value show PyBTSC is better chemosensor than PyCTSC, and the binding affinity is in the order of Cu{sup 2+}Г‹Ж’F{sup -}Г‹Ж’CN{sup −.} The detailed mechanism was investigated using DFT and TD-DFT calculations. The fluorescence quenching behaviour of receptor-F complex can be explained by PET mechanism along with ESPT process. The proton attached to the nitrogen which is adjacent to pyrene moiety is first make the hydrogen bond with fluoride ion at the excited state, which has confirmed by natural bond orbital (NBO) and potential energy surface (PES) analysis. - Graphical abstract: The newly synthesised thiocarbazone derivates used as an effective and selective colourimetric and “turn on” fluorescence sensor for copper ion and ‘turn off’ for fluoride and cyanide anion. The presence and absence of ions were considered as input signals and the corresponding absorption and emission responses were consired as output. The proton transfer from the nitrogen adjacent to pyrene moiety, and which takes place at the excited state (ESPT).

  5. Zinc ion coordination as a modulating factor of the ZnuA histidine-rich loop flexibility: A molecular modeling and fluorescence spectroscopy study

    Energy Technology Data Exchange (ETDEWEB)

    Castelli, Silvia [Department of Biology, University of Rome Tor Vergata, Via della Ricerca Scientifica, 00133 Rome (Italy); Stella, Lorenzo [Department of Chemical Sciences and Technologies, University of Roma Tor Vergata, Via della Ricerca Scientifica, 00133 Rome (Italy); Neuromed, IRCCS, Pozzilli 86077 (Italy); Petrarca, Patrizia [Department of Biology, University of Rome Tor Vergata, Via della Ricerca Scientifica, 00133 Rome (Italy); Battistoni, Andrea [Department of Biology, University of Rome Tor Vergata, Via della Ricerca Scientifica, 00133 Rome (Italy); Interuniversity Consortium, National Institute Biostructure and Biosystem (INBB), Viale delle Medaglie D' Oro 305, 00136 Rome (Italy); Desideri, Alessandro [Department of Biology, University of Rome Tor Vergata and CIBB, Center of Biostatistics and Bioinformatics, Via della Ricerca Scientifica, 00133 Rome (Italy); Interuniversity Consortium, National Institute Biostructure and Biosystem (INBB), Viale delle Medaglie D' Oro 305, 00136 Rome (Italy); Falconi, Mattia, E-mail: falconi@uniroma2.it [Department of Biology, University of Rome Tor Vergata and CIBB, Center of Biostatistics and Bioinformatics, Via della Ricerca Scientifica, 00133 Rome (Italy); Interuniversity Consortium, National Institute Biostructure and Biosystem (INBB), Viale delle Medaglie D' Oro 305, 00136 Rome (Italy)

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer Fluorescence data indicate that the His-loop of ZnuA interacts with Zn{sup +2} ions. Black-Right-Pointing-Pointer The ZnuA structural model proposed validates these spectroscopic findings. Black-Right-Pointing-Pointer It is proposed that a zinc loaded His-loop may facilitate the ZnuA-ZnuB recognition. -- Abstract: ZnuA is the soluble component of the high-affinity ZnuABC zinc transporter belonging to the ATP-binding cassette-type periplasmic Zn-binding proteins. The zinc transporter ZnuABC is composed by three proteins: ZnuB, the membrane permease, ZnuC, the ATPase component and ZnuA, the soluble periplasmic metal-binding protein which captures Zn and delivers it to ZnuB. The ZnuA protein contains a charged flexible loop, rich in histidines and acidic residues, showing significant species-specific differences. Various studies have established that this loop contributes to the formation of a secondary zinc binding site, which has been proposed to be important in the acquisition of periplasmic Zn for its delivery to ZnuB or for regulation of zinc uptake. Due to its high mobility the structure of the histidine-rich loop has never been solved by X-ray diffraction studies. In this paper, through a combined use of molecular modeling, mutagenesis and fluorescence spectroscopy, we confirm the presence of two zinc binding sites characterized by different affinities for the metal ion and show that the flexibility of the loop is modulated by the binding of the zinc ions to the protein. The data obtained by fluorescence spectroscopy have then be used to validate a 3D model including the unsolved histidine-rich loop.

  6. Determination of BPA, BPB, BPF, BADGE and BFDGE in canned energy drinks by molecularly imprinted polymer cleaning up and UPLC with fluorescence detection.

    Science.gov (United States)

    Gallo, Pasquale; Di Marco Pisciottano, Ilaria; Esposito, Francesco; Fasano, Evelina; Scognamiglio, Gelsomina; Mita, Gustavo Damiano; Cirillo, Teresa

    2017-04-01

    A new method for simultaneous determination of five bisphenols in canned energy drinks by UPLC with fluorescence detection, after clean up on molecularly imprinted polymers, is herein described. The method was validated at two concentration levels, calculating trueness, repeatability and within-laboratory reproducibility, specificity, linearity of detector response, the limits of quantifications and the limits of detection for each bisphenol. The method is specific, reliable and very sensitive, allowing for determination of bisphenol F diglycidyl ether (BFDGE), bisphenol A (BPA), bisphenol B (BPB), bisphenol F (BPF) and bisphenol A diglycidyl ether (BADGE) down to 0.50ng/mL; it was employed to determine contamination levels from these bisphenols in forty energy drinks of different brands, collected from the market in Naples. BPA was detected in 17 out of 40 samples (42.5%); in some energy drinks also BPF, BADGE and BFDGE were determined. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Near-infrared fluorescence molecular imaging of amyloid beta species and monitoring therapy in animal models of Alzheimer’s disease

    Science.gov (United States)

    Zhang, Xueli; Tian, Yanli; Zhang, Can; Tian, Xiaoyu; Ross, Alana W.; Moir, Robert D.; Sun, Hongbin; Tanzi, Rudolph E.; Moore, Anna; Ran, Chongzhao

    2015-01-01

    Near-infrared fluorescence (NIRF) molecular imaging has been widely applied to monitoring therapy of cancer and other diseases in preclinical studies; however, this technology has not been applied successfully to monitoring therapy for Alzheimer’s disease (AD). Although several NIRF probes for detecting amyloid beta (Aβ) species of AD have been reported, none of these probes has been used to monitor changes of Aβs during therapy. In this article, we demonstrated that CRANAD-3, a curcumin analog, is capable of detecting both soluble and insoluble Aβ species. In vivo imaging showed that the NIRF signal of CRANAD-3 from 4-mo-old transgenic AD (APP/PS1) mice was 2.29-fold higher than that from age-matched wild-type mice, indicating that CRANAD-3 is capable of detecting early molecular pathology. To verify the feasibility of CRANAD-3 for monitoring therapy, we first used the fast Aβ-lowering drug LY2811376, a well-characterized beta-amyloid cleaving enzyme-1 inhibitor, to treat APP/PS1 mice. Imaging data suggested that CRANAD-3 could monitor the decrease in Aβs after drug treatment. To validate the imaging capacity of CRANAD-3 further, we used it to monitor the therapeutic effect of CRANAD-17, a curcumin analog for inhibition of Aβ cross-linking. The imaging data indicated that the fluorescence signal in the CRANAD-17–treated group was significantly lower than that in the control group, and the result correlated with ELISA analysis of brain extraction and Aβ plaque counting. It was the first time, to our knowledge, that NIRF was used to monitor AD therapy, and we believe that our imaging technology has the potential to have a high impact on AD drug development. PMID:26199414

  8. Sequential determination of nickel and cadmium in tobacco, molasses and refill solutions for e-cigarettes samples by molecular fluorescence.

    Science.gov (United States)

    Talio, María Carolina; Alesso, Magdalena; Acosta, Mariano; Wills, Verónica S; Fernández, Liliana P

    2017-11-01

    In this work, a new procedure was developed for separation and preconcentration of nickel(II) and cadmium(II) in several and varied tobacco samples. Tobacco samples were selected considering the main products consumed by segments of the population, in particular the age (youth) and lifestyle of the consumer. To guarantee representative samples, a randomized strategy of sampling was used. In the first step, a chemofiltration on nylon membrane is carried out employing eosin (Eo) and carbon nanotubes dispersed in sodium dodecylsulfate (SDS) solution (phosphate buffer pH 7). In this condition, Ni(II) was selectively retained on the solid support. After that, the filtrate liquid with Cd(II) was re-conditioned with acetic acid /acetate buffer solution (pH 5) and followed by detection. A spectrofluorimetric determination of both metals was carried out, on the solid support and the filtered aqueous solution, for Ni(II) and Cd(II), respectively. The solid surface fluorescence (SSF) determination was performed at λ em = 545nm (λ ex = 515nm) for Ni(II)-Eo complex and the fluorescence of Cd(II)-Eo was quantified in aqueous solution using λ em = 565nm (λ ex = 540nm). The calibration graphs resulted linear in a range of 0.058-29.35μgL -1 for Ni(II) and 0.124-56.20μgL -1 for Cd(II), with detection limits of 0.019 and 0.041μgL -1 (S/N = 3). The developed methodology shows good sensitivity and adequate selectivity, and it was successfully applied to the determination of trace amounts of nickel and cadmium present in tobacco samples (refill solutions for e-cigarettes, snuff used in narguille (molasses) and traditional tobacco) with satisfactory results. The new methodology was validated by ICP-MS with adequate agreement. The proposed methodology represents a novel fluorescence application to Ni(II) and Cd(II) quantification with sensitivity and accuracy similar to atomic spectroscopies, introducing for the first time the quenching effect on SSF. Copyright © 2017 Elsevier B

  9. Cytomorphology of six halotolerant coccoid cyanobacteria using DAPI fluorescent and transmission electron microscopy, compared with molecular data

    Czech Academy of Sciences Publication Activity Database

    Cepák, Vladislav; Komárek, Jiří

    2010-01-01

    Roč. 10, č. 2 (2010), s. 229-234 ISSN 1802-5439 R&D Projects: GA AV ČR IAA600050704 Institutional research plan: CEZ:AV0Z60050516 Keywords : halophilic cyanobacteria * cytomorphology * molecular evaluation Subject RIV: EF - Botanics Impact factor: 0.978, year: 2010

  10. Monitoring of chlorsulfuron in biological fluids and water samples by molecular fluorescence using rhodamine B as fluorophore.

    Science.gov (United States)

    Alesso, Magdalena; Escudero, Luis A; Talio, María Carolina; Fernández, Liliana P

    2016-11-01

    A new simple methodology is proposed for chlorsufuron (CS) traces quantification based upon enhancement of rhodamine B (RhB) fluorescent signal. Experimental variables that influence fluorimetric sensitivity have been studied and optimized. The zeroth order regression calibration was linear from 0.866 to 35.800µgL(-1) CS, with a correlation coefficient of 0.99. At optimal experimental conditions, a limit of detection of 0.259µgL(-1) and a limit of quantification of 0.866µgL(-1) were obtained. The method showed good sensitivity and adequate selectivity and was applied to the determination of trace amounts of CS in plasma, serum and water samples with satisfactory results analyzed by ANOVA test. The proposed methodology represents an alternative to traditional chromatographic techniques for CS monitoring in complex samples, using an accessible instrument in control laboratories. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Investigation of molecular mechanisms of action of chelating drugs on protein-lipid model membranes by X-ray fluorescence

    International Nuclear Information System (INIS)

    Novikova, N. N.; Zheludeva, S. I.; Koval'chuk, M. V.; Stepina, N. D.; Erko, A. I.; Yur'eva, E. A.

    2009-01-01

    Protein-lipid films based on the enzyme alkaline phosphatase were subjected to the action of chelating drugs, which are used for accelerating the removal of heavy metals from the human body, and the elemental composition of the resulting films was investigated. Total-reflection X-ray fluorescence measurements were performed at the Berlin Electron Storage Ring Company for Synchrotron Radiation (BESSY) in Germany. A comparative estimation of the protective effect of four drugs (EDTA, succimer, xydiphone, and mediphon) on membrane-bound enzymes damaged by lead ions was made. The changes in the elemental composition of the protein-lipid films caused by high doses of chelating drugs were investigated. It was shown that state-of-the-art X-ray techniques can, in principle, be used to develop new methods for the in vitro evaluation of the efficiency of drugs, providing differential data on their actions.

  12. Comparing the Spectroscopic and Molecular Characteristics of Different Dissolved Organic Matter Fractions Isolated by Hydrophobic and Anionic Exchange Resins Using Fluorescence Spectroscopy and FT-ICR-MS

    Directory of Open Access Journals (Sweden)

    Morgane Derrien

    2017-07-01

    Full Text Available Despite the environmental significance of dissolved organic matter (DOM, characterizing DOM is still challenging due to its structural complexity and heterogeneity. In this study, three different chemical fractions, including hydrophobic acid (HPOA, transphilic acid (TPIA, and hydrophilic neutral and base (HPIN/B fractions, were separated from bulk aquatic DOM samples, and their spectral features and the chemical composition at the molecular level were compared using both fluorescence excitation emission matrix-parallel factor analysis (EEM-PARAFAC and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS. The HPIN/B fraction was distinguished from the two acidic fractions (i.e., HPOA and TPIA by the EEM-PARAFAC, while the TPIA fraction was discriminated by using the molecular parameters derived from the FT-ICR MS analyses. Statistical comparison suggests that the spectral dissimilarity among the three chemical fractions might result from the acido-basic properties of DOM samples, while the differences in molecular composition were more likely to be affected by the hydrophobicity of the DOM fractions. The non-metric multidimensional scaling map further revealed that the HPOA was the most heterogeneous among the three fractions. The number of overlapping formulas among the three chemical fractions constituted only <5% of all identified formulas, and those between two different fractions ranged from 2.0% to 24.1%, implying relatively homogeneous properties of the individual chemical fractions with respect to molecular composition. Although employing chemical fractionation achieved a lowering of the DOM heterogeneity, prevalent signatures of either acido-basic property or the hydrophobic nature of DOM on the characteristics of three chemical isolated fractions were not found for this study.

  13. In vivo tomographic imaging with fluorescence and MRI using tumor-targeted dual-labeled nanoparticles

    Directory of Open Access Journals (Sweden)

    Zhang Y

    2013-12-01

    Full Text Available Yue Zhang,1 Bin Zhang,1 Fei Liu,1,2 Jianwen Luo,1,3 Jing Bai1 1Department of Biomedical Engineering, School of Medicine, 2Tsinghua-Peking Center for Life Sciences, 3Center for Biomedical Imaging Research, Tsinghua University, Beijing, People's Republic of China Abstract: Dual-modality imaging combines the complementary advantages of different modalities, and offers the prospect of improved preclinical research. The combination of fluorescence imaging and magnetic resonance imaging (MRI provides cross-validated information and direct comparison between these modalities. Here, we report on the application of a novel tumor-targeted, dual-labeled nanoparticle (NP, utilizing iron oxide as the MRI contrast agent and near infrared (NIR dye Cy5.5 as the fluorescent agent. Results of in vitro experiments verified the specificity of the NP to tumor cells. In vivo tumor targeting and uptake of the NPs in a mouse model were visualized by fluorescence and MR imaging collected at different time points. Quantitative analysis was carried out to evaluate the efficacy of MRI contrast enhancement. Furthermore, tomographic images were also acquired using both imaging modalities and cross-validated information of tumor location and size between these two modalities was revealed. The results demonstrate that the use of dual-labeled NPs can facilitate the dual-modal detection of tumors, information cross-validation, and direct comparison by combing fluorescence molecular tomography (FMT and MRI. Keywords: dual-modality, fluorescence molecular tomography (FMT, magnetic resonance imaging (MRI, nanoparticle

  14. Nonlinear Optical Properties of Fluorescent Dyes Allow for Accurate Determination of Their Molecular Orientations in Phospholipid Membranes

    Czech Academy of Sciences Publication Activity Database

    Timr, Štěpán; Brabec, J.; Bondar, Alexey; Ryba, T.; Železný, M.; Lazar, Josef; Jungwirth, Pavel

    2015-01-01

    Roč. 119, č. 30 (2015), s. 9706-9716 ISSN 1520-6106 R&D Projects: GA ČR GA13-06181S; GA ČR GA13-10799S Grant - others:GA MŠk(CZ) LO1506 Institutional support: RVO:61388963 ; RVO:67179843 Keywords : two-photon polarization microscopy * molecular orientation * absorptivity tensor Subject RIV: CF - Physical ; Theoretical Chemistry ; CE - Biochemistry (UEK-B) Impact factor: 3.187, year: 2015

  15. Dual fluorescence of excited state intra-molecular proton transfer of HBFO: mechanistic understanding, substituent and solvent effects.

    Science.gov (United States)

    Yang, Wenjing; Chen, Xuebo

    2014-03-07

    A combined approach of the multiconfigurational perturbation theory with the Rice-Ramsperger-Kassel-Marcus methodology has been employed to calculate the minimum potential energy profiles and the rates of excited state intra-molecular proton transfer (ESIPT) for the WOLED material molecule of HBFO and its four meta- or para-substituted compounds in gas phase, acetonitrile and cyclohexane solvents. The kinetic control for these reactions is quantitatively determined and extensively studied on the basis of the accurate potential energy surfaces when the thermodynamic factor associated with the free energy change becomes negligible in the case of the existence of a significant barrier in the ESIPT process. These computational efforts contribute to a deep understanding of the ESIPT mechanism, dual emission characteristics, kinetic controlling factor, substituent and solvent effects for these material molecules. The white light emission is generated by the establishment of dynamic equilibrium between enol and keto forms in the charge transfer excited SCT((1)ππ*) state. The performance of white light emission is quantitatively demonstrated to be mainly sensitive to the molecular tailoring approach of the electronic properties of meta- or para- substituents by the modulation of the forward/backward ESIPT rate ratio. The quality of white light emission is slightly tunable through its surrounding solvent environment. These computational results will provide a useful strategy for the molecular design of OLED and WOLED materials.

  16. Evidence for the molecular-scale origin of the suppression of physical ageing in confined polymer: fluorescence and dielectric spectroscopy studies of polymer-silica nanocomposites

    International Nuclear Information System (INIS)

    Priestley, Rodney D; Rittigstein, Perla; Broadbelt, Linda J; Fukao, Koji; Torkelson, John M

    2007-01-01

    Fluorescence spectroscopy was used to characterize the rate of physical ageing at room temperature in nanocomposites of silica (10-15 nm diameter) nanoparticles in poly(methyl methacrylate) (PMMA). The physical ageing rate was reduced by more than a factor of 20 in 0.4 vol% silica-PMMA nanocomposites relative to neat PMMA. The molecular-scale origin of this nearly complete arresting of physical ageing was investigated with dielectric spectroscopy. The strength of the β relaxation process was reduced by nearly 50% in the nanocomposite relative to neat PMMA. This reduced strength of the β process results from dipoles (ester groups) having hindered motions or being virtually immobile on the timescale being probed at a frequency of 100 Hz. This hindered mobility results from hydrogen bonding between PMMA ester side groups and hydroxyl units on the surface of the silica nanoparticles. In contrast, no reduction in physical ageing rate was observed upon addition of silica to polystyrene, which cannot form hydrogen bonds with the silica surfaces. Thus, the molecular origin of the suppressed physical ageing in silica-PMMA nanocomposites is the interfacial hydrogen bonding, which leads to a major reduction in the strength of the β process, i.e., the β process is largely responsible for the observed physical ageing

  17. Fluorescent molecularly imprinted polymers as plastic antibodies for selective labeling and imaging of hyaluronan and sialic acid on fixed and living cells.

    Science.gov (United States)

    Panagiotopoulou, Maria; Kunath, Stephanie; Medina-Rangel, Paulina Ximena; Haupt, Karsten; Tse Sum Bui, Bernadette

    2017-02-15

    Altered glycosylation levels or distribution of sialic acids (SA) or hyaluronan in animal cells are indicators of pathological conditions like infection or malignancy. We applied fluorescently-labeled molecularly imprinted polymer (MIP) particles for bioimaging of fixed and living human keratinocytes, to localize hyaluronan and sialylation sites. MIPs were prepared with the templates D-glucuronic acid (GlcA), a substructure of hyaluronan, and N-acetylneuraminic acid (NANA), the most common member of SA. Both MIPs were found to be highly selective towards their target monosaccharides, as no cross-reactivity was observed with other sugars like N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-glucose and D-galactose, present on the cell surface. The dye rhodamine and two InP/ZnS quantum dots (QDs) emitting in the green and in the red regions were used as fluorescent probes. Rhodamine-MIPGlcA and rhodamine-MIPNANA were synthesized as monodispersed 400nm sized particles and were found to bind selectively their targets located in the extracellular region, as imaged by epifluorescence and confocal microscopy. In contrast, when MIP-GlcA and MIP-NANA particles with a smaller size (125nm) were used, the MIPs being synthesized as thin shells around green and red emitting QDs respectively, it was possible to stain the intracellular and pericellular regions as well. In addition, simultaneous dual-color imaging with the two different colored QDs-MIPs was demonstrated. Importantly, the MIPs were not cytotoxic and did not affect cell viability; neither was the cells morphology affected as demonstrated by live cell imaging. These synthetic receptors could offer a new and promising imaging tool to monitor disease progression. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Probing the binding of Cu(2+) ions to a fragment of the Aβ(1-42) polypeptide using fluorescence spectroscopy, isothermal titration calorimetry and molecular dynamics simulations.

    Science.gov (United States)

    Makowska, Joanna; Żamojć, Krzysztof; Wyrzykowski, Dariusz; Żmudzińska, Wioletta; Uber, Dorota; Wierzbicka, Małgorzata; Wiczk, Wiesław; Chmurzyński, Lech

    2016-09-01

    Steady-state and time-resolved fluorescence quenching measurements supported by isothermal titration calorimetry (ITC) and molecular dynamics simulations (MD), with the NMR-derived restraints, were used to investigate the interactions of Cu(2+) ions with a fragment of the Aβ(1-42) polypeptide, Aβ(5-16) with the following sequence: Ac-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-NH2, denoted as HZ1. The studies presented in this paper, when compared with our previous results (Makowska et al., Spectrochim. Acta A 153: 451-456), show that the affinity of the peptide to metal ions is conformation-dependent. All the measurements were carried out in 20mM 2-(N-morpholino)ethanesulfonic acid (MES) buffer solution, pH6.0. The Stern-Volmer equations, along with spectroscopic observations, were used to determine the quenching and binding parameters. The obtained results unequivocally suggest that Cu(2+) ions quench the fluorescence of HZ1 only through a static quenching mechanism, in contrast to the fragment from the N-terminal part of the FPB28 protein, with sequence Ac-Tyr-Lys-Thr-Ala-Asp-Gly-Lys-Thr-Tyr- NH2 (D9) and its derivative with a single point mutation: Ac-Tyr-Lys-Thr-Ala-Asn-Gly-Lys-Thr-Tyr- NH2 (D9_M), where dynamic quenching occurred. The thermodynamic parameters (ΔITCH, ΔITCS) for the interactions between Cu(2+) ions and the HZ1 peptide were determined from the calorimetric data. The conditional thermodynamic parameters suggest that, under the experimental conditions, the formation of the Cu(2+)-HZ1 complex is both an enthalpy and entropy driven process. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Gastrin-releasing peptide receptor-targeted gadolinium oxide-based multifunctional nanoparticles for dual magnetic resonance/fluorescent molecular imaging of prostate cancer

    Directory of Open Access Journals (Sweden)

    Cui DT

    2017-09-01

    Full Text Available Danting Cui,1 Xiaodan Lu,1 Chenggong Yan,1 Xiang Liu,1 Meirong Hou,1 Qi Xia,2 Yikai Xu,1 Ruiyuan Liu2,3 1Department of Medical Imaging Center, Nanfang Hospital, Southern Medical University, Guangzhou, People’s Republic of China; 2School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, People’s Republic of China; 3School of Biomedical Engineering, Southern Medical University, Guangzhou, People’s Republic of China Abstract: Bombesin (BBN, an analog of gastrin-releasing peptide (GRP, specifically binds to GRP receptors, which are overexpressed in human prostate cancer (PC. Here, we synthesized a BBN-modified gadolinium oxide (Gd2O3 nanoprobe containing fluorescein (Gd2O3-5(6-carboxyfluorescein [FI]-polyethylene glycol [PEG]-BBN for targeted magnetic resonance (MR/optical dual-modality imaging of PC. The Gd2O3-FI-PEG-BBN nanoparticles exhibited a relatively uniform particle size with an average diameter of 52.3 nm and spherical morphology as depicted by transmission electron microscopy. The longitudinal relaxivity (r1 of Gd2O3-FI-PEG-BBN (r1 =4.23 mM–1s–1 is comparable to that of clinically used Magnevist (Gd-DTPA. Fluorescence microscopy and in vitro cellular MRI demonstrated GRP receptor-specific and enhanced cellular uptake of the Gd2O3-FI-PEG-BBN in PC-3 tumor cells. Moreover, Gd2O3-FI-PEG-BBN showed more remarkable contrast enhancement than the corresponding nontargeted Gd2O3-FI-PEG according to in vivo MRI and fluorescent imaging. Tumor immunohistochemical analysis further demonstrated improved accumulation of the targeted nanoprobe in tumors. BBN-conjugated Gd2O3 may be a promising nanoplatform for simultaneous GRP receptor-targeted molecular cancer diagnosis and antitumor drug delivery in future clinical applications. Keywords: magnetic resonance imaging, gadolinium oxide, bombesin, gastrin-releasing peptide receptor, molecular imaging

  20. Instruments for radiation measurement in life sciences (5). ''Development of imaging technology in life science''. 9. Advantages of RI and fluorescence in imaging

    International Nuclear Information System (INIS)

    Furukawa, Takako; Jin, Zhao-Hui

    2009-01-01

    Imaging has been used as an effective research tool in many fields. In recent years, ''molecular imaging'' has come to attract a major attention as it studies molecular events in living animals and humans. Variety of modalities is used in molecular imaging, sometimes in combination, and the machines and techniques are going through rapid progress. Two of popular modalities among them are fluorescence imaging and radioisotope (RI) imaging such as positron emission tomography (PET) and single photon emission tomography (SPECT). Fluorescence imaging provides rich selection in imaging probes and the resolution can reach into sub-cellular level. RI imaging, especially PET, is superior to the others in quantitative analysis and the direct applicability to humans. In this article the two imaging modalities are overviewed comparing their characteristics. (author)

  1. Lateral distribution of NBD-PC fluorescent lipid analogs in membranes probed by molecular dynamics-assisted analysis of Förster Resonance Energy Transfer (FRET) and fluorescence quenching.

    Science.gov (United States)

    Loura, Luís M S

    2012-11-08

    Förster resonance energy transfer (FRET) is a powerful tool used for many problems in membrane biophysics, including characterization of the lateral distribution of lipid components and other species of interest. However, quantitative analysis of FRET data with a topological model requires adequate choices for the values of several input parameters, some of which are difficult to obtain experimentally in an independent manner. For this purpose, atomistic molecular dynamics (MD) simulations can be potentially useful as they provide direct detailed information on transverse probe localization, relative probe orientation, and membrane surface area, all of which are required for analysis of FRET data. This is illustrated here for the FRET pairs involving 1,6-diphenylhexatriene (DPH) as donor and either 1-palmitoyl,2-(6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] hexanoyl)- sn-glycero-3-phosphocholine (C6-NBD-PC) or 1-palmitoyl,2-(12-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]dodecanoyl)-sn-glycero-3-phosphocholine (C12-NBD-PC) as acceptors, in fluid vesicles of 1,2-dipalmitoyl-sn-3-glycerophosphocholine (DPPC, 50 °C). Incorporation of results from MD simulations improves the statistical quality of model fitting to the experimental FRET data. Furthermore, the decay of DPH in the presence of moderate amounts of C12-NBD-PC (>0.4 mol%) is consistent with non-random lateral distribution of the latter, at variance with C6-NBD-PC, for which aggregation is ruled out up to 2.5 mol% concentration. These conclusions are supported by analysis of NBD-PC fluorescence self-quenching. Implications regarding the relative utility of these probes in membrane studies are discussed.

  2. Molecular imaging: current status and emerging strategies

    International Nuclear Information System (INIS)

    Pysz, M.A.; Gambhir, S.S.; Willmann, J.K.

    2010-01-01

    In vivo molecular imaging has a great potential to impact medicine by detecting diseases in early stages (screening), identifying extent of disease, selecting disease- and patient-specific treatment (personalized medicine), applying a directed or targeted therapy, and measuring molecular-specific effects of treatment. Current clinical molecular imaging approaches primarily use positron-emission tomography (PET) or single photon-emission computed tomography (SPECT)-based techniques. In ongoing preclinical research, novel molecular targets of different diseases are identified and, sophisticated and multifunctional contrast agents for imaging these molecular targets are developed along with new technologies and instrumentation for multi-modality molecular imaging. Contrast-enhanced molecular ultrasound (US) with molecularly-targeted contrast microbubbles is explored as a clinically translatable molecular imaging strategy for screening, diagnosing, and monitoring diseases at the molecular level. Optical imaging with fluorescent molecular probes and US imaging with molecularly-targeted microbubbles are attractive strategies as they provide real-time imaging, are relatively inexpensive, produce images with high spatial resolution, and do not involve exposure to ionizing irradiation. Raman spectroscopy/microscopy has emerged as a molecular optical imaging strategy for ultrasensitive detection of multiple biomolecules/biochemicals with both in vivo and ex vivo versatility. Photoacoustic imaging is a hybrid of optical and US techniques involving optically-excitable molecularly-targeted contrast agents and quantitative detection of resulting oscillatory contrast agent movement with US. Current preclinical findings and advances in instrumentation, such as endoscopes and microcatheters, suggest that these molecular imaging methods have numerous potential clinical applications and will be translated into clinical use in the near future.

  3. Encapsulation of 3-hydroxyflavone and fisetin in β-cyclodextrins: Excited state proton transfer fluorescence and molecular mechanics studies

    Science.gov (United States)

    Banerjee, Anwesha; Sengupta, Pradeep K.

    2006-06-01

    Excited-state intramolecular proton-transfer (ESIPT) and dual emission properties (emission profile, anisotropy and decay kinetics) of 3-hydroxyflavone (a synthetic, model flavonol) and fisetin (3,7,3',4'-OH-flavone, a therapeutically active plant flavonol) have been exploited to study their encapsulation in nano-cavities comprising of natural and chemically modified β-cyclodextrins. In the presence of β-CDs, both the flavonols show significantly enhanced relative yields (along with changes in other emission parameters) of the tautomer emission. In addition, for fisetin, large blue shifts are observed for the normal emission (which has significant charge transfer character). From these we infer that the flavonols are encaged in predominantly hydrophobic micro-environments, where external hydrogen bonding perturbations (interfering with the intrinsic ESIPT), and dipolar relaxation effects, are minimized. This is further explained from results of molecular mechanics calculations which indicate selectivity in orientation of the encapsulated flavonols. Moreover, chemical modification of the β-CDs is found to profoundly influence the binding affinities of the guest flavonols.

  4. Establishment of a new human pleomorphic malignant fibrous histiocytoma cell line, FU-MFH-2: molecular cytogenetic characterization by multicolor fluorescence in situ hybridization and comparative genomic hybridization

    Directory of Open Access Journals (Sweden)

    Isayama Teruto

    2010-11-01

    Full Text Available Abstract Background Pleomorphic malignant fibrous histiocytoma (MFH is one of the most frequent malignant soft tissue tumors in adults. Despite the considerable amount of research on MFH cell lines, their characterization at a molecular cytogenetic level has not been extensively analyzed. Methods and results We established a new permanent human cell line, FU-MFH-2, from a metastatic pleomorphic MFH of a 72-year-old Japanese man, and applied multicolor fluorescence in situ hybridization (M-FISH, Urovysion™ FISH, and comparative genomic hybridization (CGH for the characterization of chromosomal aberrations. FU-MFH-2 cells were spindle or polygonal in shape with oval nuclei, and were successfully maintained in vitro for over 80 passages. The histological features of heterotransplanted tumors in severe combined immunodeficiency mice were essentially the same as those of the original tumor. Cytogenetic and M-FISH analyses displayed a hypotriploid karyotype with numerous structural aberrations. Urovysion™ FISH revealed a homozygous deletion of the p16INK4A locus on chromosome band 9p21. CGH analysis showed a high-level amplification of 9q31-q34, gains of 1p12-p34.3, 2p21, 2q11.2-q21, 3p, 4p, 6q22-qter, 8p11.2, 8q11.2-q21.1, 9q21-qter, 11q13, 12q24, 15q21-qter, 16p13, 17, 20, and X, and losses of 1q43-qter, 4q32-qter, 5q14-q23, 7q32-qter, 8p21-pter, 8q23, 9p21-pter, 10p11.2-p13, and 10q11.2-q22. Conclusion The FU-MFH-2 cell line will be a particularly useful model for studying molecular pathogenesis of human pleomorphic MFH.

  5. The use of molecular sieves to simulate hot lesions in (18)F-fluorodeoxyglucose--positron emission tomography imaging.

    Science.gov (United States)

    Matheoud, R; Secco, C; Ridone, S; Inglese, E; Brambilla, M

    2008-04-21

    We investigated the use of a kind of zeolite, the Bowie chabazite, to produce radioactive sources of different shapes, dimensions and activity concentrations that can be used for lesion simulation in positron emission tomography (PET) imaging. The (18)F-fluorodeoxyglucose ((18)F-FDG) uptake of a group of 12 zeolites was studied as a function of their weight (120-1,520 mg) and of the activity concentration of the (18)F-FDG solution (1-37 MBq ml(-1)), using a multiple linear regression model. The reproducibility, homogeneity and stability over time of the (18)F-FDG uptake were assessed. The fit of the regression model is good (r(2) = 0.83). This relation allows the production of zeolites of a desired (18)F-FDG activity using knowledge of the concentration of the soaking solution and the weight of the zeolite. The reproducibility of the (18)F-FDG uptake after heating the zeolites is elevated (CV% = 3.68). The almost complete regeneration of the zeolites allows us to reuse them in successive experiments. The stability of the (18)F-FDG uptake on zeolites is far from ideal. When placed in a saline solution the 'activated' zeolites release the (18)F-FDG with an effective half-time of 53 min. The sealing of the zeolites in plastic film bags has been demonstrated to be effective in preventing any release of (18)F-FDG. These features, together with their variable dimensions and shapes, make them ideal (18)F-FDG sources with a fixed target-to-background ratio that can be placed anywhere in a phantom to study lesion detectability in PET imaging.

  6. The use of molecular sieves to simulate hot lesions in 18F-fluorodeoxyglucose-positron emission tomography imaging

    International Nuclear Information System (INIS)

    Matheoud, R; Secco, C; Brambilla, M; Ridone, S; Inglese, E

    2008-01-01

    We investigated the use of a kind of zeolite, the Bowie chabazite, to produce radioactive sources of different shapes, dimensions and activity concentrations that can be used for lesion simulation in positron emission tomography (PET) imaging. The 18 F-fluorodeoxyglucose ( 18 F-FDG) uptake of a group of 12 zeolites was studied as a function of their weight (120-1520 mg) and of the activity concentration of the 18 F-FDG solution (1-37 MBq ml -1 ), using a multiple linear regression model. The reproducibility, homogeneity and stability over time of the 18 F-FDG uptake were assessed. The fit of the regression model is good (r 2 = 0.83). This relation allows the production of zeolites of a desired 18 F-FDG activity using knowledge of the concentration of the soaking solution and the weight of the zeolite. The reproducibility of the 18 F-FDG uptake after heating the zeolites is elevated (CV% = 3.68). The almost complete regeneration of the zeolites allows us to reuse them in successive experiments. The stability of the 18 F-FDG uptake on zeolites is far from ideal. When placed in a saline solution the 'activated' zeolites release the 18 F-FDG with an effective half-time of 53 min. The sealing of the zeolites in plastic film bags has been demonstrated to be effective in preventing any release of 18 F-FDG. These features, together with their variable dimensions and shapes, make them ideal 18 F-FDG sources with a fixed target-to-background ratio that can be placed anywhere in a phantom to study lesion detectability in PET imaging. (note)

  7. Detection of intramyocardially injected DiR-labeled mesenchymal stem cells by optical and optoacoustic tomography.

    Science.gov (United States)

    Berninger, Markus T; Mohajerani, Pouyan; Wildgruber, Moritz; Beziere, Nicolas; Kimm, Melanie A; Ma, Xiaopeng; Haller, Bernhard; Fleming, Megan J; Vogt, Stephan; Anton, Martina; Imhoff, Andreas B; Ntziachristos, Vasilis; Meier, Reinhard; Henning, Tobias D

    2017-06-01

    The distribution of intramyocardially injected rabbit MSCs, labeled with the near-infrared dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbo-cyanine-iodide (DiR) using hybrid Fluorescence Molecular Tomography-X-ray Computed Tomography (FMT-XCT) and Multispectral Optoacoustic Tomography (MSOT) imaging technologies, was investigated. Viability and induction of apoptosis of DiR labeled MSCs were assessed by XTT- and Caspase-3/-7-testing in vitro . 2 × 10 6 , 2 × 10 5 and 2 × 10 4 MSCs labeled with 5 and 10 μg DiR/ml were injected into fresh frozen rabbit hearts. FMT-XCT, MSOT and fluorescence cryosection imaging were performed. Concentrations up to 10 μg DiR/ml did not cause apoptosis in vitro (p > 0.05). FMT and MSOT imaging of labeled MSCs led to a strong signal. The imaging modalities highlighted a difference in cell distribution and concentration correlated to the number of injected cells. Ex-vivo cryosectioning confirmed the molecular fluorescence signal. FMT and MSOT are sensitive imaging techniques offering high-anatomic resolution in terms of detection and distribution of intramyocardially injected stem cells in a rabbit model.

  8. A chemosensor showing discriminating fluorescent response for highly selective and nanomolar detection of Cu²⁺ and Zn²⁺ and its application in molecular logic gate.

    Science.gov (United States)

    Fegade, Umesh A; Sahoo, Suban K; Singh, Amanpreet; Singh, Narinder; Attarde, Sanjay B; Kuwar, Anil S

    2015-05-04

    A fluorescent based receptor (4Z)-4-(4-diethylamino)-2-hydroxybenzylidene amino)-1,2dihydro-1,5-dimethyl-2-phenylpyrazol-3-one (receptor 3) was developed for the highly selective and sensitive detection of Cu(2+) and Zn(2+) in semi-aqueous system. The fluorescence of receptor 3 was enhanced and quenched, respectively, with the addition of Zn(2+) and Cu(2+) ions over other surveyed cations. The receptor formed host-guest complexes in 1:1 stoichiometry with the detection limit of 5 nM and 15 nM for Cu(2+) and Zn(2+) ions, respectively. Further, we have effectively utilized the two metal ions (Cu(2+) and Zn(2+)) as chemical inputs for the manufacture of INHIBIT type logic gate at molecular level using the fluorescence responses of receptor 3 at 450 nm. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Simultaneous optical coherence tomography and lipofuscin autofluorescence imaging of the retina with a single broadband light source at 480nm

    OpenAIRE

    Jiang, Minshan; Liu, Tan; Liu, Xiaojing; Jiao, Shuliang

    2014-01-01

    We accomplished spectral domain optical coherence tomography and auto-fluorescence microscopy for imaging the retina with a single broadband light source centered at 480 nm. This technique is able to provide simultaneous structural imaging and lipofuscin molecular contrast of the retina. Since the two imaging modalities are provided by the same group of photons, their images are intrinsically registered. To test the capabilities of the technique we periodically imaged the retinas of the same ...

  10. Order of magnitude sensitivity increase in X-ray Fluorescence Computed Tomography (XFCT) imaging with an optimized spectro-spatial detector configuration: theory and simulation.

    Science.gov (United States)

    Ahmad, Moiz; Bazalova, Magdalena; Xiang, Liangzhong; Xing, Lei

    2014-05-01

    The purpose of this study was to increase the sensitivity of XFCT imaging by optimizing the data acquisition geometry for reduced scatter X-rays. The placement of detectors and detector energy window were chosen to minimize scatter X-rays. We performed both theoretical calculations and Monte Carlo simulations of this optimized detector configuration on a mouse-sized phantom containing various gold concentrations. The sensitivity limits were determined for three different X-ray spectra: a monoenergetic source, a Gaussian source, and a conventional X-ray tube source. Scatter X-rays were minimized using a backscatter detector orientation (scatter direction > 110(°) to the primary X-ray beam). The optimized configuration simultaneously reduced the number of detectors and improved the image signal-to-noise ratio. The sensitivity of the optimized configuration was 10 μg/mL (10 pM) at 2 mGy dose with the mono-energetic source, which is an order of magnitude improvement over the unoptimized configuration (102 pM without the optimization). Similar improvements were seen with the Gaussian spectrum source and conventional X-ray tube source. The optimization improvements were predicted in the theoretical model and also demonstrated in simulations. The sensitivity of XFCT imaging can be enhanced by an order of magnitude with the data acquisition optimization, greatly enhancing the potential of this modality for future use in clinical molecular imaging.

  11. An ultrasensitive and highly selective fluorescent and colorimetric chemosensor for citrate ions based on rhodamine B and its application as the first molecular security keypad lock based on phosphomolybdic acid and citrate inputs

    Energy Technology Data Exchange (ETDEWEB)

    Tavallali, Hossein, E-mail: Tavallali@pnu.ac.ir; Baezzat, Mohammad-Reza; Deilamy-Rad, Gohar; Parhami, Abolfath; Hasanli, Nahid

    2015-04-15

    Rhodamine B (Rh{sub B}) has been developed as novel and efficient colorimetric and fluorometric chemosensor for citrate ions (Cit{sup 3−}) in an absolutely aqueous media. The UV–vis absorption and fluorescent emission titrations experiments have been employed to study the sensing process. Rh{sub B} could act as an efficient “ON–OFF” fluorescent chemosensor for phosphomolybdic acid (PMA) based on an electron transfer (ET) process. Also (Rh{sub B}{sup +}){sub 3}.PMA{sup 3−} could operate as an “OFF–ON” fluorescent chemosensor for citrate ions based on a ligand substitution process. The chemosensor Rh{sub B} shows excellent fluorescence sensitivity and selectivity toward citrate in aqueous media, and displays ON–OFF–ON type fluorescence change with alternately adding PMA and citrate to the media along with reversible association–dissociation of the complex. The (Rh{sub B}{sup +}){sub 3}.PMA{sup 3−} can be applied to the quantification of citrate with a linear ranges covering from 0.053 to 0.83 and 0.08 to 1.6 µM by detection limits of 6.0 and 9.1 nM for fluorescence and colorimetric methods respectively. The keypad lock operation is particularly important, as the output of the system depends not only on the proper combination but also on the order of input signals, creating the correct password that can be used to “open” this molecular keypad lock through strong fluorescence emission at 575 nm. As a whole, its various logic gate properties may improve its impact for the development of new-generation “intelligence” digital devices. The ionic PMA and Cit{sup 3−} inputs to (Rh{sub B}{sup +}){sub 3}.PMA{sup 3−} have been mimicked as a superimposed electronic molecular keypad lock. Also indicates that Rh{sub B} is suitable for the detection of Cit{sup 3−} ions in the biological environment. - Highlights: • Our probe is commercially available with good photostability and high quantum yield. • Both color and fluorescence change

  12. Fluorescence spectroscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2016-01-01

    Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses the foundati......Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses...

  13. TH-AB-209-01: Making Benchtop X-Ray Fluorescence Computed Tomography (XFCT) Practical for in Vivo Imaging by Integration of a Dedicated High-Performance X-Ray Source in Conjunction with Micro-CT Functionality

    International Nuclear Information System (INIS)

    Manohar, N; Cho, S; Reynoso, F

    2016-01-01

    Purpose: To make benchtop x-ray fluorescence computed tomography (XFCT) practical for routine preclinical imaging tasks with gold nanoparticles (GNPs) by deploying, integrating, and characterizing a dedicated high-performance x-ray source and addition of simultaneous micro-CT functionality. Methods: Considerable research effort is currently under way to develop a polychromatic benchtop cone-beam XFCT system capable of imaging GNPs by stimulation and detection of gold K-shell x-ray fluorescence (XRF) photons. Recently, an ad hoc high-power x-ray source was incorporated and used to image the biodistribution of GNPs within a mouse, postmortem. In the current work, a dedicated x-ray source system featuring a liquid-cooled tungsten-target x-ray tube (max 160 kVp, ∼3 kW power) was deployed. The source was operated at 125 kVp, 24 mA. The tube’s compact dimensions allowed greater flexibility for optimizing both the irradiation and detection geometries. Incident x-rays were shaped by a conical collimator and filtered by 2 mm of tin. A compact “OEM” cadmium-telluride x-ray detector was implemented for detecting XRF/scatter spectra. Additionally, a flat panel detector was installed to allow simultaneous transmission CT imaging. The performance of the system was characterized by determining the detection limit (10-second acquisition time) for inserts filled with water/GNPs at various concentrations (0 and 0.010–1.0 wt%) and embedded in a small-animal-sized phantom. The phantom was loaded with 0.5, 0.3, and 0.1 wt% inserts and imaged using XFCT and simultaneous micro-CT. Results: An unprecedented detection limit of 0.030 wt% was experimentally demonstrated, with a 33% reduction in acquisition time. The reconstructed XFCT image accurately localized the imaging inserts. Micro-CT imaging did not provide enough contrast to distinguish imaging inserts from the phantom under the current conditions. Conclusion: The system is immediately capable of in vivo preclinical XFCT

  14. TH-AB-209-01: Making Benchtop X-Ray Fluorescence Computed Tomography (XFCT) Practical for in Vivo Imaging by Integration of a Dedicated High-Performance X-Ray Source in Conjunction with Micro-CT Functionality

    Energy Technology Data Exchange (ETDEWEB)

    Manohar, N; Cho, S [UT MD Anderson Cancer Center, Houston, TX (United States); Reynoso, F [UT MD Anderson Cancer Center, Houston, TX (United States); Washington University School of Medicine, St. Louis, MO (United States)

    2016-06-15

    Purpose: To make benchtop x-ray fluorescence computed tomography (XFCT) practical for routine preclinical imaging tasks with gold nanoparticles (GNPs) by deploying, integrating, and characterizing a dedicated high-performance x-ray source and addition of simultaneous micro-CT functionality. Methods: Considerable research effort is currently under way to develop a polychromatic benchtop cone-beam XFCT system capable of imaging GNPs by stimulation and detection of gold K-shell x-ray fluorescence (XRF) photons. Recently, an ad hoc high-power x-ray source was incorporated and used to image the biodistribution of GNPs within a mouse, postmortem. In the current work, a dedicated x-ray source system featuring a liquid-cooled tungsten-target x-ray tube (max 160 kVp, ∼3 kW power) was deployed. The source was operated at 125 kVp, 24 mA. The tube’s compact dimensions allowed greater flexibility for optimizing both the irradiation and detection geometries. Incident x-rays were shaped by a conical collimator and filtered by 2 mm of tin. A compact “OEM” cadmium-telluride x-ray detector was implemented for detecting XRF/scatter spectra. Additionally, a flat panel detector was installed to allow simultaneous transmission CT imaging. The performance of the system was characterized by determining the detection limit (10-second acquisition time) for inserts filled with water/GNPs at various concentrations (0 and 0.010–1.0 wt%) and embedded in a small-animal-sized phantom. The phantom was loaded with 0.5, 0.3, and 0.1 wt% inserts and imaged using XFCT and simultaneous micro-CT. Results: An unprecedented detection limit of 0.030 wt% was experimentally demonstrated, with a 33% reduction in acquisition time. The reconstructed XFCT image accurately localized the imaging inserts. Micro-CT imaging did not provide enough contrast to distinguish imaging inserts from the phantom under the current conditions. Conclusion: The system is immediately capable of in vivo preclinical XFCT

  15. Introduction to fluorescence

    CERN Document Server

    Jameson, David M

    2014-01-01

    "An essential contribution to educating scientists in the principles of fluorescence. It will also be an important addition to the libraries of practitioners applying the principles of molecular fluorescence."-Ken Jacobson, Kenan Distinguished Professor of Cell Biology and Physiology, University of North Carolina at Chapel Hill"An exquisite compendium of fluorescence and its applications in biochemistry enriched by a very exciting historical perspective. This book will become a standard text for graduate students and other scientists."-Drs. Zygmunt (Karol) Gryczynski and Ignacy Gryczynski, University of North Texas Health Science Center"… truly a masterwork, combining clarity, precision, and good humor. The reader, novice or expert, will be pleased with the text and will not stop reading. It is a formidable account of the fluorescence field, which has impacted the life sciences so considerably in the last 60 years."-Jerson L. Silva, M.D., Ph.D., Professor and Director, National Institute of Science and Tech...

  16. Pittsburgh Compound B and AV-1451 positron emission tomography assessment of molecular pathologies of Alzheimer's disease in progressive supranuclear palsy.

    Science.gov (United States)

    Whitwell, Jennifer L; Ahlskog, J Eric; Tosakulwong, Nirubol; Senjem, Matthew L; Spychalla, Anthony J; Petersen, Ronald C; Jack, Clifford R; Lowe, Val J; Josephs, Keith A

    2018-03-01

    Little is known about Alzheimer's disease molecular proteins, beta-amyloid and paired helical filament (PHF) tau, in progressive supranuclear palsy (PSP). Recent techniques have been developed to allow for investigations of these proteins in PSP. We determined the frequency of beta-amyloid deposition in PSP, and whether beta-amyloid deposition in PSP is associated with PHF-tau deposition pattern, or clinical features. Thirty probable PSP participants underwent MRI, [ 18 F]AV-1451 PET and Pittsburgh compound B (PiB) PET. Apolipoprotein (APOE) genotyping was also performed. A global PiB standard-uptake value ratio (SUVR) was calculated. AV-1451 SUVRs were calculated for a set of Alzheimer's disease (AD)-related regions and a set of PSP-related regions. Voxel-level analyses were conducted to assess for differences in AV-1451 uptake patterns and MRI atrophy between PiB(+) and PiB(-) cases compared to 60 normal PiB(-) controls. Statistical testing for correlations and associations between variables of interest were also performed. Twelve subjects (40%) showed beta-amyloid deposition. Higher PiB SUVR correlated with older age but not with AV-1451 SUVR in the AD- or PSP-related regions. Higher AV-1451 SUVR in AD-related regions was associated with higher AV-1451 SUVR in PSP-related regions. We found little evidence for beta-amyloid related differences in clinical metrics, proportion of APOE e4 carriers, pattern of AV-1451 uptake, or pattern of atrophy. Beta-amyloid deposition occurs in a relatively high proportion of PSP subjects. Unlike in Alzheimer's disease, however, there is little evidence that beta-amyloid, and PHF-tau, play a significant role in neurodegeneration in PSP. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Correlative cryogenic tomography of cells using light and soft x-rays

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Elizabeth A.; Cinquin, Bertrand P.; Do, Myan; McDermott, Gerry [Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA (United States); National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA (United States); Le Gros, Mark A., E-mail: MALegros@lbl.gov [Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA (United States); Physical BioSciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA (United States); National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA (United States); Larabell, Carolyn A., E-mail: carolyn.larabell@ucsf.edu [Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA (United States); Physical BioSciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA (United States); National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA (United States)

    2014-08-01

    Correlated imaging is the process of imaging a specimen with two complementary modalities, and then combining the two data sets to create a highly informative, composite view. A recent implementation of this concept has been the combination of soft x-ray tomography (SXT) with fluorescence cryogenic microscopy (FCM). SXT–FCM is used to visualize cells that are held in a near-native, cryopreserved. The resultant images are, therefore, highly representative of both the cellular architecture and molecular organization in vivo. SXT quantitatively visualizes the cell and sub-cellular structures; FCM images the spatial distribution of fluorescently labeled molecules. Here, we review the characteristics of SXT–FCM, and briefly discuss how this method compares with existing correlative imaging techniques. We also describe how the incorporation of a cryo-rotation stage into a cryogenic fluorescence microscope allows acquisition of fluorescence cryogenic tomography (FCT) data. FCT is optimally suited for correlation with SXT, since both techniques image the specimen in 3-D, potentially with similar, isotropic spatial resolution. - Highlights: • We describe a new correlated imaging modality: soft x-ray tomography combined (SXT) with confocal fluorescence tomography (CFT). • Data from the two modalities are combined accurately and precisely using fiducials visible in both types of data. • Cells imaged by SXT–CFT are maintained close to their native state by cryo-preservation. • SXT–CFT is applicable to most cell types, especially cells grown in suspension. • ‘Super-resolution’ microscopes being developed for CFT data acquisition match the spatial resolution of SXT.

  18. Characterization of a unique tomaymycin-d(CICGAATTCICG)2 adduct containing two drug molecules per duplex by NMR, fluorescence, and molecular modeling studies

    International Nuclear Information System (INIS)

    Boyd, F.L.; Stewart, D.; Hurley, L.H.; Remers, W.A.; Barkley, M.D.

    1990-01-01

    Tomaymycin is a member of the pyrrolo[1,4]benzodiazepine [P(1,4)B] antitumor antibiotic group. This antibiotic is proposed to react with the exocyclic 2-amino group (N2) of guanine to form a covalent adduct that lies snugly within the minor groove of DNA. While DNA-footprinting experiments using methidiumpropyl-EDTA have revealed the favored bonding sequences for tomaymycin and related drugs on DNA, the stereochemistry at the covalent bonding site (C-11) and orientation in the minor groove were not established by these experiments. In previous studies using a combined fluorescence, high-field NMR, and molecular modeling approach, the authors have shown that for tomaymycin there are two diastereomeric species (11R and 11S) on both calf thymus DNA and d(ATGCAT) 2 . Although they were able to infer the identify of the two species on d(ATGCAT) 2 , definitive experimental evidence was lacking. They have designed and synthesized a self-complementary 12-mer [d(CICGAATTCICG) 2 ] based on the Dickerson dodecamer [d(CGCGAATTCGCG) 2 ] that bonds identically two tomaymycin molecules, each having a defined orientation and stereochemistry. The results presented in this study together with previous investigations show that the orientation of the drug molecule in the minor groove, and stereochemistry at the covalent linkage site, is dependent upon both the flanking sequence and drug structure. This conclusion mandates caution be used in rationalizing the biochemical and and biological effects of P(1,4)B bonding to DNA until precise structural information is established

  19. Computerized tomography

    International Nuclear Information System (INIS)

    Rubashov, I.B.

    1985-01-01

    Operating principle is described for the devices of computerized tomography used in medicine for diagnosis of brain diseases. Computerized tomography is considered as a part of computerized diagnosis, as a part of information science. It is shown that computerized tomography is a real existed field of investigations in medicine and industrial production

  20. Development of novel emission tomography system

    Science.gov (United States)

    Fu, Geng

    In recent years, small animals, such as mice and rats, have been widely used as subjects of study in biomedical research while molecular biology and imaging techniques open new opportunities to investigate disease model. With the help of medical imaging techniques, researchers can investigate underlying mechanisms inside the small animal, which are useful for both early diagnosis and treatment monitoring. Based on tracer principle single photon emission computed tomography (SPECT) has increased popularity in small animal imaging due to its higher spatial resolution and variety of single-photon emitting radionuclides. Since the image quality strongly depends on the detector properties, both scintillation and semiconductor detectors are under active investigation for high resolution X-ray and gamma ray photon detection. The desired detector properties include high intrinsic spatial resolution, high energy resolution, and high detection efficiency. In this thesis study, we have made extensive efforts to develop novel emission tomography system, and evaluate the use of both semiconductor and ultra-high resolution scintillation detectors for small animal imaging. This thesis work includes the following three areas. Firstly, we have developed a novel energy-resolved photon counting (ERPC) detector. With the benefits of high energy resolution, high spatial resolution, flexible detection area, and a wide dynamic range of 27--200keV, ERPC detector is well-suited for small animal SPECT applications. For prototype ERPC detector excellent imaging (˜350microm) and spectroscopic performance (4keV Co-57 122keV) has been demonstrated in preliminary study. Secondly, to further improve spatial resolution to hundred-micron level, an ultra-high resolution Intensified EMCCD (I-EMCCD) detector has been designed and evaluated. This detector consists of the newly developed electron multiplying CCD (EMCCD) sensor, columnar CsI(Tl) scintillator, and an electrostatic de-magnifier (DM) tube

  1. Hybrid system for in vivo real-time planar fluorescence and volumetric optoacoustic imaging

    Science.gov (United States)

    Chen, Zhenyue; Deán-Ben, Xosé Luís.; Gottschalk, Sven; Razansky, Daniel

    2018-02-01

    Fluorescence imaging is widely employed in all fields of cell and molecular biology due to its high sensitivity, high contrast and ease of implementation. However, the low spatial resolution and lack of depth information, especially in strongly-scattering samples, restrict its applicability for deep-tissue imaging applications. On the other hand, optoacoustic imaging is known to deliver a unique set of capabilities such as high spatial and temporal resolution in three dimensions, deep penetration and spectrally-enriched imaging contrast. Since fluorescent substances can generate contrast in both modalities, simultaneous fluorescence and optoacoustic readings can provide new capabilities for functional and molecular imaging of living organisms. Optoacoustic images can further serve as valuable anatomical references based on endogenous hemoglobin contrast. Herein, we propose a hybrid system for in vivo real-time planar fluorescence and volumetric optoacoustic tomography, both operating in reflection mode, which synergistically combines the advantages of stand-alone systems. Validation of the spatial resolution and sensitivity of the system were first carried out in tissue mimicking phantoms while in vivo imaging was further demonstrated by tracking perfusion of an optical contrast agent in a mouse brain in the hybrid imaging mode. Experimental results show that the proposed system effectively exploits the contrast mechanisms of both imaging modalities, making it especially useful for accurate monitoring of fluorescence-based signal dynamics in highly scattering samples.

  2. Molecular imaging of oncolytic viral therapy

    Directory of Open Access Journals (Sweden)

    Dana Haddad

    2014-01-01

    Full Text Available Oncolytic viruses have made their mark on the cancer world as a potential therapeutic option, with the possible advantages of reduced side effects and strengthened treatment efficacy due to higher tumor selectivity. Results have been so promising, that oncolytic viral treatments have now been approved for clinical trials in several countries. However, clinical studies may benefit from the ability to noninvasively and serially identify sites of viral targeting via molecular imaging in order to provide safety, efficacy, and toxicity information. Furthermore, molecular imaging of oncolytic viral therapy may provide a more sensitive and specific diagnostic technique to detect tumor origin and, more importantly, presence of metastases. Several strategies have been investigated for molecular imaging of viral replication broadly categorized into optical and deep tissue imaging, utilizing several reporter genes encoding for fluorescence proteins, conditional enzymes, and membrane protein and transporters. Various imaging methods facilitate molecular imaging, including computer tomography, magnetic resonance imaging, positron emission tomography, single photon emission CT, gamma-scintigraphy, and photoacoustic imaging. In addition, several molecular probes are used for medical imaging, which act as targeting moieties or signaling agents. This review will explore the preclinical and clinical use of in vivo molecular imaging of replication-competent oncolytic viral therapy.

  3. A fluorescence scanning electron microscope

    International Nuclear Information System (INIS)

    Kanemaru, Takaaki; Hirata, Kazuho; Takasu, Shin-ichi; Isobe, Shin-ichiro; Mizuki, Keiji; Mataka, Shuntaro; Nakamura, Kei-ichiro

    2009-01-01

    Fluorescence techniques are widely used in biological research to examine molecular localization, while electron microscopy can provide unique ultrastructural information. To date, correlative images from both fluorescence and electron microscopy have been obtained separately using two different instruments, i.e. a fluorescence microscope (FM) and an electron microscope (EM). In the current study, a scanning electron microscope (SEM) (JEOL JXA8600 M) was combined with a fluorescence digital camera microscope unit and this hybrid instrument was named a fluorescence SEM (FL-SEM). In the labeling of FL-SEM samples, both Fluolid, which is an organic EL dye, and Alexa Fluor, were employed. We successfully demonstrated that the FL-SEM is a simple and practical tool for correlative fluorescence and electron microscopy.

  4. Recent developments in multimodality fluorescence imaging probes

    Directory of Open Access Journals (Sweden)

    Jianhong Zhao

    2018-05-01

    Full Text Available Multimodality optical imaging probes have emerged as powerful tools that improve detection sensitivity and accuracy, important in disease diagnosis and treatment. In this review, we focus on recent developments of optical fluorescence imaging (OFI probe integration with other imaging modalities such as X-ray computed tomography (CT, magnetic resonance imaging (MRI, positron emission tomography (PET, single-photon emission computed tomography (SPECT, and photoacoustic imaging (PAI. The imaging technologies are briefly described in order to introduce the strengths and limitations of each techniques and the need for further multimodality optical imaging probe development. The emphasis of this account is placed on how design strategies are currently implemented to afford physicochemically and biologically compatible multimodality optical fluorescence imaging probes. We also present studies that overcame intrinsic disadvantages of each imaging technique by multimodality approach with improved detection sensitivity and accuracy. KEY WORDS: Optical imaging, Fluorescence, Multimodality, Near-infrared fluorescence, Nanoprobe, Computed tomography, Magnetic resonance imaging, Positron emission tomography, Single-photon emission computed tomography, Photoacoustic imaging

  5. Proton and electron impact on molecular and atomic oxygen: I. High resolution fluorescence spectra in the visible and VUV spectral range and emission cross-sections for dissociative ionisation and excitation of O2

    International Nuclear Information System (INIS)

    Wilhelmi, O.; Schartner, K.H.

    2000-01-01

    For pt.II see ibid., vol.11, p.45-58, 2000. Molecular oxygen O 2 was dissociated in collisions with protons and electrons in the intermediate velocity range (p + -energies: 17-800 keV, e - -energies: 0.2-2 keV). Fluorescence from excited atomic and singly ionised fragments and from singly ionised molecules was detected in the VUV and in the visible and near UV spectral range. Highly resolved spectra are presented for the VUV (46-131 nm) and the near UV/visible (340-605 nm) spectral range. Absolute emission cross-sections have been determined for dissociative ionisation and excitation leading to fluorescence in the VUV. Results are compared with published data. (orig.)

  6. Ratiometric fluorescent receptors for both Zn2+ and H2PO4(-) ions based on a pyrenyl-linked triazole-modified homooxacalix[3]arene: a potential molecular traffic signal with an R-S latch logic circuit.

    Science.gov (United States)

    Ni, Xin-long; Zeng, Xi; Redshaw, Carl; Yamato, Takehiko

    2011-07-15

    A ratiometric fluorescent receptor with a C(3) symmetric structure based on a pyrene-linked triazole-modified homooxacalix[3]arene (L) was synthesized and characterized. This system exhibited an interesting ratiometric detection signal output for targeting cations and anions through switching the excimer emission of pyrene from the "on-off" to the "off-on" type in neutral solution. (1)H NMR titration results suggested that the Zn(2+) center of receptor L·Zn(2+) provided an excellent pathway of organizing anion binding groups for optimal host-guest interactions. It is thus believed that this receptor has potential application in sensing, detection, and recognition of both Zn(2+) and H(2)PO(4)(-) ions with different optical signals. In addition, the fluorescence emission changes by the inputs of Zn(2+) and H(2)PO(4)(-) ions can be viewed as a combinational R-S latch logic circuit at the molecular level.

  7. In vivo tomographic imaging of lung colonization of tumour in mouse with simultaneous fluorescence and X-ray CT.

    Science.gov (United States)

    Zhang, Bin; Gao, Fuping; Wang, Mengjiao; Cao, Xu; Liu, Fei; Wang, Xin; Luo, Jianwen; Wang, Guangzhi; Bai, Jing

    2014-01-01

    Non-invasive in vivo imaging of diffuse and wide-spread colonization within the lungs, rather than distinct solid primary tumors, is still a challenging work. In this work, a lung colonization mouse model bearing A549 human lung tumor was simultaneously scanned by a dual-modality fluorescence molecular tomography (FMT) and X-ray computed tomography (CT) system in vivo. A two steps method which incorporates CT structural information into the FMT reconstruction procedure is employed to provide concurrent anatomical and functional information. By using the target-specific fluorescence agent, the fluorescence tomographic results show elevated fluorescence intensity deep within the lungs which is colonized with diffuse and wide-spread tumors. The results were confirmed with ex vivo fluorescence reflectance imaging and histological examination of the lung tissues. With FMT reconstruction combined with the CT information, the dual-modality FMT/micro-CT system is expected to offer sensitive and noninvasive imaging of diffuse tumor colonization within the lungs in vivo. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Molecular imaging and optical diagnosis from single molecule to human body

    International Nuclear Information System (INIS)

    Tamura, Mamoru

    2006-01-01

    The combination of molecular biology and optelectronics has given rise to open a new field, bio-photonics, in the 21st century. In this review, recent advances in several in vitro and in vivo single-molecule detection methods for animals are discussed. The possible applications of optical diagnosis are also included, which are optical mammography, diffuse optical tomography and fluorescence endoscopy. The potential of the light use of in diagnosis is emphasized. (author)

  9. Fluorescent molecularly imprinted polymer thin films for specific protein detection prepared with dansyl ethylenediamine-conjugated O-acryloyl L-hydroxyproline.

    Science.gov (United States)

    Inoue, Yuki; Kuwahara, Atsushi; Ohmori, Kohei; Sunayama, Hirobumi; Ooya, Tooru; Takeuchi, Toshifumi

    2013-10-15

    Protein-imprinted polymers, capable of specific transduction of protein binding events into fluorescent signal change, were designed and synthesized by using dansyl ethylenediamine-conjugated O-acryloyl L-hydroxyproline (Hyp-En-Dans). Human serum albumin (HSA) was used as a model target protein and HSA-imprinted polymers (HSA-IP) were prepared on glass substrates. Specific fluorescence change was observed for HSA binding on the imprinted polymer thin film, whereas a weaker response was observed for other proteins, including bovine serum albumin, chymotrypsin, lysozyme, and avidin. The binding specificity was found to derive from the rigid structure of the hydrogen-bondable pyrrolidine moiety. Compared with SPR measurements, the non-specific binding caused by the polymer matrix and/or randomly located fluorescent monomer residues that did not compose specific binding sites did not contribute to the observed fluorescence change. These results revealed that the proposed protein-imprinting technique using Hyp-En-Dans could provide a highly selective protein-sensing platform, in which only specific binding events would be detected by fluorescent measurements. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Validation of a simple and fast method to quantify in vitro mineralization with fluorescent probes used in molecular imaging of bone

    International Nuclear Information System (INIS)

    Moester, Martiene J.C.; Schoeman, Monique A.E.; Oudshoorn, Ineke B.; Beusekom, Mara M. van; Mol, Isabel M.; Kaijzel, Eric L.; Löwik, Clemens W.G.M.; Rooij, Karien E. de

    2014-01-01

    Highlights: •We validate a simple and fast method of quantification of in vitro mineralization. •Fluorescently labeled agents can detect calcium deposits in the mineralized matrix of cell cultures. •Fluorescent signals of the probes correlated with Alizarin Red S staining. -- Abstract: Alizarin Red S staining is the standard method to indicate and quantify matrix mineralization during differentiation of osteoblast cultures. KS483 cells are multipotent mouse mesenchymal progenitor cells that can differentiate into chondrocytes, adipocytes and osteoblasts and are a well-characterized model for the study of bone formation. Matrix mineralization is the last step of differentiation of bone cells and is therefore a very important outcome measure in bone research. Fluorescently labelled calcium chelating agents, e.g. BoneTag and OsteoSense, are currently used for in vivo imaging of bone. The aim of the present study was to validate these probes for fast and simple detection and quantification of in vitro matrix mineralization by KS483 cells and thus enabling high-throughput screening experiments. KS483 cells were cultured under osteogenic conditions in the presence of compounds that either stimulate or inhibit osteoblast differentiation and thereby matrix mineralization. After 21 days of differentiation, fluorescence of stained cultures was quantified with a near-infrared imager and compared to Alizarin Red S quantification. Fluorescence of both probes closely correlated to Alizarin Red S staining in both inhibiting and stimulating conditions. In addition, both compounds displayed specificity for mineralized nodules. We therefore conclude that this method of quantification of bone mineralization using fluorescent compounds is a good alternative for the Alizarin Red S staining

  11. Validation of a simple and fast method to quantify in vitro mineralization with fluorescent probes used in molecular imaging of bone

    Energy Technology Data Exchange (ETDEWEB)

    Moester, Martiene J.C. [Department of Radiology, Leiden University Medical Center (Netherlands); Schoeman, Monique A.E. [Department of Orthopedic Surgery, Leiden University Medical Center (Netherlands); Oudshoorn, Ineke B. [Department of Radiology, Leiden University Medical Center (Netherlands); Percuros BV, Leiden (Netherlands); Beusekom, Mara M. van [Department of Radiology, Leiden University Medical Center (Netherlands); Mol, Isabel M. [Department of Radiology, Leiden University Medical Center (Netherlands); Percuros BV, Leiden (Netherlands); Kaijzel, Eric L.; Löwik, Clemens W.G.M. [Department of Radiology, Leiden University Medical Center (Netherlands); Rooij, Karien E. de, E-mail: k.e.de_rooij@lumc.nl [Department of Radiology, Leiden University Medical Center (Netherlands); Percuros BV, Leiden (Netherlands)

    2014-01-03

    Highlights: •We validate a simple and fast method of quantification of in vitro mineralization. •Fluorescently labeled agents can detect calcium deposits in the mineralized matrix of cell cultures. •Fluorescent signals of the probes correlated with Alizarin Red S staining. -- Abstract: Alizarin Red S staining is the standard method to indicate and quantify matrix mineralization during differentiation of osteoblast cultures. KS483 cells are multipotent mouse mesenchymal progenitor cells that can differentiate into chondrocytes, adipocytes and osteoblasts and are a well-characterized model for the study of bone formation. Matrix mineralization is the last step of differentiation of bone cells and is therefore a very important outcome measure in bone research. Fluorescently labelled calcium chelating agents, e.g. BoneTag and OsteoSense, are currently used for in vivo imaging of bone. The aim of the present study was to validate these probes for fast and simple detection and quantification of in vitro matrix mineralization by KS483 cells and thus enabling high-throughput screening experiments. KS483 cells were cultured under osteogenic conditions in the presence of compounds that either stimulate or inhibit osteoblast differentiation and thereby matrix mineralization. After 21 days of differentiation, fluorescence of stained cultures was quantified with a near-infrared imager and compared to Alizarin Red S quantification. Fluorescence of both probes closely correlated to Alizarin Red S staining in both inhibiting and stimulating conditions. In addition, both compounds displayed specificity for mineralized nodules. We therefore conclude that this method of quantification of bone mineralization using fluorescent compounds is a good alternative for the Alizarin Red S staining.

  12. Does fluoride disrupt hydrogen bond network in cationic lipid bilayer? Time-dependent fluorescence shift of Laurdan and molecular dynamics simulations

    Czech Academy of Sciences Publication Activity Database

    Pokorná, Šárka; Jurkiewicz, Piotr; Vazdar, M.; Cwiklik, Lukasz; Jungwirth, Pavel; Hof, Martin

    2014-01-01

    Roč. 141, č. 22 (2014), 22D516 ISSN 0021-9606 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 ; RVO:61388963 Keywords : fluorescence sfifts * Cationic lipids * Hydrogen bond networks Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.952, year: 2014

  13. Use of the fluorescence quantum yield for the determination of the number-average molecular weight of polymers of epicatechin with 4β→8 interflavin bonds

    Science.gov (United States)

    D. Cho; W.L. Mattice; L.J. Porter; Richard W. Hemingway

    1989-01-01

    Excitation at 280 nm produces a structureless emission band with a maximum at 321-324 nm for dilute solutions of catechin, epicatechin, and their oligomers in l,4-dioxane or water. The fluorescence quantum yield, Q, has been measured in these two solvents for five dimers, a trimer, a tetramer, a pentamer, a hexamer, and a polymer in which the monomer...

  14. Time-resolved fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Gustavsson, Thomas; Mialocq, Jean-Claude

    2007-01-01

    This article addresses the evolution in time of light emitted by a molecular system after a brief photo-excitation. The authors first describe fluorescence from a photo-physical point of view and discuss the characterization of the excited state. Then, they explain some basic notions related to fluorescence characterization (lifetime and decays, quantum efficiency, so on). They present the different experimental methods and techniques currently used to study time-resolved fluorescence. They discuss basic notions of time resolution and spectral reconstruction. They briefly present some conventional methods: intensified Ccd cameras, photo-multipliers and photodiodes associated with a fast oscilloscope, and phase modulation. Other methods and techniques are more precisely presented: time-correlated single photon counting (principle, examples, and fluorescence lifetime imagery), streak camera (principle, examples), and optical methods like the Kerr optical effect (principle and examples) and fluorescence up-conversion (principle and theoretical considerations, examples of application)

  15. Positron emission tomography

    International Nuclear Information System (INIS)

    Chandrasekhar, Preethi; Himabindu, Pucha

    2000-01-01

    Positron Emission Tomography (PET) is a non-invasive nuclear imaging technique used to study different molecular pathways and anatomical structures. PET has found extensive applications in various fields of medicine viz. cardiology, oncology, psychiatry/psychology, neuro science and pulmonology. This study paper basically deals with the physics, chemistry and biology behind the PET technique. It discusses the methodology for generation of the radiotracers responsible for emission of positrons and the annihilation and detection techniques. (author)

  16. Fluorescent Protein Approaches in Alpha Herpesvirus Research

    Directory of Open Access Journals (Sweden)

    Ian B. Hogue

    2015-11-01

    Full Text Available In the nearly two decades since the popularization of green fluorescent protein (GFP, fluorescent protein-based methodologies have revolutionized molecular and cell biology, allowing us to literally see biological processes as never before. Naturally, this revolution has extended to virology in general, and to the study of alpha herpesviruses in particular. In this review, we provide a compendium of reported fluorescent protein fusions to herpes simplex virus 1 (HSV-1 and pseudorabies virus (PRV structural proteins, discuss the underappreciated challenges of fluorescent protein-based approaches in the context of a replicating virus, and describe general strategies and best practices for creating new fluorescent fusions. We compare fluorescent protein methods to alternative approaches, and review two instructive examples of the caveats associated with fluorescent protein fusions, including describing several improved fluorescent capsid fusions in PRV. Finally, we present our future perspectives on the types of powerful experiments these tools now offer.

  17. Molecular imaging in oncology

    International Nuclear Information System (INIS)

    Weber, W.A.

    2007-01-01

    Molecular imaging is generally defined as noninvasive and quantitative imaging of targeted macromolecules and biological processes in living organisms. A characteristic of molecular imaging is the ability to perform repeated studies and assess changes in biological processes over time. Thus molecular imaging lends itself well for monitoring the effectiveness of tumor therapy. In animal models a variety of techniques can be used for molecular imaging. These include optical imaging (bioluminescence and fluorescence imaging), magnetic resonance imaging (MRI) and nuclear medicine techniques. In the clinical setting, however, nuclear medicine techniques predominate, because so far only radioactive tracers provide the necessary sensitivity to study expression and function of macromolecules non-invasively in patients. Nuclear medicine techniques allows to study a variety of biological processes in patients. These include the expression of various receptors (estrogen, androgen, somatostatin receptors and integrins). In addition, tracers are available to study tumor cell proliferation and hypoxia. The by far most commonly used molecular imaging technique in oncology is, however, positron emission tomography (PET) with the glucose analog [ 18 F]fluorodeoxyglucose (FDG-PET). FDG-PET permits non-invasive quantitative assessment of the accelerated exogenous glucose use of malignant tumors. Numerous studies have now shown that reduction of tumor FDG-uptake during therapy allows early prediction of tumor response and patient survival. Clinical studies are currently underway to determine whether FDG-PET can be used to individualize tumor therapy by signaling early in the course of therapy the need for therapeutic adjustments in patients with likely non-responding tumors. (orig.)

  18. Separation and determination of citrinin in corn using HPLC fluorescence detection assisted by molecularly imprinted solid phase extraction clean-up

    Science.gov (United States)

    A liquid chromatography based method to detect citrinin in corn was developed using molecularly imprinted solid phase extraction (MISPE) sample clean-up. Molecularly imprinted polymers were synthesized using 1,4-dihydroxy-2-naphthoic acid as the template and an amine functional monomer. Density func...

  19. Analysis of Biophysical, Optical and Genetic Diversity of DoD Coral Reef Communities Using Advanced Fluorescence and Molecular Biology Techniques (Addendum)

    Science.gov (United States)

    2011-08-01

    coloration and by a decrease in photosynthetic efficiency that resembles coral bleaching . For instance, early reports referred to Vibrio infection...in coral as “bacterial bleaching ”. We examined the physiological mechanisms and fluorescence signatures of YBD using laboratory cultures of isolated...pigment content (a sign similar to coral bleaching ), cell degeneration and lysis. The exposure to Vibrio was accompanied by a marked reduction (by

  20. Thermochromic Fluorescence from B18H20(NC5H5)(2): An Inorganic-Organic Composite Luminescent Compound with an Unusual Molecular Geometry

    Czech Academy of Sciences Publication Activity Database

    Londesborough, Michael Geoffrey Stephen; Dolanský, Jiří; Cerdán, L.; Lang, Kamil; Jelínek, Tomáš; Oliva, J. M.; Hnyk, Drahomír; Roca-Sanjuan, D.; Frances-Monerris, A.; Martinčík, Jiří; Nikl, Martin; Kennedy, John David

    2017-01-01

    Roč. 5, č. 6 (2017), č. článku UNSP 1600694. ISSN 2195-1071 R&D Projects: GA ČR(CZ) GA13-09876S Institutional support: RVO:61388980 ; RVO:68378271 Keywords : boranes * energy transfer * fluorescence * solar concentrators * thermochromicity Subject RIV: CA - Inorganic Chemistry ; BH - Optics, Masers, Lasers (FZU-D) OBOR OECD: Inorganic and nuclear chemistry ; Optics (including laser optics and quantum optics) (FZU-D) Impact factor: 6.875, year: 2016

  1. Fluorescence microscopy.

    Science.gov (United States)

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

    2014-10-01

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  2. Separate and simultaneous binding effects of aspirin and amlodipine to human serum albumin based on fluorescence spectroscopic and molecular modeling characterizations: A mechanistic insight for determining usage drugs doses

    International Nuclear Information System (INIS)

    Abdollahpour, Nooshin; Asoodeh, Ahmad; Saberi, Mohammad Reza; Chamani, JamshidKhan

    2011-01-01

    The binding of aspirin (ASA) and amlodipine (AML) to human serum albumin (HSA) in aqueous solution was investigated by multiple techniques such as fluorescence quenching, resonance light scattering (RLS), three-dimensional fluorescence spectroscopy, FT-IR and zeta-potential measurements in an aqueous solution at pH=7.4. For the protein-ligand association reaction, fluorescence measurements can give important clues as to the binding of ligands to proteins, e.g., the binding mechanism, binding mode, binding constants, binding sites, etc. Fluorescence spectroscopy showed that ASA and AML could quench the HSA fluorescence spectra, and this quenching effect became more significant when both ASA and AML coexisted. The results pointed at the interaction between HSA and both drugs as ternary systems decreasing the binding constant and binding stability of the HSA-drug complex as a binary system. Therefore, by reducing the amount of drugs transported to their targets, the free drug concentration of the target would be reduced, lowering the efficacy of the drugs. It was demonstrated that there exists antagonistic behavior between the two drugs when it comes to binding of HSA. Furthermore, the fluorescence results also showed that the quenching mechanism of HSA-drug complexes as binary and ternary systems is a static procedure. The number of binding sites of HSA-ASA, (HSA-AML)ASA, HSA-AML and (HSA-ASA) AML were 1.31, 0.92, 1 and 0.93, respectively. Due to the existence of the antagonistic action between ASA and AML, the binding distance r was reduced. The results of synchronous fluorescence and three-dimensional fluorescence spectra showed that the antagonistic action between ASA and AML would alter the micro-environment around Trp and Tyr residues. Moreover, the simultaneous presence of ASA and AML during binding to HSA should be taken into account in multidrug therapy, as it induces the necessity of a monitoring therapy owing to the possible increase of uncontrolled toxic

  3. Separate and simultaneous binding effects of aspirin and amlodipine to human serum albumin based on fluorescence spectroscopic and molecular modeling characterizations: A mechanistic insight for determining usage drugs doses

    Energy Technology Data Exchange (ETDEWEB)

    Abdollahpour, Nooshin [Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of); Asoodeh, Ahmad [Department of Chemistry, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad (Iran, Islamic Republic of); Saberi, Mohammad Reza [Department of Medical Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of); Chamani, JamshidKhan, E-mail: chamani@ibb.ut.ac.ir [Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of)

    2011-09-15

    The binding of aspirin (ASA) and amlodipine (AML) to human serum albumin (HSA) in aqueous solution was investigated by multiple techniques such as fluorescence quenching, resonance light scattering (RLS), three-dimensional fluorescence spectroscopy, FT-IR and zeta-potential measurements in an aqueous solution at pH=7.4. For the protein-ligand association reaction, fluorescence measurements can give important clues as to the binding of ligands to proteins, e.g., the binding mechanism, binding mode, binding constants, binding sites, etc. Fluorescence spectroscopy showed that ASA and AML could quench the HSA fluorescence spectra, and this quenching effect became more significant when both ASA and AML coexisted. The results pointed at the interaction between HSA and both drugs as ternary systems decreasing the binding constant and binding stability of the HSA-drug complex as a binary system. Therefore, by reducing the amount of drugs transported to their targets, the free drug concentration of the target would be reduced, lowering the efficacy of the drugs. It was demonstrated that there exists antagonistic behavior between the two drugs when it comes to binding of HSA. Furthermore, the fluorescence results also showed that the quenching mechanism of HSA-drug complexes as binary and ternary systems is a static procedure. The number of binding sites of HSA-ASA, (HSA-AML)ASA, HSA-AML and (HSA-ASA) AML were 1.31, 0.92, 1 and 0.93, respectively. Due to the existence of the antagonistic action between ASA and AML, the binding distance r was reduced. The results of synchronous fluorescence and three-dimensional fluorescence spectra showed that the antagonistic action between ASA and AML would alter the micro-environment around Trp and Tyr residues. Moreover, the simultaneous presence of ASA and AML during binding to HSA should be taken into account in multidrug therapy, as it induces the necessity of a monitoring therapy owing to the possible increase of uncontrolled toxic

  4. Confocal fluorescence techniques in industrial application

    Science.gov (United States)

    Eggeling, Christian; Gall, Karsten; Palo, Kaupo; Kask, Peet; Brand, Leif

    2003-06-01

    The FCS+plus family of evaluation tools for confocal fluorescence spectroscopy, which was developed during recent years, offers a comprehensive view to a series of fluorescence properties. Originating in fluorescence correlation spectroscopy (FCS) and using similar experimental equipment, a system of signal processing methods such as fluorescence intensity distribution analysis (FIDA) was created to analyze in detail the fluctuation behavior of fluorescent particles within a small area of detection. Giving simultaneous access to molecular parameters like concentration, translational and rotational diffusion, molecular brightness, and multicolor coincidence, this portfolio was enhanced by more traditional techniques of fluorescence lifetime as well as time-resolved anisotropy determination. The cornerstones of the FCS+plus methodology will be shortly described. The inhibition of a phosphatase enzyme activity gives a comprehensive industrial application that demonstrates FCS+plus' versatility and its potential for pharmaceutical drug discovery.

  5. Use of fluorescence spectroscopy to measure molecular autofluorescence in diabetic subjects; Utilizacao da espectroscopia de fluorescencia para mensuramento de moleculas autofluorescentes em individuos diabeticos

    Energy Technology Data Exchange (ETDEWEB)

    Gomes, Cinthia Zanini

    2011-07-01

    Diabetes Mellitus (DM) comprises a complex metabolic syndrome, caused by reduced or absent secretion of insulin by pancreatic beta cells, leading to hyperglycemia. Hyperglycemia promotes glycation of proteins and, consequently, the appearance of advanced glycation end products (AGEs). Currently, diabetic patients are monitored by determining levels of glucose and glycated hemoglobin (HbA1c). The complications caused by hyperglycemia may be divided into micro and macrovascular complications, represented by retinopathy, nephropathy, neuropathy and cardiovascular disease. Albumin (HSA) is the most abundant serum protein in the human body and is subject to glycation. The Protoporphyrin IX (PpIX) is the precursor molecule of heme synthesis, structural component of hemoglobin. The in vitro and animals studies have indicated that hyperglycemia promotes a decrease in its concentration in erythrocytes. The fluorescence spectroscopy is a technique widely used in biomedical field. The autofluorescence corresponds to the intrinsic fluorescence present in some molecules, this being associated with the same structure. The aim of this study was to use fluorescence spectroscopy to measure levels of erythrocyte PpIX autofluorescence and AGE-HSA in diabetic and healthy subjects and compare them with levels of blood glucose and HbA1c. This study was conducted with 151 subjects (58 controls and 93 diabetics). Epidemiological data of patients and controls were obtained from medical records. For control subjects, blood glucose levels were obtained from medical records and levels of Hb1Ac obtained by using commercial kits. The determination of the PpIX autofluorescence was performed with excitation at 405 nm and emission at 632 nm. Determination of AGE-HSA was performed with excitation at 370 nm and emission at 455 nm. Approximately 50% of diabetic had micro and macrovascular lesions resulting from hyperglycemia. There were no significant differences in the PpIX emission intensity values

  6. RBAP, a Rhodamine B-Based Derivative: Synthesis, Crystal Structure Analysis, Molecular Simulation, and Its Application as a Selective Fluorescent Chemical Sensor for Sn2+

    Directory of Open Access Journals (Sweden)

    Xiaofeng Bao

    2014-06-01

    Full Text Available A new fluorescent chemosensor based on a Rhodamine B and a benzyl 3-aminopropanoate conjugate (RBAP was designed, synthesized, and structurally characterized. Its single crystal structure was obtained and analyzed by X-ray analysis. In a MeOH/H2O (2:3, v/v, pH 5.95 solution RBAP exhibits a high selectivity and excellent sensitivity for Sn2+ ions in the presence of many other metal cations. The binding analysis using the Job’s plot suggested the RBAP formed a 1:1 complex with Sn2+.

  7. RBAP, a Rhodamine B-Based Derivative: Synthesis, Crystal Structure Analysis, Molecular Simulation, and Its Application as a Selective Fluorescent Chemical Sensor for Sn2+

    OpenAIRE

    Xiaofeng Bao; Xiaowei Cao; Xuemei Nie; Yanyan Jin; Baojing Zhou

    2014-01-01

    A new fluorescent chemosensor based on a Rhodamine B and a benzyl 3-aminopropanoate conjugate (RBAP) was designed, synthesized, and structurally characterized. Its single crystal structure was obtained and analyzed by X-ray analysis. In a MeOH/H2O (2:3, v/v, pH 5.95) solution RBAP exhibits a high selectivity and excellent sensitivity for Sn2+ ions in the presence of many other metal cations. The binding analysis using the Job’s plot suggested the RBAP formed a 1:1 complex with Sn2+.

  8. Molecular spectroscopy

    International Nuclear Information System (INIS)

    Kokh, Eh.; Zonntag, B.

    1981-01-01

    The latest investigation results on molecular spectroscopy with application of synchrotron radiation in the region of vacuum ultraviolet are generalized. Some results on investigation of excited, superexcited and ionized molecule states with the use of adsorption spectroscopy, photoelectron spectroscopy, by fluorescent and mass-spectrometric methods are considered [ru

  9. On the mechanism of non-radiative decay of blue fluorescent protein chromophore: New insight from the excited-state molecular dynamics simulations and potential energy calculations

    Science.gov (United States)

    Zhao, Li; Liu, Jian-Yong; Zhou, Pan-Wang

    2017-11-01

    A detailed theoretical investigation based on the ab initio on-the-fly surface hopping dynamics simulations and potential energy surfaces calculations has been performed to unveil the mechanism of the photoinduced non-adiabatic relaxation process of the isolated blue fluorescent protein (BFP) chromophore in gas phase. The data analysis presents that the dominant reaction coordinate of the BFP chromophore is driven by a rotation motion around the CC double bridging bond, which is in remarkable difference with a previous result which supports a Hula-Twist rotation pattern. Such behavior is consistent with the double bond rotation pattern of the GFP neutral chromophore. In addition, the dynamics simulations give an estimated decay time of 1.1 ps for the S1 state, which is agrees well with the experimental values measured in proteins. The present work offers a straightforward understanding for the decay mechanism of the BFP chromophore and suggestions of the photochemical properties of analogous protein chromophores. We hope the current work would be helpful for further exploration of the BFP photochemical and photophysical properties in various environments, and can provide guidance and prediction for rational design of the fluorescent proteins catering for different demands.

  10. Simultaneous detection of selenium by atomic fluorescence and sulfur by molecular emission by flow-injection hydride generation with on-line reduction for the determination of selenate, sulfate and sulfite

    Energy Technology Data Exchange (ETDEWEB)

    Tyson, J.F., E-mail: tyson@chem.umass.edu [Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, MA 01003 (United States); Palmer, C.D. [Lead Poisoning Trace Elements Laboratory, Wadsworth Center, New York State Department of Health, P.O. Box 509, Empire State Plaza, Albany, NY 12201-0509 (United States)

    2009-10-12

    An inductively coupled plasma atomic fluorescence spectrometry (ICP-AFS) instrument, was modified so that it was capable of monitoring transient chromatographic or flow-injection profiles and that sulfur molecular emission and selenium atomic fluorescence could be monitored simultaneously in an argon-hydrogen diffusion flame on a glass burner. The analytes were introduced as hydrogen selenide and hydrogen sulfide, generated on a flow-injection manifold. Selenate was reduced to hydride-forming selenite by microwave-assisted on-line reaction with hydrochloric acid, and sulfate, or sulfite, was reduced to hydride-forming sulfide by a mixture of hydriodic acid, acetic acid and sodium hypophosphite. The effects of the nature of reducing agent, flow rate, microwave power and coil length were studied. The limit of detection (3 s) for selenium was 10 {mu}g L{sup -1}, and for sulfide was 70 {mu}g L{sup -1} (200-{mu}L injection volume). The calibration was linear for selenium up to 2 mg L{sup -1} and to 10 mg L{sup -1} for sulfide. The throughput was 180 h{sup -1}. The three sulfur species could be differentiated on the basis of reactivity at various microwave powers.

  11. De novo design of chiral organotin cancer drug candidates: validation of enantiopreferential binding to molecular target DNA and 5'-GMP by UV-visible, fluorescence, (1)H and (31)P NMR.

    Science.gov (United States)

    Arjmand, Farukh; Sharma, Girish Chandra; Sayeed, Fatima; Muddassir, Mohd; Tabassum, Sartaj

    2011-12-02

    N,N-bis[(R-/S-)-1-benzyl-2-ethoxyethane] tin (IV) complexes were synthesized by applying de novo design strategy by the condensation reaction of (R-/S-)2-amino-2-phenylethanol and dibromoethane in presence of dimethyltin dichloride and thoroughly characterized by elemental analysis, conductivity measurements, IR, ESI-MS, (1)H, (13)C and (119)Sn, multinuclear NMR spectroscopy and XRD study. Enantioselective and specific binding profile of R-enantiomer 1 in comparison to S-enantiomer 2 with ultimate molecular target CT-DNA was validated by UV-visible, fluorescence, circular dichroism, (1)H and (31)P NMR techniques. This was further corroborated well by interaction of 1 and 2 with 5'-GMP. Copyright © 2011 Elsevier B.V. All rights reserved.

  12. Computed Tomography (CT) -- Sinuses

    Medline Plus

    Full Text Available ... Physician Resources Professions Site Index A-Z Computed Tomography (CT) - Sinuses Computed tomography (CT) of the sinuses ... CT of the Sinuses? What is CT (Computed Tomography) of the Sinuses? Computed tomography, more commonly known ...

  13. Illustrated computer tomography

    International Nuclear Information System (INIS)

    Takahashi, S.

    1983-01-01

    This book provides the following information: basic aspects of computed tomography; atlas of computed tomography of the normal adult; clinical application of computed tomography; and radiotherapy planning and computed tomography

  14. Molecular recognition: Comparative study of a tunable host-guest system by using a fluorescent model system and collision-induced dissociation mass spectrometry on dendrimers

    DEFF Research Database (Denmark)

    Pittelkow, M.; Nielsen, C.B.; Broeren, A.C.

    2005-01-01

    Host-guest interactions between the periphery of adamantylurea-functionalized dendrimers (host) and ureido acetic acid derivatives (guest) were shown to be specific, strong and spatially well-defined. The binding becomes stronger when using phosphonic or sulfonic acid derivatives. In the present...... work we have quantified the binding constants for the host-guest interactions between two different host motifs and six different guest molecules. The host molecules, which resemble the periphery of a poly(propylene imine) dendrimer, have been fitted with an anthracene-based fluorescent probe. The two...... host motifs differ in terms of the length of the spacer between a tertiary amine and two ureido functionalities. The guest molecules all contain an acidic moiety (either a carboxylic acid, a phosphonic acid, or a sulfonic acid) and three of them also contain an ureido moiety capable of forming multiple...

  15. Optical tomography of tissues

    International Nuclear Information System (INIS)

    Zimnyakov, D A; Tuchin, Valerii V

    2002-01-01

    Methods of optical tomography of biological tissues are considered, which include pulse-modulation and frequency-modulation tomography, diffusion tomography with the use of cw radiation sources, optical coherent tomography, speckle-correlation tomography of nonstationary media, and optoacoustic tomography. The method for controlling the optical properties of tissues is studied from the point of view of increasing a probing depth in optical coherent tomography. The modern state and prospects of the development of optical tomography are discussed. (review)

  16. Application of Fluorescence Emission for Characterization of Albendazole and Ricobendazole Micellar Systems: Elucidation of the Molecular Mechanism of Drug Solubilization Process.

    Science.gov (United States)

    Priotti, Josefina; Leonardi, Darío; Pico, Guillermo; Lamas, María C

    2018-04-01

    Albendazole (ABZ) and ricobendazole (RBZ) are referred to as class II compounds in the Biopharmaceutical Classification System. These drugs exhibit poor solubility, which profoundly affects their oral bioavailability. Micellar systems are excellent pharmaceutical tools to enhance solubilization and absorption of poorly soluble compounds. Polysorbate 80 (P80), poloxamer 407 (P407), sodium cholate (Na-C), and sodium deoxycholate (Na-DC) have been selected as surfactants to study the solubilization process of these drugs. Fluorescence emission was applied in order to obtain surfactant/fluorophore (S/F) ratio, critical micellar concentration, protection efficiency of micelles, and thermodynamic parameters. Systems were characterized by their size and zeta potential. A blue shift from 350 to 345 nm was observed when ABZ was included in P80, Na-DC, and Na-C micelles, while RBZ showed a slight change in the fluorescence band. P80 showed a significant solubilization capacity: S/F values were 688 for ABZ at pH 4 and 656 for RBZ at pH 6. Additionally, P80 micellar systems presented the smallest size (10 nm) and their size was not affected by pH change. S/F ratio for bile salts was tenfold higher than for the other surfactants. Quenching plots were linear and their constant values (2.17/M for ABZ and 2.29/M for RBZ) decreased with the addition of the surfactants, indicating a protective effect of the micelles. Na-DC showed better protective efficacy for ABZ and RBZ than the other surfactants (constant values 0.54 and 1.57/M, respectively), showing the drug inclusion into the micelles. Entropic parameters were negative in agreement with micelle formation.

  17. Computed tomography

    International Nuclear Information System (INIS)

    Wells, P.; Davis, J.; Morgan, M.

    1994-01-01

    X-ray or gamma-ray transmission computed tomography (CT) is a powerful non-destructive evaluation (NDE) technique that produces two-dimensional cross-sectional images of an object without the need to physically section it. CT is also known by the acronym CAT, for computerised axial tomography. This review article presents a brief historical perspective on CT, its current status and the underlying physics. The mathematical fundamentals of computed tomography are developed for the simplest transmission CT modality. A description of CT scanner instrumentation is provided with an emphasis on radiation sources and systems. Examples of CT images are shown indicating the range of materials that can be scanned and the spatial and contrast resolutions that may be achieved. Attention is also given to the occurrence, interpretation and minimisation of various image artefacts that may arise. A final brief section is devoted to the principles and potential of a range of more recently developed tomographic modalities including diffraction CT, positron emission CT and seismic tomography. 57 refs., 2 tabs., 14 figs

  18. Doppler Tomography

    Science.gov (United States)

    Marsh, T. R.

    I review the method of Doppler tomography which translates binary-star line profiles taken at a series of orbital phases into a distribution of emission over the binary. I begin with a discussion of the basic principles behind Doppler tomography, including a comparison of the relative merits of maximum entropy regularisation versus filtered back-projection for implementing the inversion. Following this I discuss the issue of noise in Doppler images and possible methods for coping with it. Then I move on to look at the results of Doppler Tomography applied to cataclysmic variable stars. Outstanding successes to date are the discovery of two-arm spiral shocks in cataclysmic variable accretion discs and the probing of the stream/magnetospheric interaction in magnetic cataclysmic variable stars. Doppler tomography has also told us much about the stream/disc interaction in non-magnetic systems and the irradiation of the secondary star in all systems. The latter indirectly reveals such effects as shadowing by the accretion disc or stream. I discuss all of these and finish with some musings on possible future directions for the method. At the end I include a tabulation of Doppler maps published in refereed journals.

  19. DNA nanotechnology and fluorescence applications.

    Science.gov (United States)

    Schlichthaerle, Thomas; Strauss, Maximilian T; Schueder, Florian; Woehrstein, Johannes B; Jungmann, Ralf

    2016-06-01

    Structural DNA nanotechnology allow researchers to use the unique molecular recognition properties of DNA strands to construct nanoscale objects with almost arbitrary complexity in two and three dimensions. Abstracted as molecular breadboards, DNA nanostructures enable nanometer-precise placement of guest molecules such as proteins, fluorophores, or nanoparticles. These assemblies can be used to study biological phenomena with unprecedented control over number, spacing, and molecular identity. Here, we give a general introduction to structural DNA nanotechnology and more specifically discuss applications of DNA nanostructures in the field of fluorescence and plasmonics. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Nanomaterials and MRI molecular probe

    International Nuclear Information System (INIS)

    Inubushi, Toshiro

    2008-01-01

    This paper presents the current state and future prospect of enhancing probes in MRI which enable to image specific cells and molecules mainly from the aspect of cell trafficking. Although MRI requires such probes for specific imaging, it has an advantage that anatomical images are simultaneously available even during surgical operation without radiation exposure, differing from X-CT, -transillumination and positron emission tomography (PET). In the development of novel MRI molecular probes, the recent topic concerns the cell trafficking biology where cells related with transplantation and immunological therapy can be traced. Although superparamagnetic iron oxide (SPIO) has been used as a commercially available enhancer, this nanoparticle has problems like a difficulty to penetrate cell, cytotoxicity and others. For these, authors have developed the nanoparticle SPIO covered with silica shell, which can be chemically modified, e.g., by binding fluorescent pigments to possibly allow MR bimodal molecular imaging. For penetration of particles in cells, envelop of Sendai virus is used. PET-CT has been more popular these days; however, MRI is superior to CT for imaging soft tissues, and development of PET-MRI is actively under progress aiming the multi-modal imaging. At present, molecular probes for MRI are certainly not so many as those for PET and cooperative efforts to develop the probes are required in medical, technological and pharmaceutical fields. (R.T.)

  1. Capping of Silybin with β-Cyclodextrin Influences its Binding with Bovine Serum Albumin: A Study by Fluorescence Spectroscopy and Molecular Modeling

    Energy Technology Data Exchange (ETDEWEB)

    Natesan, Sudha; Sowrirajan, Chandrasekaran; Dhanaraj, Premnath; Enoch, Israel V. M. V. [Karunya Univ., Tamil Nadu (India)

    2014-07-15

    The association of silybin with β-cyclodextrin and its influence on silybin's binding with bovine serum albumin are reported. The stoichiometry, binding constant, and the structure of silybin-β-cyclodextrin inclusion complex are reported. The titrations of silybin with bovine serum albumin in the absence and presence of β-cyclodextrin are carried out and the differences in binding strengths are discussed. Molecular modeling is used to optimize the sites and mode of binding of silybin with bovine serum albumin. Forster resonance energy transfer is calculated and the proximity of interacting molecules is reported in the presence and absence of β-cyclodextrin.

  2. Capping of Silybin with β-Cyclodextrin Influences its Binding with Bovine Serum Albumin: A Study by Fluorescence Spectroscopy and Molecular Modeling

    International Nuclear Information System (INIS)

    Natesan, Sudha; Sowrirajan, Chandrasekaran; Dhanaraj, Premnath; Enoch, Israel V. M. V.

    2014-01-01

    The association of silybin with β-cyclodextrin and its influence on silybin's binding with bovine serum albumin are reported. The stoichiometry, binding constant, and the structure of silybin-β-cyclodextrin inclusion complex are reported. The titrations of silybin with bovine serum albumin in the absence and presence of β-cyclodextrin are carried out and the differences in binding strengths are discussed. Molecular modeling is used to optimize the sites and mode of binding of silybin with bovine serum albumin. Forster resonance energy transfer is calculated and the proximity of interacting molecules is reported in the presence and absence of β-cyclodextrin

  3. New developments in molecular imaging: positron emission tomography time-of-flight (TOF-PET); Nuevos desarrollos en imagen molecular: Tomografia por Emision de Positrones con Teimpo de Vuelo (TOF-PET)

    Energy Technology Data Exchange (ETDEWEB)

    Aguilar, P.; Couce, B.; Iglesias, A.; Lois, C.

    2011-07-01

    Positron Emission tomography (PET) in increasingly being used in oncology for the diagnosis and staging of disease, as well as in monitoring response to therapy. One of the last advances in PET is the incorporation of Time-of-Flight (TOF) information, which improves the tomographic reconstruction process and subsequently the quality of the final image. In this work, we explain the principles of PET and the fundamentals of TOF-PET. Clinical images are shown in order to illustrate how TOF-PET improves the detectability of small lesions, particularly in patients with high body mass index. (Author) 20 refs.

  4. Computerized tomography

    International Nuclear Information System (INIS)

    Caille, J.M.; Salamon, G.

    1980-01-01

    As X-ray Ct becomes more commonplace, other techniques of investigation using roughly comparable hardware and software have appeared. Positron-Emission Tomography already provides indispensable physiological and physio-pathological information. Similarly, in the histo-chemical field, Nuclear Magnetic Resonance seems very promising. Some of these new techniques will no doubt shortly be considered as essential as CT in establishing accurate diagnoses non-invasively. (orig./VJ) [de

  5. Computed tomography

    International Nuclear Information System (INIS)

    Andre, M.; Resnick, D.

    1988-01-01

    Computed tomography (CT) has matured into a reliable and prominent tool for study of the muscoloskeletal system. When it was introduced in 1973, it was unique in many ways and posed a challenge to interpretation. It is in these unique features, however, that its advantages lie in comparison with conventional techniques. These advantages will be described in a spectrum of important applications in orthopedics and rheumatology

  6. Multiphoton tomography of astronauts

    Science.gov (United States)

    König, Karsten; Weinigel, Martin; Pietruszka, Anna; Bückle, Rainer; Gerlach, Nicole; Heinrich, Ulrike

    2015-03-01

    Weightlessness may impair the astronaut's health conditions. Skin impairments belong to the most frequent health problems during space missions. Within the Skin B project, skin physiological changes during long duration space flights are currently investigated on three European astronauts that work for nearly half a year at the ISS. Measurements on the hydration, the transepidermal water loss, the surface structure, elasticity and the tissue density by ultrasound are conducted. Furthermore, high-resolution in vivo histology is performed by multiphoton tomography with 300 nm spatial and 200 ps temporal resolution. The mobile certified medical tomograph with a flexible 360° scan head attached to a mechano-optical arm is employed to measure two-photon autofluorescence and SHG in the volar forearm of the astronauts. Modification of the tissue architecture and of the fluorescent biomolecules NAD(P)H, keratin, melanin and elastin are detected as well as of SHG-active collagen. Thinning of the vital epidermis, a decrease of the autofluoresence intensity, an increase in the long fluorescence lifetime, and a reduced skin ageing index SAAID based on an increased collagen level in the upper dermis have been found. Current studies focus on recovery effects.

  7. Determination of ciprofloxacin in Jiaozhou Bay using molecularly imprinted solid-phase extraction followed by high-performance liquid chromatography with fluorescence detection

    International Nuclear Information System (INIS)

    Lian, Ziru; Wang, Jiangtao

    2016-01-01

    A high selective pre-treatment method for the cleanup and preconcentration of ciprofloxacin in natural seawater samples was developed based on molecularly imprinted solid-phase extraction (MISPE). The ciprofloxacin imprinted polymers were synthesized and the characteristics of obtained polymers were evaluated by scanning electron microscopy, Fourier transform infrared spectroscopy and binding experiments. The imprinted materials showed high adsorption ability for ciprofloxacin and were applied as special solid-phase extraction sorbents for selective separation of ciprofloxacin. An off-line MISPE procedure was optimized and the developed MISPE method allowed direct purification and enrichment of the ciprofloxacin from the aqueous samples prior to high-performance liquid chromatography analysis. The recoveries of spiked seawater on the MISPE cartridges ranged from 75.2 to 112.4% and the relative standard deviations were less than 4.46%. Five seawater samples from Jiaozhou Bay were analyzed and ciprofloxacin was detected in two samples with the concentrations of 0.24 and 0.38 μg L −1 , respectively. - Highlights: • Ciprofloxacin molecularly imprinted polymers (Cip-MIPs) were prepared. • The characteristics and recognition efficiency of MIPs were studied. • An off-line method for Cip was developed using MIPs as solid-phase extraction. • Cip in five seawater samples from Jiaozhou Bay of China was determined.

  8. Using Fluorescent Viruses for Detecting Bacteria in Water

    Science.gov (United States)

    Tabacco, Mary Beth; Qian, Xiaohua; Russo, Jaimie A.

    2009-01-01

    A method of detecting water-borne pathogenic bacteria is based partly on established molecular-recognition and fluorescent-labeling concepts, according to which bacteria of a species of interest are labeled with fluorescent reporter molecules and the bacteria can then be detected by fluorescence spectroscopy. The novelty of the present method lies in the use of bacteriophages (viruses that infect bacteria) to deliver the fluorescent reporter molecules to the bacteria of the species of interest.

  9. Optical-based molecular imaging: contrast agents and potential medical applications

    International Nuclear Information System (INIS)

    Bremer, Christoph; Ntziachristos, Vasilis; Weissleder, Ralph

    2003-01-01

    Laser- and sensitive charge-coupled device technology together with advanced mathematical modelling of photon propagation in tissue has prompted the development of novel optical imaging technologies. Fast surface-weighted imaging modalities, such as fluorescence reflectance imaging (FRI) and 3D quantitative fluorescence-mediated tomography have now become available [1, 2]. These technical advances are paralleled by a rapid development of a whole range of new optical contrasting strategies, which are designed to generate molecular contrast within a living organism. The combination of both, technical advances of light detection and the refinement of optical contrast media, finally yields a new spectrum of tools for in vivo molecular diagnostics. Whereas the technical aspects of optical imaging are covered in more detail in a previous review article in ''European Radiology'' [3], this article focuses on new developments in optical contrasting strategies and design of optical contrast agents for in vivo diagnostics. (orig.)

  10. Investigation of Al2O3 barrier film properties made by atomic layer deposition onto fluorescent tris-(8-hydroxyquinoline) aluminium molecular films

    International Nuclear Information System (INIS)

    Maindron, Tony; Aventurier, Bernard; Ghazouani, Ahlem; Jullien, Tony; Rochat, Névine; Simon, Jean-Yves; Viasnoff, Emilie

    2013-01-01

    Al 2 O 3 films have been deposited at 85 °C by atomic layer deposition onto single 100 nm thick tris-(8-hydroxyquinoline) aluminium (AlQ 3 ) films made onto silicon wafers. It has been found that a thick ALD-deposited Al 2 O 3 layer (> 11 nm) greatly prevents the photo-oxidation of AlQ 3 films when exposed to continuous UV irradiation (350 mW/cm 2 ). Thin Al 2 O 3 thicknesses (< 11 nm) on the contrary yield lower barrier performances. Defects in the Al 2 O 3 layer have been easily observed as non-fluorescent AlQ 3 singularities, or black spots, under UV light on the system Si/AlQ 3 /Al 2 O 3 stored into laboratory conditions (22 °C/50% Relative Humidity (RH)) for long time scale (∼ 2000 h). Accelerated aging conditions in a climatic chamber (85 °C/85% RH) also allow faster visualization of the same defects (168 h). The black spot density grows upon time and the black spot density occurrence rates have been calculated to be 0.024 h −1 ·cm −2 and 0.243 h −1 ·cm −2 respectively for the two testing conditions. A detailed investigation of these defects did show that they cannot be ascribed to the presence of a detectable particle. In that sense they are presumably the consequence of the existence of nanometre-scaled defects which cannot be detected onto fresh samples. Interestingly, an additional overcoating of ebeam-deposited SiO 2 onto the Si/AlQ 3 /Al 2 O 3 sample helps to decrease drastically the black spot density occurrence rates down to 0.004 h −1 ·cm −2 and 0.04 h −1 ·cm −2 respectively for 22 °C/50% RH and 85 °C/85% RH testing conditions. These observations highlight the moisture sensitivity of low temperature ALD-deposited Al 2 O 3 films and confirm the general idea that a single Al 2 O 3 ALD film performs as an ultra-high barrier but needs to be overprotected from water condensation by an additional moisture-stable layer. - Highlights: • Thin Al 2 O 3 films have been deposited by atomic layer deposition onto organic films.

  11. Positron emission tomography imaging of gene expression

    International Nuclear Information System (INIS)

    Tang Ganghua

    2001-01-01

    The merging of molecular biology and nuclear medicine is developed into molecular nuclear medicine. Positron emission tomography (PET) of gene expression in molecular nuclear medicine has become an attractive area. Positron emission tomography imaging gene expression includes the antisense PET imaging and the reporter gene PET imaging. It is likely that the antisense PET imaging will lag behind the reporter gene PET imaging because of the numerous issues that have not yet to be resolved with this approach. The reporter gene PET imaging has wide application into animal experimental research and human applications of this approach will likely be reported soon

  12. Investigation of Al{sub 2}O{sub 3} barrier film properties made by atomic layer deposition onto fluorescent tris-(8-hydroxyquinoline) aluminium molecular films

    Energy Technology Data Exchange (ETDEWEB)

    Maindron, Tony; Aventurier, Bernard [LETI/DOPT/SCOOP/Laboratoire des Composants pour la Visualisation, CEA-LETI, MINATEC Campus, 17 rue des Martyrs, F-38054 Grenoble Cedex 9 (France); Ghazouani, Ahlem; Jullien, Tony [LETI/DTSI/SDEP/Laboratoire Dépôt Equipe 2, CEA-LETI, MINATEC Campus, 17 rue des Martyrs, F-38054 Grenoble Cedex 9 (France); Rochat, Névine [LETI/DTSI/Service de Caractérisation des Matériaux et Composants, CEA-LETI, MINATEC Campus, 17 rue des Martyrs, F-38054 Grenoble Cedex 9 (France); Simon, Jean-Yves; Viasnoff, Emilie [LETI/DOPT/SCOOP/Laboratoire des Composants pour la Visualisation, CEA-LETI, MINATEC Campus, 17 rue des Martyrs, F-38054 Grenoble Cedex 9 (France)

    2013-12-02

    Al{sub 2}O{sub 3} films have been deposited at 85 °C by atomic layer deposition onto single 100 nm thick tris-(8-hydroxyquinoline) aluminium (AlQ{sub 3}) films made onto silicon wafers. It has been found that a thick ALD-deposited Al{sub 2}O{sub 3} layer (> 11 nm) greatly prevents the photo-oxidation of AlQ{sub 3} films when exposed to continuous UV irradiation (350 mW/cm{sup 2}). Thin Al{sub 2}O{sub 3} thicknesses (< 11 nm) on the contrary yield lower barrier performances. Defects in the Al{sub 2}O{sub 3} layer have been easily observed as non-fluorescent AlQ{sub 3} singularities, or black spots, under UV light on the system Si/AlQ{sub 3}/Al{sub 2}O{sub 3} stored into laboratory conditions (22 °C/50% Relative Humidity (RH)) for long time scale (∼ 2000 h). Accelerated aging conditions in a climatic chamber (85 °C/85% RH) also allow faster visualization of the same defects (168 h). The black spot density grows upon time and the black spot density occurrence rates have been calculated to be 0.024 h{sup −1}·cm{sup −2} and 0.243 h{sup −1}·cm{sup −2} respectively for the two testing conditions. A detailed investigation of these defects did show that they cannot be ascribed to the presence of a detectable particle. In that sense they are presumably the consequence of the existence of nanometre-scaled defects which cannot be detected onto fresh samples. Interestingly, an additional overcoating of ebeam-deposited SiO{sub 2} onto the Si/AlQ{sub 3}/Al{sub 2}O{sub 3} sample helps to decrease drastically the black spot density occurrence rates down to 0.004 h{sup −1}·cm{sup −2} and 0.04 h{sup −1}·cm{sup −2} respectively for 22 °C/50% RH and 85 °C/85% RH testing conditions. These observations highlight the moisture sensitivity of low temperature ALD-deposited Al{sub 2}O{sub 3} films and confirm the general idea that a single Al{sub 2}O{sub 3} ALD film performs as an ultra-high barrier but needs to be overprotected from water condensation by an

  13. Intensifying screens in transaxial tomography

    International Nuclear Information System (INIS)

    Debelder, M.H.; Bollen, R.H.

    1981-01-01

    This patent claim by Agfa-Gevaert relates to a method for the production of transaxial tomographs, a combination of materials therefor and X-ray intensifying screens incorporating at least one reflecting element for use in transaxial tomography, wherein the exposure of a photographic silver halide emulsion material proceeds at an angle within the range of 2 0 to 10 0 in conjunction with an X-ray fluorescent intensifying screen including an ultra-violet and/or visible radiation reflective coating or sheet to increase the radiation output of the screen and to reduce the exposure time and radiation dose e.g. in medical X-ray applications. (author)

  14. Fluorescent Probes and Fluorescence (Microscopy Techniques — Illuminating Biological and Biomedical Research

    Directory of Open Access Journals (Sweden)

    Gregor P. C. Drummen

    2012-11-01

    Full Text Available Fluorescence, the absorption and re-emission of photons with longer wavelengths, is one of those amazing phenomena of Nature. Its discovery and utilization had, and still has, a major impact on biological and biomedical research, since it enables researchers not just to visualize normal physiological processes with high temporal and spatial resolution, to detect multiple signals concomitantly, to track single molecules in vivo, to replace radioactive assays when possible, but also to shed light on many pathobiological processes underpinning disease states, which would otherwise not be possible. Compounds that exhibit fluorescence are commonly called fluorochromes or fluorophores and one of these fluorescent molecules in particular has significantly enabled life science research to gain new insights in virtually all its sub-disciplines: Green Fluorescent Protein. Because fluorescent proteins are synthesized in vivo, integration of fluorescent detection methods into the biological system via genetic techniques now became feasible. Currently fluorescent proteins are available that virtually span the whole electromagnetic spectrum. Concomitantly, fluorescence imaging techniques were developed, and often progress in one field fueled innovation in the other. Impressively, the properties of fluorescence were utilized to develop new assays and imaging modalities, ranging from energy transfer to image molecular interactions to imaging beyond the diffraction limit with super-resolution microscopy. Here, an overview is provided of recent developments in both fluorescence imaging and fluorochrome engineering, which together constitute the “fluorescence toolbox” in life science research.

  15. Fluorescence extended X-ray absorption fine structure analysis of half-metallic ferromagnet 'zinc-blende CrAs' grown on GaAs by molecular beam epitaxy

    CERN Document Server

    Ofuchi, H; Ono, K; Oshima, M; Akinaga, H; Manago, T

    2003-01-01

    In this work, geometric structures for a half-metallic ferromagnet 'zinc-blende CrAs', which showed ferromagnetic behavior beyond room temperature, were investigated using fluorescence extended X-ray absorption fine structure (EXAFS) measurement. The EXAFS measurements revealed that As atoms around Cr atoms in the 2 nm CrAs film grown on a GaAs(0 0 1) substrate were coordinated tetrahedrally, indicating formation of zinc-blende CrAs. The Cr-As bond length in the zinc-blende CrAs is 2.49 A. This value is close to that which was estimated from the lattice constant (5.82 A) of ferromagnetic zinc-blende CrAs calculated by full-potential linearized augmented-plane wave method. The EXAFS analysis show that the theoretically predicted zinc-blende CrAs can be fabricated on GaAs(0 0 1) substrate by low-temperature molecular-beam epitaxy.

  16. Fluorescence extended X-ray absorption fine structure analysis of half-metallic ferromagnet 'zinc-blende CrAs' grown on GaAs by molecular beam epitaxy

    International Nuclear Information System (INIS)

    Ofuchi, H.; Mizuguchi, M.; Ono, K.; Oshima, M.; Akinaga, H.; Manago, T.

    2003-01-01

    In this work, geometric structures for a half-metallic ferromagnet 'zinc-blende CrAs', which showed ferromagnetic behavior beyond room temperature, were investigated using fluorescence extended X-ray absorption fine structure (EXAFS) measurement. The EXAFS measurements revealed that As atoms around Cr atoms in the 2 nm CrAs film grown on a GaAs(0 0 1) substrate were coordinated tetrahedrally, indicating formation of zinc-blende CrAs. The Cr-As bond length in the zinc-blende CrAs is 2.49 A. This value is close to that which was estimated from the lattice constant (5.82 A) of ferromagnetic zinc-blende CrAs calculated by full-potential linearized augmented-plane wave method. The EXAFS analysis show that the theoretically predicted zinc-blende CrAs can be fabricated on GaAs(0 0 1) substrate by low-temperature molecular-beam epitaxy

  17. Digital communication through intermolecular fluorescence modulation.

    Science.gov (United States)

    Raymo, F M; Giordani, S

    2001-06-14

    [see reaction]. Ultraminiaturized processors incorporating molecular components can be developed only after devising efficient strategies to communicate signals at the molecular level. We have demonstrated that a three-state molecular switch responds to ultraviolet light, visible light, and H+, attenuating the emission intensity of a fluorescent probe. Intermolecular communication is responsible for the transduction of three input signals into a single optical output. The behavior of the communicating ensemble of molecules corresponds to that of a logic circuit incorporating seven gates.

  18. Imaging prostate cancer: an update on positron emission tomography and magnetic resonance imaging

    DEFF Research Database (Denmark)

    Bouchelouche, Kirsten; Turkbey, Baris; Choyke, Peter

    2010-01-01

    , and molecular imaging information. Developments in imaging technologies, specifically magnetic resonance imaging (MRI) and positron emission tomography (PET)/computed tomography (CT), have improved the detection rate of prostate cancer. MRI has improved lesion detection and local staging. Furthermore, MRI...

  19. Boronic acids for fluorescence imaging of carbohydrates.

    Science.gov (United States)

    Sun, Xiaolong; Zhai, Wenlei; Fossey, John S; James, Tony D

    2016-02-28

    "Fluorescence imaging" is a particularly exciting and rapidly developing area of research; the annual number of publications in the area has increased ten-fold over the last decade. The rapid increase of interest in fluorescence imaging will necessitate the development of an increasing number of molecular receptors and binding agents in order to meet the demand in this rapidly expanding area. Carbohydrate biomarkers are particularly important targets for fluorescence imaging given their pivotal role in numerous important biological events, including the development and progression of many diseases. Therefore, the development of new fluorescent receptors and binding agents for carbohydrates is and will be increasing in demand. This review highlights the development of fluorescence imaging agents based on boronic acids a particularly promising class of receptors given their strong and selective binding with carbohydrates in aqueous media.

  20. U-SPECT-BioFluo : An integrated radionuclide, bioluminescence, and fluorescence imaging platform

    NARCIS (Netherlands)

    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

  1. Dynamic Measurement of Tumor Vascular Permeability and Perfusion using a Hybrid System for Simultaneous Magnetic Resonance and Fluorescence Imaging.

    Science.gov (United States)

    Ren, Wuwei; Elmer, Andreas; Buehlmann, David; Augath, Mark-Aurel; Vats, Divya; Ripoll, Jorge; Rudin, Markus

    2016-04-01

    Assessing tumor vascular features including permeability and perfusion is essential for diagnostic and therapeutic purposes. The aim of this study was to compare fluorescence and magnetic resonance imaging (MRI)-based vascular readouts in subcutaneously implanted tumors in mice by simultaneous dynamic measurement of tracer uptake using a hybrid fluorescence molecular tomography (FMT)/MRI system. Vascular permeability was measured using a mixture of extravascular imaging agents, GdDOTA and the dye Cy5.5, and perfusion using a mixture of intravascular agents, Endorem and a fluorescent probe (Angiosense). Dynamic fluorescence reflectance imaging (dFRI) was integrated into the hybrid system for high temporal resolution. Excellent correspondence between uptake curves of Cy5.5/GdDOTA and Endorem/Angiosense has been found with correlation coefficients R > 0.98. The two modalities revealed good agreement regarding permeability coefficients and centers-of-gravity of the imaging agent distribution. The FMT/dFRI protocol presented is able to accurately map physiological processes and poses an attractive alternative to MRI for characterizing tumor neoangiogenesis.

  2. Patch-based anisotropic diffusion scheme for fluorescence diffuse optical tomography—part 1: technical principles

    International Nuclear Information System (INIS)

    Correia, Teresa; Arridge, Simon

    2016-01-01

    Fluorescence diffuse optical tomography (fDOT) provides 3D images of fluorescence distributions in biological tissue, which represent molecular and cellular processes. The image reconstruction problem is highly ill-posed and requires regularisation techniques to stabilise and find meaningful solutions. Quadratic regularisation tends to either oversmooth or generate very noisy reconstructions, depending on the regularisation strength. Edge preserving methods, such as anisotropic diffusion regularisation (AD), can preserve important features in the fluorescence image and smooth out noise. However, AD has limited ability to distinguish an edge from noise. In this two-part paper, we propose a patch-based anisotropic diffusion regularisation (PAD), where regularisation strength is determined by a weighted average according to the similarity between patches around voxels within a search window, instead of a simple local neighbourhood strategy. However, this method has higher computational complexity and, hence, we wavelet compress the patches (PAD-WT) to speed it up, while simultaneously taking advantage of the denoising properties of wavelet thresholding. The proposed method combines the nonlocal means (NLM), AD and wavelet shrinkage methods, which are image processing methods. Therefore, in this first paper, we used a denoising test problem to analyse the performance of the new method. Our results show that the proposed PAD-WT method provides better results than the AD or NLM methods alone. The efficacy of the method for fDOT image reconstruction problem is evaluated in part 2. (paper)

  3. Experimental station for gas phase fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Stankiewicz, M.; Garcia, E. Melero; Ruiz, J. Alvarez; Erman, P.; Hatherly, P.A.; Kivimaeki, A.; Rachlew, E.; Rius i Riu, J.

    2004-01-01

    The details of an experimental setup for gas phase atomic and molecular fluorescence measurements using synchrotron radiation are described in this article. The most significant part of the apparatus is an optical arrangement, which allows for simultaneous measurements of dispersed as well as total fluorescence intensity using an effusive gas jet and an inbuilt gas cell assembled in a convenient plug and measure configuration. The first measurements concerning fluorescence of the N 2 molecule around the N 1s edge obtained with this setup are presented

  4. Molecular sensors and molecular logic gates

    International Nuclear Information System (INIS)

    Georgiev, N.; Bojinov, V.

    2013-01-01

    Full text: The rapid grow of nanotechnology field extended the concept of a macroscopic device to the molecular level. Because of this reason the design and synthesis of (supra)-molecular species capable of mimicking the functions of macroscopic devices are currently of great interest. Molecular devices operate via electronic and/or nuclear rearrangements and, like macroscopic devices, need energy to operate and communicate between their elements. The energy needed to make a device work can be supplied as chemical energy, electrical energy, or light. Luminescence is one of the most useful techniques to monitor the operation of molecular-level devices. This fact determinates the synthesis of novel fluorescence compounds as a considerable and inseparable part of nanoscience development. Further miniaturization of semiconductors in electronic field reaches their limit. Therefore the design and construction of molecular systems capable of performing complex logic functions is of great scientific interest now. In semiconductor devices the logic gates work using binary logic, where the signals are encoded as 0 and 1 (low and high current). This process is executable on molecular level by several ways, but the most common are based on the optical properties of the molecule switches encoding the low and high concentrations of the input guest molecules and the output fluorescent intensities with binary 0 and 1 respectively. The first proposal to execute logic operations at the molecular level was made in 1988, but the field developed only five years later when the analogy between molecular switches and logic gates was experimentally demonstrated by de Silva. There are seven basic logic gates: AND, OR, XOR, NOT, NAND, NOR and XNOR and all of them were achieved by molecules, the fluorescence switching as well. key words: fluorescence, molecular sensors, molecular logic gates

  5. Riboflavin enhanced fluorescence of highly reduced graphene oxide

    Science.gov (United States)

    Iliut, Maria; Gabudean, Ana-Maria; Leordean, Cosmin; Simon, Timea; Teodorescu, Cristian-Mihail; Astilean, Simion

    2013-10-01

    The improvement of graphene derivates' fluorescence properties is a challenging topic and very few ways were reported up to now. In this Letter we propose an easy method to enhance the fluorescence of highly reduced graphene oxide (rGO) through non-covalent binding to a molecular fluorophore, namely the riboflavin (Rb). While the fluorescence of Rb is quenched, the Rb - decorated rGO exhibits strong blue fluorescence and significantly increased fluorescence lifetime, as compared to its pristine form. The data reported here represent a promising start towards tailoring the optical properties of rGOs, having utmost importance in optical applications.

  6. Photoacoustic Tomography

    Science.gov (United States)

    Wang, Lihong V.

    Photoacoustic tomography (PAT) refers to imaging that is based on the photoacoustic effect. Although the photoacoustic effect as a physical phenomenon was first reported on by Alexander Graham Bell in 1880 [1], PAT as an imaging technology was developed only after the advent of ultrasonic transducers, computers, and lasers [2-31]. A review on biomedical photoacoustics is available [32]. The motivation for PAT is to combine optical-absorption contrast with ultrasonic spatial resolution for deep imaging in the optical quasi-diffusive or diffusive regime. In PAT, the tissue is irradiated by usually a short-pulsed laser beam to achieve a thermal and acoustic impulse response (Fig. 19.1). Locally absorbed light is converted into heat, which is further converted to a pressure rise via thermo-elastic expansion. The initial pressure rise - determined by the local optical absorption coefficient (μ â ), fluence (ψ) and other thermal and mechanical properties - propagates as an ultrasonic wave, which is referred to as a photoacoustic wave.

  7. Axial tomography

    International Nuclear Information System (INIS)

    Brueckner, K.A.; Lewis, J.H.

    1979-01-01

    The invention relates to axial tomography, sometimes referred to as cross-sectional x-ray. The apparatus described may utilize the conventional x-ray or ultrasonic source and detector and scanning mechanism for producing the plurality of sets of radiation detector output signals. It has the means for storing the detector output signals in analog form with the signals of one set overlying the signals of another set so that signals resulting from radiation through a zone of the object being examined are summed at a corresponding zone in the storage device, typically an electronic storage tube. The summed signals are read from the storage device with a radially inversely proportional reader producing a second signal for storage, again typically in an electronic storage tube. These signals stored in the second storage device are read with Laplacian relation, with the resultant sigal being a video signal that may be connected to a TV monitor for display of the sectional image. In alternative embodiments, optical film systems and electrostatic systems are utilized. (JTA)

  8. Reviews in fluorescence 2010

    CERN Document Server

    Geddes, Chris D

    2011-01-01

    ""Reviews in Fluorescence 2010"", the seventh volume of the book serial from Springer, serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence and closely related disciplines. It summarizes the year's progress in fluorescence and its applications, with authoritative analytical reviews specialized enough to be attractive to professional researchers, yet also appealing to the wider audience of scientists in related disciplines of fluorescence. ""Reviews in Fluorescence"" offers an essential reference material for any lab working in the fluoresc

  9. Principles of fluorescence techniques

    CERN Document Server

    2016-01-01

    Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.

  10. Variation in radiotherapy target volume definition, dose to organs at risk and clinical target volumes using anatomic (computed tomography) versus combined anatomic and molecular imaging (positron emission tomography/computed tomography): intensity-modulated radiotherapy delivered using a tomotherapy Hi Art machine: final results of the VortigERN study.

    Science.gov (United States)

    Chatterjee, S; Frew, J; Mott, J; McCallum, H; Stevenson, P; Maxwell, R; Wilsdon, J; Kelly, C G

    2012-12-01

    Contrast-enhanced computed tomography (CECT) is the current standard for delineating tumours of the head and neck for radiotherapy. Although metabolic imaging with positron emission tomography (PET) has been used in recent years, the studies were non-confirmatory in establishing its routine role in radiotherapy planning in the modern era. This study explored the difference in gross tumour volume and clinical target volume definitions for the primary and nodal volumes when FDG PET/CT was used as compared with CECT in oropharyngeal cancer cases. Twenty patients with oropharyngeal cancers had a PET/CT scan in the treatment position after consent. Target volumes were defined on CECT scans by a consultant clinical oncologist who was blind to the PET scans. After obtaining inputs from a radiologist, another set of target volumes were outlined on the PET/CT data set. The gross and clinical target volumes as defined on the two data sets were then analysed. The hypothesis of more accurate target delineation, preventing geographical miss and comparative overlap volumes between CECT and PET/CT, was explored. The study also analysed the volumes of intersection and analysed whether there was any TNM stage migration when PET/CT was used as compared with CECT for planning. In 17 of 20 patients, the TNM stage was not altered when adding FDG PET information to CT. PET information prevented geographical miss in two patients and identified distant metastases in one case. PET/CT gross tumour volumes were smaller than CECT volumes (mean ± standard deviation: 25.16 cm(3) ± 35.8 versus 36.56 cm(3) ± 44.14; P standard deviation: CECT versus PET/CT 32.48 cm(3) ± 36.63 versus 32.21 cm(3) ± 37.09; P > 0.86) were not statistically different. Similarity and discordance coefficients were calculated and are reported. PET/CT as compared with CECT could provide more clinically relevant information and prevent geographical miss when used for radiotherapy planning for advanced oropharyngeal

  11. Reviews in fluorescence 2008

    CERN Document Server

    Geddes, Chris D

    2010-01-01

    This volume serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence spectroscopy. It summarizes the year's progress in fluorescence and its applications as well as includes authoritative analytical reviews.

  12. Fluorescent optical position sensor

    Science.gov (United States)

    Weiss, Jonathan D.

    2005-11-15

    A fluorescent optical position sensor and method of operation. A small excitation source side-pumps a localized region of fluorescence at an unknown position along a fluorescent waveguide. As the fluorescent light travels down the waveguide, the intensity of fluorescent light decreases due to absorption. By measuring with one (or two) photodetectors the attenuated intensity of fluorescent light emitted from one (or both) ends of the waveguide, the position of the excitation source relative to the waveguide can be determined by comparing the measured light intensity to a calibrated response curve or mathematical model. Alternatively, excitation light can be pumped into an end of the waveguide, which generates an exponentially-decaying continuous source of fluorescent light along the length of the waveguide. The position of a photodetector oriented to view the side of the waveguide can be uniquely determined by measuring the intensity of the fluorescent light emitted radially at that location.

  13. Safe biodegradable fluorescent particles

    Science.gov (United States)

    Martin, Sue I [Berkeley, CA; Fergenson, David P [Alamo, CA; Srivastava, Abneesh [Santa Clara, CA; Bogan, Michael J [Dublin, CA; Riot, Vincent J [Oakland, CA; Frank, Matthias [Oakland, CA

    2010-08-24

    A human-safe fluorescence particle that can be used for fluorescence detection instruments or act as a safe simulant for mimicking the fluorescence properties of microorganisms. The particle comprises a non-biological carrier and natural fluorophores encapsulated in the non-biological carrier. By doping biodegradable-polymer drug delivery microspheres with natural or synthetic fluorophores, the desired fluorescence can be attained or biological organisms can be simulated without the associated risks and logistical difficulties of live microorganisms.

  14. Dual fluorescence of single LH2 antenna nanorings

    International Nuclear Information System (INIS)

    Freiberg, A.; Raetsep, M.; Timpmann, K.; Trinkunas, G.

    2004-01-01

    A dual nature of fluorescence from LH2 pigment-protein complexes, which is a part of the light harvesting system of purple bacteria, is confirmed by fluorescence-lifetime dependence on recording wavelength and spectrally selective spectroscopy. An analysis based on the Holstein molecular crystal model, modified by allowing diagonal disorder, suggests coexistence of large- and small-radius self-trapped excitons, which serve as the origin of the dual fluorescence

  15. Algorithmic fundamentals of computerized tomography and of transverse analogue tomography

    International Nuclear Information System (INIS)

    Heckmann, K.

    1981-01-01

    Computerized tomography and transverse analogue tomography are two different approaches to the same goal, namely, transverse tomography. The algorithm is discussed and compared. Transverse tomography appears capable of further development, judging by this comparison. (orig.) [de

  16. Optimization of fluorescent proteins

    NARCIS (Netherlands)

    Bindels, D.S.; Goedhart, J.; Hink, M.A.; van Weeren, L.; Joosen, L.; Gadella (jr.), T.W.J.; Engelborghs, Y.; Visser, A.J.W.G.

    2014-01-01

    Nowadays, fluorescent protein (FP) variants have been engineered to fluoresce in all different colors; to display photoswitchable, or photochromic, behavior; or to show yet other beneficial properties that enable or enhance a still growing set of new fluorescence spectroscopy and microcopy

  17. Fluorescence optical imaging in anticancer drug delivery.

    Science.gov (United States)

    Etrych, Tomáš; Lucas, Henrike; Janoušková, Olga; Chytil, Petr; Mueller, Thomas; Mäder, Karsten

    2016-03-28

    In the past several decades, nanosized drug delivery systems with various targeting functions and controlled drug release capabilities inside targeted tissues or cells have been intensively studied. Understanding their pharmacokinetic properties is crucial for the successful transition of this research into clinical practice. Among others, fluorescence imaging has become one of the most commonly used imaging tools in pre-clinical research. The development of increasing numbers of suitable fluorescent dyes excitable in the visible to near-infrared wavelengths of the spectrum has significantly expanded the applicability of fluorescence imaging. This paper focuses on the potential applications and limitations of non-invasive imaging techniques in the field of drug delivery, especially in anticancer therapy. Fluorescent imaging at both the cellular and systemic levels is discussed in detail. Additionally, we explore the possibility for simultaneous treatment and imaging using theranostics and combinations of different imaging techniques, e.g., fluorescence imaging with computed tomography. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Synthesis and characterization of photoswitchable fluorescent silica nanoparticles

    NARCIS (Netherlands)

    Fölling, J.; Polyakova, S.; Belov, V.; van Blaaderen, A.; Bossi, M.L.; Hell, S.W.

    2008-01-01

    We have designed and synthesized a new functional (amino reactive) highly efficient fluorescent molecular switch (FMS) with a photochromic diarylethene and a rhodamine fluorescent dye. The reactive group in this FMS -N-hydroxysuccinimide ester- allows selective labeling of amino containing molecules

  19. Video-rate optical flow corrected intraoperative functional fluorescence imaging

    NARCIS (Netherlands)

    Koch, Maximilian; Glatz, Juergen; Ermolayev, Vladimir; de Vries, Elisabeth G. E.; van Dam, Gooitzen M.; Englmeier, Karl-Hans; Ntziachristos, Vasilis

    Intraoperative fluorescence molecular imaging based on targeted fluorescence agents is an emerging approach to improve surgical and endoscopic imaging and guidance. Short exposure times per frame and implementation at video rates are necessary to provide continuous feedback to the physician and

  20. Fluorescence of irradiated hydrocarbons. [. gamma. rays

    Energy Technology Data Exchange (ETDEWEB)

    Gulis, I G; Evdokimenko, V M; Lapkovskii, M P; Petrov, P T; Gulis, I M; Markevich, S V [AN Belorusskoj SSR, Minsk. Inst. Fiziko-Organicheskoj Khimii

    1977-01-01

    A visible fluorescence has been found out in ..gamma..-irradiated aqueous solutions of carbohydrates. Two bands have been distinguished in fluorescence spectra of the irradiated solution of dextran: a short-wave band lambdasub(max)=140 nm (where lambda is a wave length) at lambdasub(..beta..)=380 nm and a long-wave band with lambdasub(max)=540 nm at lambdasub(..beta..)=430 nm. A similar form of the spectrum has been obtained for irradiated solutions of starch, amylopectin, low molecular glucose. It has been concluded that a macromolecule of polysaccharides includes fluorescent centers. A relation between fluorescence and ..cap alpha..-oxiketon groups formed under irradiation has been pointed out.

  1. Fundamentals of fluorescence microscopy exploring life with light

    CERN Document Server

    Mondal, Partha Pratim

    2014-01-01

    This book starts at an introductory level and leads reader to the most advanced developments in fluorescence imaging and super-resolution techniques that have enabled the emergence of new disciplines such as nanobioimaging, multiphoton microscopy, photodynamic therapy, nanometrology and nanosensors. The interdisciplinary subject of fluorescence microscopy and imaging requires complete knowledge of imaging optics and molecular physics. So, this book approaches the subject by introducing optical imaging concepts before going deep into the advanced imaging systems and their applications. Molecular orbital theory forms the basis for understanding fluorescent molecules and thereby facilitates complete explanation of light-matter interaction at the geometrical focus. The two disciplines have some overlap since light controls the states of molecules and conversely, molecular states control the emitted light. These two mechanisms together determine essential fluorescence  factors and phenomena such as, molecular cro...

  2. Computed Tomography (CT) -- Head

    Medline Plus

    Full Text Available ... When the image slices are reassembled by computer software, the result is a very detailed multidimensional view ... Safety Images related to Computed Tomography (CT) - Head Videos related to Computed Tomography (CT) - Head Sponsored by ...

  3. What is Computed Tomography?

    Science.gov (United States)

    ... Imaging Medical X-ray Imaging What is Computed Tomography? Share Tweet Linkedin Pin it More sharing options ... Chest X ray Image back to top Computed Tomography (CT) Although also based on the variable absorption ...

  4. Computed Tomography (CT) -- Head

    Medline Plus

    Full Text Available ... Stroke Brain Tumors Computer Tomography (CT) Safety During Pregnancy Head and Neck Cancer X-ray, Interventional Radiology and Nuclear Medicine Radiation Safety Images related to Computed Tomography (CT) - ...

  5. Review of muon tomography

    International Nuclear Information System (INIS)

    Feng Hanliang; Jiao Xiaojing

    2010-01-01

    As a new detection technology, Muon tomography has some potential benefits, such as being able to form a three- dimensional image, without radiation, low cost, fast detecting etc. Especially, muon tomography will play an important role in detecting nuclear materials. It introduces the theory of Muon tomography, its advantages and the Muon tomography system developed by decision sciences corporation and Los Alamos national laboratory. (authors)

  6. Computed Tomography (CT) -- Head

    Medline Plus

    Full Text Available ... Physician Resources Professions Site Index A-Z Computed Tomography (CT) - Head Computed tomography (CT) of the head uses special x-ray ... What is CT Scanning of the Head? Computed tomography, more commonly known as a CT or CAT ...

  7. Emission computed tomography

    International Nuclear Information System (INIS)

    Ott, R.J.

    1986-01-01

    Emission Computed Tomography is a technique used for producing single or multiple cross-sectional images of the distribution of radionuclide labelled agents in vivo. The techniques of Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET) are described with particular regard to the function of the detectors used to produce images and the computer techniques used to build up images. (UK)

  8. Computed Tomography (CT) -- Sinuses

    Medline Plus

    Full Text Available ... Physician Resources Professions Site Index A-Z Computed Tomography (CT) - Sinuses Computed tomography (CT) of the sinuses uses special x-ray equipment ... story here Images × Image Gallery Patient undergoing computed tomography (CT) scan. View full size with caption Pediatric Content ...

  9. Computed Tomography (CT) -- Head

    Medline Plus

    Full Text Available ... Physician Resources Professions Site Index A-Z Computed Tomography (CT) - Head Computed tomography (CT) of the head uses special x-ray equipment ... story here Images × Image Gallery Patient undergoing computed tomography (CT) scan. View full size with caption Pediatric Content ...

  10. Positron emission tomography

    International Nuclear Information System (INIS)

    Reivich, M.; Alavi, A.

    1985-01-01

    This book contains 24 selections. Some of the titles are: Positron Emission Tomography Instrumentation, Generator Systems for Positron Emitters, Reconstruction Algorithms, Cerebral Glucose Consumption: Methodology and Validation, Cerebral Blood Flow Tomography Using Xenon-133 Inhalation: Methods and Clinical Applications, PET Studies of Stroke, Cardiac Positron Emission Tomography, and Use of PET in Oncology

  11. Fluorescent S-layer fusion proteins

    International Nuclear Information System (INIS)

    Kainz, B.

    2010-01-01

    This work describes the construction and characterisation of fluorescent S-layer fusion proteins used as building blocks for the fabrication of nanostructured monomolecular biocoatings on silica particles with defined fluorescence properties. The S-layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a was fused with the pH-dependant cyan, green and yellow variant of the green fluorescent protein (GFP) and the red fluorescent protein mRFP1. These fluorescent S-layer fusion proteins, acting as scaffold and optical sensing element simultaneously, were able to reassemble in solution and on silica particles forming 2D nanostructures with p2 lattice symmetry (a=11 ±0.5 nm, b=14 ±0.4 nm, g=80 ±1 o ). The pH-dependant fluorescence behaviour was studied with fluorimetry, confocal microscopy and flow cytometry. These fluorescent S-layer fusion proteins can be used as pH-sensor. 50% of the fluorescence intensity decreases at their calculated pKa values (pH6 - pH5). The fluorescence intensity of the GFP variants vanished completely between pH4 and pH3 whereas the chromophore of the red protein mRFP1 was only slightly affected in acidic conditions. At the isoelectric point of the S-layer coated silica particles (pH4.6 ±0.2) an increase in particle aggregation was detected by flow cytometry. The cyan and yellow fluorescent proteins were chosen to create a bi-fluorescent S-layer tandem fusion protein with the possibility for resonance energy transfer (FRET). A transfer efficiency of 20% and a molecular distance between the donor (ECFP) and acceptor (YFP) chromophores of around 6.2 nm could be shown. This bi-fluorescent ECFP-SgsE-YFP tandem fusion protein was able to reassemble on solid surfaces. The remarkable combination of fluorescence and self-assembly and the design of bi-functional S-layer tandem fusion protein matrices makes them to a promising tool in nanobiotechnology. (author) [de

  12. Evaluation of hypoxia in a feline model of head and neck cancer using 64Cu-ATSM positron emission tomography/computed tomography

    International Nuclear Information System (INIS)

    Ballegeer, Elizabeth A; Madrill, Nicole J; Berger, Kevin L; Agnew, Dalen W; McNiel, Elizabeth A

    2013-01-01

    Human and feline head and neck squamous cell carcinoma (HNSCC) share histology, certain molecular features, as well as locally aggressive and highly recurrent clinical behavior. In human HNSCC, the presence of significant hypoxia within these tumors is considered an important factor in the development of a more aggressive phenotype and poor response to therapy. We hypothesized that feline head and neck tumors, particularly HNSCC, would exhibit hypoxia and that 64 Cu-diacetyl-bis(N4-methylthiosemicarbazone) (Cu-ATSM) positron emission tomography/computed tomography (PET/CT) would permit detection of intratumoral hypoxia. 12 cats with measureable head and neck tumors were given 64 Cu-ATSM and iodinated contrast for PET/CT scan. The presence or absence of hypoxia was also assessed using an intratumoral fluorescent life-time probe to quantitate pO 2 and pimonidazole immunohistochemical staining in biopsy specimens. In two cats, intratumoral O 2 and 64 Cu-ATSM uptake was measured before and after treatment with anti-angiogenic agents to determine the effect of these agents on hypoxia. Eleven of twelve feline tumors demonstrated significant 64 Cu-ATSM uptake, regardless of malignant or benign etiology. The presence (and absence) of hypoxia was confirmed using the fluorescent O 2 detection probe in nine tumors, and using pimonidazole staining in three tumors. Squamous cell carcinomas (HNSCC) demonstrated the highest degree of hypoxia, with T max /M ratios ranging from 4.3 to 21.8. Additional non-neoplastic tissues exhibited 64 Cu-ATSM uptake suggestive of hypoxia including reactive draining lymph nodes, non-malignant thyroid pathology, a tooth root abscess, and otitis media. In two cats with HNSCC that received anti-vascular agents, the pattern of 64 Cu-ATSM uptake was altered after treatment, demonstrating the potential of the feline model to study the modulation of tumor oxygenation. Feline HNSCC serves as a clinically relevant model for the investigation of

  13. X-ray micro-modulated luminescence tomography (XMLT)

    Science.gov (United States)

    Cong, Wenxiang; Liu, Fenglin; Wang, Chao; Wang, Ge

    2014-01-01

    Imaging depth of optical microscopy has been fundamentally limited to millimeter or sub-millimeter due to strong scattering of light in a biological sample. X-ray microscopy can resolve spatial details of few microns deep inside a sample but contrast resolution is inadequate to depict heterogeneous features at cellular or sub-cellular levels. To enhance and enrich biological contrast at large imaging depth, various nanoparticles are introduced and become essential to basic research and molecular medicine. Nanoparticles can be functionalized as imaging probes, similar to fluorescent and bioluminescent proteins. LiGa5O8:Cr3+ nanoparticles were recently synthesized to facilitate luminescence energy storage with x-ray pre-excitation and subsequently stimulated luminescence emission by visible/near-infrared (NIR) light. In this paper, we propose an x-ray micro-modulated luminescence tomography (XMLT, or MLT to be more general) approach to quantify a nanophosphor distribution in a thick biological sample with high resolution. Our numerical simulation studies demonstrate the feasibility of the proposed approach. PMID:24663898

  14. Atomic-fluorescence spectrophotometry

    International Nuclear Information System (INIS)

    Bakhturova, N.F.; Yudelevich, I.G.

    1975-01-01

    Atomic-fluorescence spectrophotometry, a comparatively new method for the analysis of trace quantities, has developed rapidly in the past ten years. Theoretical and experimental studies by many workers have shown that atomic-fluorescence spectrophotometry (AFS) is capable of achieving a better limit than atomic absorption for a large number of elements. The present review examines briefly the principles of atomic-fluorescence spectrophotometry and the types of fluorescent transition. The excitation sources, flame and nonflame atomizers, used in AFS are described. The limits of detection achieved up to the present, using flame and nonflame methods of atomization are given

  15. Fluorescence of irradiated hydrocarbons

    International Nuclear Information System (INIS)

    Gulis, I.G.; Evdokimenko, V.M.; Lapkovskij, M.P.; Petrov, P.T.; Gulis, I.M.; Markevich, S.V.

    1977-01-01

    A visible fluorescence has been found out in γ-irradiated aqueous of carbohydrates. Two bands have been distinguished in fluorescence spectra of the irradiated solution of dextran: a short-wave band lambdasub(max)=140 nm (where lambda is a wave length) at lambdasub(β)=380 nm and a long-wave band with lambdasub(max)=540 nm at lambdasub(β)=430 nm. A similar form of the spectrum has been obtained for irradiated solutions of starch, amylopectin, lowmolecular glucose. It has been concluded that a macromolecule of polysaccharides includes fluorescent centres. A relation between fluorescence and α-oxiketon groups formed under irradiation has been pointed out

  16. Turbocharging Quantum Tomography

    Energy Technology Data Exchange (ETDEWEB)

    Blume-Kohout, Robin J. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Gamble, John King [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Nielsen, Erik [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Maunz, Peter Lukas Wilhelm [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Scholten, Travis L. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Rudinger, Kenneth Michael [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)

    2015-01-01

    Quantum tomography is used to characterize quantum operations implemented in quantum information processing (QIP) hardware. Traditionally, state tomography has been used to characterize the quantum state prepared in an initialization procedure, while quantum process tomography is used to characterize dynamical operations on a QIP system. As such, tomography is critical to the development of QIP hardware (since it is necessary both for debugging and validating as-built devices, and its results are used to influence the next generation of devices). But tomography suffers from several critical drawbacks. In this report, we present new research that resolves several of these flaws. We describe a new form of tomography called gate set tomography (GST), which unifies state and process tomography, avoids prior methods critical reliance on precalibrated operations that are not generally available, and can achieve unprecedented accuracies. We report on theory and experimental development of adaptive tomography protocols that achieve far higher fidelity in state reconstruction than non-adaptive methods. Finally, we present a new theoretical and experimental analysis of process tomography on multispin systems, and demonstrate how to more effectively detect and characterize quantum noise using carefully tailored ensembles of input states.

  17. High resolution IVEM tomography of biological specimens

    Energy Technology Data Exchange (ETDEWEB)

    Sedat, J.W.; Agard, D.A. [Univ. of California, San Francisco, CA (United States)

    1997-02-01

    Electron tomography is a powerful tool for elucidating the three-dimensional architecture of large biological complexes and subcellular organelles. The introduction of intermediate voltage electron microscopes further extended the technique by providing the means to examine very large and non-symmetrical subcellular organelles, at resolutions beyond what would be possible using light microscopy. Recent studies using electron tomography on a variety of cellular organelles and assemblies such as centrosomes, kinetochores, and chromatin have clearly demonstrated the power of this technique for obtaining 3D structural information on non-symmetric cell components. When combined with biochemical and molecular observations, these 3D reconstructions have provided significant new insights into biological function.

  18. Synthesis and characterization of photoswitchable fluorescent silica nanoparticles.

    Science.gov (United States)

    Fölling, Jonas; Polyakova, Svetlana; Belov, Vladimir; van Blaaderen, Alfons; Bossi, Mariano L; Hell, Stefan W

    2008-01-01

    We have designed and synthesized a new functional (amino reactive) highly efficient fluorescent molecular switch (FMS) with a photochromic diarylethene and a rhodamine fluorescent dye. The reactive group in this FMS -N-hydroxysuccinimide ester- allows selective labeling of amino containing molecules or other materials. In ethanolic solutions, the compound displays a large fluorescent quantum yield of 52 % and a large fluorescence modulation ratio (94 %) between two states that may be interconverted with red and near-UV light. Silica nanoparticles incorporating the new FMS were prepared and characterized, and their spectroscopic and switching properties were also studied. The dye retained its properties after the incorporation into the silica, thereby allowing light-induced reversible high modulation of the fluorescence signal of a single particle for up to 60 cycles, before undergoing irreversible photobleaching. Some applications of these particles in fluorescence microscopy are also demonstrated. In particular, subdiffraction images of nanoparticles were obtained, in the focal plane of a confocal microscope.

  19. Fluorescence lifetime assays: current advances and applications in drug discovery.

    Science.gov (United States)

    Pritz, Stephan; Doering, Klaus; Woelcke, Julian; Hassiepen, Ulrich

    2011-06-01

    Fluorescence lifetime assays complement the portfolio of established assay formats available in drug discovery, particularly with the recent advances in microplate readers and the commercial availability of novel fluorescent labels. Fluorescence lifetime assists in lowering complexity of compound screening assays, affording a modular, toolbox-like approach to assay development and yielding robust homogeneous assays. To date, materials and procedures have been reported for biochemical assays on proteases, as well as on protein kinases and phosphatases. This article gives an overview of two assay families, distinguished by the origin of the fluorescence signal modulation. The pharmaceutical industry demands techniques with a robust, integrated compound profiling process and short turnaround times. Fluorescence lifetime assays have already helped the drug discovery field, in this sense, by enhancing productivity during the hit-to-lead and lead optimization phases. Future work will focus on covering other biochemical molecular modifications by investigating the detailed photo-physical mechanisms underlying the fluorescence signal.

  20. single photon emission tomography and positron emission tomography - Part 1 (October 2012), Part 2 (October 2010)

    International Nuclear Information System (INIS)

    Buvat, Irene

    2010-10-01

    The objective of this lecture is to present the single photon emission computed tomography (SPECT) and the positron emission tomography (PET) imaging techniques. Part 1 Content: 1 - Introduction: anatomic, functional and molecular imaging; 2 - Radiotracers: chemical and physical constraints, gamma photon emitters, positon emitters, radioisotopes production, emitters type and imaging techniques; 3 - Gamma cameras; 4 - Quantification in emission tomography: attenuation, scattering, un-stationary spatial resolution; 5 - Synthesis and conclusion. Part 2 content: 1 - Positon emitters; 2 - Positons detection: Coincidence detection (electronic collimation, PET detectors with gamma cameras, dedicated PET detectors, spectrometry); PET detectors type; time-of-flight PET; 2D PET; 3D PET; 3 - Quantification in emission tomography: detected events, attenuation, scattering, fortuitous coincidences, standardisation; 4 - Common SPECT and PET problems: partial volume effect, movement, tomographic reconstruction, calibration, dead time; 5 - Synthesis and conclusion

  1. Projection neuron circuits resolved using correlative array tomography

    Directory of Open Access Journals (Sweden)

    Daniele eOberti

    2011-04-01

    Full Text Available Assessment of three-dimensional morphological structure and synaptic connectivity is essential for a comprehensive understanding of neural processes controlling behavior. Different microscopy approaches have been proposed based on light microcopy (LM, electron microscopy (EM, or a combination of both. Correlative array tomography (CAT is a technique in which arrays of ultrathin serial sections are repeatedly stained with fluorescent antibodies against synaptic molecules and neurotransmitters and imaged with LM and EM (Micheva and Smith, 2007. The utility of this correlative approach is limited by the ability to preserve fluorescence and antigenicity on the one hand, and EM tissue ultrastructure on the other. We demonstrate tissue staining and fixation protocols and a workflow that yield an excellent compromise between these multimodal imaging constraints. We adapt CAT for the study of projection neurons between different vocal brain regions in the songbird. We inject fluorescent tracers of different colors into afferent and efferent areas of HVC in zebra finches. Fluorescence of some tracers is lost during tissue preparation but recovered using anti-dye antibodies. Synapses are identified in EM imagery based on their morphology and ultrastructure and classified into projection neuron type based on fluorescence signal. Our adaptation of array tomography, involving the use of fluorescent tracers and heavy-metal rich staining and embedding protocols for high membrane contrast in EM will be useful for research aimed at statistically describing connectivity between different projection neuron types and for elucidating how sensory signals are routed in the brain and transformed into a meaningful motor output.

  2. Membranes and Fluorescence microscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2009-01-01

    Fluorescence spectroscopy-based techniques using conventional fluorimeters have been extensively applied since the late 1960s to study different aspects of membrane-related phenomena, i.e., mainly relating to lipid-lipid and lipid-protein (peptide) interactions. Even though fluorescence...

  3. Multimodal fluorescence imaging spectroscopy

    NARCIS (Netherlands)

    Stopel, Martijn H W; Blum, Christian; Subramaniam, Vinod; Engelborghs, Yves; Visser, Anthonie J.W.G.

    2014-01-01

    Multimodal fluorescence imaging is a versatile method that has a wide application range from biological studies to materials science. Typical observables in multimodal fluorescence imaging are intensity, lifetime, excitation, and emission spectra which are recorded at chosen locations at the sample.

  4. Azadioxatriangulenium: exploring the effect of a 20 ns fluorescence lifetime in fluorescence anisotropy measurements

    Science.gov (United States)

    Bogh, Sidsel A.; Bora, Ilkay; Rosenberg, Martin; Thyrhaug, Erling; Laursen, Bo W.; Just Sørensen, Thomas

    2015-12-01

    Azaoxatriangulenium (ADOTA) has been shown to be highly emissive despite a moderate molar absorption coefficient of the primary electronic transition. As a result, the fluorescence lifetime is ~20 ns, longer than all commonly used red fluorescent organic probes. The electronic transitions in ADOTA are highly polarised (r 0  =  0.38), which in combination with the long fluorescence lifetime extents the size-range of biomolecular weights that can be detected in fluorescence polarisation-based experiments. Here, the rotational dynamics of bovine serum albumin (BSA) are monitored with three different ADOTA derivatives, differing only in constitution of the reactive linker. A detailed study of the degree of labelling, the steady-state anisotropy, and the time-resolved anisotropy of the three different ADOTA-BSA conjugates are reported. The fluorescence quantum yields (ϕ fl) of the free dyes in PBS solution are determined to be ~55%, which is reduced to ~20% in the ADOTA-BSA conjugates. Despite the reduction in ϕ fl, a ~20 ns intensity averaged lifetime is maintained, allowing for the rotational dynamics of BSA to be monitored for up to 100 ns. Thus, ADOTA can be used in fluorescence polarisation assays to fill the gap between commonly used organic dyes and the long luminescence lifetime transition metal complexes. This allows for efficient steady-state fluorescence polarisation assays for detecting binding of analytes with molecular weights of up to 100 kDa.

  5. Computed tomography for radiographers

    International Nuclear Information System (INIS)

    Brooker, M.

    1986-01-01

    Computed tomography is regarded by many as a complicated union of sophisticated x-ray equipment and computer technology. This book overcomes these complexities. The rigid technicalities of the machinery and the clinical aspects of computed tomography are discussed including the preparation of patients, both physically and mentally, for scanning. Furthermore, the author also explains how to set up and run a computed tomography department, including advice on how the room should be designed

  6. Meaning of Interior Tomography

    Science.gov (United States)

    Wang, Ge; Yu, Hengyong

    2013-01-01

    The classic imaging geometry for computed tomography is for collection of un-truncated projections and reconstruction of a global image, with the Fourier transform as the theoretical foundation that is intrinsically non-local. Recently, interior tomography research has led to theoretically exact relationships between localities in the projection and image spaces and practically promising reconstruction algorithms. Initially, interior tomography was developed for x-ray computed tomography. Then, it has been elevated as a general imaging principle. Finally, a novel framework known as “omni-tomography” is being developed for grand fusion of multiple imaging modalities, allowing tomographic synchrony of diversified features. PMID:23912256

  7. Simultaneous optical coherence tomography and lipofuscin autofluorescence imaging of the